Monthly Archives: May 2022

Autotrophic and Mixotrophic Growth of an Ammonia Gas-tolerant Bacterium, Paenibacillus lentus NH33 in Ammonia Gas

DOI: 10.31038/GEMS.2022423

Abstract

The ammonia gas-tolerant bacterium Paenibacillus lentus NH33 (hereafter referred to as NH33) was isolated using minimal medium containing glucose in the presence of 1300 ppm of ammonia gas. NH33 showed no growth in the absence of gaseous ammonia (undissociated ammonia), indicating that it is an obligate requiring gaseous ammonia for growth. We investigated how NH33 uses gaseous ammonia. It also grew on minimal agar medium with or without glucose in the presence of 1350 ppm of ammonia gas, indicating that it was also capable of both mixotrophic and chemolithoautotrophic growth. Under the mixotrophic condition, NH33 used gaseous ammonia as a nitrogen source, but did not use dissociated ammonia, nitrite, or nitrate; thus, it appears to use gaseous ammonia as its sole nitrogen source. Under the autotrophic condition, NH33 was capable of using gaseous ammonia as its sole energy source, but not dissociated ammonia. The ammonia gas concentration that was optimal for the growth of NH33 was determined to be 1350 and 170 ppm for the mixotrophic and autotrophic growth conditions, respectively. These growth characteristics indicated that NH33 is a novel nitrifying bacterium that uses gaseous ammonia as its sole energy and nitrogen source.

Keywords

Gaseous ammonia, Ammonia-oxidizing bacteria, Chemolithoautotrophic growth, Mixotrophic growth, Paenibacillus lentus, Nitrite

Introduction

Odors are generally attributed to the production of various volatile compounds arising from the anaerobic decomposition of fecal matter. The number of complaints regarding odors from outdoor toilets, compost facilities, and livestock farms is increasing year-by-year [1]. Deodorization technologies can be classified into three categories, i.e., technologies employing chemical [2], physical, or biological treatment methods [3]. An important advantage of biological treatment methods over physical and chemical treatment methods is that biological processes can be conducted at moderate temperatures and atmospheric pressure. We have been developing biological deodorization techniques by applying the functions of microbes, but this approach is challenging as it requires the isolation of microorganisms capable of growing in the presence of gaseous ammonia. We have isolated promising microorganisms that decrease the odor of ammonia using a newly developed culture method called the closed gaseous ammonia-exposing culture method. The isolated microorganisms were all bacterial species that could be classified into two groups: 1) hypergaseous ammonia-tolerant bacteria and 2) gaseous ammonia-requiring bacteria, which are obligates requiring gaseous ammonia as the sole nitrogen source. These newly discovered bacteria had the ability to eliminate toxic ammonia, or conversely, to assimilate toxic ammonia, which has not been seen before in bacteria. An ammonia gas-tolerant bacterium, Paenibacillus lentus NH33 (hereafter referred to as NH33), was capable of growing in minimal agar medium containing glucose in the presence of gaseous ammonia (1350 ppm), but not in the absence of gaseous ammonia. This indicated that NH33 is a special organism that requires only gaseous ammonia as the nitrogen source. NH33 had a high gaseous ammonia-absorbing rate of 2.20 mmol/10¹⁰ cells. The ammonia-eliminating property of NH33 may be useful for biological deodorization technology [4]. The forms of ammonia include undissociated ammonia (NH₃), which is the gaseous form, and dissociated ammonia (NH₄⁺), which is the ionic form [5]. Ammonia is the preferred nitrogen source for most bacteria and fungi, and plants also require ammonia from the soil as a nitrogen source. Microorganisms also use nitrite and nitrate as nitrogen sources, and these nutrients are reduced to ammonia by reductase [6]. However, ammonia is a paradoxical nutrient: although ammonia is required for growth, undissociated ammonia has a strong cytotoxic effect. Undissociated ammonia dissolves easily through the lipid barrier and across the cytoplasmic membrane, and has a detrimental effect on the growth and metabolism of organisms [7]. Well-known organisms that use ammonia are chemolithoautotrophic ammonia-oxidizing bacteria, which are among a select group of microbes that have the ability to use ammonia as the sole source of energy and reductant for growth [8]. Ammonia-oxidizing bacteria extract energy from a single inorganic source (NH₃), assimilate inorganic substrates (e.g., CO₂ and NH₃), and use them to synthesize all of the biochemicals needed for growth. The product of ammonia oxidation is nitrite. On the other hand, anaerobic ammonium oxidation (anammox) bacteria were discovered in wastewater sludge in the early 1990s; they have the unique metabolic ability to combine ammonium and nitrite or nitrate to form nitrogen gas [9]. Nitrification is catalyzed in two steps by ammonia-oxidizing bacteria and nitrite-oxidizing bacteria that are phylogenetically not closely related, and it was thought that no organism could oxidize both substrates. However, recently, complete ammonia oxidizers (comammox) that are capable of converting ammonia to nitrate within a single organism were unexpectedly discovered within the genus Nitrospira [10].

Ammonia has nutritional roles both as a nitrogen source and an energy source [11]. Ammonia-oxidizing bacteria use ammonium ions not only as an energy source, but also as a nitrogen source. NH33 is a previously unknown obligate that specifically requires gaseous ammonia for growth. In this study, to understand how NH33 utilizes gaseous ammonia, we investigated the autotrophic and mixotrophic growth properties of NH33 [12,13], including its growth in the presence of gaseous ammonia, and its energy and nitrogen sources.

Materials and Methods

Mixotrophic Culture

To prepare minimal medium containing glucose (MMG) agar plate, 15 mL of agar (Wako Pure Chemical Industries, LTD., Osaka, Japan) solution (1.5% final concentration) was sterilized by autoclaving, then (10x) minimal medium concentrate solution and (10x) glucose concentrate solution were added, and the agar was poured into Petri dish. This medium contained no nitrogen source. One liter of MMG (pH7.0) contained 2 μg of biotin, 400 μg of calcium pantothenate, 2 μg of folic acid, 2000 μg of inositol, 400 μg of niacin, 200 μg of p-aminobenzoic acid, 400 μg of pyridoxine hydrochloride, 200 μg of riboflavin, 400 μg of thiamin hydrochloride, 500 μg of boric acid, 40 μg of copper sulfate, 100 μg of potassium iodide, 200 μg of ferric chloride, 400 μg of manganese sulfate, 200 μg of sodium molybdate, 400 μg of zinc sulfate, 1.0 g of monopotassium phosphate, 0.5 g of magnesium sulfate, 0.1 g of sodium chloride, 0.1 g of calcium chloride, and 5.0 g of glucose. One mg of NH33 was added to 1 mL of sterilized water and suspended until homogenous (NH33 suspension). The NH33 suspension (50 μL/plate) was spread onto MMG agar plates without lids. The one plate was placed in an airtight plastic chamber (7.0 L) with ammonium hydrogen carbonate powder (0.178 g), and incubated at 40°C for 3 to 12 days (Figure 1). The ammonium hydrogen carbonate had completely sublimated after 2 to 3 h, and the ammonia gas concentration in the chamber reached 1350 ppm.

Figure 1: Photograph of the closed gaseous ammonia exposure culture system. A MM agar plate for chemolithoautotrophic growth or a MMG agar plate for mixotrophic growth was placed in the 7-L chamber with ammonium hydrogen carbonate powder ((NH₃)HCO₃). The chamber was tightly closed, and incubated at 40^oC. The ammonium hydrogen carbonate had completely sublimated during the 2.5- to 3-h incubation period to provide a gaseous mixture including gaseous ammonia. The amount of ammonium hydrogen carbonate powder in the chamber was adjusted to change the concentration of gaseous ammonia for the experiments.

Autotrophic Culture

To prepare minimal medium (MM) agar plate, 15 mL of agar solution (1.5% final concentration) was sterilized by autoclaving, then (100x) minimal medium concentrate solution was added, and the agar was poured into Petri dish. The MM contained no nitrogen and carbon source. One liter of MMG (pH7.0) contained 2 μg of biotin, 400 μg of calcium pantothenate, 2 μg of folic acid, 2000 μg of inositol, 400 μg of niacin, 200 μg of p-aminobenzoic acid, 400 μg of pyridoxine hydrochloride, 200 μg of riboflavin, 400 μg of thiamin hydrochloride, 500 μg of boric acid, 40 μg of copper sulfate, 100 μg of potassium iodide, 200 μg of ferric chloride, 400 μg of manganese sulfate, 200 μg of sodium molybdate, 400 μg of zinc sulfate, 1.0 g of monopotassium phosphate, 0.5 g of magnesium sulfate, 0.1 g of sodium chloride, and 0.1 g of calcium chloride. The NH33 suspension (50 μL/plate) was spread onto MM agar plates without lids. The plates were placed in an airtight plastic chamber (7.0 L) with ammonium hydrogen carbonate powder (0.178 g), and incubated at 40°C for 3 to 12 days. The ammonium hydrogen carbonate had completely sublimated after 2 to 3 h, and the ammonia gas concentration in the chamber reached 1350 ppm.

Transmission Electron Microscopy (TEM) Observations

The cultured cells on a small patch of agar medium were fixed with 2.0% paraformaldehyde and 2.5% glutaraldehyde in 0.1 M phosphate buffer (pH 7.4) at 4°C for 120 min. The samples were subsequently washed three times with 0.1 M phosphate buffer (pH 7.4), then incubated in 1% OsO₄ at 4°C for 120 min. The samples were dehydrated in a series of increasing ethanol concentrations as follows: 10 min in 50% ethanol at 4°C; 10 min in 70% ethanol at 4°C; 10 min in 80% ethanol at 4°C; 10 min in 90% ethanol at room temperature (RT), 10 min in 95% ethanol at RT, 10 min in 99.5% ethanol at RT twice, 10 min in 100% ethanol at RT twice, and 15 min in propylene oxide at RT twice. Then, the cells were incubated for 60 min in a 1:2 mixture of EPON:propylene oxide, 60 min in a 1:1 mixture of EPON:propylene oxide, 60 min in a 2:1 mixture of EPON:propylene oxide, and overnight in 100% EPON. Ultrathin sections (60 to 80 nm) were cut parallel to the bacterial layer, collected on single-slot Formvar-coated copper grids, and counterstained with 0.5% uranyl acetate at 20°C for 30 min, and 3% lead citrate at 20°C for 7 min [14,15]. Bacteria were imaged using a HT7700 transmission electron microscope (Hitachi, Tokyo, Japan) at an electron voltage of 80 kV.

Optimal Gaseous Ammonia Concentration for Mixotrophic and Autotrophic Growth

The NH33 suspension (50 μL/plate) was spread onto MM or MMG agar plates without lids, placed in an airtight plastic chamber (7.0 L) with ammonium hydrogen carbonate powder (0 to 0.337 g), and incubated at 40°C for 72 h. The ammonium hydrogen carbonate had completely sublimated after 2 to 3 h, and the ammonia gas concentration in the chamber reached 0 to 2560 ppm. The gaseous ammonia concentration was measured with an ammonia gas sensor (New Cosmos Electric Co. Ltd., Osake, Japan). The amount of gaseous ammonia was calculated using the ideal gas law [14].

Usable Nitrogen Sources in Mixotrophic Culture

The NH33 suspension was spread onto MMG agar plate containing different nitrogen sources (10 mM of (NH₄)SO₄, NaNO₂, or KNO₃), and cultured under air at 40°C for 72 h. To investigate the usefulness of gaseous ammonia as a nitrogen source, NH33 was inoculated onto MMG agar plate without lids, then the plate was placed in an airtight plastic chamber (7.0 L) with ammonium hydrogen carbonate powder (0.178 g; initial gaseous ammonia concentration of 1350 ppm), and incubated at 40°C for 72 h.

Usable Energy Sources in Autotrophic Culture

The NH33 suspension was spread onto MM agar plate containing ammonium (10 mM of (NH₄)₂SO₄ or NH₄Cl), and incubated at 40°C for 72 h under air. In addition, NH33 was inoculated onto MM agar plate, and incubated at 40°C for 72 h under 1350 ppm of gaseous ammonia.

Rate of Stable Nitrogen Isotope Uptake in NH33 Cells

A lidless MM agar plate inoculated with NH33 was placed in the 7-L chamber in the presence of 0.35 mL of 3.4% ¹⁵N-labeled ammonia solution (Kao Co. Ltd., Tokyo, Japan) or unlabeled ammonia solution (3.4%), and incubated at 40°C for 72 h. Initially, the concentration of the gaseous ammonia that had vaporized from the ammonia solution was 170 ppm. Cultured cells were recovered with distilled water and centrifuged. The recovered cells were dried at 100°C for 6 h. The rate of ¹⁵N stable isotope uptake (calculated as the ratio of ¹⁵N/total N) in the dried samples was measured by a combined system with a mass spectrometer (DELTA^plus Advantage, Thermo Fisher Scientific, Bremen, Germany) and an elemental analyzer (FLASH 2000, Thermo Fisher Scientific, Bremen, Germany). The accuracy of this analysis system is ±0.3% of the measured value. As the standard samples, International Atomic Energy Agency (IAEA)-N-1 and IAEA-N-2 were used [16].

Detection of Nitrite in NH33 Cells

The NH33 suspension was spread onto MM agar plate and incubated at 40°C for 72 h in the presence of gaseous ammonia at 170 ppm. Cultured cells were recovered with distilled water, washed with a 0.8% NaCl solution, and centrifuged. The cells were then resuspended in 0.8% NaCl solution, and grounded by a bead grinder at 3200 rpm for 30 s. The nitrite concentration in the supernatant after centrifugation was measured by the naphthylethylenediamine hydrochloride spectrophotometric method [17]. The reaction time and temperature was 120 s and 30°C, respectively. The absorbance of the reaction solution was immediately measured by a spectrophotometer at 540 nm.

Biomass Estimation

Biomass production in wet weight was estimated for agar plate cultures by collecting 2.0 ml sterilized distilled water and streak bar. The collected cells were centrifuged (7,740 x g, 10 min) in micro tubes. The cell pellet was then gently washed with 1.5 mL of sterilized distilled water to remove culture medium salts. The sample was again pelleted (7,740 x g, 10 min), the supernatant was carefully removed and 1.5 mL of sterilized distilled water used to resuspend the pellet into pre-weighed 1.5 mL micro tubes. Cells were again pelleted at 9,000 x g for 5 min, and the supernatant discarded. Samples were semi-dried at 40˚C until visible water evaporated. Tubes were weighed on a precision balance (Sartorius CP324S) to estimate the weight of biomass. The wet weight of cells was quantified by subtraction the weight of tube from the total weight.

Results

Mixotrophic Growth

NH33 was inoculated onto MMG agar plate (mixotrophic medium), and incubated at 40°C for 6 days in the presence of 1350 ppm of gaseous ammonia (Figure 1). As shown in Figure 2A, opal colonies with a mucoid morphology were observed [18,19]. Compared to the growth on the MM agar plate, there was more growth on the MMG agar plate, and the colonies had larger diameters that reached over 10 mm. This suggested that active motility [20] or swarming behavior [21] occurred when NH33 was grown on mixotrophic medium. NH33 is an obligate that specifically requires gaseous ammonia for growth, as demonstrated by its inability to grow on MMG agar medium without gaseous ammonia.

Autotrophic Growth

NH33 was inoculated onto MM agar plate (autotrophic medium), and incubated at 40°C for 6 days in the presence of 1350 ppm of gaseous ammonia. As shown in Figure 2B, opal single colonies with diameters of 6 to 7 mm formed on the MM agar plate. NH33 was able to grow on MM agar plate, which did not contain carbon and nitrogen sources, indicating that it is a chemolithoautotrophic bacterium that could gain energy by the oxidation of gaseous ammonia [22].

TEM Observations

The TEM images of the mixotrophically and autotrophically grown NH33 cells are shown in the right panel of Figure 2. NH33 had a bacilliform morphology, and measured 5.0 to 6.5 µm in length and 0.5 to 0.7 µm in width. Under the two different culture conditions, the cells had a cell wall, and a previously unknown white substance which shows low electron dense was observed in the cytoplasm. In particular, the low electron dense area filled most of the cytoplasm in the autotrophically grown cells. Many of the cells grown on the autotrophic medium had an irregular shape, but this was not observed in the cells grown on the mixotrophic medium.

Figure 2: The colony morphology (left) and a transmission electron microscopic image (right) of Paenibacillus lentus NH33. P. lentus NH33 was cultured for 6 days at 40^oC on MMG agar plate in the presence of gaseous ammonia at 1350 ppm for mixotrophic growth (A), and MM agar plate in the presence of gaseous ammonia at 1350 ppm for autotrophic growth (B).

Optimal Gaseous Ammonia Concentration for Mixotrophic and Autotrophic Growth

We investigated the optimal gaseous ammonia concentration for the growth of NH33 on MMG agar plate. As shown in Figure 3A, the gaseous ammonia concentration at 1350 ppm (initial concentration) was optimal for cell growth (relative growth rate: 100). Gaseous ammonia at 2560 ppm repressed the growth of NH33 to a relative growth rate of 10, indicating that an excessively high concentration of gaseous ammonia had a toxic effect on NH33 growth. No growth of NH33 was observed on MMG agar plate without gaseous ammonia.

Figure 3: Optimal gaseous ammonia concentration for mixotrophic and autotrophic growth. Paenibacillus lentus NH33 was cultured on MMG agar plate (A) and MM agar plate (B) in the presence of different concentrations of gaseous ammonia. The values indicate the relative wet weight of the cells.

With the initial gaseous ammonia concentration set to 1350 ppm, we measured the changes in the gaseous ammonia concentration and the growth of NH33 on MMG agar plate (Figure 4A). When no bacteria were inoculated, no change was seen in the gaseous ammonia concentration in the chamber during 170 h of incubation. On inoculated plates, NH33 continued to grow until the 80-h time point, when the gaseous ammonia concentration had decreased to 900 ppm, and the cells stopped growing. These results demonstrated that NH33 assimilated gaseous ammonia into the cells. A plot of the wet weight of cells versus the amount of assimilated gaseous ammonia (Figure 4a) indicated that mixotrophically grown NH33 required 0.62 mmol of gaseous ammonia to produce a biomass of 1.0 g (wet weight). The optimal gaseous ammonia concentration for the autotrophic growth of NH33 on MM agar plate (autotrophic medium) was investigated (Figure 3B). The optimal growth of NH33 on MM agar plate was observed (relative value: 100) when incubated in the presence of gaseous ammonia at 170 ppm. Gaseous ammonia at 2560 ppm decreased the growth rate to a relative value of 5. NH33 was more sensitive to gaseous ammonia when grown under the autotrophic growth condition than under the mixotrophic growth condition. No growth of NH33 was observed under the autotrophic condition without gaseous ammonia. These results indicated that NH33 used gaseous ammonia as an energy source. The increase in the biomass of NH33 grown on MM agar plate and the change in the gaseous ammonia concentration from the initial concentration of 150 ppm in the closed chamber were evaluated (Figure 4B). No change in the gaseous ammonia concentration was observed in the closed chamber during 100 h of incubation of MM agar plate without bacterial inoculation. The biomass continuously increased during 75 h of incubation. During this incubation period, the gaseous ammonia concentration decreased from 150 ppm to 90 ppm. This result demonstrated that NH33 assimilated gaseous ammonia to grow. Figure 4b shows a plot of the amount of assimilated gaseous ammonia versus the amount of biomass (wet weight). Autotrophically growing NH33 cells required 0.64 mmol of gaseous ammonia to gain 1.0 g of biomass (wet weight).

Figure 4: Growth curves of Paenibacillus lentus NH33 on mixotrophic (A) and autotrophic (B) growth media and the changes in the gaseous ammonia concentration. The wet weight of P. lentus NH33 cells and the changes in the gaseous ammonia concentration are indicated by empty squares and circles, respectively. The changes in the gaseous ammonia concentration when using agar medium without P. lentus NH33 inoculation are indicated by black circles. The values indicate the means ± standard deviation of three independent experiments. The cell yield of P. lentus NH33 versus the amount of assimilated gaseous ammonia was plotted for the mixotrophic (a) and autotrophic (b) growth conditions.

Usable Nitrogen Sources in Mixotrophic Growth

NH33 could grow well on MMG agar plate in the presence of 1350 ppm of gaseous ammonia (specific growth rate: 2.21 ± 1.18), but not in the absence of gaseous ammonia (Table 1). Replacing gaseous ammonia with ammonium sulfate, sodium nitrite, or potassium nitrate did not enable the growth of NH33 (Table 1). These results indicated that NH33 could not use dissociated ammonia (i.e., ammonium ions), nitrite, or nitrate as a nitrogen source. NH33 required only undissociated ammonia (gaseous ammonia) as a nitrogen source.

Table 1: Usable nitrogen sources for Paenibacillus lentus NH33 growth on MMG agar plate (mixotrophic medium).

Medium	Nitrogen source		Cell yield (mg wet weight/h/plate)
MMG agar	Gaseous ammonia	1350 ppm	2.21 ± 1.18
MMG agar	None		–
MMG agar	(NH₄)₂SO₄	10 mM	–
MMG agar	NaNO₂	10 mM	–
MMG agar	KNO₃	10 mM	–

-, no growth

Usable Energy Sources in Autotrophic Culture

NH33 could grow on MM agar plate in the presence of 1350 ppm of gaseous ammonia (specific growth rate: 0.11 ± 0.01; Figure 2B, Table 2), but it could not grow on MM agar plate containing dissociated ammonia, i.e., 10 mM (NH₄)₂SO₄ or NH₄Cl, instead of gaseous ammonia (Table 2). These results suggested that NH33 could specifically use undissociated ammonia, i.e., gaseous ammonia, but not dissociated ammonia as an energy source. In addition, NH33 did not grow on MM agar medium without carbon and nitrogen sources (Table 2).

Table 2: Usable energy sources for Paenibacillus lentus NH33 growth on MM agar plate (autotrophic medium)

Medium	Energy source		Cell yield (mg wet weight/h/plate)
MM agar	Gaseous ammonia	1350 ppm	0.11 ± 0.01
MM agar	(NH₄)₂SO₄	10 mM	–
MM agar	NH₄Cl	10 mM	–
MM agar	None		–

-, no growth

Rate of Stable Nitrogen Isotope Uptake in NH33 Cells

NH33 was cultured in the presence of stable nitrogen isotope-labeled [¹⁵N] gaseous ammonia (170 ppm) or conventional gaseous ammonia (170 ppm) at 40°C for 72 h. No difference in the growth rate was observed between the two cultures. The rate of stable nitrogen isotope uptake (calculated as the ratio of ¹⁵N/total N) in the recovered cells was analyzed. As shown in Table 3, all of the nitrogen molecules in the cells grown in the presence of the stable nitrogen isotope-labeled [¹⁵N] gaseous ammonia were replaced by the stable nitrogen isotope. The rate of stable nitrogen isotope uptake (¹⁵N/total N) in the cells grown in the presence of the conventional gaseous ammonia was 0.0036, which is similar to the value of the natural content of ¹⁵N [23,24]. These results confirmed that NH33 took up and assimilated nitrogen derived from the gaseous ammonia. Thus, NH33 is a novel organism that uses gaseous ammonia as both an energy source and nitrogen source.

Table 3: Stable ¹⁵N isotope content in Paenibacillus lentus NH33 grown in 170 ppm of ¹⁵N ammonia gas

Growth type	¹⁵N atoms/total N atoms
Cells grown in ¹⁵NH₃ gas	0.99999
Cells grown in NH₃gas	0.00367
Natural content of ¹⁵N	0.00366

Detection of Nitrite in NH33 Cells

NH33 cells that were cultured on MM agar plate in the presence of 170 ppm of gaseous ammonia at 40°C for 72 h (autotrophic growth cells) were collected and physically ground. The naphthylethylenediamine visual colorimetric method was used to detect nitrite. Figure 5 shows the absorbance spectrum of nitrite solution. The single peak at 540 nm indicates the presence of nitrite, and the maximum absorbance value of the peak reflects the amount of nitrite. As shown in Figure 5, line C, the absorbance peak at 540 nm was observed in the supernatant of the ground cells. The absorbance value at 540 nm indicated that 0.1 µg of nitrite was extracted from 100 mg (wet weight) of NH33 cells, which suggested that gaseous ammonia was oxidized to nitrite by NH33. No nitrite was detected in the agar medium, indicating that NH33 accumulates nitrite intracellularly, and does not eliminate it.

Figure 5: Detection of nitrite in the cell-free extract from autotrophically grown Paenibacillus lentus NH33 cells. The absorbance spectra of 0.5 mg/L (line A), 0.2 mg/L (line B), and 0.05 mg/L (line D) nitrite standard solutions were determined by a spectrophotometer. The spectral peak indicates the nitrite content. The specific peak (line C) was also detected in the cell-free extract.

Discussion

NH33 was first isolated from soil using minimal medium (MM) agar plate containing 28 mM glucose in the presence of gaseous ammonia at 1350 ppm [4]. This bacterium could not grow on MM agar plate containing glucose without gaseous ammonia, indicating that it cannot carry out aerobic nitrogen fixation. Bacillus subtilis str. 168 [25,26] and Escherichia coli K-12 [27] could not grow on nutrient agar plate in the presence of gaseous ammonia at 1350 ppm. NH33 is not only a gaseous ammonia-requiring microbe, but also a gaseous ammonia-tolerant microbe. This report showed for the first time that NH33 is capable of growing on MM agar plate in the presence of gaseous ammonia. This growth property indicated chemolithoautotrophic growth, in which energy is gained through ammonia oxidization. NH33 could grow on MM with or without organic substrates in the presence of gaseous ammonia, indicating mixotrophic growth. Up until now, many researchers have thought that ammonia-oxidizing bacteria do not require an organic carbon source, and that autotrophic growth using ammonia is inhibited by the presence of organic carbons [28]. However, that notion was put into question when a pure culture of a mixotrophic ammonia-oxidizing bacterium (NH33) was established. Ammonia usually has a dual nutritional role: it serves simultaneously as a source of nitrogen and energy [29] for some chemolithoautotrophic bacterial species. It was suggested that NH33 uses glucose as an energy and carbon source, and gaseous ammonia mainly as a nitrogen source, when grown on MMG agar plate. When NH33 is grown on MM agar plate, it uses gaseous ammonia as both an energy and nitrogen source, and carbon dioxide as a carbon source. Microorganisms take up extracellular ammonia, nitrate, urea, and amino acids as nitrogen substrates, and assimilate them [30]. The most reductive form of nitrogen (ammonia) is a favorable nitrogen source as it is energetically efficient. When NH33 was grown on MMG agar plate in the presence of gaseous ammonia, it used gaseous ammonia (undissociated ammonia) as the main nitrogen source. However, NH33 grown on MMG was unable to use ammonium (protonated ammonia), nitrite, and nitrate as a substitute for gaseous ammonia. To introduce ionic substrates intracellularly, organisms require the active transporter (channel protein) system [31,32]. NH33 may lack gene sets involved in the active transporter system for nitrogen species uptake or express them at extremely low levels. Undissociated substances, such as gaseous ammonia, permeate through the cell membrane to the cytoplasm by passive diffusion [33]. The cell surface of NH33 has a high affinity for gaseous ammonia, and can directly assimilate gaseous ammonia intracellularly. NH33 likely gains energy by the oxidation of gaseous ammonia during chemolithoautotrophic growth (growth on MM agar plate in the presence of gaseous ammonia). Ammonia is a stable compound consisting of a nitrogen atom and three hydrogen atoms. It is well-known that the unshared electron pair in the ammonia molecule readily undergoes protonation for the formation of an ammonium ion, as indicated by the following formula: NH₃ + H⁺⇄ NH₄⁺. Ammonia in solution is present as NH₃ and NH₄⁺, and the ratio of NH₃/NH₄⁺ depends on the pH, as defined by the Henderson-Hasselbalch equation [34]. For example, under physiological conditions with a blood pH of 7.4, more than 98% of ammonia is found in the NH₄⁺ form. NH33 may use dissociated ammonia (ammonium ion) as an energy source by dissolving gaseous ammonia in the agar medium. However, NH33 was incapable of growth on MM agar plate (pH 7.0) containing ammonium ion (ammonium sulfate or ammonium chloride). NH33 could be defined as a nitrifying bacterium that uses only undissociated ammonia, but not dissociated ammonia as an energy source. Therefore, NH33 is an obligate that requires gaseous ammonia as both a nitrogen source and an energy source to support its chemolithoautotrophic metabolism.

NH33 is a chemolithoautotrophic bacterium that uses gaseous ammonia as the sole energy source. It extracts energy from a single inorganic source (NH₃), assimilates inorganic substrates (e.g., CO₂ and NH₃), and uses them to synthesize all of the biochemicals necessary to support growth. Ammonia-oxidizing bacteria are incapable of growing mixotrophically [35]. In this study, rather than inhibiting growth, organic compounds, such as glucose, enhanced the growth of NH33. It was considered that NH33 preferentially uses glucose before gaseous ammonia when grown in the presence of gaseous ammonia and glucose simultaneously. Lactose and fructose promoted cell growth more than glucose (data not shown). This mixotrophic growth property suggested that NH33 likely expresses beta-galactosidase and glucose isomerase [36,37]. NH33 is the first reported ammonia-oxidizing bacterium to show mixotrophic growth properties. During mixotrophic growth on MMG agar plate, NH33 mainly used ammonia gas as a nitrogen source. The optimum gaseous ammonia concentration for mixotrophic growth was 1350 ppm. During autotrophic growth on MM agar plate, NH33 used gaseous ammonia as both an energy and nitrogen source. The optimum ammonia gas concentration for autotrophic growth was 170 ppm. Organic substances, such as glucose, were thought to promote the formation of mucoid-like colonies [38] and enhance ammonia gas resistance. The amount of gaseous ammonia required to obtain 1.0 g of cells (wet weight) was 0.62 to 0.64 mmol in both the mixotrophic and autotrophic growth conditions.

NH33 was cultured under autotrophic growth conditions in the presence of gaseous ammonia containing stable isotope nitrogen (¹⁵N), and the stable isotope nitrogen ratio in the cells was subsequently examined. All of the nitrogen in the cells was ¹⁵N, confirming that the gaseous ammonia in the air had been assimilated into NH33 cells. Autotrophic ammonia-oxidizing bacteria are among a specific group of microbes that are able to use ammonia as the sole energy source and reductant for growth. They extract energy from a single inorganic source (NH₃), assimilate inorganic substrates (CO₂ and NH₃), and use them to synthesize all of the biochemicals necessary to support growth. In ammonia-oxidizing bacteria, ammonia is first oxidized to hydroxylamine by ammonia monooxygenase [39]. The hydroxylamine is then oxidized to nitrite by hydroxylamine oxidoreductase. The predominant nitrogen oxide produced in the process of ammonia oxidation is nitrite [40]. Nitrite was detected in the NH33 cells grown autotrophically in the presence of 170 ppm gaseous ammonia. The final substance of gaseous ammonia oxidation in NH33 was nitrite, suggesting that NH33 has an ammonia oxidation mechanism similar to that of conventional ammonia-oxidizing bacteria. However, it remains unclear whether nitrate can be detected in NH33. It is necessary in future studies to determine whether NH33 can completely oxidize ammonia. Chemolithoautotrophic ammonia-oxidizing bacteria are responsible for the rate-limiting step of nitrification in a wide variety of environments, and are thus very important in the global cycling of nitrogen. NH33 is an autotrophic bacterium capable of both mixotrophic and autotrophic growth. Under autotrophic conditions, it uses only gaseous ammonia as an energy source and a nitrogen source. These findings considerably challenge the currently accepted role of microbial communities in global nitrogen cycling. Due to the importance of this functional group of bacteria, understanding of their ecology and physiology has become a subject of intense research in recent years.

Conclusion

Previous studies of ammonia-oxidizing bacteria have reported ammonium ions in a dissociated state as an energy source. Many researchers are unaware of the existence of organisms that can grow in the presence of gaseous ammonia, such as the Paenibacillus lentus NH33 examined in this study. P. lentus NH33 is a previously unknown ammonia-oxidizing bacterium that uses gaseous ammonia as its sole nitrogen source for both assimilation and dissimilation. Understanding and systematizing gaseous ammonia-requiring bacteria in detail will contribute to the elucidation of yet-unknown roles of ammonia in the biosphere and details of the nitrogen cycle.

Acknowledgement

This research was supported by a Grant-in-Aid from the Japan Science and Technology Agency (Grant Number: MP28116808363).

References

Senanu BM, Boakye P, Oduro-Kwarteng S, Sewu DD, et al. (2021) Inhibition of ammonia and hydrogen sulphide as faecal sludge odour control in dry sanitation toilet facilities using plant waste materials. Scientific Reports 11: 1-13. [crossref]
Bertone E, Chang C, Thiel P, O’Halloran K (2018) Analysis and modelling of powdered activated carbon dosing for taste and odour removal. Water Res 139: 321-328.
Kalemba K, Kasperczyk D, Urbaniec K, Kozik V (2017) Biological methods for odor treatment-A review. J Clean Prod 152: 223-241.
Tada S, Itoh Y, Kiyoshi K, Yoshida N (2021) Isolation of ammonia gas-tolerant extremophilic bacteria and their application to the elimination of malodorous gas emitted from outdoor heat-treated toilets. J Biosci Bioeng 131: 509-517. [crossref]
Myszograj S, Płuciennik-Koropczuk E (2020) COD and nitrogen compounds balance in mechanical-biological wastewater treatment plant with sludge treatment. Desalination Water Treat 186: 443-449.
Reitzer L (2003) Nitrogen assimilation and global regulation in Escherichia coli. Annu Rev Microbiol 57: 155-176. [crossref]
Dasarathy S, Mookerjee RP, Rackayov V, Thrane VR, Balasubramaniyan V, et al. (2017) Ammonia toxicity: from head to toe?. Metab Brain Dis 32: 529-538. [crossref]
Arp DJ, Stein LY (2003) Metabolism of inorganic N compounds by ammonia-oxidizing bacteria. Crit Rev Biochem Mol Biol 38: 471-495. [crossref]
Kuenen JG (2008) Anammox bacteria: from discovery to application. Nat Rev Microbiol 6: 320-326.
Hu HW, He JZ (2017) Comammox-a newly discovered nitrification process in the terrestrial nitrogen cycle. J Soils Sediments 17: 2709-2717.
Nicol GW, Schleper C (2006) Ammonia-oxidising Crenarchaeota: important players in the nitrogen cycle?. Trends Microbiol 14: 207-212.
Steinmüller W, Bock E (1976) Growth of Nitrobacter in the presence of organic matter. I. Mixotrophic growth. Arch Microbiol 108: 299-304. [crossref]
Pflug IJ, Smith GM, Christensen R (1981) Effect of soybean casein digest agar lot on number of Bacillus stearothermophilus spores recovered. Appl Environ Microbiol 42: 226-230. [crossref]
Haar L, Gallagher JS (1978) Thermodynamic properties of ammonia. J Phys Che Ref Data 7: 635-792.
Wenzel M, Dekker MP, Wang B, Burggraaf MJ, Wilbert B, et al. (2021) A flat embedding method for transmission electron microscopy reveals an unknown mechanism of tetracycline. Commun Biol 4: 1-13. [crossref]
Tayasu I, Hirasawa R, Ogawa NO, Ohkouchi N, et al. (2011) New organic reference materials for carbon-and nitrogen-stable isotope ratio measurements provided by Center for Ecological Research, Kyoto University, and Institute of Biogeosciences, Japan Agency for Marine-Earth Science and Technology. Limnology (Tokyo) 12: 261-266.
Ridnour LA, Sim JE, Hayward MA, Wink DA, et al. (2000) A spectrophotometric method for the direct detection and quantitation of nitric oxide, nitrite, and nitrate in cell culture media. Anal Biochem 281: 223-229. [crossref]
Payment P, Coffin E, Paquette G (1994) Blood agar to detect virulence factors in tap water heterotrophic bacteria. Appl Environ Microbiol 60: 1179-1183. [crossref]
Chiarelli A, Cabanel N, Rosinski-Chupin I, Zongo PD, Thierry N, et al. (2020) Diversity of mucoid to non-mucoid switch among carbapenemase-producing Klebsiella pneumoniae. BMC Microbiol 20: 1-14.
Park SY, Kim R, Ryu CM, Choi SK, Choong HL et al. (2008) Citrinin, a mycotoxin from Penicillium citrinum, plays a role in inducing motility of Paenibacillus polymyxa. FEMS Microbiol Ecol 65: 229-237. [crossref]
Fünfhaus A, Göbel J, Ebeling J, Knispel H, et al. (2018) Swarming motility and biofilm formation of Paenibacillus larvae, the etiological agent of American Foulbrood of honey bees (Apis mellifera). Sci Rep 8: 1-12.
Chain P, Lamerdin J, Larimer F, Regala W, et al. (2003) Complete genome sequence of the ammonia-oxidizing bacterium and obligate chemolithoautotroph Nitrosomonas europaea. J Bacteriol 185: 2759-2773. [crossref]
Naidu K, Maseko S, Kruger G, Lin J (2020) Purification and characterization of α-amylase from Paenibacillus D9 and Escherichia coli recombinants. Biocatal Biotransformation 38: 24-34.
Pethybridge H, Choy CA, Logan JM, Allain V et al. (2018) A global meta-analysis of marine predator nitrogen stable isotopes: Relationships between trophic structure and environmental conditions. Glob Ecol Biogeogr 27: 1043-1055.
Zolfaghari Emameh R, Kazokaitė J, Yakhchali B (2021) Bioinformatics analysis of extracellular subtilisin E from Bacillus subtilis. J Biomol Struct Dyn 4: 1-8. [crossref]
Zeigler DR, Prágai Z, Rodriguez S, Chevreux B, et al. (2008) The origins of 168, W23, and other Bacillus subtilis legacy strains. J Bacteriol 190: 6983-6995. [crossref]
Sandler SJ, Leroux M, Windgassen TA, Keck JL (2021) Escherichia coli K‐12 has two distinguishable PriA‐PriB replication restart pathways. Mol Microbiol 116: 1140-1150. [crossref]
Krümmel A, Harms H (1982) Effect of organic matter on growth and cell yield of ammonia-oxidizing bacteria. Microbiol. 133: 50-54.
Kowalchuk GA, Stephen J (2001) Ammonia-oxidizing bacteria: a model for molecular microbial ecology. Annu Rev Microbiol 55: 485-529. [crossref]
Ouyang Y, Norton JM, Stark JM, Reeve JR, et al. (2016) Ammonia-oxidizing bacteria are more responsive than archaea to nitrogen source in an agricultural soil. Soil Biol Biochem 96: 4-15.
Islam S, Islam R. Kandwal P, Khanam S, et al. (2020) Nitrate transport and assimilation in plants: a potential review. Arch Agron Soil Sci 68: 1-18.
Li C, Ariga I, Mikami K (2019) Difference in nitrogen starvation-inducible expression patterns among phylogenetically diverse ammonium transporter genes in the red seaweed Pyropia yezoensis. Am J Plant Sci 10: 1325-1349.
Keerio HA, Bae W, Park J, Kim M (2020) Substrate uptake, loss, and reserve in ammonia-oxidizing bacteria (AOB) under different substrate availabilities. Process Biochem 91: 303-310.
Thurston RV, Russo RC, Vinogradov GA (1981) Ammonia toxicity to fishes. Effect of pH on the toxicity of the unionized ammonia species. Environ Sci Technol 15: 837-840.
Liu X, Lin J, Tian K, Yan W (2008) Metabolism of organic compounds by extremely acidophilic, obligately chemolithoautotrophic Thiobacilli: a review. Chin J biotechnol 24: 1-7.
Kim KH, Seo YL, Baek JH, Jin HM, et al. (2021) Paenibacillus agri nov., isolated from soil. Int J Syst Evol Microbiol 71.
Soni R, Nanjani S, Keharia H (2021) Genome analysis reveals probiotic propensities of Paenibacillus polymyxa Genomics 113: 861-873.
Daane LL, Harjono I, Barns SM, Launen L, et al. (2002) PAH-degradation by Paenibacillus and description of Paenibacillus naphthalenovorans sp. nov., a naphthalene-degrading bacterium from the rhizosphere of salt marsh plants. Int J Syst Evol Microbiol 52: 131-139. [crossref]
Hommes NG, Sayavedra-Soto LA, Arp DJ (2001) Transcript analysis of multiple copies of amo (encoding ammonia monooxygenase) and hao (encoding hydroxylamine oxidoreductase) in Nitrosomonas europaea. J Bacteriol 183: 1096-1100. [crossref]
Sepehri A, Sarrafzadeh MH (2019) Activity enhancement of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria in activated sludge process: metabolite reduction and CO₂ mitigation intensification process. Appl Water Sci 9: 1-12.

Autism Diagnostic Criteria, Symptoms, Child Diagnosis and Evaluation Scales

Leave a reply

DOI: 10.31038/ASMHS.2022641

Abstract

Introduction: Autistic children have not withdrawn from reality due to some mental illness, rather they have failed to come out in reality due to a serious, and to a large extent, developmental disorder. As a result, autism is not currently considered psychosis, but is classified as a pervasive developmental disorder. This means that it is classified along with the other developmental difficulties rather than the mental illnesses themselves.

Aim: The aim of this study was to present an organized context of autism, its symptoms and clinical picture, its diagnostic criteria as well as child rating scales.

Method: The recent literature was reviewed with keywords: autism, symptoms, diagnostic criteria, evaluation, assessment of autism.

Conclusions: Autistic Disorder or autism is a severe form of Diffuse Developmental Disorder and has been described as a “spectrum disorder”, which means that the clinical picture of autism is not homogeneous, but ranges from milder forms (with minimal and mild form). autistic elements and normal intelligence) to more severe forms (with multiple autistic elements accompanied by severe mental retardation).

Keywords

Autism, Symptoms, Diagnostic criteria, Evaluation, Assessment of autism

Introduction

Autism is a disorder that is affecting more and more people and has been described as a “spectrum disorder” which means that the clinical picture of autism is not homogeneous, but ranges from milder forms (with minimal and mild autistic features and normal intelligence)up to more severe forms (with multiple autistic elements accompanied by severe mental retardation). Autism has a reduction in the following areas [1]:

Mutual social transactions
Communication
General behavior (stereotyped and divisive reactions occur)
Interests
Activities

In the areas mentioned above, people with Autism not only differ due to developmental delays, but also have divergent reactions, which are not usually found in children with normal development.

Diagnostic Criteria

The diagnosis of Autistic Disorder even today is based on the characteristics of the child’s behavior and not on medical examinations, although it is widely accepted that the causes of autism are due to organic rather than environmental factors [1].

It is true that Kanner himself fueled these orientations considerably by referring to “cold rational” parents and the upper social class, in conjunction with his claim that autistic children have potentially high intelligence. We need a brief reference to these issues because they are involved in the definition of autism in a variety of ways [2-7]:

a) Are the points that were mainly refuted in the course of Kanner’s initial presentation,

b) The data on them are also indirectly answered in the hypothesis for a psychogenic definition of autism, essentially excluding it and

c) From here the research went further, which It was already focusing on the specifics of autism, thus leading to modern perceptions of this disorder but also to highlighting its complexity. Kanner concluded that children should maintain normal intelligence, indirectly: from the islands of potential that he found to maintain among their difficulties, such as:

Their “excellent” memory. Many studies have shown that the majority of autistic children actually have some degree of mental retardation, which is usually not severe. In fact, it remains and is still seen later, even if their social performance improves as they grow older [8-11].

The other thing that is clear here and important for the definition of autism is that, although mental retardation and autism often coexist, they are not identical. A few points from reliable studies can show interesting differences:

Autism can coexist with illnesses and conditions that usually cause mental retardation but the association does not seem to be as strong: it seems to go particularly well with low sclerosis, which is a rare condition, but much less so with Down syndrome or cerebral palsy, which are much more common causes of mental retardation [1].
Epileptic seizures, common in children with mental retardation but usually beginning in early and middle childhood, typically first appear in autism (in ¼% of cases) around or near adolescence, or later [12,13].
Characteristic differences are presented by retarded children and in particular aspects of cognitive emotional and social function. As in the immediate recognition of the leaf, in the recognition of emotions as they appear in human faces and in the emotional understanding
Kanner’s first study of autism highlights a number of traits that he found to be characteristic of all the children he saw. These features are as follows [2]:

Excessive Autistic Loneliness

Children failed to interact smoothly with people, and seemed to be overjoyed when left alone. This lack of social response appeared, according to Kanner, very early in life, as shown by the autistic infant’s failure to reach out to the parent who would take him or hug him.

Anxious Depressive Desire to Maintain Similarity

Children were overly irritated by changes in routine or their surroundings. A different school routine, a rearrangement of furniture, could be the cause of an explosion, and the child may not be able to calm down until the familiar order is restored.

Excellent Memory

The children Kanner saw showed an ability to recall large and insignificant amounts of material (eg an encyclopedia content page) that did not keep pace with their clear learning difficulties or mental retardation in other cases.

Delayed Hoarseness

Children repeated the language they were hearing but failed to use words to communicate on topics beyond their immediate needs. The echo may explain the reversal of Kanner’s pronouns – that children use the “You” when referring to themselves and the “I” to the other person. This use would be an immediate repetition of another speaker’s observation. Similarly, autistic children commonly use an entire question to ask for something that follows (eg “Do you want a sweet?”, Meaning ” I want a sweet”).

Hypersensitivity to Stimuli

Kanner noted that many of the children he saw reacted strongly to certain noises and objects, such as vacuum cleaners, elevators, and even blowing air. Some even have problems or fantasies with food.

Limitation to Spontaneous Activity Diversity

This is evident in children’s repetitive movements in their verbal expressions and their interests. Kanner, however, realized that children often seemed to have a good relationship with objects revealing an amazing skill in spinning things or completing puzzles.

Good Cognitive Abilities

Kanner believed that the outstanding memory and skill shown in some of his cases reflected a superior intelligence, in addition to the fact that many children were considered to have severe learning disabilities.This strong impression on intelligence often observed in parents and teachers. A good memory is especially torturous – it leads everyone to form the impression that only if it turned to some practical use the child would learn well. An impression of intelligence is still given by the complete lack of obvious features in most cases of autism. Unlike children, who have many types of severe learning disabilities (eg Down syndrome), autistic children usually have a “normal” appearance. Kanner pointed out his “intelligent features”.

Highly Intelligent Families

Kanner pointed out that all of his cases had intelligent parents, which may have facilitated the referral of their children to a specialist – so Kanner’s sample is unlikely to be representative. Kanner also described parents as cold, although in his first article he was far from a psychogenic theory. On the contrary, he considers that “those in the world with an innate lack of ability to form a normal, biologically determined, emotional contact with people. The areas of speech that are particularly affected by autistic disorder, both comprehension and expression, are those that are related to the dialectical elements of speech, its prosody (tone of voice, volume, strength, intonation) [14], with its non-verbal elements (posture, distance, facial expressions etc.), the factual nature of speech and its use in everyday communication practice [10].

Kanner isolated only two of these key elements of autism: “excessive isolation and depressive obsession with maintaining similarity.” He considered that the other symptoms are either secondary or caused by these two elements (eg communication deficiencies), or that they do not correspond to autism (eg stereotypes) [12].

Symptoms

The symptoms of autism can generally be divided into “deficiencies” and “surpluses” of behavior. The shortcomings concern the main areas of development, such as [2]:

Attention

a) Avoid eye contact

b) Distraction

c) Minimal or excessive preoccupation with certain objects

d) Expression of concern

Speaking

a) Hoarseness

b) Incomprehensible joint

c) Inappropriate hoarseness of voice

d) Inappropriate volume of voice

e) Incoherent reason

f) Repetitive speech

Social and Emotional Events

a) Avoidance or refusal of physical contact

b) Avoidance or refusal of social contact / communication

c) Lack of interest in peers

d) General lack of interest in people

e) Lack of initiative and response to social transactions

f) Apathy to stimuli that cause fear

g) Excessive fear response to stimuli that usually do not elicit a phobic response

h) Apathy or excessive reaction to separation from the mother

i) Apathetic or inappropriately emotional expressions

j) Lack of empathy

Play

a) Peculiar use of toys abstaining from symbolic or representational play

b) Abstention from binary or group play with peers

c) Excessive attachment to certain games

d) Interest in a very limited number of games

Sensory Processing

a) Idiosyncratic processing of visual stimuli (eg plain gaze)

b) Indifference or excessive reaction to auditory stimuli (eg does not react to a loud click, while closing his ears to the sound of the vacuum cleaner)

c) Indifference or excessive reaction to tactile stimuli (eg apathetic pain remains, while caressing is not tolerated)

d) Hypersensitivity to certain flavors and odors

Selective Attention to Certain Characteristics of Environmental Stimuli or Overselectivity

It tends to focus on individual characteristics of environmental stimuli, such as color or shape, resulting in processing them piecemeal rather than spherically. This fragmentary or over-selective treatment exacerbates the difficulty for the person with autism to distinguish and recognize objects or symbols and to generalize their skills with new stimuli and in new conditions [5,6]. For example, if he has learned to name the person depicting a photograph, he recognizes it only from that photograph. He does not realize that the person he depicts in different photos is the same. This difficulty is due to the fragmentary observation of an element or elements of the photograph, such as the clothes of the person depicted or the landscape, instead of globally observing all the facial features that are the main stimulus for recognizing people [4].

Cognitive Functions

a) Mental retardation

b) Unstable learning

c) Developmental gaps in cognitive areas

d) Learning regression

“Surpluses” in the behavior of people with autism are mainly associated with maladaptive and stereotypical reactions and can be classified into the following categories:

Dissociative Behavior

a) Anger attacks

b) Disobedience

c) Aggression

d) Self-injuries

Stereotypical Reactions

a) In tactile stimuli (eg visual surveillance with half-closed eyes, monitoring of objects holding and twisting them like a whirlpool)

b) In speech (eg incoherent sounds or continuous phrases)

c) In the smell (eg smells persistently of objects or people)

d) In taste (eg holds food in the mouth for a long time) e) in touch (eg rubs fingers together)

e) In motion (eg shakes his fingers, hands or feet for a long time, tightens his muscles)

f) Rituals (eg placing objects in a straight line)

Special Abilities

a) Excellent memory and parrot ability

b) High arithmetic ability

c) Hyperlexia

d) Dexterity in puzzles and games with complex mechanisms

Diagnosis Scales and Evaluation of Children with Autism

The use of questionnaires and scales that assess only behavioral problems as well as the limited interests and repetitive and stereotypical behavior of children with ASD is useful not only for diagnostic purposes, but because it is a guide for prioritizing the goals of therapeutic intervention. That is, we decide to start the therapeutic intervention from the most serious problems of the child and the family (that is, the problems that most affect the life of the child and his family as well as those that greatly impede learning) and then we intervene in the least serious problems (those that least affect the child’s life and learning). According to the DCM-III-R we can classify the diagnostic criteria of autism into three categories: social behavior, communication and deviant behavior. In order to diagnose autism, it is necessary to add up a total of six or more “points” from the three categories mentioned above. In particular, the presence of at least two “points” from the first category and at least one point from the second and third category is necessary.

A. Diagnostic criteria for autism disorder by DCM-III-R

At least eight of the following traits are present and include two traits from group A, one from group B and one from group C [1].

Note: Consider a criterion only if the behavior is abnormal to the individual’s level of development.

A. Qualitative Deficiency in Mutual Social Interaction as Manifested by the Following

(The examples in parentheses are arranged so that what is mentioned first is more likely to suit smaller or more disadvantaged individuals and the latter to larger or less disadvantaged individuals with this disorder).

Clear lack of vigilance regarding the existence or feelings of others (eg treats one person as if it were a piece of furniture, does not perceive another person’s anguish, obviously does not perceive the need for others’ privacy).
Lack or abnormal pursuit of relief in times of distress (eg does not seek relief even when ill, wounded or tired, seeks relief in a stereotypical way, eg says “cheese, cheese, cheese »Whenever he is injured).
Lack or inadequate imitation (eg does not shake his hand when saying “hello”, does not copy the mother’s activities at home, mechanical imitation of actions without content) Lack or abnormal play with others (does not actively participate in simple toys, prefers solitary play activities, engages in other children’s play only as “mechanical assistance”).
Clear lack of ability to make friends with peers (no interest in making friends with peers, despite the interest in making friends shows a lack of understanding of the habits that govern social transaction (for example, reads a phone book to non-interested peers).

B. Quality Deficiency in Verbal and Non-verbal Communication and Creative Imagination Activities, as Manifested by the Following

(The numbered features are arranged so that those presented first are more likely to apply to smaller or more disadvantaged individuals and the latter to larger or less disadvantaged individuals with this disorder).

No means of communication, such as the use of articulate communication screams, facial expressions, gestures, imitations or spoken language.
Clear abnormal non-verbal communication on topics such as a research look, facial expression, posture or gestures to start or arrange a social transaction (eg does not anticipate being held in the arms, does not bend his body when held, does not look at the person or laugh when he takes a social approach, does not shake hands with his parents or visitors, has a steady plain look at social situations).
Lack of creative imagination activity, such as playing the roles of an adult, fictional person or animal, lack of interest in fictional storytelling.
Clear abnormalities in speech production, which includes characteristics such as volume, color, agony, proportion, rhythm and tone (eg monotonous speech, question-like melody, or high color).
Clear anomalies in the form or content of speech, which includes stereotyped and repetitive use of speech (eg direct echo or mechanical repetition of TV commercials), use of “you” instead of “I” (eg uses: “Do you want a nut?” To mean:”I want a nut”), temperamental use of words or phrases (eg uses: “Go on the green aisle” to mean: “I want to go to the cot”), or frequent irrelevant remarks (eg begins to talks about train timetables during a sports discussion).
Clearly inadequate ability to initiate or maintain a conversation with others beyond adequate speech (eg it results in long monologues that deal with a topic regardless of its connection to other topics).

C. Clearly Limited Repertoire of Activities and Interests

Stereotypical body movements, e.g. hitting – twisting – knitting of the hands, strong banging of the head, complex movements of the whole body.
Persistent bias towards parts of objects (eg smell of objects, repetitive sensual contact with materials, rotation of toy car wheels) or attachment to unusual objects (eg insists on rotating a piece of rope).
Clear anxiety about changes in insignificant aspects of the environment, e.g. when a jar is moved from the usual position.
Reckless obsession with routines with precise detail, e.g. obsession to always and exactly follow the same shopping habit.
Clearly limited range of differences and a bias with a narrow interest, e.g. is interested in putting objects in order, collecting meteorological facts or pretending to be a fictional person.

D. Beginning during Infancy or Childhood

Identify the onset of childhood (after 36 months of age).

Categories with Indicative Examples of Grading Scale Classification of Autism (CARS, Childhood Autism Rating Scale, 1988)

Because people with Autism, in addition to psychoeducational needs, have additional problems due to deviant behavior, specially weighted scales and questionnaires are particularly useful to assess the behavioral problems, anxiety, and other disorders that these individuals experience. The most well-known and widespread of these questionnaires and scales are: the Autism Behavior Checklist (ABC) [3], which helps determine if a student should be referred for an autism test in the first place. , Autism Diagnostic Interview- Revised (ADI-R) [8] (also used to diagnose autism) and the Childhood Autism Rating Scale (CARS) [3], perhaps the most common scale for autism, used to determine the severity of symptoms. The score on this scale classifies autistic disorder as mild, moderate, and severe. CARS groups autism-related symptoms into 15 categories and each symptom is classified based on 7 scores (1, 1.5, 2, 2.5, 3, 3.5 and 4). The grade of the symptoms ranges from grade 1 (which means the manifestation of behavior at a normal level for his chronological age) to grade 4 (which means the manifestation of a symptom at a level that indicates a serious disorder). Below are the 15 categories of symptoms with examples [15-17].

1. Interpersonal Relationships

E.g. the grade 2.5-3 is given when the child sometimes does not perceive the presence of adults. It sometimes takes persistent effort to get his attention. Takes minimal communication initiatives.

2. Imitation

E.g. Grade 4 is given when the child rarely imitates sounds, words or movements, even at the urging of adults.

3. Emotional Manifestations

E.g. grade 1 is given when the child expresses his feelings with the quality and intensity dictated by social situations. Emotional expression is judged by the child’s facial expression, posture and manners.

4. Movement of the Body

E.g. Grade 2 is given when the child shows minor peculiarities in movement, such as awkwardness, repetitive or uncoordinated movements and when he is very short he makes particularly unusual movements.

5. Use of Objects

For example, grade 4 is given when the child has little interest in toys or other objects, or uses them in very peculiar ways. Absorbs dealing with elements or parts of the game that are not important and is impressed by the reflection of light on these objects. Repeatedly shakes a part of the object or plays exclusively with a single object. When a child becomes addicted to the above reactions, it is very difficult to distract him from them.

6. Adaptability to Change

E.g. Grade 2 is given when the child is still engaged in the same activity or the same material, despite the efforts of an adult to distract him.

7. Visual Reactions

E.g. grade 1 is given, if the child has normal for his age reactions to visual stimuli. It uses its sight, like its other senses, to process new stimuli.

8. Acoustic Reactions

E.g. grade 3 is given when the child’s reactions to auditory stimuli vary. The child sometimes systematically ignores some sounds and other times, scares and closes his ears if he hears normal sounds of the environment.

9. Reactions related to Taste, Smell and Touch

E.g. grade 2 is given when the child insists on putting in his mouth, smelling or trying to eat non-edible objects. The same degree is given when he is either completely ignorant or overly intolerant of mild pain, for which the average child would simply show little discomfort.

10. Stress Reactions

E.g. Grade 4 is given when the child repeatedly shows phobias about situations and objects that are harmless, while it is very difficult to alleviate his fear. The child may expose himself to dangers that children his age have learned to avoid.

11. Verbal Communication

For example, grade 4 is given when the child does not use communication language. It can make babies cry, strange sounds that sound like animal cries, complex noisy sounds that characterize articulated words or phrases in a repetitive and strange way.

12. Non-Verbal Communication

E.g. Grade 3 is given when the child cannot express his needs and desires using ways of non-verbal communication and does not understand the expressive communication expressions of others.

13. Mobility Level

E.g. Grade 1 is given when the child shows no deviation from children of his age in motor activities. The child does not show hyperactivity or hyperactivity in relation to children of his age.

14. Level of Stability in Cognitive Skills

E.g. Grade 3 is given when the child in general does not have the same cognitive skills as children of his age, but in one or two areas shows normal development.

15. General Impressions

The score ranges from 1 (the child does not show symptoms of autism) to 4 (the child has many symptoms or autism in severe form).

References

Rutter M (1990) Infant Autism, Athens, Greek Letters.
Frith U (1999) Autism, Athens: Greek Letters.
Happe F (1008) Autism, Athens: Gutenderg.
Stamatis S (1987) Fortified Silence, Athens: Gull.
Kathleen Ann Quill. Scientific Supervision: A. Messini G Antoniadis (2000), Teaching autistic children, published by Ellin, Athens.
Karantanos C (2001) Autism – Europe, Hellenic Society for the Protection of Autistic People.
Grandin T, Scariano M (1995) Diagnosis: Autism, Athens: Greek Letters.
Genna (2002) Autism and diffuse developmental disorders, Published by Ch. Zacharopoulos, Athens.
Notas S, Brown W, Cohen I (2003) Conference Proceedings. Larissa: XE Seminar Athens, November 18 (2002) Therapeutic Approaches to Diffuse Developmental Disorders. Society for the Mental Health of Children and Adolescents.
Upgrading and expansion of the institution of education of people with special needs (disabled) in Primary and Secondary Education, (2003) ACT OP. E.A. EC: 1.1.4.a ACTION B ‘: DEVELOPMENT OF CURRICULUM STUDIES, RESPONSIBLE:Markakis Emmanouil, Honorary Special Assistant Special Education Pedagogical Institute
Kuder S Jay (2003) Teaching Students with Language and Communication Disabilities (2nd Edition). Boston: Allyn & Bacon.
Raymond D Kent (2004) The MIT Encyclopaedia of Communication Disorders, MIT Press.
Reed A Vicki (1993) An Introduction to Children with Language Disorders, Prentice Hall (a Pearson Education company).
Faherty C (2003) What does it mean to me? Athena. Greek letters.
Scariano M (1995) Diagnosis: Autism, Athens, Greek Letters.
Andy Bondy, Kate Dickey, Diane Black, Sarah Buswell (2002) The Pyramid Approach to Education: Lesson Plans for Young Children, USA, Pyramid Educational Products Inc.
Lennard-Brown (2004) Autism, Translation: Nikolakaki, Maria, Ed. Savallas.

Risk Factors for Postoperative Nausea and Vomiting, Some Helpful Hints

Leave a reply

DOI: 10.31038/ASMHS.2022634

Introduction

Recovery following anesthesia is usually complicated by nausea, vomiting, and retching. PONV (postoperative nausea and vomiting) is a significant concern; people frequently regard PONV as terrible than postoperative pain [1]. PONV normally clears up or is treated without repercussions, although it may need an unexpected admittance to the hospital and cause a delay in discharge [2,3].

In this article, we’ll go over a few points concerning PONV risk factors that you should be aware of.

Pathophysiology

Nausea and vomiting are mediated by five neurotransmitter receptors: neurokinin 1 (NK1) – substance P, dopamine D2, muscarinic M1, 5-hydroxytryptamine (HT)-3 serotonin, and histamine H1. PONV might be prevented or treated by targeting any of these receptors [4].

Central mechanisms – Higher centers of cortex connecting with the central pattern generator (previously known as the center of vomiting) in the medulla can cause nausea and emesis. Anxiety, pain, conditioned nausea which is triggered by environmental cues, fear, and vestibular system stimulation are all major stimuli that can elicit nausea and vomiting during the perioperative period. During tympanoplasty, for example, surgical activation of the vestibular system via the H1 histamine and M1 cholinergic receptors may cause severe PONV [5].
Mechanisms in the periphery – Direct stimulation of the stomach from gastric injuries, bleeding, or toxins causes enterochromaffin cells to produce substance P and serotonin, activating the 5-HT3 receptors in the vagal and splanchnic nerves [6].
Toxins and Drugs – The chemical and neurological processes by which medications and toxins elicit nausea and vomiting, including anesthetics and opioids, are complicated and poorly understood [7]. Both opioids and inhalation anesthetics can cause nausea and vomiting by activating the area postrema directly beneath of the fourth ventricle in the medulla. The postrema then transmits dopamine and serotonin to the central pattern generator, which causes the vomiting reflex to be triggered [8,9].

Risk Factors

PONV develops in roughly 30% of infants and adults following anesthesia without prophylaxis [10]. The risk of PONV varies greatly from patient to patient; in high-risk individuals, the rate of PONV might be 80% [11]. The risk of PONV varies depending on the patient, the anesthesia used, and even the type of operation.

Patient Risk Factors

Nausea and vomiting before surgery – PONV might be the outcome of a pre-surgery ailment that caused nausea and vomiting.
Female gender – Female gender is the most accurate patient-specific predictor of PONV [12,13].
Prior to puberty, female children do not have an increased risk of PONV [14,15].
Patients who have already had PONV or motion sickness – Previous PONV and/or motion sickness increase the risk of PONV in adults [16].
A parent or sibling’s history of PONV or postoperative vomiting (POV), as well as a parent or sibling’s history of PONV or POV, increases the risk of POV/PONV in children [17].
Being a nonsmoker – Being a nonsmoker is a risk factor for PONV in and of itself [12,13,16].
Age – The majority of research have found a minor, gradual decline in PONV in people as they get older [10,12]. The age of 50 was found to be a risk factor for PONV in a prospective trial of over 2200 patients who received general anesthesia (PDNV) [18].
Young age seems to be protective in children. POV is uncommon in youngsters under the age of three, and it becomes more common as they become older, with puberty it reduces again [14].
Chemotherapy-induced nausea and vomiting – A background of chemotherapy-induced nausea and vomiting may aggravate PONV (CINV) [19].

Anesthetic Factors

Anesthetic technique – When compared to simply regional anesthesia, general anesthesia is linked to a greater rate of PONV [16].
Total intravenous anesthesia versus Volatile anesthetics – The use of volatile anesthetics is a major factor in the development of PONV [16,20].
Intravenous (IV) anesthetics – At dosages routinely used for induction of anesthesia, etomidate does not raise PONV on its own [21]. Low-dose ketamine in the perioperative period has been shown to minimize PONV, as well as postoperative pain and opioid needs [22,23].
Nitrous oxide (N₂O) – When compared to anesthesia without N₂O, N₂O may slightly increase the incidence of PONV, particularly in children and high-risk individuals [24].
Duration of anesthesia – Anesthesia for longer periods of time using volatile anesthetics may raise the incidence of PONV [12,13,25].
Opioid administration and decrease – Several studies have found that perioperative opioid administration increases the risk of PONV in a dose-dependent way [11,16,26].
Neostigmine versus Sugammadex for reversal agent – As shown in a meta-analysis of 10 randomized studies including 933 individuals, combining neostigmine with either atropine or glycopyrrolate did not significantly enhance the incidence of overall nausea or vomiting [27].
It’s uncertain how common PONV occurs once sugammadex is used to reverse neuromuscular blocking medications.

Type of Surgery

When compared to other general surgical operations, the best data shows that cholecystectomy, laparoscopic, and gynecologic surgeries are linked with a moderately elevated risk of PONV [12].

In children, strabismus operation is a significant, and arguably the most critical, predictor of POV [14,17]. POV also happens in up to 70% of children who undergo adenotonsillectomy without prophylaxis [28], 60% of kids who have otoplasty [29], and 40% of children who have inguinal scrotal or penile surgeries [30].

In conclusion, PONV is a condition with distinct characteristics that can be avoided by recognizing preventive causes. In our review, we have covered several noteworthy elements.

References

Macario A, Weinger M, Carney S, Kim A (1999) Which clinical anesthesia outcomes are important to avoid? The perspective of patients. Anesthesia and Analgesia 89: 652-658. [crossref]
Fortier J, Chung F, Su J (1998) Unanticipated admission after ambulatory surgery–a prospective study. Canadian Journal of Anaesthesia 45: 612-619. [crossref]
Hill RP, Lubarsky DA, Phillips-Bute B, Fortney JT, Creed MR, et al. (2000) Cost-effectiveness of prophylactic antiemetic therapy with ondansetron, droperidol, or placebo. Anesthesiology 92: 958-967. [crossref]
Bashashati M, McCallum RW (2014) Neurochemical mechanisms and pharmacologic strategies in managing nausea and vomiting related to cyclic vomiting syndrome and other gastrointestinal disorders. European Journal of Pharmacology 722: 79-94. [crossref]
Becker DE (2010) Nausea, vomiting, and hiccups: a review of mechanisms and treatment. Anesthesia Progress 57: 150-156. [crossref]
Spiller R (2007) Recent advances in understanding the role of serotonin in gastrointestinal motility in functional bowel disorders: alterations in 5-HT signalling and metabolism in human disease. Neurogastroenterology and Motility 2: 25-31. [crossref]
Horn CC, Wallisch WJ, Homanics GE, Williams JP (2014) Pathophysiological and neurochemical mechanisms of postoperative nausea and vomiting. European Journal of Pharmacology 722: 55-66. [crossref]
Bountra C, Gale JD, Gardner CJ, Jordan CC, Kilpatrick GJ, et al. (1996) Towards understanding the aetiology and pathophysiology of the emetic reflex: novel approaches to antiemetic drugs. Oncology 1: 102-109.
Mitchelson F (1992) Pharmacological agents affecting emesis. A review (Part I). Drugs 43: 295-315. [crossref]
Cohen MM, Duncan PG, DeBoer DP, Tweed WA (1994) The postoperative interview: assessing risk factors for nausea and vomiting. Anesthesia and Analgesia 78: 7-16. [crossref]
Apfel CC, Läärä E, Koivuranta M, Greim CA, Roewer N (1999) A simplified risk score for predicting postoperative nausea and vomiting: conclusions from cross-validations between two centers. Anesthesiology 91: 693-700. [crossref]
Apfel CC, Heidrich FM, Jukar-Rao S, Jalota L, Hornuss C, et al. (2012) Evidence-based analysis of risk factors for postoperative nausea and vomiting. British Journal of Anaesthesia 109: 742-753. [crossref]
Sinclair DR, Chung F, Mezei G (1999) Can postoperative nausea and vomiting be predicted? Anesthesiology 91: 109-118. [crossref]
Eberhart LHJ, Geldner G, Kranke P, Morin AM, Schäuffelen A, et al. (2004) The development and validation of a risk score to predict the probability of postoperative vomiting in pediatric patients. Anesthesia and Analgesia 99: 1630-1637. [crossref]
Eberhart LH, Morin AM, Guber D, Kretz FJ, Schäuffelen A, et al. (2004) Applicability of risk scores for postoperative nausea and vomiting in adults to paediatric patients. British Journal of Anaesthesia 93: 386-392. [crossref]
Koivuranta M, Läärä E, Snåre L, Alahuhta S (1997) A survey of postoperative nausea and vomiting. Anaesthesia 52: 443-449. [crossref]
Kranke P, Eberhart LH, Toker H, Roewer N, Wulf H, et al. (2007) A prospective evaluation of the POVOC score for the prediction of postoperative vomiting in children. Anesthesia and Analgesia 105:1592-1597. [crossref]
Apfel CC, Philip BK, Cakmakkaya OS, Shilling A, Shi YY, et al. (2012) Who is at risk for postdischarge nausea and vomiting after ambulatory surgery? Anesthesiology 117: 475-486. [crossref]
DA Silva HB, Sousa AM, Guimarães GM, Slullitel A, Ashmawi HA (2015) Does previous chemotherapy-induced nausea and vomiting predict postoperative nausea and vomiting? Acta Anaesthesiologica Scandinavica 59: 1145-1153. [crossref]
Apfel CC, Kranke P, Katz MH, Goepfert C, Papenfuss T, et al. (2002) Volatile anaesthetics may be the main cause of early but not delayed postoperative vomiting: a randomized controlled trial of factorial design. British Journal of Anaesthesia 88: 659-668. [crossref]
St Pierre M, Dunkel M, Rutherford A, Hering W (2000) Does etomidate increase postoperative nausea? A double-blind controlled comparison of etomidate in lipid emulsion with propofol for balanced anaesthesia. Eur J Anaesthesiol 17: 634-641. [crossref]
Laskowski K, Stirling A, McKay WP, Lim HJ (2011) A systematic review of intravenous ketamine for postoperative analgesia. Canadian journal of anaesthesia. Journal Canadien d’anesthesie 58: 911-923. [crossref]
Subramaniam K, Subramaniam B, Steinbrook RA (2004) Ketamine as adjuvant analgesic to opioids: a quantitative and qualitative systematic review. Anesthesia and Analgesia 99: 482-495. [crossref]
Tramèr M, Moore A, McQuay H (1996) Omitting nitrous oxide in general anaesthesia: meta-analysis of intraoperative awareness and postoperative emesis in randomized controlled trials. British Journal of Anaesthesia 76: 186-193. [crossref]
Apfel CC, Kranke P, Eberhart LH, Roos A, Roewer N (2002) Comparison of predictive models for postoperative nausea and vomiting. British Journal of Anaesthesia 88: 234-240. [crossref]
Roberts GW, Bekker TB, Carlsen HH, Moffatt CH, Slattery PJ, et al. (2005) Postoperative nausea and vomiting are strongly influenced by postoperative opioid use in a dose-related manner. Anesthesia and Analgesia 101: 1343-1348. [crossref]
Cheng CR, Sessler DI, Apfel CC (2005) Does neostigmine administration produce a clinically important increase in postoperative nausea and vomiting? Anesthesia and Analgesia 101: 1349-1355. [crossref]
Ferrari LR, Donlon JV (1992) Metoclopramide reduces the incidence of vomiting after tonsillectomy in children. Anesthesia and Analgesia 75: 351-354. [crossref]
Paxton D, Taylor RH, Gallagher TM, Crean PM (1995) Postoperative emesis following otoplasty in children. Anaesthesia 50: 1083-1085. [crossref]
Villeret I, Laffon M, Duchalais A, Blond MH, Lecuyer AI, et al. (2002) Incidence of postoperative nausea and vomiting in paediatric ambulatory surgery. Paediatric Anaesthesia 12: 712-717. [crossref]

Effect of Nitrogen and Phosphorus Fertilizer Rates on Yield and Yield Components Sorghum (Sorghum bicolor L. Moench) at Kersa Woreda of Oromia Region

Leave a reply

DOI: 10.31038/AFS.2022434

Abstract

Soil fertility is among the most important constraints that threaten sorghum production in Jimma Zone in Oromia Region. Therefore, a field experiment was carried out at Kersa woreda for four consecutive cropping seasons from 2013/14 to 2016/17 to evaluate the response of various levels of nitrogen (N) and phosphorus (P) fertilizer using sorghum. The treatments consisted of factorial combinations of four rates each of N (0, 23, 46 and 69 kg N ha⁻¹) and P (0, 11.5, 23 and 34.5 kg P₂O₅ ha⁻¹) laid down in a randomized complete block design (RCBD) with three replications. For data analysis, correlation coefficient and ANOVA were used. The result showed that the yield and yield components of the sorghum crop were highly significant response to impacts of inorganic N-P fertilizer nutrients. Parameters, such as, plant height, head weight, grain yield, biomass yield and stover yield, were statistically significantly different by nitrogen and phosphorus different fertilizer rates. Also, these parameters were significant and positive correlation to each other. This result revealed that the highest (4.14 t/ha) grain yield was obtained from 69 kgh^-1 N and 23 kgha^-1 P₂O₅inorganic fertilizer, whereas the lowest (1.37 t/ha) grain yield was recorded from control treatment. Compared to the control treatment, the highest rate of N/P (69/23 kg ha^-1) increased sorghum grain yield by about 202.2%. It is concluded that nitrogen and phosphorus at the rate of 69 kgh^-1 N and 23 kgha^-1 P₂O₅has the best performance in obtaining maximum grain yield of sorghum crop. Therefore, N-P at the rate of 69 kgh^-1 N and 23 kgha^-1 P₂O₅is highly recommended for optimum grain yield of sorghum crop in the study area.

Keywords

fertilize, Grain yield, Nitrogen, phosphorus, sorghum

Introduction

Globally, sorghum (Sorghum bicolor L. Moench) is the fifth most vital rice, wheat, barley and cereal crop after maize [1,2]. It is an important crop staple food crop in the semi-arid tropics of Africa, south Asia and Central America [3]. In Ethiopia, sorghum is a major staple food crop, ranking second after maize in total production. It ranks third after wheat and maize in productivity per hectare, and after Teff and maize in area cultivated. It is grown in almost all regions, covering a total land area of 1.8 million ha [4]. Sorghum grain is as nutritious as other cereal grains; contains about 11% water, 340 k/cal of energy, 11.6% protein, 73% carbohydrate and 3% fat by weight [5,6].

Despite the large-scale production and various merits, Sorghum production and productivity have been far below the potential. Currently the average regional productivity is 2.1 t ha^-1 but, the study area productivity is below 1.3 t ha^-1 which is very low as compared to other small grain cereals grown in Ethiopia [4]. Low productivity of crops has been attributed to abiotic stress for drought and low soil fertility and biotic stress for the disease, insect and weeds [7,8].

Soil fertility is one of the major production constraints in the Ethiopia. Deficiency of nitrogen and phosphorus is the main factor that severely reduces the yield of sorghum [9]. According to [10], although soil fertility status is dynamic and variable from locality to locality, and it is difficult to end up with a blanket recommendation invariably, some soil amendment studies were undertaken at different times and places. In addition to fertilizer rates, soil acidity also affects the productivity of the land by affecting availability of nutrients and hindering the activity of microorganisms. Also nitrogen is commonly the most limiting nutrient for crop production in the major world’s agricultural areas and therefore adoption of good N management strategies often result in large economic benefits to farmers. The various factors accounting for the poor soil fertility include topography, soil erosion, deforestation, population pressure, and continuous cultivation without proper soil fertility maintenance [11-13]. Among the major constraints on increased production of sorghum are poor soil fertility, limited supply of production inputs, low prices for the produce, and undeveloped markets [7]. The application of inorganic P fertilizer increased the efficient utilization of inorganic N fertilizer by the plants in grain yield and total biomass production; also, P nutrition of soils is critical for the efficient use of inorganic N fertilizer, thus deficiency in soil P limited the efficient use of applied N by the plant [14]. There was limited research conducted concerning fertilizers rates as a result of this fact, the farmers rely on traditional practices. Most of the farmers in experiment site do not use NP combination above the recommended rate. Therefore, there is a need to study the effect of different NP rates on the yield and yield components of sorghum.

Materials and Methods

Description of the Study Area

The experiment was conducted at Kersa woreda, Jimma zone of the Oromia national state for four consecutive cropping seasons from 2013/14 to 2016/17. The site is located at about 28 km east from Jimma town and 7° 40′ 0″ N latitude and 36° 50′ 0″ E longitude at an average elevation of 1740 to 2660 m amsl and average maximum and minimum temperature is 28.8°C and 11.8°C respectively and reliably receives good rains, ranging from 1,200 – 2,800 mm per annum cropping season. The middle and high altitude soils are less rich in nutrients due to the fact that they have been under human land use for long. The zone is one of the major coffee growing areas of Oromia region well-endowed with natural resources contributing significantly to the national economy of the country. The major crops grown, other than coffee, are maize, teff, sorghum, barley, pulses (beans and peas), root crops (enset-false banana and potato) and fruits.

Description of the Experimental Materials

Plant Materials

In the present study, Sorghum varieties Abamelko which adapted to the agro-ecology of the area were used. Varieties Abamelko is the most promising released by Jimma Agricultural Research Centre. It has wider adaptability and grows well at altitudes ranging from 1740 to 2660 meters above sea level with annual rain fall of 1,200 – 2,800 mm.

Experimental Design and Treatments

The experiment was conducted in Kersa district of Oromia Region in 2013/204 to 2016/2017 cropping seasons. The treatments consisted of four N levels (0, 23, 46 and 69 kg N ha^-1) and four P levels (0, 11.5, 23 and 34.5 kg P₂O₅ ha^-1). The experiment was arranged in randomized complete block design (RCBD) with three replications in a factorial arrangement. The plot size was 14.25 m² (3.75 m × 4 m) and consisted of 5 rows. A distance of 0.5 m and 1 m were left between plots and blocks, respectively. The spacing of 75 × 15 cm was used between rows and plants, respectively and there were 26 plants planted per row with a total of 133 plants per plot. Urea and Triple supper phosphate (TSP) were used as a source nitrogen and phosphorus fertilizer, respectively. Nitrogen fertilizer was applied by split; application method in the form of urea half at planting and the remaining 45 days after planting. Phosphorus was applied once in the form of TSP at the time of planting. Agronomic practices such as weeding and cultivation were done uniformly for all treatments as per need.

Data Collection and Measurement

Plant height (cm) was measured at physiological maturity from the ground level to the tip of head from ten randomly taken plants and was averaged on per plant basis. Head weight (g): samples of ten heads were weighed after harvesting and sun drying to determine weight per head. Weight was adjusted to 12.5% moisture level. Grain and stover yields (t/ha) were determined by harvesting the entire net plot area and converted into tons per hectare. Grain yield was adjusted to 12.5% moisture level; whereas stover yield was weighed after leaving it in open air for 7 days. The biomass yield (t/ha) was calculated as the sum of the grain and stover yields.

Data Analysis

Collected data were subjected to analysis of variance (ANOVA) by using Statistical Analysis System Software Version 9.3 [15] and significant treatment means was separated using Least Significance Difference (LSD) test and correlation coefficient within and between yield and agronomic parameters was done.

Results and Discussion

Effect NP Fertilizer Combination on Plant Growth, Yield and Yield Components Sorghum

Plant Height

The mean of plant height and the analysis of variance are shown in Table 1. There were highly significant variations (p ≤ 0.001) among the fertilizers types on sorghum height. The Application of NP fertilizer linearly and significantly increased plant height as compared to the control (no application any fertilizer). Similarly, the highest mean value (197.42 cm) was obtained from the application of 69 kg ha^-1N and 34.5 kg ha^-1P₂O, while the lowest mean value (157.57 cm) was recorded from the control treatment (Table 1). The treatment increased mean value of plant height by about 25.29% as compared with unfertilized plots. Plant height was significant and positive correlation with stover yield (Table 2). This result suggests that Plant height increased with increasing application of NP fertilizer rates.

Table 1: Effect nitrogen and phosphorus fertilizer rates on sorghum height

Treatment (NP)	Plant height (cm)
Treatment (NP)	2013/2014	2014/2015	2015/2016	2016/2017	Over year
0/0	171^f	1.32^e	149^h	177.93^e	157.57ⁱ
0/11.5	178^ef	135^e	156^h	179.33^e	162.16ⁱ
0/23	184^edf	140^de	165^g	197.80^cab	171.72^h
0/34.5	190^cedb	153^cdb	169g^f	181.38^de	173.51^hg
23/0	186^cedf	146^cde	168^g	188.80^cde	172.12^h
23/11.5	192^cedb	159^cba	169^gf	195.07^cb	178.57^hfg
23/23	200^cadb	155^cdb	170g^ef	197.71^cab	180.54^defg
23/34.5	206^ab	156^cdba	178^dec	200.53^cab	185.02^debfc
46/0	190^cedb	160^cba	176^def	192.78^cdb	179.56^efg
46/11.5	202^cab	154^cdb	177^def	194.89^cb	181.78^defc
46/23	202^cab	167^ab	185^bc	190.67^cdbe	186.08^debc
46/34.5	206^ab	163^ab	181^dbc	198.56^cab	186.93^dbc
69/0	203^cab	164^ab	184^dbc	200.36^cab	187.76^bc
69/11.5	207^ab	166^ab	188^ba	187.67^cde	187.16^dbc
69/23	209^a	167^ab	189^ba	202.62^ab	191.99^ba
69/34.5	213^a	171^a	196^a	209.78^a	197.42^a
LSD (0.05)	17.4	16.03	6.18	13.332	7.1082
CV (%)	5.32	6.18	2.81	4.13	2.36

Values followed by the same letter within a column are not significantly different at P<0.05

Table 2: Pearson correlation coefficients between selected sorghum plant parameters

Sorghum Plant Parameters	PH	HW	GY	TBMY	STY
PH	1
HW	0.39*	1
GY	0.46**	0.91**	1
TBMY	0.37*	0.94**	0.90**	1
STY	0.35*	0.91**	0.85*	0.97**	1

**P ≤ 0.001; *P ≤ 0.05; PH: Plant Height; HW: Head Weight; GY: Grain Yield; TBMY: Total Biomass Yield

The possible reason for the increased plant height of sorghum over the control in response to the mixed application of the fertilizers might be attributed to the released major nutrients and improved soil physical property in enhancing plant growth owing to their contribution to enhanced cell division, stem elongation, promotes leaf expansion and vegetative growth of plant. This result is in agreement with [16] who reported that plant height of sorghum was increase with increasing rates of NP from 0/0 to 69/46 kg ha^-1. This study is in agreement with [17] who reported that application of NP fertilizer at rate 49/46 kgha^-1 significantly increases the plant height of sorghum, and height of sorghum was 18.21 cm greater than control treatment at rate 69 kg N and 46 P₂O₅ha^-1fertilizer. It can be concluded the increased rates of N, and P increased the plant growth and biomass, and also the increased amounts of N-P increases the production of sorghum crop [18].

Head Weight

Head weight was highly significantly affected (p≤ 0.001) by different rates of NP application (Table 3). The results indicated that head weight was linearly increased with increasing levels of NP fertilization from 0/0 to 69/23 kg NP₂O₅ ha^-1. The maximum head weight (93.17 gm) was recorded from application of 69/23 kg NP₂O₅ ha^-1, whereas zero the minimum (29.13 gm) was obtained zero application, which was significantly lower than the effect of other rates. Thus, the combination of N and P at 69/23 kg ha⁻¹ resulted in about 219.7% higher head weight compared with the application of no fertilizer (Table 3).

Table 3: Effect nitrogen and phosphorus rates on head weight of sorghum

Treatment (NP)	Head Weight (t/ha)
Treatment (NP)	2013/2014	2014/2015	2015/2016	2016/2017	Over year
0/0	25.43^j	21.1^d	43.16^g	26.85^e	29.13^e
0/11.5	33.13^ij	21.7^cd	46.01^fg	34.76^de	33.90^e
0/23	35.19^ihj	41.44^cbd	47.74^fg	48.26^cde	43.16^de
0/34.5	39.48^ihfgj	51.91^cabd	62.28^efg	59.87^cdbe	53.39^dc
23/0	38.31^hgj	55.18^cabd	67.80^ed	52.83^cde	53.53^dc
23/11.5	41.99^iehfg	56.42^cabd	64.64^efd	57.76^cdbe	55.20^dc
23/23	48.07^ehfgd	60.04^ab	82.50^cd	72.77^cba	65.84^bc
23/34.5	53.84^ed	60.77^ab	81.40^ecd	63.65^cdba	64.91^bc
46/0	44.15^iehfgd	55.79^cabd	97.31^cb	74.35^cba	67.90^bc
46/11.5	50.67^ecfgd	60.21^ab	88.84^c	57.94^cdbe	64.41^bc
46/23	82.89^a	79.79^a	116.41^ab	90.35^ba	92.36^a
46/34.5	57.29^cd	60.93^ab	115.88^ab	75.57^cba	77.42^ba
69/0	56.79^cd	58.79^cab	82.57^cd	67.9^cdba	66.51^bc
69/11.5	52.90^ecfd	80.15^a	116.59^ab	79.23^cba	82.22^ba
69/23	75.79^ab	83.27^a	118.58^a	95.06^a	93.17^a
69/34.5	62.40^ab	83.41^a	89.72^c	73.95^cba	77.37b^a
LSD (0.05)	14.31	37.524	19.321	33.431	18.565
CV (%)	17.2	38.67	14.02	31.11	17.45

Values followed by the same letter within a column are not significantly different at P< 0.05

This might be attributed to large quantities of nitrogen and phosphorus is translocated from the other plant parts to the grain as it develops heads. This current result is in agreement with [19] who indicated that Sorghum with headings increased consistently with increasing rates of application of inorganic N-P fertilizers from 58 to 121.67. In the same way, head weight was significant and positive correlation with grain yield (Table 2). The present study is similar to the findings of [20] who reported that head weight had the highest direct effect on grain yield sorghum, and also there is significant and high positive correlation between grain yield and head weight (r=0.976).

Grain Yield

Application of nitrogen and phosphorus fertilizers highly significantly (p ≤ 0.001) influenced grain yield. Mean values of the data showed that maximum grain yield (4.14 t/ha) was produced by the treatments of 69/23 kg ha^-1NP inorganic fertilizers. However, the control plots resulted in minimum grain yield (1.37 t/ha) (Table 4). Compared to the control treatment, the highest rate of N/P (69/23 kg ha^-1) increased sorghum grain yield by 202.2%. Hence, application of 69/23 N and P fertilizer rate gave the highest sorghum yield all years (Table 5). Also, grain yield was significant and positive correlation growth parameters, yield and yield components of sorghum (Table 2).

Table 4: Effect nitrogen and phosphorus rates on grain yield of sorghum

Treatment (NP)	Grain Yield (t/ha)
Treatment (NP)	2013/2014	2014/2015	2015/2016	2016/2017	Over year
0/0	1.40^j	1.42^h	1.56ⁱ	1.30^f	1.37^h
0/11.5	1.88^ij	1.50^h	1.74ⁱ	1.41^ef	1.47^h
0/23	2.01^ij	1.51^h	2.33^h	1.63^def	1.90^g
0/34.5	2.28^hifjg	2.35^g	2.58^hg	2.50^cadbef	2.55^fe
23/0	2.21^hijg	2.63^gf	2.85^fg	1.93^cdef	2.46^f
23/11.5	2.44^heifg	2.63^gf	2.98^f	2.57^cdbe	2.76^fed
23/23	2.82^hefdg	2.66^gfe	3.01^fe	2.68^cadb	2.81^fed
23/34.5	3.19^ecd	3.03^dfe	3.07^fe	2.26^cdbef	2.83^ed
46/0	2.58^heif	3.10^cdfe	3.09^fe	2.62^cadbe	2.95^d
46/11.5	2.99^ecfdg	3.22^cdbe	3.29^de	2.33^cdbef	2.98^d
46/23	5.01^a	3.68^ba	4.31^a	3.40^ab	3.95^ba
46/34.5	3.40^cd	3.52^cdba	3.46^dc	3.04^cab	3.53^c
69/0	3.37^cd	3.52^cdba	3.59^c	2.97^cab	3.35^c
69/11.5	3.13^ecfd	3.43^cdba	4.14^ba	3.26^cab	3.52^c
69/23	4.57^ab	3.80^a	4.33^a	3.56^a	4.14^a
69/34.5	3.72^cb	3.66b^a	4.01^b	2.62^cadb	3.61^bc
LSD (0.05)	0.9013	0.5747	0.2877	1.2097	0.3636
CV (%)	18.39	12.07	5.48	28.96	7.55

Values followed by the same letter within a column are not significantly different at P<0.05

Similar result was observed in [17] that there is significant increase in grain yield of sorghum when supplied with higher rates of NP fertilizer. This result is in agreement with [19] who reported that application of 92/30 kg ha^-1NP fertilizer rate increased the grain yield of sorghum. Also, this study is in agreement with the finding of [14] who stated that increasing N rates significantly increased grain and total dry biomass production, whereas the application of inorganic P fertilizer increased the efficient utilization of inorganic N fertilizer by the plants in grain yield and total biomass production.

Total Biomass Yield

The total biomass yields of sorghum were highly significantly (p≤ 0.001) affected by inorganic N-P fertilizers rates (Table 5). Total biomass yield was increased with applying NP fertilizers. Accordingly, the maximum mean total biomass yield (10.27 t/ha) of sorghum was recorded from application of 69/23 NP fertilizer, while the lowest mean total biomass yield (3.52 t/ha) of sorghum was recorded from control without application of any fertilizer (Table 5). The current result indicated that NP fertilizer increased the biomass yield by 191.76% over control treatments.

Table 5: Effect nitrogen and phosphorus rates on total biomass yield of sorghum

Treatment (NP)	Total Biomass Yield (t/ha)
Treatment (NP)	2013/2014	2014/2015	2015/2016	2016/2017	Over year
0/0	4.30ⁱ	3.45^f	5.90ⁱ	3.17^c	3.52^g
0/11.5	5.07hⁱ	4.06^ef	6.18ⁱ	3.65^c	4.07^fg
0/23	5.89^ghfi	4.27^def	6.95^h	6.42^bc	5.07^efg
0/34.5	6.61^ghf	6.15^dec	7.78^g	7.68^bc	6.13^edf
23/0	6.38^ghfi	6.42^dc	7.87^g	6.75^bc	6.00^edf
23/11.5	6.42^ghfi	6.92^bc	8.52^f	8.80^ba	6.69^ed
23/23	7.79^gdfe	7.26^abc	9.73^e	9.05^ba	7.26^edc
23/34.5	10.09^cab	7.71^abc	10.04^de	7.70^ba	7.64^dbc
46/0	5.763^ghi	7.83^abc	10.14^de	9.13^ba	6.99^edc
46/11.5	6.793^ghe	7.90^abc	10.56^dc	7.47^ba	6.91^edc
46/23	12.20^a	8.79^ab	12.82^a	10.60^ba	9.70^ab
46/34.5	8.85^cdbe	8.92^ab	10.88^c	9.38^ba	8.36^adbc
69/0	7.94d^fe	8.93^ab	10.89^c	9.08^ba	8.17^adbc
69/11.5	8.033^dfe	8.92^ab	12.06^a	12.92^a	9.23^abc
69/23	10.323^ab	9.40^ab	13.10^a	13.34^a	10.27^a
69/34.5	9.61^db	9.16^ab	13.07^a	9.81^ba	9.12^abc
LSD (0.05)	2.154	2.335	0.4632	36.17	2.4202
CV (%)	16.93	19.30	2.84	5.0874	20.17

Values followed by the same letter within a column are not significantly different at P<0.05

The maximum mean total biomass yield recorded from 69/23 NP over the control might be due to more nutrients gained from both NP combined fertilizers. The result was in line with [17] who reported that application of NP fertilizer at rate 69/46 kg ha^-1 increases the grain yield of sorghum significantly. In the same way, study conducted by [19] observed that NP fertilizer application at rate 92/30 kgha^-1 increases the total biomass yield of sorghum by 68.21% advantage over control treatment. This current study is supported by [14] who reported that the application of inorganic P fertilizer increased the efficient utilization of inorganic N fertilizer by the plants in grain yield and total biomass production; also, P nutrition of soils is critical for the efficient use of inorganic N fertilizer, thus deficiency in soil P limited the efficient use of applied N by the plant.

Stover Yield

The stover yield of sorghum was showed highly significant (p≤ 0.001) difference due to treatments (Table 6). Accordingly, the maximum stover yield (8.05 t/ha) of sorghum was recorded from application of 69/23 kgha^-1 NP inorganic fertilizer, while the lowest stover yield (3.58 t/ha) of sorghum was obtained from the unfertilized plot. This 69/23 kgha^-1 NP treatment gave stover yield advantage of 4.47 t/ha (124.86%) compared to unfertilized plots (Table 6).

Table 6: Effect nitrogen and phosphorus rates on stover yield of sorghum

Treatment (NP)	Stover Yield (t/ha)
Treatment (NP)	2013/2014	2014/2015	2015/2016	2016/2017	Over year
0/0	6.47^d	2.03^d	4.33ⁱ	1.87^e	3.58^g
0/11.5	6.55^cd	2.56^cd	4.44ⁱ	2.24^de	3.85^fg
0/23	6.71^cd	2.75^cbd	4.62ⁱ	4.78^cde	4.61^feg
0/34.5	6.80^cd	3.80^cabd	5.20^hg	5.18^cde	5.13^fdeg
23/0	6.80^cd	3.79^cabd	5.02^h	4.82^cde	4.99^fdeg
23/11.5	6.74^cd	4.28^cab	5.53^g	6.23^cdba	5.57^cfde
23/23	6.78^cd	4.60ab	6.73^f	6.37^cdba	5.97^cdeb
23/34.5	6.94^cd	4.68^ab	6.97^fe	5.44^cdbe	5.85^cdeb
46/0	6.76^cd	4.72^a	7.06^fed	6.51^cba	6.10^cdeb
46/11.5	6.84^cd	4.68^ab	7.27^ed	5.14^cde	5.82^cdeb
46/23	8.22^cab	5.11^a	8.51^b	7.20^cba	7.07^cab
46/34.5	8.10^cabd	5.40^a	7.42^d	6.34^cdba	6.64^cadb
69/0	7.06^cd	5.42^a	7.30^ed	6.11^cdba	6.31^cadeb
69/11.5	7.14^cbd	5.48^a	7.92^c	9.66^ba	7.37^ab
69/23	8.85^ab	5.61^a	8.76^ba	9.78^a	8.05^a
69/34.5	9.05^a	5.50^a	9.06^a	7.19^ba	7.49^ab
LSD (0.05)	1.7274	1.9371	0.3709	4.2371	1.7831
CV (%)	14.31	26.39	3.35	42.86	18.12640

Values followed by the same letter within a column are not significantly different at P<0.05

This indicates that presence of greater stover yield of sorghum might be due to the fact that crops supplied with adequate nutrients have more vegetative growth, longer linear growth rate and more dry matter accumulation which directly related to an increment in stover yield. This result is supported by [17] who reported that application of nitrogen with phosphorus increase on plant height, grain yield and biomass yield of sorghum. The increase in stover yield with higher N levels might be due to the increase in grain and total dry biomass of maize with higher N rates [14].

Correlation between Growth Parameters, Yield and Yield Components of Sorghum

A summary of the correlation coefficients of the relationships between selected growth parameters, yield and yield components of sorghum in kersa woreda farmers farm, are presented in Table 2. Accordingly, the result indicates all selected parameters of sorghum were significant and positive correlation with one another. Hence, this result suggests that, sorghum grain yield, plant height, head weight, total biomass yield and stover yield is highly influenced NP inorganic fertilizer. The current study is in agreement with [20] who revealed that there was significant and positive correlation between grain yields, head weight and 1000 grain mass, whereas significant but negative correlation exists between panicle count and panicle length.

Conclusion

Overall, the present study showed that the yield and yield components of the sorghum crop were significant response to impacts of inorganic N-P fertilizer nutrients. Parameters, such as, plant height, head weight, grain yield, biomass yield and stover yield, were statistically significantly different by nitrogen and phosphorus different fertilizer rates. Also, these parameters were significant and positive correlation to each other. Therefore, N and P fertilizers are very vital nutrients in limiting the growth, development and the production of the crops in the study area.

This result revealed the highest (4.14 t/ha) grain yield was obtained from 69 kgh^-1 N and 23 kgha^-1 P₂O₅fertilizer, whereas the lowest (1.37 t/ha) grain yield was recorded from control treatment. Compared to the control treatment, the highest rate of N/P (69/23 kg ha^-1) increased sorghum grain yield by about 202.2%. It is concluded that nitrogen and phosphorus at the rate of 69 kgh^-1 N and 23 kgha^-1 P₂O₅has the best performance in obtaining maximum grain yield of sorghum crop. Therefore, N-P at the rate of 69 kgh^-1 N and 23 kgha^-1 P₂O₅is highly recommended for optimum grain yield of sorghum crop in the study area.

References

FAO (2014) Food and Agriculture Organization of the United Nations.
El-Mageed TAA, Mohammed SA, El-Samnoudi IM, Ibrahirm AEM (2018) Interactive effects of soil salinity and water table depth on soil properties and sorghum (Sorghum Bicolor L. Moench) production. Archives of Agriculture and Environmental Science 3: 15-24.
Kumar V, Chopra AK (2013) Response of sweet sorghum after fertigation with sugar mill effluent in two seasons. Sugar Tech 15: 285-299.
CSA (2015) Area and production of major crops. Addis Ababa.
Thimmaiah SK (2002) Effect of salinity on biochemical composition of sorghum (Sorghum bicolor L.) seeds. Indian Journal of Agricultural and Biochemistry 15: 13-15.
Taylor J, Schober TJ, Bean SR (2006) Novel food and non-food uses for sorghum and millets. Journal Cereal Science 44: 252-271.
Olanite JA, Anele UY, Arigbede OM, Jolaosho AO, Onifade OS (2010) Effect of plant spacing and nitrogen fertilizer levels on the growth, dry-matter yield and nutritive quality of columbus grass (Sorghum Almum Stapf) In Southwest Nigeria. Grass Forage Science 65: 369 375.
Mbwika J, Odame H, EN (2011) Feasibility study on striga control in sorghum.
Ashiono GB, Gatuiku S, Mwangi P, Akuja TE (2005) Effects of nitrogen and phosphorus application on growth and yield of dual purpose sorghum in the dry highlands of Kenya. Asian Journal of Plant Sciences 4: 379-382.
Alemu Lelago, Tekalign Mamo, Wassie Haile and Hailu Shiferaw (2016) Assessment and Mapping of Status and Spatial Distribution of Soil Macronutrients in Kambata Tembaro Zone, Southern Ethiopia. Advances in Plants and Agriculture Research 4: 144.
International Food Policy Research Institute (IFPRI) (2010) Fertilizer and Soil Fertility Potential in Ethiopia. IFPRI, Washington DC, USA.
Kehali Jembere, Tekalign Mamo, Kibebew Kibret (2017) Characteristics of agricultural landscape features and local soil fertility management practices in Northwestern Amhara, Ethiopia. Journal of Agronomy 16: 180-195.
Mohammed Kedir, Taye Kufa, Bayu Dume (2021) Assessment of Soil Chemical Properties and Coffee Leaf Analysis in Goma Woreda of Oromia Region. Agriculture, Forestry and Fisheries 10: 93-101.
Benedicta Y, Fosu‑Mensah, Michael Mensah (2016) The effect of phosphorus and nitrogen fertilizers on grain yield, nutrient uptake and use efficiency of two maize (Zea mays L.) varieties under rain fed condition on Haplic Lixisol in the forest‑savannah transition zone of Ghana. Fosu‑Mensah and Mensah Environ Syst Res 5: 22.
SAS Institute, Inc (2012) The SAS System for Window Release 9.3; SAS Institute, Inc. Cary, NC, USA.
Legesse H, Gobeze L (2015) Growth and grain yield response of sorghum (Sorghum Bicolor L. Moench) varieties to moisture conservation practices and NP Fertilizer at moisture stress area of Amaro, Southern Ethiopia. Ashese Journal of Agricultural Science 1: 001-005.
Sebnie W, Mengesha M (2018) Response of nitrogen and phosphorus fertilizer rate for sorghum (Sorghum bicolor L. Moench) production in Wag-Lasta area of Ethiopia. Archives of Agriculture and Environmental Science 3: 180-186.
Bayu W, Rethman NFG, Hammes PS, Alemu G (2006) Effects of Farmyard Manure and Inorganic Fertiliz- ers on Sorghum Growth, Yield, and Nitrogen Use in a Semi-Arid Area of Ethiopia. Journal of Plant Nutrition 29: 391- 407.
Masebo N, Menamo M (2016) The effect of application of different rate of N-P Fertilizers rate on yield and yield components of sorghum (Sorghum Bicolor): Case of Derashe Woreda, Snnpr, Ethiopia. Journal of Natural Sciences Research 6: 88-94.
Ezeaku and Mohammed (2006) Character association and path analysis in grain sorghum. African Journal of Biotechnology 5: 1337-1340.

Detection of Wheat Stem Rust (Puccinia graminis f.sp. tritici) Physiological Races from Major Wheat Producing Regions of Ethiopia

Leave a reply

DOI: 10.31038/AFS.2022433

Abstract

Wheat is important crop in globally both in production and productivity. It is sourced as food for global population. Due to un-matched wheat production and population growth it is not able to meet the feed. The low production and productivity it is due to fungal disease specially rust (stem rust) disease. The objective of the experiment was to identify and detect Pgt across surveyed areas. The experiment was conducted in 2019/20 main rainy season East Shewa zone, North Shewa zone and Hadiya Zones which has high wheat production potential with suitability environment for the disease development. A total of fifty five infected wheat stem rust samples were collected from the three regions. Only 35 isolates have given produced infectious pustules. Races: TKTTF, TTKTF, TTTTF, TKPTF, TKKTF and TTKTT have been identified. TTTTF, TKTTF and TKKTF are found at all assessed regions of Ethiopia. The collected samples were analyzed at Ambo Agricultural Research Center (AmARC) laboratory for race identification.

Keywords

Distribution, Cultivars, Diversity, Incidence, Races, Rust and severity

Introduction

Wheat is a cereal grains produced and consumed globally [1]. It is one of strategic crop for food security and a source of livelihood in emerging countries [2]. Wheat also remains a major source of dietary calories and proteins [2]. As a result, yield and its cultivation area of wheat have been increased to contribute to total production increase [3]. Ethiopia is experienced and largest wheat producer in sub-Saharan Africa with about 0.75 million hectare. Ethiopia can produce both bread and durum wheat cultivated in the highlands of the country largely in the areas like North West, East and Central parts [4]. However, the relations of yield and production in Ethiopia were not well understood in quantitative terms. At present, wheat is produced both in rain fed and irrigated conditions with an average yield of 2.764 t/ha in 2019 [5], but the yield is still lower the global wheat production about 3.56 t/ha. Wheat stem rust is the major constraints for wheat production in the world and in the Eastern part of Africa particularly Ethiopia and Kenya. Yield loss of stem rust reaches 100% in conducive environment and susceptible varieties during year of epidemics. Countries such as Kenya and Ethiopia experience recurrent epidemics of stem rust due to evolution of new stem rust races [6]. The alternate hosts of P. graminis include Berberis spp., Mahonia spp., P. recondita and Clematis [7]. The sexual cycle produces a great genetic diversity with a large number of virulence genes [8]. Evidence points to the recombination of wheat stem rust and the scabrum rust (P. graminis f. sp. secalis) [9, 10]. Puccinia graminis tritici, evolves and mutates, the popularly grown wheat varieties remain at constant stake of losing their resistance to break the strongest of resistant genes [11]. The first virulent stem rust race which was designated as Ug99 in Uganda in 1999 has threatened wheat production globally [12]. The emergence of new virulent races in East Africa are continue to pose a threat to global wheat production and food security [13]. Most highlands of Ethiopia are considered as a hot spot for the development of stem rust complex [14]. Stem rust race are dominant and widely distributed in different regions with high frequency [15]. Several historical events were happened in parts of Ethiopia in recent stem rust caused great losses: stem rust epidemics in 1975 on variety Laketch; in 1992/93, on variety Enkoy; in 1994, on variety Kubsa; and, in 2013, on variety Digelu. The epidemics are due to the appearance of new races as a result of mutation and sexual recombination. To minimize the threat of epidemics, it is important to characterize the race composition of the pathogens and the appearance of new races in the Ethiopia. So it is aimed to identify the Puccinia graminis f.sp. tritici physiological races distribution across different areas and identify seedling resistance test.

Materials and Methods

The field assessment was carried out during 2019/20 main cropping season in three major wheat growing areas such as East Shewa zone, North Shewa zone and Hadiya Zones which are selected based on wheat production potential and highly suitable environment for the disease development. During assessment farmer’s field, Farmer’s Training Center (FTC) and agricultural research wheat stations with different crop growth stages based on Zadoks growth stage (0-9) key (Table 1).

Table 1: Agro-ecological descriptions of areas

Zone	Districts	Coordinates		Altitude (m.a.s.l)	Temperature (°C)		RF (mm)
Zone	Districts	N	E	Altitude (m.a.s.l)	Min.	Max.	RF (mm)
East shewa zone	Ada’a	08°44′′	38°58′′	1950	8°C	28°C	851
	Gimbichu	08°58′′	39°06′′	2450	9°C	29°C	1200
	Lume	08°12′′	39°17′′	1900	9.2°C	29.3°C	951
Hadya zone	Lemo	07°30′′	37⁰55′′	2001	13°C	26°C	1150
	Misha	07°56’′	38°52′′	2143	10.5°C	22.5°C	869
	Duna	07°20′′	37°39′′	2453	12°C	24°C	932
North shewa zone	Moret ena Jiru	09°36′′	39°38′′	2828	6.1°C	24°C	890
	Basona werana	10°41′′	39°47′′	2828	13.5°C	21.5°C	1000
	Minjar	08°45′′	39°15′′	2120	13°C	29°C	854

Stem rust severity and incidence was made at five points along the two diagonals (in an “W” pattern) of the field using 1m x 1m (1m²) quadrant and used to calculate average values. The stem rust incidence was calculated using the number of infected plants and expressed as a percentage of the total number of plants assessed.

The disease severity was measured as a percentage of stem area infected by rust disease according to Modified Cobb’s scale (Figure 1). The severity of the disease was examined on randomly selected five plants in quadrant.

Figure 1: Rust severity estimation on leaves and stem of wheat. A. Percentage occupied by uredinia. B. Rust severities by Peterson et al. (1948). Source: Roelfs et al. (1992)

In addition to the disease parameters, agronomic and geographical data were recorded. Data on geographical information including latitude, longitude and elevation of each field were recorded using Garmin 600 model GPS.

Sample Collection, Isolation and Multiplication of Single-pustules Pathogen

Infected wheat stem sheath and leaf were cut into pieces of 5-10 cm in length using scissors then placed in paper bags. The collected samples were labeled with the zone, district, variety and date of collection then transported to Ambo Agricultural Research Center (AmARC) laboratory for race identification. Sterilized soil composed of three growing Medias; sand, soil and farmyard manure mixed at the ratio of 2:1:1 by volume were used. “McNair” seeds were raised in 5cm diameter pots by spreading the seeds on filter paper in Petri dishes, moisten with water to allow the radicles sprout and sprouted seeds were planted in to growing pots.

Spores collected from rust infected sample after suspension then inoculated onto a week old McNair seedlings [16]. The inoculated seedlings were placed on a table for 30 minutes until the Soltrol evaporate then the seedling is moistened with fine droplets of distilled water and placed in the incubation chamber in a dark at 18-22°C followed by exposure to light for 3-4 hours to facilitate infection. The humidifier switched on for about 1:30 hours, so the seedlings have enough moisture for the whole dark period to condition facilitate the initial infection. Then after, the seedlings were transferred to glass compartments in the greenhouse a temperature of 18-25°C and relative humidity of 60-70%.

Inoculation of Wheat Stem Rust Differential Host Lines

Five seeds of the twenty wheat stem rust differentials with known resistance genes and susceptible variety McNair were grown in 5 cm diameter pots. The required amount 4 mg of spore was prepared in 1ml lightweight mineral oil suspension and inoculated onto a week old seedlings of the differentials. Inoculated plants were moistened with fine droplets of distilled water and placed in an incubation chamber 18-22°C and 98-100% RH. Inoculated seedlings were placed in separate glass compartments greenhouse temperature adjusted with 18°C and 25°C. Natural daylight was supplemented with an additional 4 hrs/day that emitted by cool white fluorescent tubes arranged directly above plants (Figure 2) [7].

Figure 2: Picture captured during the sample collection on severely infected wheat varieties

Determination of Stem Rust Races

Seedling infection types (ITs) were scored 14 days after inoculation using 0 to 4 scoring scale described by [17]. The IT readings of 3 (medium-size uredia with/without chlorosis) and 4 (large uredia without chlorosis or necrosis) were regarded as susceptible. Other readings, i.e. 0 (immune or fleck), 1 (small uredia with necrosis) and 2 (small to medium uredia with chlorosis or necrosis) were resistant (Figure 3).

Figure 3: Infection types of Pgt and host response. Source: Stackman et al. (1962)

Race identification and designation was done using the North American’s nomenclature system for Pgt and grouped the differential lines into five subsets (Table 2). Each isolate was assigned five letter of designation code of [7]. Based on low IT′s isolate that produces on 20 differential lines; the race was designated with a five letter race code BBBBB. Conversely, an isolate that produces a high IT on the 20 differential lines races coded as TTTTT. If an isolate produces a low IT on Sr31 and Sr24, but high infection type on the remaining 18 differential lines, the race was designated as TTTTF.

Table 2: North American’s nomenclature system for Pgt differential wheat lines

*Wheat Pgt* gene differential sets and infection phenotype coding**
Set	Differential lines identified by Pgt resistance gene
Set 1	5	21	9e	7b
Set 2	11	6	8a	9g
Set 3	36	9b	30	17
Set 4	9a	9d	10	Tmp
Set 5	24	31	38	McN
gt-code	Infection phenotype: High: virulent (susceptible); low=avirulent (resistant).
B	Low	Low	Low	Low
C	Low	Low	Low	High
D	Low	Low	High	Low
F	Low	Low	High	High
G	Low	High	Low	Low
H	Low	High	Low	High
J	Low	High	High	Low
K	Low	High	High	High
L	High	Low	Low	Low
M	High	Low	Low	High
N	High	Low	High	Low
P	High	Low	High	High
Q	High	High	Low	Low
R	High	High	Low	High
S	High	High	High	Low
T	High	High	High	High

Source: [7]

Disease Data Collection

Seedling infection types (ITs) data obtained from the race analysis study was used for the identification of races using the North American′s nomenclature system for distribution of Pgt races percentage across the study area (zones), altitude ranges and cultivated varieties was analyzed using descriptive statistics by using Microsoft excel. Infection type data obtained from the seedling resistance analysis of the greenhouse experiment was used to group cultivars under different resistance categories according to [17]. Percentage of durum wheat cultivars having resistant vs. susceptible reaction to selected stem rust races was computed using descriptive statistics in by using Microsoft excel.

Result and Discussion

A total of fifty five infected wheat stem rust samples were collected from the three regions. Of those samples collected, 35 isolates have given produced infectious pustules, but 20 samples didn’t yield viable isolates. Of these isolates, 6 races namely TKTTF, TTKTF, TTTTF, TKPTF, TKKTF and TTKTT were identified (Figure 3). TTTTF, TKTTF and TKKTF are found at all selected regions of Ethiopia while; race TTKTF is only found at North shewa and Hadiya zone of wheat potential production area (Figure 4).

Figure 4: Distribution of Puccinia graminis f.sp. tritici races in assessed areas. Prevalence of puccinia graminis f.sp. tritici races

The diversified distributions of races were prevalent at northern shewa. Out of the samples 35 viable stem rust samples collected; TKKTF race is identified from 17 isolates at about 48.57%; while TTTTF is isolated 8 times. This implies that that TKKTF and TTTTF are the dominantly diversified races in the areas. The race variation at different location wheat varieties grown and environmental conditions [16, 18, 19]. TKKTF was identified form Hidase variety which is wiped out by this race in 2017/18. From the study indicated that race TTTTF was the second most diversified races in the study locations. The second most everyday identified races were TTTTF and TKTTF at a rate of 14.81% each. Race TKKTF was found at all surveyed locations except Ada’a district (Table 3).

Table 3: Amount of races isolated from assessed fields related to cultivated varieties

Race	Field inspected	Percentage of races	Variety
TKKTF	17	30.91%	Mangudo, Kubsa, Kakaba, Land races and Hidase
TTTTF	8	14.55%	Hidase, Kubsa, Mangudo and Tesfaye
TKTTF	4	7.27%	Mangudo and kubsa
TTKTF	3	5.46%	Kakaba and Kubsa
TTKTT	2	3.64%	Kubsa and Mangudo
TKPTF	1	1.82%	Uknown

Altitudinal Variation Influence on Pgt Races Distribution

Altitudinal ranges have influence on distribution of stem rust races diversity. The result revealed that there are variations in stem rust incidence, severity levels and races as well. Much more in number about 22 races with the percentage of 62.86% were identified from high altitudes ranging from 2300-2863 m.a.s.l, conversely; lower number of races was identified at low altitudes ranging from 1500-2300 m.a.s.l. Wide ranges of incidence 0-100% was recorded at higher altitude between 2300-2863 m.a.s.l while the narrow incidence 20-100% was recorded at low altitude between 1500-2300 m.a.s.l (Table 4).

Table 4: Altitudinal variation influence on Pgt races distribution in 2018/2019

Altitude (m.a.s.l)	No. of identified races	Percentage %	Incidence range	Severity range	Host response
1500-2300	13	37.14%	20-100	5-70	MS-S
2300-2863	22	62.86%	0-100	0-70	MS-S
Mean	17.5	50	0-100	0-70	MS-S

Effect of Growth Stage on Pgt Races Distribution

Wheat stem rust distribution is affected by wheat growth and maturity stage depending on food accumulated and prepared in the host plant. Races about 21 in number were identified were detected at dough full growth stage in enough food accumulated (Table 5). Wider ranges of disease incidence 0-100% were recorded at dough stage, while narrow range disease incidence 15-80% was at milk stage. Higher rust severities were recorded at maturity stage 15-70%, but lower severity 5-40% was at milk stage. This implies that wheat growth and maturity stage have influences on rust epidemics.

Table 5: Effect of growth stage on Pgt races distribution in 2018/2019

Maturity stage	No. of identified races	Percentage %	Incidence range	Severity range	Host response
Milk	9	25.71	15-80	5-40	MS-S
Dough	21	60.00	0-100	0-70	MS-S
Maturity stage	5	14.29	100	15-70	MS-S
Mean	11.67	33.33	0-100	0-70	MS-S

As shown in the graph below more races were detected on kubsa with high percentage record. Races about 14 times were identified sample collected kubsa bread wheat variety. Hidase is one of the popular varieties which is one of the severely infected currently about four races with a percentage of 11.43% (Figure 5) have been recorded on samples collected. One of detected variety is kakaba about 7 races has been identified with percentage of 20%. Among this lesser race about 1 and 2.86% was identified on Tesfaye variety. Mangudo is found to be the second variety infected by nine races with percentage 25.71% among others (Figure 5).

Figure 5: Number of races identified and recorded percentage

Summary and Conclusion

Thirty five stem rust isolates were analyzed on twenty stem rust differentials and six stem rust races namely: TKTTF, TTKTF, TTTTF, TKPTF, TKKTF and TTKTT are identified. Race TKPTF was prevailed only at East Shewa zone Ada’a districts, while races TKKTF and TTTTF were dominant isolate from all assessed zones of regions and collected samples which receive the first and second ranks of samples. This implies that TKKTF and TTTTF are the dominantly diversified races at surveyed fields at a rate of 48.15 and 14.55% respectively. Race TKKTF was found at all surveyed locations except Ada’a district. Much more in number about 22 races with the percentage of 62.86% were identified from high altitudes ranging from 2300-2863 m.a.s.l, conversely; lower number of races was identified at low altitudes ranging from 1500-2300 m.a.s.l. Wide ranges of incidence 0-100% was recorded at higher altitude between 2300-2863 m.a.s.l while the narrow incidence 20-100% was recorded at low altitude between 1500-2300 m.a.s.l. In addition to this agro-ecological variation and types of wheat varieties cultivated has greater influence in the presence of races. Survey and surveillance is required to clearly indicate the available Pgt race available in Ethiopia. To develop management methods for each races the breeder is intended to upgrade research on resistance protocols.

Acknowledgment

I would like to thank Ethiopian Institute of Agricultural Research and Debre Zeit Agricultural Research Center for the release of Budget.

Conflict of interest

The author has no declared any conflict of interest

References

Igrejas G, Branlard G (2020) The importance of wheat. In Wheat quality for improving processing and human health.(1-7). Springer, Cham.
Shiferaw B, Smale M, Braun HJ, Duveiller E, Reynolds M, et al. (2013) Crops that feed the world 10. Past successes and future challenges to the role played by wheat in global food security. Food Security5: 291-317.
Tesfaye B, Mesfin K, Tolera A, Gebresilasie H, Gebreyes G, et al. (2019) Some maize agronomic practices in Ethiopia: A review of research experiences and lessons from agronomic panel survey in Oromia and Amhara regions. African journal of agricultural research 14: 1749-1763.
Addis Ababa Chamber of Commerce and Sectoral Associations (AACCSA) (2017) Value chain study on wheat industry in Ethiopia by afro-universal consultant and general trading P.L.C. final report Addis Ababa, January, 2017: 12-59.
CSA (2019) Report on area and production of major crops (For private peasant holding, Meher Season Volume 1). Addis Ababa, Ethiopia: Central Statistical Agency.
Eshete BB (2018) Status and challenges of wheat stem rust (Puccinia graminis F. sp. Tritici) and threats of new races in Ethiopia. Int J For Hortic 4: 22-31.
Jin Y, Szabo LJ, Pretorius ZA, Singh RP, Ward R, et al. (2008) Detection of virulence to resistance gene Sr24 within race TTKS of Puccinia graminis f. sp. tritici. Plant Disease 92: 923-926. [crossref]
Roelfs AP (1992) Rust diseases of wheat: concepts and methods of disease management. Cimmyt.
Burdon JJ, Marshall DR, Luig NH (1981) Isozyme analysis indicates that a virulent cereal rust pathogen is a somatic hybrid. Nat 293: 565-566.
Luig NH, Watson I (1972) The role of wild and cultivated grasses in the hybridization of formae speciales of Puccinia graminis. Aust J Biol Sci 25: 335-342.
Ambika R, Meenakshi D (2018) Wheat Stem Rust Race Ug99: A Shifting Enemy. Int J Curr Microbiol App Sci 7: 1262-1266.
Pretorius ZA, Singh RP, Wagoire WW, Payne TS (2000) Detection of virulence to wheat stem rust resistance gene Sr31 in Puccinia graminis f. sp. tritici in Uganda. Plant Dis 84: 203. [crossref]
Olivera P, Newcomb M, Szabo LJ, Rouse M, Johnson J, et al. (2015) Phenotypic and genotypic characterization of race TKTTF of Puccinia graminis sp. triticithat caused a wheat stem rust epidemic in southern Ethiopia in 2013–14. Phytopathology 105: 917-928. [crossref]
Abebe T, Woldeab G, Dawit W (2012) Distribution and Physiologic Races of Wheat Stem Rust in Tigray, Ethiopia. Journal of Plant Pathology and Microbiology 3: 142.
Hailu A, Woldeab G, Dawit W, Hailu E (2015) Distribution of wheat stem rust (Puccinia Graminis F. Sp. Tritici) in West and Southwest Shewa Zones and identification of its physiological races. Advances in Crop Science and Technology 3: 189.
Roelfs AP (1992) Rust diseases of wheat: concepts and methods of disease management. Cimmyt.
Stakman EC, Stewart DM, Loegering WQ (1962) Identification of physiologic races of Puccinia graminis var. tritici. Washington: USDA 5-50.
Tesfaye L (2016) Seedling resistance to stem rust (Puccinia graminissp. tritici) and molecular marker analysis of resistance genes in some wheat cultivars. plant 6: 16-23.
Peterson RF, Campbell AB, Hannah AE (1948) A diagrammatic scale for estimating rust intensity of leaves and stem of cereals. Can J Res Sect C 26: 496-500.

Breeding and Culture of Macrobrachium rosenbergii; Giant Freshwater Prawn (Scampi), Practiced along the Coast of Kerala, India

Leave a reply

DOI: 10.31038/AFS.2022432

Abstract

The Giant freshwater prawn, Macrobrachium rosenbergii (De Man, 1879), commonly known as Scampi, is one of the most important freshwater prawn species widely cultured in several tropical and sub-tropical countries around the world. It has several attractive attributes as a candidate species viz., fast growth rate, compatibility to grow under poly-/mixed-culture, hardy nature, high market price and demand in both domestic and export markets. Besides, it can also be cultured in low saline brackish water areas (salinity < 10 ppt). It is an indigenous species of India and is naturally occurring in most of the river systems along both coasts of India. It can be cultured alone or with compatible fish species such as Catla (Catla catla) and Rohu (Labeo rohita). It is also a suitable species for incorporating in paddy-cum-fish culture (rice-prawn farming) system. Culture of Scampi can be carried out in earthen ponds, cement tanks and pens.

Keywords

Macrobrachium rosenbergii, Broodstock culture, Seed production, Management

Introduction

The species is characterized by the overlapping of pleura of second abdominal segment over those of first and third segment [1]. It can easily be identified by its large second pair of thoracic legs in male. Rostrum is long and is bent in the middle and upturned distally. The rostral teeth formula is 12-13/11-13 (most common). There are distinct black bands on the dorsal side at the junction of all abdominal segments. In the juveniles, on the lateral sides of the carapace, several horizontal blue/black bands are characteristics of this species (Figure 1) [2-14].

Figure 1: External appearance of Macrobrachium rosenbergii

Distribution

Macrobrachium rosenbergii, a tropical species, is widely distributed in the Indo-Pacific region, ranging from the Indus River Delta through India, Shri Lanka, Bangladesh, Myanmar, Malaysia, Thailand, Vietnam, Indonesia and the Philippines, to Australia and New Guinea. Natural distribution of the species is limited to estuarine and freshwater zones of river mouths and backwaters having temperature usually ranging from 25-34°C and salinity from 0-20 ppt. The species is distributed in the lower stretches of most of the river systems of both the coast of India. It has been introduced in many parts of the world for commercial farming [13].

Habit and Habitat

It is benthic in its habit, sluggish by nature and hides under shades and shelters in the shallow areas of rivers, canals, lakes and ponds during day time to avoid direct sunlight and is very active during night time. It moves slowly and continuously and with slight disturbance jerks backwards and retreats. It is omnivorous, becomes cannibalistic when hungry and has territorial instincts [1,10].

Life Cycle

The giant freshwater prawn has five distinct phases in its life cycle: egg, larva (zoea), post-larva, Juvenile and adult. In nature, juvenile to adult stages are spent in freshwater habitat. Attainment of maturity and mating takes place in freshwater/habitat [1,6]. The egg-bearing (berried) females migrates to brackishwater environment for incubation of fertilized eggs and embryonic development. Hatching and growth of larvae through eleven stages, till they metamorphose to post-larvae, takes place in brackishwater environment. The post-larvae/juveniles ascend to the freshwater zones of the rivers, backwaters, lakes, canals, etc., which are subjected to the tidal influence.

Materials and Methods

Hatchery Production of Seed

Good quality seed is the single most critical input in successful prawn farming as the survival, growth and overall production depends on it. Due to the obligatory requirement of brackish water for hatchery operations, most of the prawn hatcheries are located nearer to the coast. Inland hatcheries mostly use diluted brine (concentrated seawater transported from salt-pans) or synthetic salts to prepare artificial brackish water. After the breakthrough in closing the life cycle of the species in captivity by Dr. SW Ling in 1962, several researchers have developed different types of larval rearing techniques for hatchery production of post-larvae (PL). The most widely used method is clear water technique originally developed by AQUACOP [2-5,8].

ICAR-CIFA has developed and standardized a semi-closed two-phase clear water technology for larval rearing of Scampi. In this technique larval rearing is carried out in two phases (9). In the first phase high density (>200-300 larvae/l) rearing is carried out in smaller tanks (500-1000 L) for 10 to 12 days. In second phase low density (50-60 larvae/l) rearing is carried out in larger tanks (>2000 L) till the entire batch metamorphoses to post-larvae (in 20-25 days). During the first phase the larvae are fed exclusively on live feed Artemia nauplii, whereas during the second phase the dominant feed item is the inert feed such as egg custard, while live-feed is a minor component. Water is exchanged at 50% once every alternate day to maintain water quality. This technique is simple to operate and helps in optimum utilization of space and feed and gives good results with PL output of >35/l.

Steps Involved in Establishing a Hatchery

The following section gives a brief account of the steps involved in establishing a Scampi Hatchery.

Site Selection

A careful selection of site is essential for the successful operation of hatchery in a particular locality. It is also equally important to consider the following essential factors to ensure success in achieving the production target.

Climatic Conditions

Temperature is a key environmental factor for successful operation of hatchery as Scampi is a tropical species. Since the optimum temperature range required during seed production is 28-31°C, the hatchery should be located in tropical or sub-tropical zones. Area selected should have temperature near the optimum range over a minimum period of eight months for profitable operation of hatchery. Besides temperature, rainfall, sunlight, humidity and wind speed at the site are also considered before selecting it for hatchery. Areas vulnerable to natural calamities such as floods, cyclones and earthquakes are not suitable for hatchery construction.

Topography

Assessment of transects, evaluation of slope and determination of the most economic way of constructing hatchery are important. Flat or slightly sloping lands are good and slope close to 2% minimizes the construction cost for broodstock ponds associated with hatcheries. In addition, gravity water-filling and draining from the pond becomes cost-effective and easy.

Soil

Soils that sustain biological activities and have water retention capacity apart from structural stability are considered suitable.

Availability of Adequate Freshwater and Seawater

The hatchery site should preferably be near the coastal areas. Seawater used in the hatchery should be free from pollutants. Seawater can be pumped from surface of the sea or estuary during high tide phases through an in situ filter bed [5]. Saltwater also can be drawn from underground source by sinking deep tube-well fitted with suitable pumps. Freshwater can be drawn from a river/canal/shallow groundwater source. Un-contaminated freshwater is essential for hatchery operations, mainly for broodstock management, for diluting seawater (larval medium) and for general use.

Good Physical Access to the Site

Site should have good all-weather approach-road for facilitating easy and low-cost transportation of construction material, pond and hatchery inputs and for marketing seed.

Uninterrupted Power Supply

Adequate power supply is most important consideration during hatchery activity. Therefore the site should have good proximity to uninterrupted power source.

Hatchery Facilities

The following section give a brief account of the facilities required for a Scampi Hatchery:

Hatchery Building

A proper building or shed based on the scale of operation to house the larval rearing tanks, post-larval holding tanks and Artemia tanks is essential for the successful operation of the hatchery. Small hatcheries may be set up in a shed made up of palmyrah trunk and leaves, or a bamboo framework, but large hatcheries are to be constructed in permanent shed. A low-cost permanent shed should have side walls of brick and cement and flooring with proper drainage facility and should have a mix of asbestos- and translucent fibre-sheets fitted over the roof. The translucent sheets meant for good light penetration should cover around 15% of the total roof area. A common drain of around 24’’ to 30’’ wide and 15’’ to 20’’ deep may be constructed to drain water from all the tanks by gravity.

Water Storage Tanks

Separate cement tanks for storing seawater, freshwater and mixed water (larval rearing water) are required. The tanks for the first two types of water may be constructed outside the hatchery shed, whereas the tank(s) for larval rearing water (salinity 12 ppt) are better located either under a temporary shed or even inside the hatchery proper (in small hatchery) to get water of ambient temperature in the larval rearing tanks. The size and capacity of above three types of tanks will depend on the overall production capacity of the hatchery. Huge quantity of larval rearing water is normally required in a flow through hatchery. Larval rearing medium of about 12-times the total volume of rearing tanks are required for each seed production cycle. The hatchery should have water storage facility of at least 3-times the volume of its larval rearing tanks to allow for adequate water storage, treatment and mixing time for preparation of larval medium. To minimize the costs, tanks are better constructed at ground-level and provision for pumping the water at required place should be made.

Broodstock Holding Unit

Broodstok holding facility may comprise of FRP or cement tanks, the size depends on the capacity of hatchery. These tanks inside the hatchery are for keeping both mature male and female prawns for breeding and for final maturation of eggs, or keeping berried females collected from the wild/grow-out facilities for acclimatization to hatchery conditions. This facility should be separate and away from the larval rearing unit as prawns collected from outside may be infected and need to be given prophylactic treatments.

Larval Rearing Unit

Larval rearing unit should be established as a separate unit so that it is safe and free from any likely outside infection. In large hatcheries, several such small units may be established instead of making a single very large unit to prevent spreading of infection. Larval rearing unit may comprise of large number of tanks made up of FRP, ferro-cement, or cement. Tanks can be circular, rectangular or cylindrico-conical in shape. Usually rectangular tanks of 2 to 10 t capacity are preferred. All right-angled corners should be rounded off to facilitate cleaning and to prevent algal growth. The tank bottom should preferably be ‘U-shaped’ and have sufficient slope so as to drain completely. The interior of the tanks should be painted with several layers of dark coloured pure epoxy resin to prevent leaching of toxic chemicals and to provide smooth surface. The depth of the tanks should be approximately 1.0 m and the water column not more than 0.9 m. The number of larval rearing tanks (LRTs) depends on the hatchery capacity.Tanks should be provided with vigourous aeration from a grid of air-blowers and pipes. The air-stones of all the aeration points should be close to the tank bottom. The tanks should have provision for inlets to receive larval rearing water through pipeline from the larval water storage tanks [6,7,11].

Post-larval Holding Tanks

Rectangular cement or concrete tanks of 2 to 10 t capacity are suitable for holding post-larvae till disposal. The number of such tanks depends on the hatchery capacity. Post-larval Rearing Tanks (PLRTs) also can function as broodstock holding tank. These tanks are better placed outside the main hatchery building to reduce and offset the construction cost, but should be provided all around with green shade-netting (commonly used in green-houses) and covered over a pipe framework fitted at a height of approximately 8-10 feet. Such arrangement will keep tank water free from algal growth and also from dust. The tanks should be provided with separate inlets for freshwater supply and an aeration system.

Artemia Cysts Hatching Tanks

Artemia or Brine Shrimp cysts are a source of pathogens and hence there should be a separate unit for their hatching. The size of the tanks depend on the overall requirement of Artemia nauplii (AN) per day in the hatchery. Cylindro-conical shaped tanks are better from operational point of view. They should have transparent bottom where nauplii could be easily attracted by artificial light (lamp) and drained from bottom outlet. Cylindrico-conical fibreglass reinforced plastic tanks of 100 to 500 litre capacity with a central drain and water control structure can be used as Artemia cysts hatching tanks.

Aeration System

A reliable 24-hour oil-free aeration system is essential for hatchery in order to maintain dissolved oxygen levels in excess of 5 ppm in various units of the hatchery. The air supply is essential in all the tanks used for broodstock holding, hatching, larval rearing, post-larval rearing and Artemia hatching. Although majority of the units require mild aeration, it should be vigorous or rather bumping particularly in larval rearing and Artemia hatching tanks. Three-phase electrically operated air-blowers both roots-type and fan-type can be used but fantype provide better aeration and create relatively less noise. Diesel operated air-blowers could also be used where power supply is either not available or there is frequent failure. The capacity of air-blower should depend on the overall requirement of air in different units or the size of the hatchery. A 200 CFM (5.66 m3/minute) air-blower is sufficient to supply air for a hatchery capable of producing 20 million post-larvae/year. The air-blower or a set of air-blowers should be used at a stretch for a maximum period of 8 hours and switched off, followed by supply from a second set of similar blowers. This practice reduces losses from early wear and tear and avoids sudden aeration failure in the hatchery. The aeration pipeline grid should be installed in such a way and place that it cannot breakdown due to movement of people and other hatchery items. It is better to arrange pipelines overhead and each tank may be provided separate pipeline of smaller diameter pipe of 0.5 to 1.0 inch PVC pipe (dropping from the main pipeline). Each dropping pipeline in turn ought to be provided with small holes for fixing plastic joints and fixing aeration tubings of 1/8 inch diameter, which are provided with air-stones and dropped into the tanks for aeration.

Water Supply System

A separate pipeline both for freshwater and larval-rearing water is essential in the hatchery. Freshwater is required in almost all the hatchery sections, i.e. broodstock holding tanks, hatching tanks, larval-rearing tanks and post-larval tanks for use as media and also for washing the tanks, whereas, larval-rearing water of 12 ppt salinity is required only in the larval tanks (LRTs). Accordingly, separate pipeline for both types of waters would be required with provision for separate inlet near one side of the tank. The pipeline and all other fittings including ball valves should be made up of PVC rigid pipe as metal pipes and fittings are likely to be corroded by saline water and leaching of metal ions may take place in the tanks which may harm the prawn larvae.The diameter of water pipeline shall depend on the volume of water required every day in the hatchery. As a thumb rule, initially from pump side, it may start with 3-6 inch diameter pipe and subsequently reduced to 1-2 inches at the tank inlet. Complete pipe line is to be laid inside the hatchery building so that temperature fluctuations are minimized.The water system is simple and all the storage tanks should be sufficiently elevated above the larval rearing tanks (LRTs) so that brackish water can be introduced by gravity.

Power Back-up System

The hatchery needs round the clock power supply for the operation of aeration and water grids. Power breakdown even for a short duration may cause mortality of hatchery live-stock. Therefore, a back-up power system of sufficient capacity is essential for the hatchery. The diesel generators can support power back-up for sufficient duration. The generators are to be installed at a suitable place slightly away from the main hatchery building to minimize sound and air pollution.

Laboratory

A small laboratory, having working platform for keeping equipment/chemicals/glassware/plasticware, should be established possibly within the main hatchery building for easy approach. The laboratory should be provided with necessary equipment and facilities like refrigerator, salinity refractometer, pH meter, dissolved oxygen (DO) meter, weighing scales (chemical/digital/dial/spring balances), hand lens, different types of microscopes (field/dissection/low-power binocular/compound), pressure cooker, mixie, necessary glassware, plastic-ware and chemicals for estimation of DO, hardness, alkalinity, etc.

Broodstock Management

Scampi broodstock may be procured both from wild and grow-out ponds, in later case, care should be taken that the stock is not under severe inbreeding depression. Raising healthy brooders in the close vicinity or at the hatchery site is ideal. If reared at the hatchery site, the stocking density should be <10,000/ha. Half of the feed ration may be substituted with the equivalent amount of pieces of fresh feeds, such as mussels flesh, cut to the appropriate size, at least twice per week. 1 kg of wet feed is roughly equivalent to 200 g of pellet diet. The feed ration should be given in two equal portions, normally early in the morning and late evening. The pond water should maintain optimum water conditions with partial exchange (30-40%) every fortnightly in case of earthen ponds.

Only berried females in an advance stage of egg-incubation (those carrying grey egg-mass) should be brought to the hatchery for hatching eggs so as to minimize cost of maintenance at the brood holding tanks. The berried females having entire egg mass should be selected and stocked in these tanks .The size of the brood prawn should preferably be 60-100 g in weight. It should be apparently healthy and free from diseases particularly from epibiont fouling, lesions, spots, infected appendages, etc. Brooders should be procured and transported with utmost care so that it does not lead to injury and loss of egg mass. Transport of berried females over shorter duration can be undertaken in buckets or tubs containing water of the same pond. For two to three hours journeys, the broodstock can be transported in open containers having water of the same pond along with some aquatic weeds. The container(s) may be provided aeration from a battery operated portable aerator. For long distance transportation (>12 hours), brooders may be packed in 9 inches (23 cm) long 50 mm dia slotted PVC pipes, tide on both ends with meshed cloth. 3-5 such pipes may then be kept in one polythene bag having 5-6 litre of water and packed with oxygen and transported in a carton. It is recommended to transport the bags in insulated containers to avoid temperature fluctuations and movement. The temperature should be maintained at 25-27°C. The rostrum of each prawn should be blunted with scissors or a rubber cap should be placed on it so that the polythene bag does not get punctured. For transporting in PVC tanks with aeration, a maximum stocking rate of one prawn per 40 litre of water should be maintained.

The berried females should be handled with utmost care after their arrival in the hatchery and also while shifting from one tank to the other. The female should be caught under water using a bucket and keep them immersed in water while shifting to the other tanks. Catching with hands or scoop net result in shedding of egg mass and injury to the female and hence poor hatching performance. The female should be disinfected with formalin (@ 50 ppm) under vigorous aeration for 8-10 hours followed by complete change of tank water for the control of epizoan parasites before putting them in the hatching tank.

Hatchery Operation

Operation of hatchery involves different activities starting from preparation of water till post-larval disposal. The following section briefly describes the steps involved in hatchery operation.

Preparation of Larval Rearing Water

Seawater for larval rearing should preferably be collected from a sea coast having little pollution impact. For transportation of seawater, plastic barrels or FRP tanks are desirable. Transporting by truck-tankers having tank made up of iron may increase iron contents in the water and hence should be avoided. Seawater need to be disinfected for probable pathogens by active chlorine and potassium permanganate @ 5 ppm and 2 ppm respectively after shifting into the treatment tanks under vigorous aeration. Good quality freshwater is also required for preparing larval rearing water of 12-13 ppt salinity from seawater. The prepared mixed water should be disinfected with active chlorine @ 5 ppm under vigorous aeration for at least for 48 hours and the residual chlorine may be removed by adding sodium thiosulphate. The water should then be filtered with 5µ bolting silk cloth bag and used in the larval rearing tanks.

Larval Production and Rearing

The usual practice followed in most commercial hatcheries is stocking a large number of berried females of similar egg colour for hatching in a large tank. However, this is unsafe for many reasons particularly heterogenous size of larvae (zoea), disease spreading, mixing of healthy and unhealthy larvae that would cause problems at later stages. Hence only few berried females required to supply enough larvae for larval rearing tanks should be kept in each hatching tank for minimising chances of spreading pathogens and for production of healthy batch of larvae [11,12].

Hatching tanks should be provided aeration round the clock. Hatching generally takes place in the night and hence freshly hatched larvae (length about 2 mm) are to be harvested as soon as possible through siphoning as the female may consume them if kept for prolong period. Fresh or low salinity water (salinity 3-4 ppt) having pH below 8.3 and temperature 28-3°C should be used in these tanks. The larvae should be disinfected with 15 ppm formalin for 5-10 minutes before shifting to larval rearing tanks.

Larval rearing tanks (LRTs) should be thoroughly cleaned and disinfected with bleaching powder at least two days prior to larvae stocking. The tanks are filled with filtered larval water (salinity 1213 ppt) prepared at least 48 hours before use preferably in indoor conditions of the hatchery. The tanks should be provided with vigorous aeration throughout the tank area and care should be taken to minimize dead ends. This practice helps in uniform circulation of food particles in the tank for easy feeding by the larvae as well as to reduce larval cannibalism due to continuous movement. The stocking density of larvae in the tanks will depend on the rearing methodology adopted. In single stocking method in which zoea larvae are reared to post-larvae, they are stocked at 50-60/litre; in two-phase stocking method, initial larval stocking density is 100 larvae/litre, which is reduced to 50-60/litre, by thinning/shifting, when they reach Stage-V Zoea. Freshly hatched Artemia nauplii (AN) should be fed all the time to all the eleven Zoea Stages (I-XI); however, feeding exclusively nauplii may be cut down after Stage-V Zoea, when egg custard is incorporated in the diet. The quantity of Artemia nauplii and egg custard should be given according to the area of the tank and not by the number of larvae being reared. This is essential because the larvae prey on the food by touching and not by seeing.

The larval tanks should be cleaned daily to remove accumulated debris, left over feed and faecal matter through siphoning. Around 50% of the tank water should be replaced with fresh larval water after cleaning. The larval tanks may be provided with 5-10 g of EDTA (Ethylene diamine tetra acetic acid) per tonne of water for chelation of heavy metals after every 3-5 days. Antibiotics should not be used in the larval tanks and instead use of probiotics is considered ideal. Both live and formulated diets are used in the hatchery for feeding larvae and post-larvae. The live feed used in prawn hatchery are Artemia nauplii and Moina macrura. The later is used only in few hatcheries, where it is cultured in the pure form separately. The formulated diet comprises of egg custard.

Preparation of Live Feed (Artemia nauplii)

Artemia, commonly known as Brine Shrimp, is a small crustacean living in salt pans and high saline water bodies. During unfavourable conditions they produce hard shelled cysts (fertilized eggs). These cysts hatch when provided with favourable conditions. Newly hatched microscopic free-swimming larvae are called nauplii. They form a highly nutritious live diet containing more than 50% crude protein and 12% lipid. The size of nauplii is important for proper use in the larval tanks. Nauplii of Artemia salina of San Francisco Bay and Great Salt Lake (USA) are comparatively very small (~400 µm in length) and considered best for use in the prawn hatchery. Artemia cysts are sold in the market in tin packs of generally one pound weight (454 g). Based on the hatching rates, it is categorized generally into three qualities ranging from 70-95%. Better the hatching rates, less the chances of contamination. Artemia cyst tins are to be stored in deep freezer immediately after the procurement otherwise its nutritional and hatching quality get deteriorated.

Artemia cysts are usually contaminated with bacteria, fungal spores, other micro-organisms and organic impurities that may infect the water of larval rearing tanks if not treated properly. Hence, cysts need to be disinfected before stocking for hatching. The disinfection eliminates the chances of infection. The number of Artemia per unit weight depends on the type of artemia. On an average, Artemia of Great Salt Lake may yield 2.7 lakh and that of San Francisco Bay 3.2 lakh nauplii per gram weight. Find out the hatching rate of Artemia from instructions written on the tin for the first time and subsequently after observing the hatching percentage by manual counting. The Artemia cysts are stocked in the hatching tanks @ 2 g/l seawater, where they hatch out between 12-24 hours. After harvesting, nauplii need to be acclimatized to the salinity of larval rearing water by gradually mixing freshwater. The nauplii should also be treated with 15 ppm formalin for disinfection. Freshly hatched nauplii are to be fed to the prawn larvae as they are rich in nutritional contents in the beginning which gradually reduces with time.

Artemia Enrichment

The nutritional quality and physical size of nauplii vary enormously from source to source and even between individual batches from a single source. Of particular importance is the level of essential polyunsaturated fatty acids, eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), which depends on the composition of primary food available to the brine shrimp in the locations where they originate and is generally found low. In order to provide sufficient quantity of these essential fatty acids, the nauplii are to be enriched with both EPA and DHA. There are various enrichment products available in the market, such as Super Selco, DHA Selco (INVE, Aquaculture), Super artemia (Catvis BV., 5222 AE, Netherland), Super HUFA (Salt Creek Inc, USA), Algamac-2000, Algamac-3050 (Biomarine Inc., USA). The methodology for enrichment is provided with these products.

Egg Custard

Egg custard is provided to advanced larvae (Zoea Stage-V and above). A good quality egg custard can be prepared by mixing whole egg, skimmed milk powder, corn flour/wheat flour, mussel/shrimp/prawn/squid meat, yeast, agar, cod liver oil and vitamin-mineral mixture. All the ingredients are blended in a mixer-grinder and cooked under steam in a pressure cooker for maximum of 15-20 minutes. It should not be over cooked as it will lose its flavour and nutritional quality. The egg custard should be used within 4-5 days of its preparation and the left over portion to be kept in a refrigerator. A measured quantity of egg custard is seived through strainers of different mesh sizes.

The mesh of sieve may be selected from fine to coarse depending on the mouth size of the larvae. Smaller larvae need smaller particles whereas larger larvae require bigger particles of egg custard and accordingly selection of sieve is done. The egg custard is then pressed through the sieve held in some water, just sufficient to accommodate the sieve mesh. Water is then drained leaving particles of egg custard in the container. The particulated custard is again washed with freshwater so that finer particles are drained off completely. Vitamin-mineral mixture is added to the egg custard mash and feed to prawn larvae. Egg custard should be fed 3-4 times during day time and left over particles should be drained out in the evening so that they do not foul the water during long stay in the tank.

Collection of Post-larvae from Larval Tanks

The first post-larva (PL) could be seen in the tank (LRT) after around 18 days of rearing; however, majority of them appear after 25 days depending on the water temperature. When sufficient numbers of Zoea Stage-XI larvae (say 40%) metamorphose to post-larvae they should be harvested and stocked in the post-larval tanks. Delay in harvesting results in cannibalism of the larvae by the post-larvae which are highly cannibalistic in nature. Post-larvae have to be harvested from the larval rearing tanks whenever sufficient numbers appear in LRTs. For collection of postlarvae from the larval tanks, aeration in the larval tanks is to be stopped. By doing this, all the larvae will come to the surface and post-larvae being photo-negative will settle at the bottom from where they could be siphoned out with the help of a flexible tubing. Post-larvae have to be checked for any probable infection at the time of harvesting and if stock is found free from any infection, they should be released in the post-larval tanks. After harvesting from the larval tanks, the postlarvae need to be acclimatized to freshwater conditions gradually.

Post larvae (PLs) should be reared in freshwater tanks (PLRTs) till they attain the desired size/age. They may be fed on commercial prawn starter diet specially prepared for them. Around 50% tank water is to be changed daily and all leftover feed, faecal matter and debris to be removed at the same time. The growth of post-larvae may be checked by observing their moulting on a regular basis. Harvesting of post-larvae should be done when they attain the desired size in terms of length. The post-larvae of size above 10 mm is considered ideal for harvesting and stocking in the nursery ponds. The seed of more or less same size should be supplied for stocking in the nursery pond. Post-larvae can be packed in polythene bags containing 4-5 litre water and oxygen and kept in cardboard cartons for transport. For long distance transportation of >12 hours, a sachet of cooling gel is placed in each seed pack container for maintaining temperature.

Hatchery Hygiene and Prophylactics

Prawn larvae are highly susceptible to pathogens and lot of mortalities are often observed in the hatcheries.

Therefore, strict surveillance is needed to avoid entry of pathogens that come both from outside and inside. As a first and foremost control measures, the entry of people in the hatchery should be restricted only to the workers of the hatchery. Soak pits to be constructed at all the entry points which are cleaned daily and filled with water containing disinfectants like bleaching powder. Similarly, wash basins containing sanitizers should also be available at the entry points. Everyone should wash their hands and feet before entering the hatchery. Lot of tools like hapa, hand nets, sieves, cloth pieces, etc, which are used in the hatchery tanks should be washed and disinfected before using in other tanks. The entry of such items should be limited to one unit of the hatchery only and should not be allowed to be used in the other unit. All the tanks of the hatchery should be washed before and after use with clean freshwater and disinfected with bleaching powder or iodophor substances. The components and equipment used in the hatchery should be washed and disinfected after every use.

V.Grow-out Culture

Scampi can be cultured either alone (mono culture) or in combination with compatible fishes like Carps, Tilapia, etc (polyculture). Culture can be carried out by direct stocking of post-lavae or stocking juveniles after a nursery phase of 45-60 days. Incorporating a nursery phase has shown improved survival and production during grow-out culture (Figure 2).

Figure 2: Grow out culture ponds with paddle wheel aerators

Nursery Phase

Nursery is the intermediate phase between hatchery and grow-out of freshwater prawn. It involves rearing of the delicate 25-day or older post-larvae (10-20 mm), obtained from hatcheries, in well prepared earthen ponds (0.01 to 0.1 ha) or concrete tanks for a period ranging from 45-60 days till they grow to juveniles (1-2 g). Stocking density ranges from 20 to 50/m2. Higher stocking densities would require aeration or water exchange. Stocking nursery reared larger juvenile prawns in grow-out ponds gives better yield and predictable production than direct stocking of post-larvae. Hence nursery rearing phase is always recommended prior to grow-out culture. Pond preparation and management are similar to that of grow-out ponds except that hide-outs are not provided in nursery ponds. Floating aquatic plants such as Eichhornia sp. may be introduced in a floating bamboo or PVC frame to cover 5-10% of pond surface area. The dense root system of these plants provides shade, shelter and food to growing post-larvae. Good quality commercial pellet feed (Starter-I) is recommended for feeding the post- larvae twice daily. If it is not available, then powdered oilcake and ricebran mixture can also be fed to post-larvae at 100% biomass per day for the first 10 days and slowly reducing the quantity as the prawns grow. Survival rates of 75 to 80% can be achieved during nursery phase under good management practices.

Grow-out Phase

Grow-out phase follows nursery phase where the juveniles harvested from nursery ponds are stocked in well prepared earthen grow-out ponds at a stocking density of 3/m2. As stocking density shows a strong negative relationship with growth, lower stocking densities are preferred if the farmer wishes to harvest larger prawns. Higher stocking densities (>5/m2) will lead to smaller prawns at harvest. The prawns are fed daily with formulated pellet diet (2-3 mm) at 10% of the biomass initially and then reduced to 3% of the biomass at the end of culture period. Monitoring important water quality parameters such as dissolved oxygen, pH and temperature is recommended to prevent loss of stock due to poor water quality. Regular monthly sampling needs to be carried out to assess the growth and health of the prawns as well as to revise the feed ration. After four months, marketable size prawns (>40 g) may be harvested by using large mesh net and this selective harvesting should continue once every 3-4 weeks for another 3-4 months and finally the prawns can be harvested by completely dewatering the pond.

Site Selection

The selected site should have warm climate for nearly 6-8 months (temperature >20°C). It should have a supply of good quality, pollution free freshwater or brackish water (<7 ppt) for at least 6 months. It should have soil with good water retention capacity.

Pond Construction

Ponds should preferably be embankment-type that can be fully drained by gravity.
Ponds should have an inlet and outlet.
Pond bottom should have a gradient/slope (1:200) towards the outlet.
Pond bund should have a suitable slope (1:2, 1:3).
Water control structure should be installed at inlet and outlet to aid water exchange.
Pond size – 0.2 to 1.0 ha (preferably 0.2-0.5 ha).
Rectangular shaped ponds with the long axis oriented in the direction of prevailing wind are most suitable.
Soil – clayey loam, sandy loam. Depth – 2 m.

Eradication of Competitors and Predators

This step may not be necessary in newly constructed ponds but in old ponds, all unwanted species such as predatory fishes, weed fishes and aquatic weeds should be removed.
Drying and exposing the pond bottom until cracks developed is the best way of eradicating predators and competitors.
Drying and exposing the pond bottom also kill pathogenic microbes and helps in oxidizing the pond bottom.
Poisons of plant origin such as mahua oil cake, tea seed cake or derris root powder may be applied in un-drainable ponds to kill predators and unwanted fishes.

Liming

Liming is an important step in pond preparation and is done after drying the pond by spreading the lime uniformly on the pond bottom.
The rate of application varies with soil pH; to a pond having soil pH above 6 agricultural lime (Calcium carbonate) is applied @ 200250 kg/ha.
Application of lime helps to correct pH; increases the buffering capacity of water; disinfects the pond bottom as well as acts as a source of calcium which is important for exoskeleton formation in prawns.

Fertilization

After liming, the pond is filled with water up to 1-2 feet and manure or fertilizers are applied for development of plankton.
Surface waters from rivers, canals or reservoirs or groundwater from bore-well may be used for culturing freshwater prawns.
A fine-mesh net should be used to screen the inlet water to prevent entry of eggs and larvae of predatory and weed fishes that may colonize the pond and lead to poor growth and survival of the stocked prawn juveniles.
Cow dung @1000 kg/ha or poultry manure @500 kg/ha and super phosphate @100 kg/ha may be applied to initiate plankton development.
The pond can be filled up to the desired level (4-5 feet) after initial manuring.
Manures or fertilizers helps in development of phytoplankton which in turn prevents development of benthic algae and rooted vegetation.
It also helps in development of bottom living animals on which the prawn feeds.

Provision of Hideout and Bird Netting

Prawn needs shelter/hideout during moulting to avoid predation by other prawns. Hence cut branches of trees, nylon screen, earthen pipes etc can be provided as hideout. Hideout materials also provide more surface area for the prawns.
Birds are one of the major predators and can cause significant reduction in survival, so tying nylon ropes or large mesh gill net above the water surface provide some protection from bird predation.

Stocking the Pond

Ponds can be stocked with post-larvae or juveniles after preparing and laying hideouts.
Prawn seed from hatchery needs to be acclimatized at the farm site by floating the transport bag in the pond for 15 minutes. After opening the bag, pond water should be allowed to flow into the bag and post-larvae/juveniles should be slowly released into the pond.
Stocking should be done early morning or late evening which is the ideal period.
A stocking density of 3/m² is desirable, which however may be reduced to 50% in polyculture pond with compatible fish species such as Catla, Rohu, Silver Carp and Grass Carp.

Water Quality Management

Visibility/transparency and colour of the pond is an important indicator of the health of pond ecosystem. In unproductive ponds the visibility can be up to the bottom which will lead to growth of bottom algae that adversely affect the growth and survival of prawns. Low visibility (<10 cm) indicate high blooming or turbidity that could cause problem of oxygen depletion and mortality of stocks. Ideally, the visibility should be maintained in the range of 30-40 cm to avoid water quality deterioration.
Daily monitoring of water quality parameters such as dissolved oxygen, pH and temperature is recommended to prevent loss of stock due to poor water quality. Loss of prawn is usually associated with low dissolved oxygen level in the pond. Therefore it is essential to maintain dissolved oxygen at optimum level of >4 ppm at all times. Provision of aerators (paddle wheel or any other such devices) is recommended especially during the final 2-3 months when the biomass in the pond is high. When the oxygen level in the pond is critically low, the prawns come to the surface along the periphery which indicates the need for taking immediate remedial measures such as water exchange or operation of aerators to avoid mortality of stock.
Water should be free of pollutants and toxic chemicals and the optimum ranges for a few most important water quality parameters for freshwater prawn culture are as follows (Table 1)

Table 1: Optimum Water Quality Parameters for Scampi Farming

Water Parameter	Optimum Range
Temperature (°C)	28-31
Salinity (ppt)	Freshwater/low-saline (<7 ppt)
pH	7.0-8.5
Dissolved Oxygen (ppm)	>4.0
Total Hardness (ppm)	40-100

Feed Management

Freshwater prawns are omnivorous and feed on both animal and plant materials, found on the pond bottom, such as algae, aquatic insects and their larvae, worms, crustaceans, small mollusks, etc.
Farmers may use commercial pellet feed having good water stability or farm made feed. Most commonly used ingredients for farm made feed includes ricebran, broken rice, groundnut oil cake, tapioca powder, fishmeal, apple-snail meat, etc.
Prawns are fed daily at 25% of the biomass during the first two months which is gradually reduced to 3% of the biomass at the end of culture period.
Although feed is usually broadcasted around the periphery of the pond in shallow area, providing of feed in checktrays kept in different areas of the pond will help in determining the quantum of feed required per day.
Feeding should be done during late evening or early morning since prawns are more active during night time.
Feed rate should be revised once every three weeks depending on the average body weight obtained during monthly sampling. Weight dependent feeding rates is given in table as follows (Table 2)
Regular sampling of prawns using cast net or small mesh seine net at 3-4 week interval is essential to assess the growth of prawns. Feed rate is revised after every sampling based on the body weight and estimated biomass in the pond.

Table 2: Optimum Feeding Rate for Scampi in Grow-out Pond

Body Weight of Prawns (g)	Feeding Rate (% Prawn Biomass)
< 2	> 25
2-5	10
5-10	8
10-15	6
15-20	4
20-25	2.5
25-30	2
> 30	1

Health Management

Diseases in freshwater prawn grow-out culture are usually found to be associated with poor rearing conditions (over-feeding, water shortage, silting etc).
Bacteria and fungus are the most common disease causing organisms. Loss of appendages, brown or black colouration of exoskeleton, fouling on the body are some of the symptoms seen in diseased prawns.
If disease symptoms are noted, water should be replaced, water quality should be tested and necessary steps should be taken. Immediate consultation of experts will help in avoiding loss of stock due to diseases.
Following good rearing practices mentioned below will help avoid diseases to a great extent:

– Use good quality seed and avoid high density stocking.

– Use good quality pellet feed, monitor the feeding using check-tray and avoid overfeeding.

– Dry out the ponds between production cycles so that the pond bed can be reoxidized.

– Water exchange (30-50%) helps in rinsing the pond and induces moulting.

– Regular monitoring of water quality especially dissolved oxygen is essential.

Yield and Production Cost

Good quality post-larvae stocked at moderate density 3/m2 and fed with good quality pellet diet will grow to an average size of 50-60 g in 6-8 months.
Periodic harvesting of prawn is always suggested due to heterogeneous growth among prawns. Large prawns (>40 g) may be harvested using seine net of suitable mesh size after four months of culture, which should continue once every 3-4 weeks thereafter for the next 3-4 months.
Final harvesting of the prawns may be done after 8 months of culture by complete dewatering and the pond should be freshly prepared for the next production cycle.
A survival rate of 65 to 70% is expected and prawn yield may range from 800 to 1000 kg/ha (320 to 400 kg/acre).
The cost of production per kg of prawn may range from Rs.150 to Rs.175/-. Major components of cost of production include cost of seed, pellet feed, energy and labour (Figure 3).

Figure 3: Post-harvest quality Macrobrachium rosenbergii brooders

Post-harvest Handling

Processing yield (tail weight percentage) of freshwater prawns (<50%) is less than that of marine shrimps (>60%) and decreases with the increase in size of the prawn and is better for females than males.
Prawns are sold either head-on or head-less. Sometimes they are sold live also. Ice-chilled uncooked prawns have a short shelf life (3 days) before they become mushy. ‘Kill chilling’ by dipping prawns in iced water prior to blanching at 65°C for 15-20 sec before icing for transport to market, significantly improves quality.
Usually harvested prawns are washed and iced immediately to prevent quality deterioration. In the processing plants they are removed from ice and washed again. The washed and drained prawns are weighed and sent for de-heading.
The iced headless prawns are then size-graded by weight.
After size-grading the product then goes for further value addition according to the requirement of the buyer, such as ‘peeled and deveined’ (PD) and ‘peeled deveined tail-on’ (PDTO). Most of the Giant Freshwater Prawn farmed in India is exported in a headless tail-on style.
Prawns are either bulk frozen or individually quick frozen at -40°C. Packed material is finally stored at -20°C.
Removal of head and intensive washing decreases initial microbial load and improves the post-storage quality of prawns which can be stored frozen for up to six months without any deterioration of flavour.

Results and Discussion

Constraints – Pitfalls and Precautions

The major problem during freshwater prawn culture is size heterogeneity in harvested crop, which demands additional effort to market them.
Tail yield of freshwater prawns (40-50%) is less than that of marine shrimp (60%)and freshwater prawns require more care in processing than marine shrimp.
There are reports of reduced growth rate in grow-out phase from some parts of India which has been attributed to ‘inbreeding depression’.
Freshwater prawns are very sensitive to low dissolved oxygen levels and mortality of stock due to low levels of oxygen in the pond is one of the major reasons of low yield.
Body weight of this prawn shows a very strong negative relationship with stocking density. Therefore, this species cannot be stocked at higher densities and moreover the price is sizedependent.
Low seed quality from hatcheries has resulted in low production.

Polyculture

Freshwater prawns can be easily integrated with existing carp culture bringing additional income to farmers without much additional cost.
Macrobrachium rosenbergii (Scampi) can be cultured with compatible fish species such as Catla (Catla catla), Rohu (Labeo rohita), Silver Carp (Hypophthalmichthys molitrix) and Tilapia.
Polyculture of carp and prawn has the advantage that both prawn and carp can utilize different food niches in the pond efficiently.
Polyculture of prawn without bottom feeders like Common Carp and Mrigal allows the prawns to obtain their share of the pellet feed that will sink to the bottom. In addition, it allows the prawn to graze on bacterial films on the bottom substrate which results in better growth performance of prawn. Further, polyculture improves the ecological balance of the pond water, preventing the formation of massive algal blooms.
Polyculture of Scampi can be carried out in earthen ponds and pens of varying sizes.
Stocking size of prawn should preferably be 2-5 g for better yield and income.
Stocking density of prawn recommended in polyculture range from 10,000 to 15,000/ha or 1 -1.5/m² and that of fish range from 6,000 to 8,000/ha.
Fish can be fed with traditional feed (mash feed of ricebran and oilcake) or floating pellet feed. The prawns need not be fed separately as they will consume the left over feed that finally sink to the pond bottom.
Monitoring of important water quality parameters such as dissolved oxygen, pH and temperature is recommended to prevent loss of stock due to poor water quality especially during last 3 to 4 months of culture.
After four months, marketable size prawns (>40 g) may be harvested by using large mesh cast net or bag net and this selective harvesting can be continued once every 3-4 weeks for another 3-4 months.
Fish can be harvested by netting after 8-10 months and finally the prawns can be harvested by complete dewatering.
At 8,000/ha stocking density the average final size of fish after 10 months of culture would range from 800 g, to 1 kg at a survival rate of 70-75%. The expected production of fish would be 5,000 kg/ha.
At 1/m² stocking density the average final size of Scampi after 8 months of culture would be 50 to 80 g if good quality scampi seed are used. Final survival rate of 60 to 70% is expected and the production of Scampi may range from 480 to 600 kg/ha (200 to 250 kg/acre).

Acknowledgements

Author wish to thank the family members( Mr. Sali V A, Mrs. Girija K R, Mr. Ambady V S, Mr. Anandu VS) for giving the essential facilities, and their thoughtful support and direction throughout the investigation.

References

FAO (2007) The next frontier in genetic research: under water FAO News Room.
Jhingran VG (1956) The capture fishery of river Ganga at Buxar (Bihar, India) in the years 1952- 1954. Indian Journal of Fisheries 1: 197-215.
Kanaujia DR, Mohanty AN (1992) Breeding and large scale seed production of the Indian river prawn Macrobrachium malcolmsonii (H. M. Edwards). J Aqua 2: 7-16.
Kanaujia DR, Pani KC, Mohanty AN (1996) Seed production of Macrobrachium malcolms onii (H.M. Edwards) in synthetic sea-water. Aqua. Trop 11: 259-260.
Kanaujia DR, Mohanty AN, Das KM (1998) Recycling of used water for the seed production of Macrobrachium malcolmsonii (H. M. Edwards). J Aqua Trop 13: 223-232.
Kanaujia DR, Das BK, Mohanty AN (1998) Mass larval mortalities in Indian river prawn Macrobrachium malcolmsonii under hatchery conditions and their control by application of antibiotics. J Aqua Trop 13: 171-179.
Kewalramani HG, Sankolli KN, Shenoy SS (1971) On the larval life history of Macrobrachium malcolmsonii (H. M. Edwards) in captivity. J Indian Fish. Assoc 1: 1-25.
Maria Lourdes A, Cuvin-Aralar, Manuel Laron A, Emiliano V, Aralar, et al. (2011) Aquaculture Extension Manual, Breeding and seed production of the giant freshwater prawn (Macrobrachium r osenbergii) Southeast Asian Fisheries Development Center Aquaculture Department Tigbauan, Iloilo, Philippines 33.
New MB, Singholka S (1985) Freshwater Prawn Farming. A Manual for the Culture of Macrobrachium ro senbergii. FAO Fisheries Technical Paper 1: 118.
Rajyalakhsmi T (1980) Comparative study of the biology of the freshwater prawn Macrobrachium malcolmsonii of Godavari and Hooghly river system. Proc Indian Nat. Acad., B 46: 72-89.
Saeed Ziaei-Nejad, Gholamreza Rafiee, Mehdi Shakouri (2009) Culture and breeding of fresh water prawn Macrobrachium ro senbergii as an exotic species in Iran, present status and future perspective, Department of Fisheries Natural Resources Faculty Behbahan high educational complex Behbahan, Iran, Asian Pacific Aquaculture.
Tiwari KK (1949) On a new species of Palaemon from Banaras with a note on Palaemon lanchesteri (de Man). Records Indian Museum 45: 333-345.
Tiwari KK (1955) Distribution of the Indo-Burmese freshwater prawns of the genus Palaemon (Fabr) and its bearing on the Satpura hypothesis. Bulletin National Institute Sciences India 7: 230-239.
Tiwari KK, Holthuis LB (1996) The identity of Macrobrachium gangeticum Bate, 1868 (Decapoda, Caridea, Palaemonidae). Crustaceana 69: 922-925.

Reaching a Meaningful Agreement among Diverse Parties: The Potential Contribution of Mind Genomics to an Iterated, Optimal Policy

Leave a reply

DOI: 10.31038/MGSPE.2022221

Abstract

Mind Genomics was used to assess the response of ordinary people to different prospective strategies involved with the nuclear deal with Iran, in 2016. Each respondent read a unique set of 25 small vignettes comprising systematically varied messages about the nuclear deal, rating each on likelihood for an agreement, and expected emotional response from Iran. From the set of 20 elements only seven elements performed strongly, but not among the total panel, only among emergent mind-sets. These were MS1 (Focus on military aspects, specifically prevention, n=29 respondents), MS2 (Focus on economic development, n=45), and MS 3 (Focus on effective negotiations and diplomacy, n=11). Most of the emotional reactions were negative. The paper suggests that Mind Genomics be used as an iterative, low cost, rapid fashion, to identify strong negotiating points, base upon the mind of the average citizen. The iterations each lasting 3-4 hours, with several iterations possible in a day at low cost, and with deep learning may radically change the process of negotiation. Mind Genomics identifies what specific messages ‘work’. The process can evolve to a joint effort by both parties to the disagreement, and by so doing craft an agreement attractive to both sides, an agreement emerging from the positive responses of the citizens of both sides.

Introduction

The world of US policy the domain of the three branches of the government, and in practice the domain of a host of consultants and others helping to formulate the policy. Often the policy seems well thought out, other times the policy seems to be either poorly thought out, or more of concern, the influence of various parties which dictate aspects of policy for their own interest.

The topic of this paper is the introduction of a tool, Mind Genomics, to help formulate policy by understanding the ‘mind’ of the average citizen, in a way that could tap into the ‘wisdom of the crowd’, and become an iterative, affordable, rapid tool to help policy formulation.

We illustrate the approach by a study run five years ago on responses to policy about Iran. The objective of the study was to demonstrate the potential of what one could learn in a matter of two days, a time that would be shortened to period of 2-4 hours as of this writing (Fall, 2021). The topic of what to do with the fractious government of Iran continues to rear its head. At the original time of the experiment, the last months of the Obama administration, the issue was raised as to what could be done to deal effectively with Iran. Donald Trump was in the midst of pre-election efforts. The research was done to identify key issues and what people wanted as support for the Republican party.

Formulation of Public Policy with the Aid of Polls

Public policy is often announced by a spokesperson for the committee putting forward that policy. It is obvious from the reports both before, and during the birth of the policy, that the policy was ‘crafted’ by a group, and that often the group is bipartisan. There is the phrase ‘horse-trading’ to discuss the back-and-forth negotiations.

At the same time, in the world of politics, whether for candidates or for political issues there are two worlds intertwined. One world is the world of experts, such as individuals from so-called think tanks, who come up with the recommendations. In the United States these individuals are disparaging called ‘Beltway Bandits’, because are housed near Washington. The experts are highly paid to work with the lawmakers and policy makers, to give advice. Occasionally, scientists enter the process as well because the issue is technical [1,2].

At the same time there are the pollsters, who measure public opinion, attitude. The emphasis here is on accurate measurement. Occasionally these pollsters might be asked to consult on policy, but their expertise is accurate measurement. The measurement may occur with well conducted local and national polls, focus groups, individual depth interviews, perhaps coupled with their own observations of what is happening at the time they are doing the research [3].

There are two languages in policy, the language of artisanship in the creation, and the language of statistics and measurement in people’s response to the creating. The language of policy creation is the language of the artisan shop, where the policy is ‘crafted,’ ‘hammered out’, etc., through the interactions and efforts of the individuals involved. The policy is ‘created’ by those tasked with the job. We can contrast this policy of ‘artisanship’ with the language used in measuring responses to the policy, the language of statistics, polls, degree of confidence, measurement of trends, and assignment of reasons for specific patterns of people’s response to the policy. Furthermore, he two languages do not overlaps. There is not much published in terms of scientifically guided iterations in the development of the artisan-crafted policy. The two worlds are different, creation and measurement.

In contrast to the above is the world of product design, especially the world of software design, but engineering in general. The product may be created by an artisan, but that product is special, one time. The true effort is to create products which work, products that have been created by iterations, with the creation coming first, then the testing, then the revision, and the testing again [4]. The key word is ‘testing as part of the iteration,’ something that is not heard publicly in the world of government policy.

A search through the literature reveals a moderate number of papers on policy, but almost none on measurement during the course of policy development in the way that one might iterate in the creation of software. We might we be in different worlds. Policy again and again seems to be crafted as a one-tine reaction, rather than being quickly evolved from iterations and testing propositions in the policy. It is that opportunity, creation and optimization through iteration, which constitutes the contribution of this paper.

Beyond Polls to Experimentation

The notion of experimentation in political science seems at first strange, simply because one thinks of the political order as an emergent, resulting from the confluences of forces and the ‘Zeitgeist,’ the spirit of the times. Philosophers have debated the nature of the political orders, the classes of political orders, and of course both the assumed ‘original political order of man’ (if there ever was one), and the most appropriate political order for a society. The important thing to note is that political order is so critical that it begs for study, whether for itself or knowledge of which allows one to achieve one’s goals.

At the same time, during the past decades there has emerge a notion of experimentation, and the idea of an experimental political science, perhaps of the same type as experimental psychology. The difference is where the material is published, and the nature of the published material. Experimental psychology began to emerge in Germany almost two centuries with the publication of Ebbinghaus’ book ‘On Memory’. The book was filled with the results of experiments, with data that could be studied, reanalyzed, challenged, and ultimately replicated or not.

We can contrast the early beginnings of experimental psychology with the beginnings of experimental political science, whose material appears in book after book, as points of view, substantiated with one or two experiments, or better rethinking of data [5,6]. There are no standard experiments in political science, experiments which constitute the basis of the science. Rather, there is talk, philosophical point of view, the need and from time-to-time re-presenting data, cast in this new light of experimentation. In other words, experimental political science is very much alive, but as hope for the future, not as a daily, simple, scalable system for producing data and knowledge. We are just not ready although the interest is certainly real, as shown by the intellectual vibrancy of the topic, a ‘must’ for breaking through into new territory [7-9].

The Mind Genomics Approach

Mind Genomics is an emerging science with roots in experimental psychology, marketing research and public polling. The fundamental nature of Mind Genomics is of a science of experimentation which discovers the mind of people with respect to a specific micro-topic. The key word is micro-topic, a focus on easy-to-understand ideas. The objective is to quantify decision making from the bottom up, and identify coherent groups, ‘mind-genomes’, based upon different, recurring patterns describing how individuals make judgments about the world of the everyday [10,11].

The part of Mind Genomics emerging from experimental psychology is the focus on the measurement of ideas, the inner psychophysics as it was called by modern day psychophysics pioneer, S.S. Stevens of Harvard University. Psychophysics itself is the search for lawful relations between physical stimuli and subjective responses, so-called outer psychophysics. It is the aspects of psychophysics to which most scientists familiar with psychology and referring to when they refer to psychophysics. Inner psychophysics, Stevens’ dream, was to apply metrics to ideas, to measure ideas.

The part of Mind Genomics emerging from consumer is the use of mixtures of test stimuli which simulate real world stimuli have cognitive meaning. One of the tools of consumer research, coincidentally developed by experimental psychologists Luce and Tukey is ‘conjoint measurement,’ the evaluation of mixtures of stimuli, and the estimation of the contribution of each element in the mixture to the overall response. In the world of commerce, mixtures are importance. They are the substance of which products and services are composed. We don’t buy single ideas, but rather combinations of features and benefits embedded in a product or a service.

The Seven Steps

Mind Genomics follows a templated process comprising seven steps. The steps begin with the creation of raw material, and finish with the identification of strong performing elements, among defined groups of respondents, including new-to-the-world groups of respondents who can be shown to think alike on this topic. The output of the Mind Genomics study may find use in driving a better program of communication of one’s product, or part of an academic effort to create the ‘wiki of the mud for a set of related issues’

Step 1: Define the Problem, Create the ‘Raw Material’, Defined as a Set of ‘Questions’, and a Specified Number of Answers to Each Question

The Mind Genomics effort is an experiment, rather than a questionnaire, although Mind Genomics has often been defined in public terms as a survey’.

The essence of Mind Genomics is to measure responses to defined stimuli, viz., combinations of messages, and these combinations called vignettes. The vignettes are combinations of statements about the topic, in our case policy towards Iran. As a consequence, the Mind Genomics process prescribes the raw material, namely the topic (Iran), a set of ‘questions’ or ‘categories’ which in sequence describe or tell a story, and for each question or category, an equal number of ‘answers.’

The approach for finding the raw material may range from sheer expertise and ‘off the cuff’ to serious research into what is in published. With the growing interest in Mind Genomics as a fast, iterative process, the movement is towards simple, superficial ideas, some based upon what has been seen or read in public sources, the others based upon one’s own ideas, or the ideas of a creative group, thinking about the topic.

Table 1 shows the list of elements. The structure of the table, four questions, five answers per question, is based on the one of the designs of the Mind Genomics system. The elements were created by author Bitran based upon his on strategic analysis work with his program, Enterprizer(r). It is important to keep in mind that Mind Genomics is a tool which puts the elements to a hard test, as we will see below. The iterative nature of Mind Genomics will allow strong elements to emerge. At the same time, however, the Mind Genomics system is not ‘creative’. And so, a good knowledge of the topic is helpful but not a requirement.

Table 1: The five questions (categories) and four answers (elements) for each question

Step 2: Create Short Vignettes Using Experimental Design

The world of science works by identifying a phenomenon of interest, defining aspects of the phenomenon to be studies, and when possible, isolating those aspects of interest, and measuring them. The aim is to determine the nature of the variable of interest. Doing so means reducing the haze around the variable, the random variation which hides that nature of the variable. The variability itself is unwanted and eliminated through research. The two strategies are to isolate the variable, eliminating extraneous forces which lead to variation, or measure the variable many times, under different situations, and average out the unwanted variation.

When we deal with issues of foreign policy and break out the issues into elements such as those shown in Table 1, the typical research strategy would be to polish each element so that each element is as clear as possible, and as simple to understand as possible. That corresponds to the first effort, measuring the variable which has been made as simple as possible, so other factors do not affect the results. The second is to test that single idea with hundreds of people, one idea at a time with each of the hundreds of people. Averaging the results from the large group should provide a stable measure of the response to the variable.

The one-at-a-time method dictates that the researcher presents the respondent with each of the elements, one element at a time as the phrase says. The respondent is instructed to maintain the same criterion, and with that one criterion rate the element. It does matter whether the element is positive, negative, deals with peace, deals with conflict; the respondent is to use the same rating scale all the time.

An ongoing problem in the on-at-time research is the unnaturalness of single elements. There is no context. The rating is easier when all of the test stimuli, the elements, are of the same type, such as military alliances, or economic alliances, educational strategies, and so forth. The respondent reads the elements, all of the same time, and has no problem evacuating the elements themselves. They are commensurate with each other. The problem arises when the elements are different. The differences may be vast, such as economic policy versus military policy. Although the researcher can instruct the respondent to use the same criterion, it is not clear that the respondent can actually do so.

A better approach, one which removes some of the artificiality of the one-at-a-time method, works by creating combinations of ideas. This is the approach used by Mind Genomics. Rather than forcing the respondent to maintain the same criterion with palpably different types of statements, Mind Genomics puts together the ideas or statements into small easy to read combinations, such as that shown in Figure 1. There is no effort to polish the combination, or to create connectives so that the combination is even more natural looking, appearing like the paragraphs that the respondent is comfortable evaluating. Although the critic might aver that the combination is not polished, that there are no connectives, that some of the laws of grammar are violated, the reality is that the combination forces the respondent to adopt one criterion and keep it because it is impossible in a Mind Genomics experiment to continue to shift judgment criteria to match what ends up seeming to be an ever-changing set of random combinations of ideas. The easiest way is to maintain one’s judgment criteria in the face of ever-changing combinations.

Figure 1: Example of a four-element vignette. Each respondent evaluates 25 unique vignettes. The vignettes for each respondent differ from each other

The combinations themselves may appear to the respondent to be utterly random. Nothing can be further from the truth. The combinations are created according to an experimental design [12]. The experimental design comprises specific combinations, allowing the variables to interact, but making sure that the 20 elements in this particular case are presented so that they are statistically independent of each other. That statistical independence is accomplished by the specific combinations. The design comprises 25 combinations or vignettes. Each vignette has a specified number of elements, at most one element or answer from any question. The vignette structure is:

Two elements in the vignette – 2 of 25 vignettes

Three elements in the vignette – 4 of 25 vignettes

Four elements in the vignette – 11 of 25 vignettes

Five elements in the vignette – 8 of 25 vignettes.

Although some critics might aver that the vignette has to be complete, with one element from each of the five categories, the reality is that respondent has no problem dealing with the sparser vignettes. The problem is the attitude of the researcher who wants completeness.

The basic design of 20 element embedded in 25 vignettes is a very efficient design. The breakthrough is design came around 1998, when the notion emerged of a permutable design. That is, one could create the basic mathematical structure of the design, specifying the combinations, and so forth. Once this was done, I was simple and straightforward to create a basic design, and then permute it, changing the elements, but maintaining the design structure. That meant renumbering the elements but keeping the elements in the same category. Thus, A1 would become A2, A5 would become A4, and so forth. The renumber would be done for all elements. This strategy, described in detail by Gofman & Moskowitz (2010), maintained the structural integrity of the experimental design, but recrafted the design slightly to cover many more of the possible combinations.

Figure 1 shows an example of a four-element vignette. The physical layout is simple, one phrase atop the other. There is no indication of categories or questions, simply a combination of the elements. No effort is made to connect the combinations.

Step 3: Execute the Study (viz., Experiment) Online

The actual study was executed through an on-line panel provider, specializing in recruiting respondents and providing them for these studies. The company, Luc.id Inc., in Louisiana, USA, is an aggregator of respondents from various panels. Working with a panel provider such as Lucid. makes the process easy. Over the past two decades it has become increasingly difficult to recruit one’s own panelists, especially for interview or experiments lasting 10+ minutes. The refusal rate has skyrocketed. As a consequence, the panel providers can deliver a group of respondents, generally filling easy specifications, for a reasonable price.

The respondents were invited to participate. The respondents were shown the following orientation. Note that a link was given for further reading about the JCPOA

By way of background Wikipedia as this this writing (Fall 2020) presents a background to the JCPOA, the Joint Comprehensive Plane of Action, which was signed in 2015.

Under the JCPOA, Iran agreed to eliminate its stockpile of medium-enriched uranium, cut its stockpile of low-enriched uranium by 98%, and reduce by about two-thirds the number of its gas centrifuges for 13 years. For the next 15 years, Iran will only enrich uranium up to 3.67%. Iran also agreed not to build any new heavy-water facilities for the same period of time. Uranium-enrichment activities will be limited to a single facility using first-generation centrifuges for 10 years. Other facilities will be converted to avoid proliferation risks. To monitor and verify Iran’s compliance with the agreement, the International Atomic Energy Agency (IAEA) will have regular access to all Iranian nuclear facilities. The agreement provides that in return for verifiably abiding by its commitments, Iran will receive relief from the U.S., European Union, and United Nations Security Council nuclear-related sanctions. https://en.wikipedia.org/wiki/Joint_Comprehensive_Plan_of_Action

The study was complete by 85 respondents, recruited by Luc.id. The base size of 85 suffices for a simple but often deep focus on the topic. The reason for the need for fewer than the hundreds of respondents in conventional survey work is that the research is searching for patterns, not for a precise measure of one point.

Step 4: Prepare the Data for Analysis by Creating New Binary Variables

The Mind Genomics exercise produces a great deal of data, since each of 85 respondents evaluated 25 different vignettes on two types of attributes, a degree of belief in the meaningful agreement (1=Definitely No … 9=Definitely yes) and a selection of the emotion that would be felt by Iran, if Iran were a person.

Our goal is to link the 20 elements to the ratings and the emotions. We do that in the next section. in this first section we transform the 9-point rating to a no/yes scale. Managers find it easier to work with binary scale, rather than to talk in percentages. Following the convention of previous efforts with Mind Genomics and the 9-point scale, we recode ratings of 1-6 to 0 (low probability), and ratings of 7-9 to 100 (high probability). The recoding could be made more stringent or less stringent. There is no ‘right answer,’ just appeal to previous processes. We do the same type of recoding for the emotions. We recode emotions as positive) negative). (Positive: Happy, Relieved, Victorious; Negative: Defeated, Fearful).

Thus, each vignette ends up with three numbers. One for the binary recode for probability of meaningful agreement, one for a positive emotion, and the complement for a negative emotion. The numbers are either 0 or 100. When it comes to the positive versus negative emotion, one of the two variables will take on the value 100, and the other by definition will take on the value 0.

Finally, vanishingly small random number is added to every newly created binary scale. this random number does not affect the results but does avoids a problematic statistical issue emerging from OLS (ordinary least0squares) regression occurring when the respondent selects all ratings for question 1 (meaningful agreement) either 1-6 or 7-9 (all 0’s or all 100’s across the 25 vignettes) or select all emotions as positive or all emotions as negative.

Step 5: Compute Means for to Better Understand the Patterns of Responses

By Step 5 we have already put the data into a form that makes it easy to compare average ratings (the focus of this step), and to link the elements to response (focus of Step 6).

We can explore the quality of the data by computing averages, considering both the number of elements in a vignette as a stratifying variable, and considering the order of testing as a stratifying variable. Even without knowing which elements are present in a vignette, we can ask whether there are any differences in the average ratings assigned to vignettes of 2,3,4 or 5 elements respectively, in terms of the binary transform of likelihood of agreement (TOP 3), and for the average Positive and average Negative emotions.

To answer the foregoing questions means simply to identify each vignette in two ways, first by the number of elements (2, 3, 4 or 5, respectively), and second by the position in the respondent’s sequence (first third, second third, final third).

Table 2 that there is no difference by position in terms of how it covaries with either likelihood to reach agreement (Q1) of emotion selected.

Table 2 also shows no effect of number of elements in terms of affecting the likelihood to reach agreement. There is, however, a quite strong and inverse covariation between the number of elements in the vignette and the selection of a positive emotion. Shorter vignettes are perceived as more likely to generate a positive emotional response by Iran, perhaps because the terms are defined, and the permission is direct. That is, shorter vignettes may leave less ‘wiggle room’, ‘and less ‘fine print’ in the agreement.

Table 2: Average values for TOP3 (likelihood of an agreement), and emotions selected (positive, negative) versus order of testing, and the number of elements in a vignette

The final topic of our surface is analysis is to get a sense of how the respondents feel about what they are reading. Question 1 allows us a sense of whether respondents feel optimistic about the process, viz., that it will happen, or feel pessimistic. Question 2 gives us a sense of their emotions. Let us average the ratings from their reactions to their own 25 vignettes, independent of what is in the vignettes. (Although, we know that each element appears equally often in the 25 vignettes; it’s just the combinations which vary).

Figure 2 shows a scatterplot of the average score for ‘reach agreement’ (% rating 7-9) vs. the average percent of selections of a positive emotion. Figure 2 shows a concentration of respondents on the left, with low average value of TOP3. We conclude from this that the individual respondents, on average, feel that the agreement will not be reached. There is no sense, however of a preponderance of emotions. Respondents simply do not seem to be able to figure out what the feelings of the Iranians will be a finding which should not surprise. Response can feel strongly about the outcome but not feel strongly about the expected feelings emerging from that outcome.

Figure 2: Scatterplot showing the average ratings for reach agreement (abscissa, TOP3) versus the percent of times that a positive emotion will be experienced by the Iranians

Step 6: Relate the Elements to the Ratings

As of today’s state-of-the-art, the pinnacle of the analysis is the ability to relate the presence / absence of the 20 elements to the response, whether the response be the TOP3 (strong likelihood of that there will be an agreement), or the selection of a positive emotion, and finally the selection of a negative emotion. Mathematically, the selection of positive versus the selection of negative emotions is complements of each other. We will be dealing with both, because in our presentation of data will look only at strong performing elements driving positive emotions, and strong performing emotions driving negative emotions, and in turn NOT presenting data from elements which do not strongly engage of or the other.

The experimental design allows us to create both group models and individual-level models relating the presence/absence of the 20 elements to the response. The original design was set up to allow a simple regression equation to describe the data: Response=k₀ + k₁(A1) + k₂(A2) … k2₀(E4). Recall that each respondent evaluated a unique set of 25 vignettes, comprising a permuted variation of the original design, a variation known to ‘work’, viz., to mathematically identically to the original design.

The first analysis created models relating the presence/absence of the elements to the actual rating of Question 1 on the 9-point scale. Although we will be looking at a transformed variable (TOP3 instead of the 9-point rating), it is instructive to see the degree to which our 85 respondents generate data which is consistent. We measure consistency by estimating the equation, and computing the goodness of fit, the multiple R, the multiple correlation. The multiple R goes from 0.00 (no fit of the variables to the ratings; totally inconsistent results) to +1.00 (perfect fit of the variables to the ratings, totally consistent results which trace the ratings precisely to the presence/absence of the elements).

Figure 3 shows the distribution of the 85 ratings. We can feel confident about the data. Even though most respondents feel that they are ‘guessing,’, that they cannot figure out the ‘correct answer,’ our estimation of consistency suggests that the results are reasonably consistent.

Figure 3: Consistency of the results for the 85 respondents, shown by the Multiple R statistic estimated from the individual-level multiple linear regressions

Step 6: Divide the Respondents by the Pattern of the Coefficients to Create Mind-sets

Our last analysis divides the respondents by the pattern of their coefficients. For each respondent we create a model or equation whose dependent variable is TOP3, previously defined as taking on one of two values. The values depend upon the original rating of Q1, the probability of reaching an agreement. Recall that ratings of Q1 1-6 were coded 0, ratings of 7-9 were coded 100.

The database generated from the individual-level regressions comprises 85 rows, one row corresponding to each respondent. Each row comprises 21 columns, one column for the additive constant, and 20 columns for the 20 coefficients. The objective of clustering is to divide this group of 85 ‘objects,’ viz respondents into a limited number of non-overlapping groups, the clusters or mind-sets, based upon a mathematical criterion. The criterion does not require the researcher to know the ‘meaning’ of the measures, viz., in this case the coefficients, but simply to have each object quantified on each measure. Thus, we have 85 objects (people) on 20 measures (coefficients). We do not consider the additive constant in the process.

The clustering program is a heuristic. There are many different clustering programs. The program used here is k-means [13], with the objective of putting the 85 people into either two groups (analytic pass 1) or three groups (analytic pass 2). The criteria are that the profiles of the 20 averages (one per coefficient A1-E4) should be ‘far away from each other’, and the distance between the objects or people in a cluster should be as small as possible. The criterion for distance is (1-Pearson Correlation Coefficient, R). The Pearson R shows the strength of a linear relation between two variables, taking on the value +1 (viz., Distance=0) when they are perfectly linearly related, and taking on the value -1 (viz., distance=2) when they are perfectly inversely related.

Our criteria for choosing the ‘best’ number of clusters combines a desire for parsimony (fewer clusters are better than more clusters), and interpretability (the clusters must tell a coherent story, and the stories of the clusters must differ from one another).

The two-cluster solution, although parsimonious, seemed too jumbled. There was no clear story. The three-cluster solution seemed a bit better. A four-cluster solution was virtually no different in types of groups than the three-cluster solutions. That is, two of the clusters in the four-cluster solution seemed quite similar. The decision was to work with a three-cluster solution.

In the language of Mind Genomics, the cluster becomes a mind-set, a way of responding to a limited set of related stimuli. The min-sets are constructed from the patterns of the coefficients form the 85 respondents who participated in this study. Over the years, the mind-sets which emerge from these focused, quite small studies, continue to repeat. The repetition comes about because when we abstract the type of individual based upon the pattern of responses, we end up with just a few really quite different groups. The psychologists called the ‘archetypes’, but the archetypes emerging from Mind Genomics are based on small, single-focus studies. Yet, again and again, these mind-sets continue to appear in many different ways. The great anthropologist, Joseph Campbell [14], would call this the ‘hero with a thousand faces.’

Step 7: The Total Panel and the Mind-sets

The Mind Genomics effort naturally brings with it many numbers for the study 21 numbers for each group, or 84 numbers for the combination of total panel and the three mind-sets. The objective of these studies is to find patterns, and not to overwhelm ourselves with numbers which may end up disguising the patterns in the dense undergrowth of numbers. To counteract the death by wall of numbers were show only positive coefficients of 8or higher. These strong performer in a Mind Genomics study. We may be losing some information by this stringent cutoff, but a coefficient of +8 or higher is strongly significant from the regression modeling, with a t statistic approaching 2.0.

Table 3 shows the total panel and the three mind-sets, created for the results from Question 1, on the likelihood of an agreement. The cluster uses the coefficient emerging when TOP3 is the dependent variable. The table shows base size first, then the additive constant, and then the strong performing elements for each mind-set.

Table 3: Performance of the strong performing elements for total panel and three emergent mind-sets. Only the seven elements with coefficients of +8 or higher are shown

The additive constants are 32-38 meaning that without additional information, but just knowing that there are negotiations, about one in three responses to the vignettes are 7-9. We know this because the additive constant tells us the likelihood of a rating of 7-9 in the absence of elements, and is a purely theoretical, computed value. Nonetheless, the additive constant gives us a good sense of basic response. It is remarkable that all three mind-sets agree so well. This is unusual. The agreement means we are dealing with specifics.

When we look at the column for total panel, we find NO strong performing elements that disappointing finding does not mean that we failed in this attempt, although it might mean failure. Our success in the study comes after we deconstruct the total panel into the three groups, based upon patterns of coefficients, not upon magnitude of coefficients. That is, our three mind-sets would have emerged if all of the coefficients were equally reduced by 20 points. In such a case three mind-sets would emerge from the patterns, but NO elements would emerge as being strong.

Before we go into the three mind-sets, which is now quite simple, its worth remarking that we began with 20 elements, the best guesses from people involved. Yet, only seven of the 20 elements emerged as strong, no elements emerged as strong for total, and surprisingly, each strong performing element appeared strong only in one of the three mind-sets.

The min-sets are easy to describe. One simply looks at the strongest element.

Mind-Set 1=Focus on military aspects (prevention) – 29 of the 85 respondents

Mind-Set 2 Focus on economic development – 45 of the 85 respondents

Mind-Set 3 – Focus on effective negotiations and diplomacy – 11 of the 85 respondents.

We move now to the elements which drive strong positive and strong negative responses. The coefficients in Table 4 emerge from six regressions. The six regression comprised three regressions for the selection of a net positive emotion, and three regressions for the selection of a net negative, in both cases two regressions for each mind-set, respectively. The regression model was run without the additive constant, because of the previously conventions in Mind Genomics practice, that emotions and other selections emerging from the nominal scales are estimated without coefficients.

This time we look only the elements which drive a percent selection of 16% or more, for either a positive or a negative emotion. Table 4 shows us that only one mind-set, MS1 (focus on military aspects, prevention) feel that there will a strong positive response. All three mind-sets feel that there will be a strong negative emotion from Iran.

Table 4: Strong positive and negative emotions selected by the respondents from the three mind-sets as they think about the feeling emerging from Iran, as driven by the element. Only coefficients of +16 or higher are shown

Discussion and Implications

When this study was executed in 2016, Mind Genomics was just beginnings its broader application to international relations, having begun in 2012 with studies of the Israeli Palestinian conflict. The realization at that time, confirmed by many subsequent studies in a variety of areas, is the relative paucity of solid information about the mind of the citizen in the world of social issues, the mind of the customer in the world of commerce, the mind of the patient in medicine, the mind of the client in legal and business issues, and so forth. There were dozens of polls, dozens of learned volumes on key issues, the ongoing broadcasting, and increasing ‘natter’ of the media with ‘talking head’ proclaiming the same new, spun one or another way.

A cursory content analysis of the literature, of the media, and so forth brings out facts, histories, opinions, and the voice of the citizen. The voice of the citizen, however, appears to be limited to simple factoids, statements, voting on issues. Furthermore, there seemed to be a desire to compare changes, and by that comparison to get a sense of where things were going. In other words, the focus was on the macro, with little content, and the depth was assumed to emerge by observing the path of the macro trends over time, perhaps with an effort to see how the trend covaries with exogenous factors, like world order world economics, and so forth. And perhaps even the world’s ‘Zeitgeist’ although Zeitgeist might be more the bias of the analyst than the reality of the items. There are examples of iterated efforts, such as China’s policy [15], but these iterations are large-scale, in the manner of iterating products, rather than ideas.

Enter Mind Genomics, here presented as the first experiment on international relations, at a time when Mind Genomics was conceived of as a one-off process, requiring a lot of thinking, a great deal of expertise for choosing the ‘right material’, and the careful efforts which accompany a scientific project. There were 85 respondents, rather than the customary hundreds of respondents, but that is not a problem. the problem here is the fact that the Mind Genomics study at that time was considered as a final effort, a one time ‘deep dive’ into the mind of the citizen. And the results are what they were, pointing to different mind-sets, but with remarkably few elements performing strongly, either in terms of driving agreement or driving emotions.

The methods of Mind Genomics have been proven again and again, in the legal, medical and commercial realms [16-18]. In those realms, the efforts of Mind Genomics have evolved from one-off, large-scale studies with 36 elements down to the current size of 16 elements (four questions and four answers to each question). The notion of the ‘final experiment’ has given way to Mind Genomics as a fast, iterative, learning=based process. Within that world-view, this study would be updated by a series of short studies, each requiring about 60 minutes to set up on publicly available program (www.BimiLeap.com), and then executed with 50-100 respondents automatically with 60-90 minutes, and the entire data set totally analyzed 10 minutes, and returned to the researcher. One might imagine the use of the iteration as a way both to arrive at good ideas, acceptable to both sides, as well as a consensus-building method, wherein both sides cooperate, and thus build good will.

In the evolution of political science, and the evolution of knowledge of people, these early studies by Mind Genomics of political issues show the potential of a systematic exploration of a topic. When that exploration becomes inexpensive, quick, easy to execute on the internet, and most importantly, ITERATIVE, we have the potential a new political science, one based upon data, extending across many countries, many people, over time, and many topics [19-21]. What was one study in 2016 could well generate a wiki of the mind for the topic of dealing with Iran, that ‘wiki’ filled with data, topic-related, and searchable for specific results and for general patterns.

Acknowledgments

The author would like to acknowledge the help of four associates who helped to design the study.

Joseph Bitran of Enterprizer®

Alice Duggan

Tawfik Hamid

Richard Sciacca

References

Garrison JA (2003. Foreign policy analysis in 20/20: a symposium. International Studies Review. 5: 155-202.
Rametsteiner E, Pülzl H, Alkan-Olsson J, Frederiksen P (2011). Sustainability indicator development—Science or political negotiation? Ecological Indicators. 11: 61-70.
Gadarian SK (2010) The politics of threat: How terrorism news shapes foreign policy attitudes. The Journal of Politics. 72: 469-483.
Wynn DC and Eckert CM (2017) Perspectives on iteration in design and development. Research in Engineering Design 28: 153-184
Druckman JN, Greene DP, Kuklinski JH, Lupia A (2011). Cambridge Handbook of Experimental Political Science. Cambridge University Press.
Morton RB, Williams KC (2010) Experimental political science and the study of causality: From nature to the lab. Cambridge University Press.
Kinder DR, Palfrey TR (1993) On behalf of an experimental political science. In: Experimental Foundations of Political Science 1-39.
Kittel B, Luhan W, Morton R (2012). Experimental political science: Principles and practices. Springer.
McDermott R (2002) Experimental methods in political science. Annual Review of Political Science. 5: 31-61.
Gere A, Radvanyi D, Moskowitz H (2017). The Mind Genomics Metaphor-From Measuring the Every-Day to Sequencing the Mind. International Journal of Genomics Data Minin. IJGD
Radványi D, Gere A, Moskowitz HR (2020) The Mind of Sustainability: A Mind Genomics Cartography. International Journal of R&D Innovation Strategy (IJRDIS). 2: 22-43.
Moskowitz HR, Gofman A, Beckley J, Ashman H (2006) Founding a new science: Mind Genomics. Journal of Sensory Studies, 21: 266-307.
Likas A, Vlassis N, Verbeek JJ (2003) The global k-means clustering algorithm. Pattern Recognition 36: 451-461.
Campbell J (1949). The Hero with a Thousand Faces, New York, Pantheon
Leutert W (2021) Innovation through iteration: Policy feedback loops in China’s economic reform. World Development. 138: 105-173.
Wren JE and Williams TC (2009) Selling blue elephants to the jury: Potential application of rule developing experimentation in litigation. Baylor Law. 61: 1
Gabay G, Moskowitz HR (2019) “Are we there yet?” Mind-Genomics and data-driven personalized health plans. In: The Cross-Disciplinary Perspectives of Management: Challenges and Opportunities. Emerald Publishing Limited.
Milutinovic V, Salom J (2016) Mind Genomics: A Guide to Data-Driven Marketing Strategy. Springer.
Kaarbo J (2003) Foreign policy analysis in the twenty-first century: back to comparison, forward to identity and ideas. International Studies Review. 5: 156-202.
Horiuchi Y, Smith DM, Yamamoto T (2018) Measuring voters’ multidimensional policy preferences with conjoint analysis: Application to Japan’s 2014 election. Political Analysis. 26: 190-209.
Rapport A (2017) Cognitive Approaches to Foreign Policy Analysis. In Oxford Research Encyclopedia of Politics.

The Effect of Preoperative Cardiopulmonary Rehabilitation on Pulmonary Infection after Cardiac Surgery

Leave a reply

DOI: 10.31038/JCCP.2022515

Abstract

In recent years, the incidence of heart disease remains high, which is closely related to people’s diet and living habits, and in cardiac surgery due to tracheal intubation, bed rest and other problems easy to cause postoperative pulmonary infection, we all know that cardiopulmonary rehabilitation training can improve the cardiopulmonary function of patients, our article on preoperative cardiopulmonary rehabilitation training on pulmonary infection after cardiac surgery to review.

Cardiovascular disease has become an important disease endangering the health of Chinese people, and its mortality rate accounts for 67.1% of the total number of cardiovascular disease deaths. In recent years, with the progress of medical technology such as various cardiac surgeries, the mortality rate has gradually decreased, but due to the inability to recover postoperative cardiopulmonary function, it is still unable to live normally or be weaned from the ventilator, and the quality of life has decreased.

A large number of evidence shows that cardiopulmonary rehabilitation can effectively improve the cardiopulmonary function of patients, mainly through respiratory rehabilitation and exercise training to improve the overall cardiopulmonary function of patients, and can improve the mental status of patients and improve exercise capacity, and cardiopulmonary rehabilitation also allows us to rehabilitation of heart disease from treatment to prevention, this paper mainly from the overview of cardiopulmonary rehabilitation and exercise methods to prevent pulmonary infection in patients after cardiac surgery.

Overview of Cardiopulmonary Rehabilitation

Definition of Cardiac Rehabilitation

Cardiac rehabilitation is to relieve the clinical symptoms of patients through comprehensive rehabilitation medical treatment, improve the daily life ability of patients, improve the quality of life, return to normal social life, and prevent the recurrence of cardiovascular disease. It is currently an important means of treating the chronic phase of the heart. The contents of cardiac rehabilitation include regular medication, exercise therapy, psychotherapy, diet therapy, and behavior therapy [1-14].

Definition of Pulmonary Rehabilitation

Pulmonary rehabilitation is an intervention for patients with discomfort symptoms and reduced daily activities, or decreased activities of daily living, chronic respiratory diseases. Pulmonary rehabilitation intervention modalities are comprehensive assessment and rehabilitation programs and implementation strategies involving multidisciplinary teams on the basis of evidence-based medicine. Pulmonary rehabilitation can stabilize and reverse the systemic manifestations of the disease, reduce symptoms, optimize functional status, increase activities of daily living and social participation, reduce the rate of acute onset and rehospitalization, and reduce the cost of medical care. Pulmonary rehabilitation has two main goals, to maximize physical, psychological, and social function, to educate patients how to improve mobility and self-care ability in daily life, to improve quality of life, and to reduce dependence on hospitalization.

Cardiopulmonary Rehabilitation Exercise Mode

Respiratory Training

Abdominal respiratory training quiet, supine, the abdomen put a sandbag of moderate weight, self-control of the thorax through the abdominal undulation uniform respiratory training.

Inspiratory resistor breathing training to control inspiratory volume and time.

It is required that after deep inspiration, the expiration is as thin and slow as a whistle, the airflow is gradually exhaled, and respiratory training should be done, and the training volume should be grasped, generally less than ten seconds each time, about ten times, and three groups should be done continuously, and the speed should be slow.

Passive training, requiring manual training by professional therapists, such as thorax, back, and scapular extrusion.

Aerobic Exercise and Resistance Training

Aerobic training belongs to the training of long-distance endurance, also known as “cardiopulmonary function training”. It is through continuous and repeated activities, and in a certain period of time, with a certain speed and a certain training intensity, it is required to complete a certain amount of exercise, so that the heartbeat rate is gradually increased to the specified highest and lowest safe heartbeat range. Common training methods are brisk walking, jogging, Tai Chi and bicycle.

Resistance training, also known as resistance training, is a movement against resistance, the main purpose is to train the muscles of the human body, and traditional resistance training includes push-ups, dumbbells, barbells and other items.

Effect of Cardiopulmonary Rehabilitation on Pulmonary Infection after Cardiac Surgery

Study on the effect of cardiopulmonary rehabilitation on pulmonary function after cardiac surgery found that respiratory training can enhance the muscle strength and endurance of respiratory muscles, improve pulmonary ventilation, improve pulmonary function, but also increase the blood flow and activity of diaphragm, while reducing the standby time, can pull out the endotracheal tube as soon as possible, transfer out of ICU, return to the general ward and get out of bed as soon as possible.

Effect of cardiopulmonary rehabilitation on cardiac function after cardiac surgery Aerobic exercise enhances the ability of the cardiovascular system to deliver oxygen to muscles, allowing the body to adapt to higher intensity and longer lasting exercise, while reducing heart rate at rest, improving cardiac function, and improving quality of life.

Effect of cardiopulmonary rehabilitation on quality of life. Cardiac rehabilitation can enhance exercise tolerance and improve quality of life in patients. Moreover, exercise training can promote patients with coronary heart disease to maintain a positive and optimistic attitude to life, respiratory and exercise-based cardiopulmonary rehabilitation can shorten the length of hospital stay, promote patients to return to normal life as soon as possible, and improve the quality of life.

Conclusion

Respiratory training and exercise training are the core contents of cardiopulmonary rehabilitation, respiratory training can improve pulmonary ventilation, improve pulmonary function, in order to reduce hypoxia, and regular exercise training can increase myocardial oxygen supply, improve the work ability of the heart and arterial blood flow reserve capacity, and improve the quality of life of patients with disease. As rehabilitation workers, we should carry out detailed evaluation for patients and intervene in precise treatment after evaluation, so as to reduce the probability of postoperative pulmonary infection, reduce the cost and hospital stay, and return to the family as soon as possible.

References

Ashtonre, Tew GA, Aning JJ, et al. (2020) Effects of shortterm, medium-term and long-term exercise training on cardio-metabolic health outcomes in adults: systematic review with meta-analysis [J]. Br J Sports Med 54: 341-348. [crossref]
Zhang H, ChangR (2019) Eflects of Exercise after Percutaneous Coronary Intervention on Cardiac Function and Cardiovascular Adverse Events in Patientswith Coronary Heart Disease: SystematicReview and Meta-Analysis [J]. Journal of sports once& medicine 18: 213-222. [crossref]
Rezaei-Hachesu P, Oliyaee A, Safaie N, et al. (2017) Comparison of coronary artery disease guidelineswith extracted knowledge from data mining [J]. J Cardiovasc ThoracRes 9: 95-101.
Lanz J, Wyss D, R ber L, et al. (2019) Mechanical complications in patients with ST-segment elevation myocardial infarction: a single centre experience [J]. PLoS One 14: 209502. [crossref]
Gomes Neto M, Duraes AR, Conceicao LSR, et al. (2018) High intensity interval training versus moderate continuous intensity training on exercise capacity and quality of life in patients with heart failure with reduced ejection fraction: a systematic review and meta-analysis [J]. Int J Cardiol 261: 134-141. [crossref]
Wu Yan, Wang Xiujuan, Kang Xiaoyan, et al. (2018) Effect of preoperative rehabilitation training on cardiopulmonary function in patients with early stage cancer undergoing thoracoscopic surgery [J]. Journal of Clinical Pulmonary Medicine 23: 1217-1221.
Gao Zhenzhen, Ji Peng, Xia Yueqing, et al. (2015) Effects of different intensity aerobic exercise on cardiac function and exercise tolerance in patients after percutaneous coronary intervention [J]. Chinese Journal of Rehabilitation Medicine 30: 344-348.
Li Xue, Shanyan, Jiang Tingting, et al. (2021) Research progress of resistance exercise in nursing care of maintenance hemodialysis patients complicated with sarcopenia [J]. Nursing Research 35: 643-647.
Wang Lei, Gao Zhenzhen, Pan Huaping (2015) Effect of individualized aerobic exercise on heart rate recovery and exercise capacity in patients with coronary heart disease [J]. Chinese Journal of Rehabilitation Medicine 30: 242-246.
Zhang Lingyun, Bai Chuanchuan, Shu Bin (2018) Research progress on the protective effect of exercise training on heart after myocardial infarction and its mechanism [J]. Chinese Journal of Rehabilitation 33: 53-55.
Jiao Haixu (2020) Research progress on cardiac rehabilitation of patients with cardiovascular disease by resistance exercise [J]. Journal of Qiqihar Medical College 41: 1140-1141.
McMahon SR, Ades PA, Thompson PD (2017) The role of cardiac rehabilitation in patients with heart disease [J]. Trends Cardiovasc Med 27: 420-425. [crossref]
SIMON M, KORN K, CHO L, et al. (2018) Cardiac rehabilitation: A class1 recommendation [J]. Cleve Clin J Med 85: 551-558. [crossref]
Anderson L (2014) Cardiac rehabilitation for peoplewith heart disease: an overview of Cochrane systematic reviews. Cochrane Database Syst Rev 2014: 011273 [crossref]

Appraisal of Abnormal Movement Disorders among Aged Schizophrenics: A Pilot Study

Leave a reply

DOI: 10.31038/ASMHS.2022633

Abstract

Introduction: Tardive dyskinesia (TD) includes involuntary choreiform or athetoid movements of the jaw, lower face, tongue, and extremities, developing in association with the use of an antipsychotic medication, and may develop in about 20 to 40 percent of patients who require long-lasting hospitalization. In the present study, the prevalence of this condition has been measured among an elderly group of schizophrenic patients.

Methods: One hundred and one elderly schizophrenic patients, who were hospitalized in the chronic section of a community psychiatric hospital, were selected for the present cross-sectional study. Abnormal Involuntary Movement Scale (AIMS) was employed to screen for patients with schizophrenia who also had TD. Scale for Assessment of Positive Symptoms, Scale for Assessment of Negative Symptoms, Schedule for Assessment of Insight, and Clinical Global Impressions – Severity of illness, as well, had been used as ancillary scales for evaluation of severity of general psychopathology of schizophrenia, and comparing the TD patients with the group of patients without TD, for probing the intervening parameters.

Results: While abnormal movements were clear in 38.61% (n=39) of elderly schizophrenic patients, only seven of them (6.93 %) could be diagnosed as TD, based on the above-mentioned criteria. All of them were using conventional antipsychotic medications, accompanied with anticholinergic medications. Among TD patients, three cases had only abnormal facial and oral movements, one patient had atypical facial and oral movements as well as anomalous extremity movements, one patient had irregular facial and oral movements in addition to unusual trunk movements, and lastly, two patients had nonstandard extremity movements. In addition, around 71% of patients with TD were aware of their unusual movements. Between-group analysis did not show any significant difference between patients with TD and patients without TD in age, duration of illness, positive symptoms, negative symptoms, insight, and general psychopathology.

Conclusion: According to the findings of the present study, the prevalence of Tardive Dyskinesia among elderly schizophrenic patients, who were using typical antipsychotic medications, is very lower than what has been indicated thus far.

Keywords

Schizophrenia; Typical antipsychotic drug; Atypical antipsychotic drug; Extrapyramidal symptoms; Tardive Dyskinesia; Medication induced movement disorder

Introduction

Tardive dyskinesia (TD) includes involuntary choreiform or athetoid movements of the jaw, lower face, tongue, and extremities, developing in association with the use of an antipsychotic medication for at least a few months, though symptoms may appear after a shorter period in older persons. In some patients, movements of this kind may appear after cessation, or after alteration or decrease in dosage of antipsychotic drugs. Tardive syndrome includes other forms of movement complications, such as akathisia or dystonia, which are distinguished by their late appearance in the course of management and their potential perseverance for months to years, even despite antipsychotic discontinuation or dosage lessening [1]. TD can appear in various ways. Initial clinical symptoms are primarily messy movements in the facial areas, mouth and tongue, which seem uncontrollable and repetitive. Also, TD symptoms can affect movement of the torso, limbs, head and neck, and patients with severe disorders may also suffer from vague speech, abnormal postures and problematic swallowing [2]. The severity of the movements may range from slight to obviously incapacitating. TD is worsened by stress and vanishes during sleep [3]. TD develops in about 10 – 20 percent of patients who are treated for more than a year. About 20 – 40 percent of patients who require long-standing hospitalization have TD [4,5]. The occurrence of TD can also depend on whether the antipsychotic drug is atypical or typical, with around 13.1% incidence with atypical antipsychotic medications and about 32.4% rate with typical antipsychotic drugs [6,7]. Females, children, patients who are more than fifty years of age, and patients with brain injury or affective disorders are at higher risk [3,9]. Increased antipsychotic medication exposure (particularly typical antipsychotics), African-American ethnicity, cognitive disturbance, alcohol or substance abuse, early occurrence of drug-induced parkinsonism, diabetes, and HIV, as well, have been accounted as other risk factors for the development of TD [10,11]. Moreover, drugs used to treat Parkinson’s disease can cause TD [12,13] (Table 1). Up to now, sustained D2 receptor blockade resulting in receptor hypersensitivity is the most common theory explaining the development of TD. Besides, genetic studies have indicated a possible relationship with polymorphisms in the DA2 receptor, DA3 receptor, dopamine transporter (DAT1), and the serotonin 2A receptor genes. Oxidative stress and cell demise secondary to augmented glutamatergic neurotransmission triggered by blockade of presynaptic dopamine receptors is also hypothesized [14-18]. Although proof proposes a genetic susceptibility to TD [19], evidence suggests that a genetic protection against TD exists [20]. Furthermore, some studies suggest that D3, D4, and D5 receptors are also involved in the pathogenesis of TD [21,22]. Since anticholinergic agents are also associated with TD, an imbalance of acetylcholine and dopamine is likely involved in TD pathogenesis [23]. While usage of adjunctive agents, like vitamin B6, procholinergic agents (e.g., donepezil [Aricept]), Ondansetron (Zofran), a selective 5-HT3 receptor antagonist, cyproheptadine (Periactin), a 5-HT and histamine antagonist, and levetiracetam (Keppra) is of limited benefit [24] (Table 1), the use of deep brain stimulation for severe and refractory TD offers hope to those who are rigorously incapacitated [24]. Among patients with schizophrenia, the life quality of patients with TD may drop radically [25], and leads to a decline in patients’ social functioning, and may affect quality of life and treatment compliance meaningfully [26]. Additionally, TD can increase the difficulty in handling the primary disorder, thus increasing the economic burden on the patient’s family [27]. Currently, appropriate proof for effectiveness exists for two Vesicular monoamine transporter 2 (VMAT2) inhibitors, valbenazine and deutetrabenazine [28,29]. Among them, Valbenazine was the first drug that has been approved for TD in the United States [30,31].

Table 1: Medications that may induce or alleviate TD

Medications That Can Induce TD	Medications and Supplements Used to Treat TD
Antipsychotic Drugs	Cholingergic Agents
Anticholinergic Agents	Clozapine, Quetiapine, Olanzapine
Antidepressants (Trazodone, doxepin, clomipramine, and amitriptyline, Fluoxetine, sertraline, Selegiline, Rasagiline, Phenelzine)	Apomorphine Vesicular monoamine transporter 2 (VMAT2) inhibitor
Antiemetics	[Tetrabenazine, Tetrabenazine Analogs
Anticonvulsants (Phenytoin carbamazepine and lamotrigine)	(Valbenazine and Deutetrabenazine)] Clonazepam
Antihistamines	Propranolol
Decongestants (Phenylpropanolamine)	Amantadine
Antimalarials	Branched-Chain Amino Acids
Antiparkinson Agents	Ginkgo Biloba
Anxiolytics (Barbiturates, meprobamate benzodiazepines)	Antioxidant Medications and Supplements (zonisamide, yi gan san (a Chinese herb),
Biogenic Amines(Tyramine) Mood Stabilizers (Lithium) Stimulants	Levetiracetam, melatonin, omega-3 fatty acids, piracetam, resveratrol, vitamin B6, and vitamin E)

Methods

One hundred and one elderly patients (≥55 years old), who received a diagnosis of schizophrenia, according to the ‘Diagnostic and Statistical Manual of Mental Disorders, 5th edition (DSM-5)’ [1], and were hospitalized in the chronic section of a community psychiatric hospital in south of Tehran, had been selected for the present cross-sectional study, which had been performed in June 2006. While the study was carried out consistent with the ‘Declaration of Helsinki and Ethical Principles for Medical Research Involving Human Subjects’ [32], the patients were informed about the procedure, and a signed informed consent was received from those who were interested in participating in the study or from a legal guardian or representative. Abnormal Involuntary Movement Scale (AIMS) was employed to screen for patients with schizophrenia who also had TD. AIMS is a 12-item tool developed at the National Institute of Mental Health (NIMH) and has been utilized by clinicians to give a numeric measure to the observed atypical movements in various sections of the body [33,34]. The AIMS has a global rating of severity, a rating of incapacitation because of the irregular movements, and an evaluation of the patient’s attentiveness to the atypical movements. Moreover, it can be used to measure anomalous movements in different types of patients, including adults, children, and adolescents. The entire test can be completed in about 10-15 minutes. These items are rated on a five-point scale of severity from 0–4. The scale is rated between 0 (none), 1 (minimal), 2 (mild), 3 (moderate), and 4 (severe). Two of the 12 items denote dental care. The remaining 10 items denote body movements. Test-retest reliability at 6-8 weeks ranges from 0.40-0.82 for each item and is 0.71 for overall severity. The scale can be done as part of a physical-neurological examination by a trained clinician. The AIMS does not make the diagnosis of the disorder causing the movement abnormality unless some criteria have been established already to ease the diagnostic process. The Schooler–Kane research criteria are commonly used to find probable antipsychotic-induced TD, and need that three criteria are met: [1] symptoms occur after at least 3 months of treatment with an antipsychotic, [2] abnormal, involuntary movements must occur in 2 or more body regions if mild, or 1 body region if moderate to severe, as determined by a rating scale such as the AIMS, and [3] there are no other conditions that may have caused the abnormal movement patterns [35]. So, in the present appraisal, patients whose abnormal involuntary movements were induced by medical or neurological diseases were excluded. Scale for Assessment of Positive Symptoms (SAPS) [36], Scale for Assessment of Negative Symptoms (SANS) [37], Schedule for Assessment of Insight (SAI) [38], and Clinical Global Impressions – Severity of illness (CGI-S) [39], as well, had been used as ancillary scales for evaluation of severity of general psychopathology of schizophrenia, and comparing the TD patients with the group of patients without TD, for probing the intervening parameters.

Statistical Analysis

Baseline characteristics were compared by ‘t tests’ for continuous variables. Between-group analysis, too, with respect to ancillary scales, like SANS, SAPS, SAI, and CGI-S, was performed by ‘t tests’. Statistical significance is defined as P-value ≤0.05. ‘Med-Calc’ statistical software, version 15.2, was the statistical software tool for analysis.

Results

While abnormal movements were clear in 38.61% (n=39) of elderly schizophrenic patients, only seven of them (6.93 %) could be diagnosed as TD, based on the above-mentioned criteria. Medication induced Extrapyramidal adverse effects, like Parkinsonism or tremor, and other abnormal movements like Tic, Chorea, Myoclonus, Ballismus and rigidity, which were present either before initiation of illness or in advance of prescription of neuroleptic, or were generated later due to comorbid medical or neurological ailments, constituted the rest of abnormal movements in the present sample of aged schizophrenic patients. Among the said group with TD, and based on the assessment by AIMS, three patients had only abnormal facial and oral movements with minimal to mild severity (code: 1-2), one patient had atypical facial and oral movements in addition to odd extremity movements with mild to moderate (code: 2-3) and minimal to mild severity (code: 1-2), respectively, one patient had anomalous facial and oral movements in addition to irregular trunk movements with minimal to mild (code: 1-2) and mild to moderate severity (code: 2-3), respectively, and lastly two patients had unusual extremity movements with minimal to moderate severity (code: 1-3). While two of them, one patient with abnormal extremity movements and the other patient with atypical facial and oral movements, had problems with teeth, the rest of the patients usually wore dentures. In addition, in the present survey, around 71% of patients with TD were aware of their unusual movements. In this regard, while two patients with only anomalous facial and oral movements had no awareness of their odd movements (code: 0), one of the patients in the said cluster was aware of mild distress (code: 2). Two patients with irregular extremity movements were aware of the uniqueness of their atypical movements, with mild distress in one of them (code: 2) and no distress in the other one (code: 1). The patient with abnormal facial and oral movements plus atypical trunk movements was aware of moderate distress (code: 3). The same was applicable, as well, for the patient with nonstandard facial and oral movements in addition to strange extremity movements. Nevertheless, none of them could be considered as severely incapacitated due to the said abnormal movements. All of them were using conventional antipsychotic medications, like chlorpromazine, haloperidol, perphenazine and trifluoperazine [Mean ± SD mg/d Chlorpromazine equivalent = 464.28 ± 118.66], in companion with anticholinegic medications (biperiden or trihexyphenidyl). While between-group analysis did not show any significant difference between patients with TD and patients without TD about some demographic parameters, like age and duration of illness, no significant difference, as well, was evident between them with respect to measuring positive symptoms, negative symptoms, insight, and general psychopathology, which had been assessed by the said ancillary scales (Table 2).

Table 2: Comparative Analysis of Demographic and Psychopathologic Parameters

Variables	Patients with TD (n=7)	Patients without TD (n=94)	T	P	CI
Age (y/o)	66.14 ± 4.99	66.01 ± 7.36	0.046	0.96	-5.50, 5.76
Duration of illness (years)	33.57 ± 3.06	31.79 ± 6.93	0.672	0.50	-3.47, 7.03
Mean (sd) mg/d Chlorpromazine equivalent	464.28 ± 118.66	431.96 ± 159.03	0.526	0.60	-89.64, 154.28
SAPS	63.71 ± 9.62	57.44 ± 10.16	1.580	0.11	-1.60, 14.14
SANS	55.38 ± 6.40	49.96 ± 8.37	1.674	0.09	-1.00, 11.84
SAI	7.61 ± 2.97	8.83 ± 3.11	1.004	0.31	-3.63, 1.19
CGI-S	4.10 ± 2.35	3.22 ± 1.16	1.776	0.07	-0.10, 1.86

Abbreviations: SAPS: Scale for Assessment of Positive Symptoms; SANS: Scale for Assessment of Negative Symptoms; SAI: Schedule for Assessment of Insight; CGI-S: Clinical Global Impressions – Severity of illness; TD: Tardive Dyskinesia

Discussion

Medication-induced TD is a complex and distinctive neurologic condition [40]. While the reported incidence of TD seems to be reduced with the usage of atypical antipsychotic drugs, the risk of developing TD remains with these medications. Furthermore, several other medication classes have a high prevalence of TD and yet are not commonly considered to be TD-inducing [41-43]. Drug-induced Parkinsonism and TD are stigmatizing movement disorders linked with exposure to dopamine receptor blocking agents such as antipsychotic drugs, but they differ in their pathophysiology and clinical management. Treatment for one may exacerbate the other, and there are important diagnostic signs that help in making a precise evaluation and founding a sensible treatment strategy. On the other hand, since the presentation differs greatly among people, it often goes undiagnosed or can be easily misdiagnosed [27]. Though movement disorders were once thought to be associated with conventional antipsychotic medications, increasing attention is being given to the possibility of induction of movement disorders by most atypical antipsychotics [44]. On the other hand, some researchers believe that published prevalence rates of TD may be falsely low [45]. This is probably due to the insidious development of TD [46]. Back to our discussion and along with the findings of the current evaluation, the frequency of TD among our sample was lesser than what had been indicated by Koning et al. [4], Waln et al. [5], Kim et al. [6], Carbon et al. [7], Ward et al. [25], Saltz et al. [47], and Huang et al. [27], though it was slightly comparable to the finding of the last study [27]. Findings of Kim et al. [6], as well, were a bit comparable with the outcomes of the present assessment, though it was about atypical antipsychotic medications. On the other hand, maybe the presence of only male patients in the current estimation has altered the results adversely, which could be greater by the addition of female patients, especially when it has been declared that women are more likely to be affected than men [3,5]. Also, while the elderly schizophrenic patients shaped the present sample and increasing age is known as a risk factor for the development of TD [7], other similar studies are mostly about the prevalence of TD among adult patients aged between 20 and 70 [27]. Nevertheless, once more, the present outcome is remarkably less than the indicated measurements [3]. But, the present conclusion is somewhat similar to the findings of Go et al. [48], who found that patients of Filipino and Asian descent had a lower frequency of TD compared to patients of Caucasian descent, even though the Filipino and Asian patients consistently took a daily dose of 700 mg chlorpromazine for at least 5 years. Moreover, in contrary to the findings of Huang et al. [27], in the present survey no significant relationship was clear between TD and dosage of antipsychotic medication, scores of negative symptoms, and severity of symptoms or age. But in the current survey, too, the occurrence of movement disorders in the facial and oral areas of chronic schizophrenic patients with TD was the most frequent finding [2,27]. This outcome is consistent with the earlier reports that the anomalous involuntary movements in head and facial zones, whose classic symptom is the mouth-tongue-cheek triple sign, are seen the most in patients with TD [49]. Also, in the existing appraisal, abnormal extremity movements were more prevalent than abnormal trunk movements, and the proportion of TD patients with multiple affected areas in comparison with TD patients with a single affected area was higher, outcomes which were comparable to the conclusions of Huang et al. [27]. But, the proportion of TD patients with self-awareness about their abnormal movements in the present assessment was remarkably higher than what has been recounted by Huang et al. [27]. Anyway, as said by some scholars, we do not have a deep understanding of this disorder due to its vague etiology, various clinical symptoms, many affected areas and wide variation in demonstration by patients [27]. So, the uncertain pathophysiology of TD remains to be a problem for the effective treatment of this ailment [40], particularly, by taking into consideration that TD may also occur in never-medicated patients with schizophrenia [3]. Accordingly, the best strategy against TD is prevention. Prevention of drug-induced TD is focused around clinical considerations for pharmacologic physiognomy [40]. Therefore, healthcare staff are liable for teaching themselves and their patients about the risks associated with antipsychotic drugs and other TD-inducing prescriptions and following up the patients’ compliance, and only allow patients to stay on these drugs for long periods if absolutely compulsory [40]. On the other hand, in many low- and middle-income countries there is also a lack of mental health resources, which results in a poorer ratio of medical staff to patients. In clinical practice, this may cause less time obtainable for each patient and hence a later recognition and diagnosis of TD [27]. Consequently, the APA has recommended monitoring patients with schizophrenia for the development of TD every 3-12 months, depending on the patient’s risk factors and the class of antipsychotic drug. Principles include every six months for patients on a typical antipsychotic drug to every twelve months for patients on an atypical antipsychotic medication [50]. Though implementation of the study in the senior group of schizophrenic patients could be accounted for as an advantage on behalf of the present valuation, small sample size, male gender, and lack of control group were among the weaknesses of the current appraisal, which could prevent generalization of the conclusion and thus confirm it as a pilot study. Further methodical studies in future with larger sample sizes and a broader spectrum of oldness may bring about more apposite results and will probably make the existing state of affairs brighter.

Conclusion

According to the finding of the present study, prevalence of Tardive Dyskinesia among elderly schizophrenic patients, who were using typical antipsychotic medications, is very lower than what has been indicated thus far.

Acknowledgement

The author acknowledges physicians and personnel of Razi Psychiatric Hospital for their valuable support and assistance.

References

American Psychiatric Association (APA) (2013) Diagnostic and Statistical Manual of Mental Disorders. 5th ed. Washington, DC: American Psychiatric Association.
Stacy M, Jankovic J (1992) Tardive tremor. Mov Disord 7: 53-57.
Janicak PG, Hussain K (2017) Medication-Induced Movement Disorders In: Sadock BJ, Sadock VA, Ruiz P, eds. Kaplan & Sadock’s Comprehensive Textbook of Psychiatry. 10th ed. Philadelphia: Wolters Kluwer 2941-2942.
Koning JP, Tenback DE, van Os J, Aleman A, Kahn RS, et al. (2010) Dyskinesia and parkinsonism in antipsychotic-naive patients with schizophrenia, first-degree relatives and healthy controls: a meta-analysis. Schizophr Bull 36: 723-731. [crossref]
Waln O, Jankovic J (2013) An update on tardive dyskinesia: from phenomenology to treatment. Tremor Other Hyperkinet Mov 12: 3. [crossref]
Kim J, Macmaster E, Schwartz TL (2014) Tardive dyskinesia in patients treated with atypical antipsychotics: case series and brief review of etiologic and treatment considerations. Drugs Context 9: 212259. [crossref]
Carbon M, Hsieh CH, Kane JM, Correll CU (2017) Tardive dyskinesia prevalence in the period of second-generation antipsychotic use. J Clin Psychiatry 78: 264-278. [crossref]
Boland RJ, Verduin ML, Ruiz P, eds. (2021) Tardive Dyskinesia. Kaplan & Sadock’s Synopsis of Psychiatry. 12^th Philadelphia: Lippincott Wolters Kluwer 1893-1896.
Aquino CC, Lang AE (2014) Tardive dyskinesia syndromes: current concepts. Parkinsonism Relat Disord 20: 113-117. [crossref]
Solmi M, Pigato G, Kane JM, Correll CU (2018) Clinical risk factors for the development of tardive dyskinesia. J Neurol Sci 389: 21-27. [crossref]
Jankelowitz SK. Treatment of neurolept-induced tardive dyskinesia. Neuropsychiatr Dis Treat 2013; 9: 1371-1380. [crossref]
Tenback DE, Bakker PR, Van Harten PN (2015) Risk factors for tardive movement disorders in schizophrenia [in Dutch]. Tijdschr Psychiatr 57: 120-124. [crossref]
Wonodi I, Adami HM, Cassady SL, Sherr JD, Avila MT, et al. (2004) Ethnicity and the course of tardive dyskinesia in outpatients presenting to the motor disorders clinic at the Maryland psychiatric research center. J Clin Psychopharmacol 24: 592-598. [crossref]
Janicak PG, Marder S, Pavuluri M (2011) Principles and Practice of Psychopharmacotherapy. 5th ed. Philadelphia, PA: Lippincott Williams & Wilkins 65-180.
Pappa S, Dazzan P (2009) Spontaneous movement disorders in antipsychotic-naive patients with firstepisode psychoses: a systematic review. Psychol Med 39: 1065-1076. [crossref]
Rana AQ, Chaudry ZM, Blanchet PJ (2013) New and emerging treatments for symptomatic tardive dyskinesia. Drug Des Devel Ther 7: 1329-1340. [crossref]
Kulkarni SK, Naidu PS (2003) Pathophysiology and drug therapy of tardive dyskinesia: current concepts and future perspectives. Drugs Today (Barc) 39: 19-49. [crossref]
Jensen N, Oliveira JRM (2014) Basal ganglia vulnerability to oxidative stress. Front Neurosci 8: 80. [crossref]
Gałecki P, Pietras T, Szemraj J (2006) Manganese superoxide dismutase gene (MnSOD) polimorphism in schizophrenics with tardive dyskinesia from central Poland [in Polish]. Psychiatr Pol 40: 937-948. [crossref]
Fedorenko OY, Loonen AJM, Lang F, Toshchakova VA, Boyarko EG, et al. (2014) Association study indicates a protective role of phosphatidylinositol-4-phosphate-5-kinase against tardive dyskinesia. Int J Neuropsychopharmacol 18: 67-73. [crossref]
Liao DL, Yeh YC, Chen HM, Chen H, Hong CJ, et al. (2001) Association between the Ser9Gly polymorphism of the dopamine D3 receptor gene and tardive dyskinesia in Chinese schizophrenic patients. Neuropsychobiology 44: 95-98. [crossref]
Zai CC, Tiwari AK, Basile V, Luca VD, Muller DJ, et al. (2009) Association study of tardive dyskinesia and five DRD4 polymorphisms in schizophrenia patients. Pharmacogenomics J 9: 168-174. [crossref]
Clayton AH (1995) Antidepressant-induced tardive dyskinesia: review and case report. Psychopharmacol Bull 31: 259-264. [crossref]
Chang EF, Schrock LE, Starr PA, Ostrem JL (2010) Long-term benefit sustained after bilateral pallidal deep brain stimulation in patients with refractory tardive dystonia. Stereotact Funct Neurosurg 88: 304-310. [crossref]
Ward KM, Citrome L (2018) Antipsychotic-Related Movement Disorders: Drug-Induced Parkinsonism vs. Tardive Dyskinesia-Key Differences in Pathophysiology and Clinical Management. Neurol Ther 7: 233-248. [crossref]
Browne S, Roe M, Lane A, Gervin M, Morris M, et al. (1996) Quality of life in schizophrenia: relationship to sociodemographic factors, symptomatology and tardive dyskinesia. Acta Psychiatr Scand 94: 118-124. [crossref]
Huang Y, Pan L, Teng F, Wang G, Li C, et al. (2017) A Cross-Sectional Study on the Characteristics of Tardive Dyskinesia in Patients with Chronic Schizophrenia. Shanghai Archives of Psychiatry 29: 295-303.
Kane JM, Correll CU, Liang GS, Burke J, O’Brien CF (2017) Efficacy of Valbenazine (NBI-98854) in Treating Subjects with Tardive Dyskinesia and Schizophrenia or Schizoaffective Disorder. Psychopharmacol Bull 47: 69-76. [crossref]
Anderson KE, Stamler D, Davis MD, Factor SA, Hauser RA, et al. (2017) Deutetrabenazine for treatment of involuntary movements in patients with tardive dyskinesia (AIM-TD): a double-blind, randomised, placebo-controlled, phase 3 trial. Lancet Psychiatry 4: 595-604. [crossref]
Hauser RA, Factor SA, Marder SR, Knesevich MA, Ramirez PM, et al. (2017) KINECT 3: a phase 3 randomized, double-blind, placebo-controlled trial of valbenazine for tardive dyskinesia. Am J Psychiatry 174: 476-484. [crossref]
Factor SA, Remington G, Comella CL, Correll CU, Burke J, et al. (2017) The effects of valbenazine in participants with tardive dyskinesia. J Clin Psychiatry 78: 1344-1350.
World Medical Association (2013) World Medical Association Declaration of Helsinki: ethical principles for medical research involving human subjects. JAMA 310: 2191-2194. [crossref]
Lane RD, Glazer WM, Hansen TE, Berman WH, Kramer SI (1985) Assessment of Tardive Dyskinesia using the Abnormal Involuntsry Movement Scales. J Nerv Ment Dis 173: 353-357. [crossref]
Munetz MR, Benjamin S (1988) How to Examine Patients Using the Abnormal Involuntary Movement Scale. Hosp Community Psychiatry 39: 1172-1177. [crossref]
Schooler NR, Kane JM (1982) Research diagnoses for tardive dyskinesia. Arch Gen Psychiatry 39: 486-487. [crossref]
Andreasen N (1984) The Scale for Assessment of Positive Symptoms (SAPS), University of Iowa, Department of Psychiatry, Iowa City, Iowa.
Andreasen N (1981) The Scale for Assessment of Negative Symptoms (SANS), University of Iowa, Department of Psychiatry, Iowa City, Iowa.
David AS (1990) Insight and Psychosis. Br J Psychiatry 156: 798-808. [crossref]
Clinical Global Impressions, “ECDEU Assessment manual for psychopharmacology, Guy W, ed, Rockville: U.S Department of Health , Education , and Welfare, 1976, DHEW Publication NO. (ADM) 76-338.
Cornett EM, Novitch M, Kaye AD, Kata V, Kaye AM (2017) Medication-Induced Tardive Dyskinesia: A Review and Update. Ochsner Journal 17: 162-174. [crossref]
Khouzam HR (2015) Identification and management of tardive dyskinesia: A case series and literature review. Postgraduate Medicine 127: 726-737. [crossref]
Ward KM, Citrome L (2018) Antipsychotic-Related Movement Disorders: Drug-Induced Parkinsonism vs. Tardive Dyskinesia-Key Differences in Pathophysiology and Clinical Management. Neurol Ther 7: 233-248. [crossref]
Woods SW, Morgenstern H, Saksa JR, Walsh BC, Sullivan MC, et al. (2010) Incidence of tardive dyskinesia with atypical versus conventional antipsychotic medications: a prospective cohort study. J Clin Psychiatry 71: 463-474. [crossref]
Kane JM, Woerner M, Lieberman J (1988) Tardive dyskinesia: prevalence, incidence, and risk factors. J Clin Psychopharmacol 8: 52-56. [crossref]
Tarsy D, Baldessarini RJ (2006) Epidemiology of tardive dyskinesia: is risk declining with modern antipsychotics? Mov Disord 21: 589-598. [crossref]
Weiden PJ, Mann JJ, Haas G, Mattson M, Frances A (1987) Clinical nonrecognition of neuroleptic-induced movement disorders: a cautionary study. Am J Psychiatry 144: 1148-1153. [crossref]
Saltz BL, Woerner MG, Kane JM, Lieberman JA, Alvir JM, et al. (1991) Prospective study of tardive dyskinesia incidence in the elderly. JAMA 266: 2402-2406.
Go CL, Rosales RL, Caraos RJ, Fernandez HH (2009) The current prevalence and factors associated with tardive dyskinesia among Filipino schizophrenic patients. Parkinsonism Relat Disord 15: 655-659. [crossref]
Caroff SN, Hurford I, Lybrand J, Campbell EC (2011) Movement disorders induced by antipsychotic drugs: Implications of the CATIE schizophrenia trial. Neurol Clin 29: 127-148. [crossref]
Lehman AF, Lieberman JA, Dixon LB, McGlashan TH, Miller AL, et al. (2004) Practice guideline for the treatment of patients with schizophrenia, second edition. Am J Psychiatry 161: 1-56.