Monthly Archives: July 2019

Late Rise Human Chorionic Gonadotropin after Embryo Transfer: Causality and Significance! A mini review

DOI: 10.31038/IGOJ.2019233

Abstract

The reliable detection of hCG in maternal blood usually coincides with the embryonic implantation phase which is around 7 days post fertilization. It is believed that hCG levels taken post embryo transfers have a diagnostic and prognostic value when it comes to reproductive outcomes. Very low initial hCG levels predict a higher risk of chemical pregnancies, miscarriages and ectopic pregnancies. A review of the literature was performed so as to understand the mechanisms leading to late hCG rises as well as significance of such findings.

Introduction

Human chorionic gonadotropin (hCG) is produced by the placental syncytio-trophoblasts as early as 7–8 blastomere stage of the embryonic development i.e. before actual implantation takes place (M.-L.Bonduelle et al. 1988). The reliable detection of hCG in maternal blood usually coincides with the embryonic implantation phase which is around 7 days post fertilization (Ahmed et al. 1983). While serial hCG levels aren’t usually monitored in spontaneous pregnancies, women undergoing assisted reproductive technologies (ART) treatments usually necessitate such an approach especially after embryo transfer (ET). It is believed that hCG levels taken post ET have a diagnostic and prognostic value when it comes to miscarriages, ectopic pregnancies, predicting multiple gestations as well as live births (Schmidt et al., 1994, McCoy et al. 2009).

Discussion

Despite the lack of consensus on the hCG cutoff values that correlate with the best ART outcomes, a bulk of the studies use a value of 70 mIU/ml on day 14 post ovum pickup as an acceptable reference value (Sung et al. 2016). Values equivalent to 5 mIU/mL or below are judged as negative pregnancy tests (Sung et al. 2016, Maslow et al. 2016). It is believed that the amount of hCG produced reflects the mass of the trophoblast tissue as well as it’s function (Porat et al. 2007). Despite the discrepancies in the literature, there is a certain agreement that very low initial hCG levels are associated with adverse pregnancy outcomes and intra-uterine growth restriction. This can be explained by the small placental mass with a suboptimal function thus preventing normal fetal growth (Haddad et al. 1999, Krantz et al. 2004, Porat et al. 2007). A possible explanation for the latter might be that some embryos have a division lag, thus a later or abnormal implantation due to variances in trophoblast differentiation (invasive/extravillous versus hCG-producing/ villous phenotype) in an endometrium of decreased receptivity (Bolton et al. 1989, Woodward et al. 1993, Smith et al. 2004, Morse et al. 2016). Jukic et al. found out that smoking status and age at menarche affected the time of implantation by almost 24 hours and thus a late hCG rise. Current active or passive smoking status was significantly associated with delayed implantation. Younger age at menarche (younger than 12 years of age) was also found to be associated with a slow initial hCG rise (Jukic et al.2011). On another note, low initial hCG levels from 1.0 to 5.0 mIU/mL might be due to a false negative result related to laboratory methodology used for the hCG titration (Maslow et al. 2016). It s worth mentioning that it’s not only the initial hCG value but the doubling time as well as the hCG- rise curve is more correlated with the pregnancy outcome (Shamonki et al. 2009, Maslow et al. 2016, Morse et al. 2016). A doubling time of 2 days has been set as the best predictor of live birth rate although an increase rate as low as 53% can also predict a viable pregnancy (Shamonki et al. 2009, Seeber et al. 2012). Initial hCG value post ET is important to diagnose a possible pregnancy, however it doesn’t correlate alone with the possibility of a live birth. Serial hCG levels are important especially when the initial values are lower than the cut-off value.

References

  1. Ahmed, A. G., and A. Klopper. “Diagnosis of early pregnancy by assay of placental proteins.” BJOG: An International Journal of Obstetrics & Gynaecology 90.7 (1983): 604–611.
  2. Bolton VN, Hawes SM, Taylor CT, Parsons JH. Development of spare human preimplantation embryos in vitro: an analysis of the correlations among gross morphology, cleavage rates, and development to the blastocyst. J In Vitro Fert Embryo Transf 1989; 6: 30–5
  3. Bonduelle, M-L., et al. “Chorionic gonadotrophin-β mRNA, a trophoblast marker, is expressed in human 8-cell embryos derived from tripronucleate zygotes.” Human Reproduction 3.7 (1988): 909–914.
  4. Haddad, Bassam, et al. “Predictive value of early human chorionic gonadotrophin serum profiles for fetal growth retardation.” Human Reproduction 14.11 (1999): 2872–2875.
  5. Jukic, A. M. Z., et al. “The association of maternal factors with delayed implantation and the initial rise of urinary human chorionic gonadotrophin.” Human reproduction 26.4 (2011): 920–926.
  6. Krantz, David, et al. “Association of extreme first-trimester free human chorionic gonadotropin-β, pregnancy-associated plasma protein A, and nuchal translucency with intrauterine growth restriction and other adverse pregnancy outcomes.” American Journal of Obstetrics & Gynecology 191.4 (2004): 1452–1458.
  7. Maslow, Bat-Sheva L., et al. “Occult abnormal pregnancies after first post–embryo transfer serum beta-human chorionic gonadotropin levels of 1.0–5.0 mIU/mL.” Fertility and sterility 105.4 (2016): 938–945.
  8. McCoy, Travis W., Steven T. Nakajima, and Henry CL Bohler. “Age and a single day-14 β-HCG can predict ongoing pregnancy following IVF.” Reproductive biomedicine online19.1 (2009): 114–120.
  9. Morse, Christopher B., et al. “Association of the very early rise of human chorionic gonadotropin with adverse outcomes in singleton pregnancies after in vitro fertilization.” Fertility and sterility 105.5 (2016): 1208–1214
  10. Porat, Shay, et al. “Early serum β-human chorionic gonadotropin in pregnancies after in vitro fertilization: contribution of treatment variables and prediction of long-term pregnancy outcome.” Fertility and sterility 88.1 (2007): 82–89.
  11. Schmidt, Lila L., et al. “The predictive value of a single beta human chorionic gonadotropin in pregnancies achieved by assisted reproductive technology.” Fertility and sterility 62.2 (1994): 333–338.
  12. Seeber, Beata E. “What serial hCG can tell you, and cannot tell you, about an early pregnancy.” Fertility and sterility 98.5 (2012): 1074–1077.
  13. Shamonki, Mousa I., et al. “Logarithmic curves depicting initial level and rise of serum beta human chorionic gonadotropin and live delivery outcomes with in vitro fertilization: an analysis of 6021 pregnancies.” Fertility and sterility 91.5 (2009): 1760–1764.
  14. Sung, Nayoung, et al. “Serum hCG-β levels of postovulatory day 12 and 14 with the sequential application of hCG-β fold change significantly increased predictability of pregnancy outcome after IVF-ET cycle.” Journal of assisted reproduction and genetics 33.9 (2016): 1185–1194.
  15. Woodward BJ, Lenton EA, Turner K. Human chorionic gonadotrophin: embryonic secretion is a time-dependent phenomenon. Hum Reprod 1993;8: 1463–8.

Hydatid Disease and Pregnancy: A Short Note

DOI: 10.31038/IGOJ.2019232

Short Commentary

Tapeworm is the common name of the cestode worm, Echinococcus granulosus, which is the causal agent of hydatid disease in humans. This helminth infection is also known as cystic echinococcosis and hydatidosis, and is included in the zoonoses group. According to WHO [1]: (i) more than 1 million people are affected with echinococcosis at any one time; (ii) infection is globally distributed and found in every continent except Antarctica; (iii) robust surveillance is fundamental in order to show burden of disease and to evaluate progress and success of control programmes. However, as for other neglected diseases which are focused in underserved populations and remote areas, data is especially scarce and will need more attention if control programmes are to be implemented and measured; (iv) ultrasonography imaging is the technique of choice for diagnosis of cystic echinococcosis; (v) cysts can be incidentally discovered by radiography. Specific antibodies are detected by different serological tests and can support the diagnosis. Biopsies and ultrasound-guided punctures may also be performed for differential diagnosis of cysts from tumors and abscesses; (vi) cystic echinococcosis are often expensive and completed to treat, sometimes requiring extensive surgery and/or prolonged drug therapy.

As to transmission: (i) a number of herbivorous and omnivorous animals act as intermediate hosts of E. granulosus. They become infected by ingesting the parasite eggs in contaminated food and water, and the parasite then develops into larval stages in the viscera; (ii) carnivores act as definitive hosts for the parasite, and host the mature tapeworm in their intestine. They are infected through the consumption of viscera of intermediate hosts that harbor the parasite; (iii) humans act as so- called accidental intermediate hosts in the sense that they acquire infection in the same way as other intermediate hosts, but are not involved in transmitting the infection to the definitive host; (iv) several distinct genotypes of E. granulosus are recognized some having distinct intermediate hosts preferences. Some genotypes are considered species distinct from E. granulosus. Not all genotypes cause infections in humans. The genotype causing the  great majority of cystic echinococcosis infections in humans is principally maintened in a dog-sheep-dog cycle, yet several other domestic animals may also be involved, including goats, swine, cattle, camels and yaks; (v)  human infection with E. granulosus leads to the development of one or more hydatid cysts located most often in the liver and lungs, and less frequently in the bones, kidney, spleen, muscles, central nervous system and eyes; (vi) abdominal pain, nauseas and vomiting are commonly seen when hydatids occur in the liver. If the lung is affected, clinical signs include chronic cough, chest pain and shortness of breath. Other signs depend on the location of the hydatid cysts and the pressure exerted on the surrounding tissues.

These results of the parasitism for tapeworms in humans show that it may have direct and indirect consequences in human health, in general.  cause devastating morbidity with severe consequences in general and in female reproductive health.  Here our objective is to alert medical practitioners of gynecology and obstetrics for hydatid cyst in pregnancy.

The authors: (2) report that “Incidence of hydatid disease in pregnancy ranges from 1 in 20,000 to 1 in30,000. The diagnosis of liver hydatid cyst is not difficult but management during pregnancy is problematic. Both medical and surgical treatments are available but there is no consensus and each case has to be individualized”; (3) report “we present a 32-years old multigravida at 25 weeks of pregnancy in whom splenic and liver cysts were diagnosed by ultrasonography and magnetic resonance imaging (MRI). The splenic cyst was removed and a healthy baby was delivered vaginally at term”; (4) report “Case report A 25-year-old, gravida 1, para 0 pregnant female with history of liver cyst was admitted to the obstetric outpatient clinic for checkup. Patient who did not have any complaints was redirected to the radiology clinic for a second trimester ultrasound scan. Abdominal ultrasound revealed lesions of type-1, hydatid cyst 84×67 mm in size at the right lobe fifth segment, and 67×64 mm in size at the seventh segment of the liver, with millimetric echogenic female cysts and well-defined thickwalls. In addition, a type-2 hydatid cyst with membrane dissociation, 55×52 mm in size, was observed at the caudate lobe. An intrauterine fetus with fetal heart rate was observed with a bipariental diameter concordant with 21 weeks of gestation. Besides, there was a cystic lesion with fine septations surrounding the uterus. The patient was admitted to the obstetric care service with hydatid cyst hydatidosis in the vicinity of the uterus; (5) report “A young, apparently healthy from a rural area in South Africa presented in the third trimester of pregnancy with a symptomatic abdominal mass between the uterine fundus and liver. The etiology was established to be an echinococcus cyst of the liver and medical treatment was initiated. The fetal outcome was good but the mother died 3 days postpartum due to an unusual but devastating complication of the hydatid cyst; (6)  report the management of hydatid disease in pregnancy, and  they write that “in this review, we have attempted to summarize the presentation and available management approaches to hydatid disease, and have suggested evidence-based guidelines for its management during pregnancy; (7)  report  2 cases of hydatid disease of liver during pregnancy, and they also   made a review of literature; (8) report “A huge primary hydatid cyst of uterus” and they made also a review of literature.

Final conclusion: 1 – we think that it was here demonstrated the importance of the hydatid cyst as an underlying cause of negative effects in pregnancy; 2 – surgery in patients with hydatid cysts needs special care before, during or after surgery. Effectively, opening of a hydatid cyst requires special care not to spill the contents into the peritoneal cavity or tissues, since this may result in an anaphylactic reaction to the spilt fluid or dissemination and implantation of the immature scolices contained in the “sand” in the fluid. Incomplete removal of viable germinal epithelium from the liming of a hydatid cyst results in the formation of multiple cysts: 3 – we are in agreement with the conclusions of the authors (9): E. granulosus can affect any organ in the body from head to toe, and high suspicion of this disease is justified in endemic regions.

Keywords

Helminths; Cestodes; Tapeworms; Echinococcus granulosus; Hydatid Disease; Pregnancy; Gynecology; Obstetrics.

References

  1. WHO (18 February 2018) Echinococcosis Key facts.
  2. Ghosh JK, Goyal SK, Behera MK, Dixit VK, Jain AK. 2014 Hydatid cyst of liver presented as obstructive jaundice in pregnancy; Managed by PAIR. Journal of Clinical and Experimental Hepatology, https://doi.org/10.1016/j.jceh.2014.11.002
  3. Can D, Oztekin O, Oztekin O, Tinar S, Sanci M. Hepatic and splenic hydatid cyst during pregnancy: a case report. Archives of Gynecology and Obstetrics, August 2003, 268(3): 239–240.
  4. Tekin AF, Yilmaz H, Kara T, Seçkin E, Aybay MN, Alkan E, Case report A very rare case hydaid bcyst surrounding uterus and magnetic resonance imaging findings in the pregnant patient, 2018 Available online at www.sciencedirect.com.
  5. Robertson M, Geerts L, Gebhardt S. A case of hidatid cyst associated with postpartum maternal dead. Ultrasound Obstet Gynecol 2006; 27: 693–696.
  6. Rodrigues G, Seetharam P. Management of hydatid disease (echinococcosis) in pregnancy. Obstet Gynecol Surv, 2008 Feb; 63 (2): 116–123.
  7. Demirel E, Ekmekci E, Izmirkatip C., Keeckci S, Gencdal S. Hydatid disease of liver during pregnancy: Report of two cases and Review of literature. 2018, Jacobspublishers.com, email: erem.dr@hotmail.com
  8. Slimane NN, Taieb M, Khiali R, Rabehi H, Bekhouuche R, et al. A huge primary hydatid cyst of uterus. A case report and review of literature. J Univer Surg 2018; 6(2): 12 (5pags)
  9. Sachar S, Goyal S, Sangwan S. Uncommon locations and presentations of hydatid cyst. Ann Med Health Sci Res 2014; 4: 447–452.

Does Rider Weight have a Measurable Effect on the Horse’s Back Muscle Longissimus dorsi?

DOI: 10.31038/IJVB.2019315

Abstract

The topic of rider weight in relation to a horse´s body weight, the so called Body Weight Ratio (BWR), has been discussed widely with regard to both health and performance by the equestrian community. However, direct measurements of the effects of a riders weight on the back muscle of horses is lacking. This study uses non-invasive multi-frequency bioimpedance (mfBIA) and Acoustic Myography (AMG) to measure the health and performance of m.Longissimus dorsi in 10 horses and with three groups of riders; Light-weight (BWR 17%), Medium (BWR 19%) and Heavy-weight (BWR 27%). mfBIA values obtained from the horses prior to being ridden at the walk and trot, revealed information about muscle mass, swelling, resting tension and metabolic activity. AMG values revealed not only the real-time efficiency/coordination of the muscle, but also it’s spatial- and temporal-summation during periods of physical activity. The results revealed no significant effect of rider weight on the mfBIA parameters of the horses in this study, and AMG values were likewise not significantly different across the three rider groups. It is tentatively concluded, that rider weight, within the limits of this study, does not appear to affect back muscle health or performance.

Introduction

Despite a lack of scientific basis for the introduction of prescribed loading capacities for horses, the Japanese Riding for the Disabled Association describes a loading capacity of between 16–17% of the body weight of the horse, whilst Hadrill in their volume entitled “Horse Healthcare”, suggests a limit of 33 to 50% of the horse´s body weight [1,2].

Interestingly, Matsuura and colleagues published findings that short and wide horses are better suited to riding with disabled individuals, so called therapeutic riding, although their gait may be disrupted when riders are too heavy [3]. It has also been reported that horses carrying loads of 25–30% of their body weight have both elevated heart rates and respiration rates [4].

Then recently, a study of 8 Icelandic horses confirmed the findings of Powell and colleagues [4], by showing that body-weight ratios of 20–35% result in an increase in heart rate, an elevated frequency of breathing as well as a rise in rectal temperature, all physiological changes that one might expect with an elevated level of physical exertion [5]. More interestingly though, and in keeping with the results of Matsuura and colleagues [3], this same research group subsequently published findings that show that the stride length of horses becomes significantly shorter and more frequent with increasing rider weight [6]. This is not altogether surprising, as anyone who has walked whilst carrying something heavy will have noticed that they do not walk with long strides, but rather short and frequent movements of their feet.

Whilst these and other studies investigating the influence of the rider on the horse have evaluated weight and riding technique [7–9], there is still a lack of direct methodology, evaluating the effect of loading on the back muscles of ridden horses.

This study has therefore chosen to measure the changes in the back muscle Longissimus dorsi of horses ridden by riders of different body-to-weight ratios (Light-weight, Medium and Heavy-weight) using the non-invasive techniques of multi-frequency bioimpedance (mfBIA), and Acoustic Myography (AMG). mfBIA as a method, can be used to directly assess changes in muscle tension, metabolic status and cellular health, whilst AMG enables the real-time assessment of muscle contraction (coordination, spatial summation, temporal summation) [10–12]. The hypothesis being, that rider weight per se, does not affect muscle health or performance.

Materials and Methods

Subjects

Ten healthy horses were used for data collection. The population consisted of 2 mares and 8 geldings, of which there was 1 Danish Warmblood, 1 Oldenburg, 4 Icelandic horses, 1 OX Arabian, 1 Appaloosa and 2 Fjord horses. The mean age of the horses was 10.4 ± 2.5 years. The body weights of the horses in this study ranged from 334–732 kg.

In order to investigate muscle health and function of the ten subjects, mfBIA measurements and AMG recordings were conducted on m. Longissimus dorsi. The measurements were carried out at the respective home of the subjects, in order to avoid any stress or environmental interference. Furthermore, the riders were recommended not to exercise their horse for between 24–48 hours before the recordings were due to be made, in order to achieve the best possible mfBIA and AMG results. These measurements, which were non-invasive, were taken with the full informed consent of the owners and riders.

Equipment and Measurements

Acoustic Myography

A gel pad was placed under the saddle (Acavallo Gel; Lonato del Garda, IT), with the AMG recording unit (CURO-diagnostics ApS, Bagsværd, DK) attached to the pad behind the saddle. CURO sensors (CURO-diagnostics ApS) were placed on both the left and right sides of the horse at the region of m. Longissimus dorsi. The muscle sites measured were prepared with acoustic gel (CURO-diagnostics ApS), which was thoroughly rubbed into the overlying hair to ensure a good connection with the skin above the muscle. Similarly, the two sensors were prepared with acoustic gel and attached to the horse using flexible self-adhesive bandage (Animal Polster, Snögg Industry AS, Kristiansand, NO). Smaller pieces of self-adhesive bandage were used to secure sensor cables to avoid errors caused by irritation of the horse and contact between the cables and sensors. The sensors were then connected to the CURO unit. Recordings were made to both an iPad in real-time, as well as directly to the CURO unit itself, in the form of a WAV file. Data collection was made during walk and trot on both left- and right-hand circles. The subjects were ridden by their usual riders and with their usual saddle and riding equipment. For further details see [10,12].

Multi-frequency bioimpedance

For mfBIA measurements, the horse was restrained in a calm standing position. The region of m. Longissimus dorsi of interest was prepared by the application of conductive paste (Ten20; Weaver and Company, Aurora, Colorado USA), followed by placement of four pure platinum electrodes (1 x 3cm; made by AH) on to the prepared muscle. The mfBIA unit (ImpediVET BIS 1, Pinkenba, AU) providing a current of 1000 μA, was subsequently attached to the electrodes. Recordings were carried out at 256 frequencies ranging from 3 kHz to 850 kHz and repeated six times with a one second interval. By repeating the recordings, it was possible to avoid any slight movement artifacts or changes in the R or Xc values due to cable movement or change in body stance (Elbrønd et al., 2015). Throughout the recordings, the R-, Xc- and the full Cole-Cole plot was assessed for normality, in order to validate the strength and accuracy of the recordings. For further details see [11].

AMG data processing and analysis

Recorded data, stored on the CURO, was analyzed for its efficiency (E-score), amplitude (S-score; spatial summation) and frequency (T-score; temporal summation), using the CURO System Software (CURO-diagnostics ApS, Bagsværd, DK). The analysis was carried out with a maximum frequency (max T) of 160 Hz and a maximum amplitude of 0,99 (max S) equivalent to approx. 1V. Analysis was carried out for both sides of m. Longissimus Dorsi while riding on both circles.

The E-, S- and T-values from the two sides of the muscle were pooled and the means ± standard deviations calculated for walk and trot within the groups; Light-weight, Medium and Heavy-weight.

mfBIA data processing and analysis

The mfBIA data were analyzed using the ImpediVET software. At the time of recording, the Cole-Cole plots were assessed for a normal distribution and the R and Xc values and plots were examined to ensure precise recordings. Subsequently, a detailed analysis was performed at 50 kHz, where the parameters Z, R, Xc, fc, Re, Mc and Ri were obtained for each subject. The Phase Angle (PA) was calculated as (arctan Xc/R). The mean ± standard deviation for each group; Light-weight, Medium and Heavy-weight were calculated.

Statistical analysis

AMG and mfBIA data were initially assessed for normal distribution using a D’Agostino & Pearson normality test in GraphPad Prism 7 for Windows (La Jolla, CA, USA). Due to the small sample size (n=10), the normality test had very little power to discriminate between normal and non-normal distributions. Nevertheless, the majority of the tested data turned out to be normally distributed.

Results

AMG results

The AMG data for the measured horses during both walking and trotting revealed a non-significant difference for the E-, S- and T-scores for all three weight groups; Light-weight (BWR = 17%; n=2), Medium (BWR = 19%; n=6) and Heavy-weight (BWR = 27%; n=2) (see Figs 1 & 2).

At the walk, a very similar E-score and T-score were found for all three rider groups, indicating an identical degree of muscular efficiency/coordination and temporal summation (approx. 75Hz). Moreover, despite an apparently higher S-score for the Light-weight and Heavy-weight groups compared with the Medium group, there was no significant difference between the values in terms of spatial summation. This serves to indicate that muscle function for all three groups was not statistically different at this gait.

IJVB 2019-107 - Adrian Denmark_F1

Figure 1. The mean ± SD of the E-, S- and T-scores recorded during walking for m. Longissimus Dorsi. Values obtained from both sides have been pooled within the three groups. Red = Heavy-weight (n=4; 2 horses x 2 muscles), Blue = Light-weight (n=4; 2 horses x 2 muscles), Black = Medium groups (n=12; 6 horses × 2 muscles).

IJVB 2019-107 - Adrian Denmark_F2

Figure 2. The mean ± SD of the E-, S- and T-scores recorded during trotting for m. Longissimus Dorsi. Values obtained from both sides have been pooled within the three groups. Red = Heavy-weight (n=4; 2 horses x 2 muscles), Blue = Light-weight (n=4; 2 horses × 2 muscles), Black = Medium groups (n=12; 6 horses × 2 muscles).

At the trot, a very similar E-score, S-score and T-score was found for all three rider groups, indicating an identical degree of efficiency/coordination, spatial summation and temporal summation (approx. 40Hz). This likewise serves to indicate that muscle function for all three groups was not statistically different at this gait.

However, when comparing the AMG data from the two gaits, the E- and S-scores were found to be consistently higher during the walk for all three groups, whereas the highest T-score values were found during trotting.

.mfBIA results

No consistent body-weight-ratio patterns were noted for either the Light-weight, Medium or Heavy-weight groups, nor were any significant differences noted. When the Light-weight mfBIA values were plotted against those obtained for the Heavy-weight group (see Fig 3), it was found that values for muscle mass (Z; PA) were comparable, as was the indicator of resting tension (fc). Likewise, there were no signs of dehydration or inflammation (R; Re) between the two extremes. There was an elevated value for (Ri), which has been shown to be correlated with VO2-max at rest in the Heavy-weight cf Light-weight group, but this was not found to be significant.

IJVB 2019-107 - Adrian Denmark_F3

Figure 3. mfBIA values for the two extreme body weight groups, measured for m. Longissimus Dorsi. Red = Heavy-weight (n=2), and Blue = Light-weight groups (n=2). Values are mean ± SD.

Discussion

Whilst these findings represent a limited data set, they appear to suggest that the weight of the rider to the horse´s weight per se does not affect muscle function or muscle health, as documented by the AMG and mfBIA measurements.

In terms of possible confounding factors, it should be noted that whilst two horses from the Medium group had been competing in the days up to the study, a factor that might have influenced the accuracy of the measurements, this level of physical activity does not appear to have had any negative effect.

The mfBIA and AMG data reveal a very similar picture, that is one in which the back muscle Longissimus dorsi is quite relaxed (low fc value) and very comparable across the two extremes of Light-weight and Heavy-weight groups. The mfBIA data further reveals a very comparable muscle mass (Z, PA) for these two extreme groups, and there are no signs of swelling or inflammation (R, Re). Indeed, it can be concluded that this back muscle is fit and healthy for both extreme groups, as well as for the Medium group of riders, being comparable with previously published values [11].

The AMG data likewise, reveal fit and healthy scores for m. Longissimus dorsi. It is interesting to note a very similar E-score for the three groups, suggestive of a similar level of training and performance, as this value increases with highly trained horses, and falls with inactivity and convalescence. What is striking is that the T-score remains very consistent between the groups. One could have anticipated that some degree of muscle soreness was present in the Heavy-weight group of horses, and it is known that soreness/pain affects temporal summation, raising the firing frequency of afflicted muscles [13]. However, this was not observed either at the walk or the trot.

It is interesting though that the change in gait from walk to trot is reflected in the E,S,T-scores as has been reported previously in dogs [14]. It can be seen that there is a fall in the E-score as the muscle becomes more active with the change from walk to trot, contracting more of the time (E-score approx. 3 to 1–2). At the same time the S-score falls a little from 6–8 at the walk to approx. 5 at the trot, reflecting an increase in fibre recruitment (spatial summation). In contrast though, the T-score increases from approx. 5.5 at the walk (approx. 75Hz) to 7.5 at the trot (approx. 40Hz). This improvement in the T-score, which represents a drop in firing frequency, is very comparable with values for dogs and m.longissimus lumborum, as they change from walk to trot [14].

In conclusion, whilst these data do not reveal any suggestion that high rider-to-horse body weight ratios have an adverse effect on muscle health or function, as assessed by the non-invasive techniques of mfBIA and AMG, there is now a great need for a more detailed study in this field.

Acknowledgement

The authors are indebted to the horse owners for allowing us to measure them and their horses.

Conflicts of Interest

AH is in the process of forming a company with the aim of commercializing the AMG equipment.

Reference

  1. Matsuura A, Sakuma S, Irimajiri M, Hodate K (2013) Maximum permissible load weight of a Taishuh pony at a trot. Journal of Animal Science 91: 3989–3996.
  2. Hadrill D (2002) Horse Healthcare. 1st Edition. ITDG Publishing, London.
  3. Matsuura A, Ohta E, Ueda K, Nakatsuji H, Kondo, S (2008) Influence of equine conformation on rider oscillation and evaluation of horses for therapeutic riding. Journal of Equine Science 19: 9–18.
  4. Powell DM, Bennett-Wimbush K, Peeples A, Duthie M (2008) Evaluation of indicators of weight-carrying ability of light riding horses. Journal of Equine Veterinary Science 28: 28–33.
  5. Stefánsdóttir GJ, Gunnarsson V, Roepstorff L, Ragnarsson S, Jansson A (2017) The effect of rider weight and additional weight in Icelandic horses in tölt: part I. Physiological responses. Animal 11: 1558–1566.
  6. Gunnarsson V, Stefánsdóttir GJ, Jansson A, Roepstorff L (2017) The effect of rider weight and additional weight in Icelandic horses in tölt: part II. Stride parameters responses. Animal 11: 1567–1572.
  7. Clayton HM, Lanovaz JL, Schamhardt HC, Van Wessum R (1999) The effect of a rider´s mass on ground reaction forces and fetlock kinematics at the trot. Equine Veterinary Journal 30: 218–221.
  8. Roepstorff L, Egenvall A, Rhodin M, Byström A, Johnston C, et al (2009) Kinetics and kinematics of the horse comparing left and right rising trot. Equine Veterinary Journal 41: 292–296.
  9. De Cocq P, Duncker AM, Clayton HM, Bobbert MF, Muller M, et al (2010) Vertical forces on the horse´s back in sitting and rising trot. Journal of Biomechanics 43: 627–631.
  10. Riis KH, Harrison AP, Riis-Olesen K (2013) Non-invasive assessment of equine muscular function: A case study. Open Veterinary Journal 3: 80–84.
  11. Harrison AP, Elbrønd VS, Riis-Olesen K, Bartels EM (2015) Multi-frequency bioimpedance in equine muscle assessment. Physiological Measurements 36: 453–464.
  12. Jensen A-M, Ahmed W, Elbrønd VS, Harrison AP (2018) The efficacy of intermittent long-term bell boot application for the correction of muscle asymmetry in equine subjects. Journal of Equine Veterinary Science 68: 73–80.
  13. Graven-Nielsen T, Kendall SA, Henriksson KG, Bengtsson M, Sörensen J, et al (2000) Ketamine reduces muscle pain, temporal summation, and referred pain in fibromyalgia patients. Pain 85: 483–491.
  14. Fenger C, Harrison AP (2017) The application of acoustic myography in canine muscle function and performance testing. SOJ Veterinary Science 3: 1–6.

Zinc Supplements – Any Benefit in Diabetes?

DOI: 10.31038/EDMJ.2019342

Abstract

Zinc is a divalent cation mainly present intracellularly and exerts several indispensible effect therefore maintaining zinc homeostasis is essential. Zinc has a pivotal role in the insulin production, storage and pathways and in carbohydrate metabolism. Plays also an important function in the development of diabetes mellitus and diabetic complications as well. Based on preclinical and clinical studies here we present the most important contributions of zinc to diabetic state and briefly demonstrate why zinc supplementation is beneficial for diabetic patients.

Keywords

Zinc, Supplementation, Diabetes mellitus

Introduction

Zinc (Zn) is the third abundant bivalent cation in the human body, after calcium. Zn is found 99,8% intracellularly. It is part of several hundreds of enzymes and of ca. two thousand cofactors. Stored mainly in skeletal muscle and brain neurones. Zn plays very important role in cognitive functions and learning, in cell proliferation, in immun system, the physical development of infants and children, the skin, the metabolic processes, etc. Unfortunately, plasma zinc is insensitive to early zinc deficit and detection of hypozincemia is not a routin process. Zinc deficiency produce a myriad of symptoms but non of them is typical for the Zn deficit. However, as an ubiquitous trace elemet, zinc has a role in the carbohydrate metabolism and its regulation as well. Diabetes mellitus (DM) is a disease based on absolute or relative insulin deficiency with subsequent disturbances in carbohydrate metabolism. DM exists in two basic forms like type I and type II diabetes (T1DM and T2DM, respectively). Although the two types of diabetes are totally different from ethiology point of view there are numerous well defined and similar Zn effects in both T1DM and T2DM. Hypozincemia and hyperzincuria is part of the typical clinical symptoms of both DM forms therefore control of Zn deficiency in diabetic patients is necessary [1].

In the present compilation we briefly demonstrate some facts and rational considerations about use of zinc supplementation in diabetes patients.

1. Preclinical Experiences

Pancrease Beta-Cell Stimulation

Zinc exert a dual effect in beta-cells: via direct effect Zn is increasing of free insulin concentration near to beta cells besides it has an inhibitory effect on glucose stimulated insulin secretion. Transient elevation of se-glucose concentration increase free cytosolic Zn levels facilitating insulin storage, but chronic high intracellular zinc concentration resuling in beta cell dysfunction, in the extreme also cell death. Metabolic stress and hypoxia downregulate zinc-importer ZnT8 protein with subsequent decrease in the intracellular/intragranular* Zn concentration. Research group of Liu demonstrated that also ZnT6 and ZnT7 transporters are needed to defend insulin secretion in beta-cells and the insulin storage [2].

Zn in Insulin

Pancreatic beta cells have exceptionally high zinc content. Zinc is prerequisit for the hexamerization (ie. three dimer structure of insulin), which is the storage form of insulin (Fig 1.) in the beta-cell granules [3]. But Zn is also necessary to the transformation from proinsulin to insulin. Therefore to maintain normal function of the production and storage of insulin zinc is indispensible [4].

EDMJ _Telessy Istvan Hungary_F1

Figure 1. Insulin hexamer with stabilizer Zn++ in the middle.

Insulin Mimetic Function

Among others also trace elements (eg. vanadium) may have insulin-like properties. In vitro (cell culture) zinc administration is able to activate insulin signaling pathway [5]. Several mechanisms of action were assumed, eg. rat experiments supported that insulin-mimetic actions of zinc are mediated via inhibition of glycogen synthase kinase 3beta. In vivo ZnO nanoparticles – in mice and rat model – improve glucose tolerance, decrease blood glucose levels, increase expression of DM related gens, eg. that of insulin and insulin receptors’, GLUT2 and glucokinase, furthermore elevate blood insulin concentration [6]. Zn also augment insulin-binding on insulin-receptors.

Modification of Carbohydrate Metabolism

Zinc hinders carbohydrate absorption by inhibiting alpha-glucosidase activiy (direct binding resulting in conformation-change) in the small intestine. The low zinc intake increases glucose utilization in rats [7]. It increases phosphofructokinase and pyruvate kinase enzymes thus support glycolysis and the subsequent lactate production. Zinc overcome the inactivation of pyruvate kinase by glucagon, too. This ion also enhances glucose transport in adipocytes and blocks glycogen synthesis by indirect enzyme inhibition [8]. Noteworthy, Zn improve lipogenesis and support expression of PPARgamma, one of the regulators of fatty acid storage and glucose metabolism, furthermore insulin sensitizer.

*ZnT8 transmembrane protein is essential component of plasma mambrane of insulin secretory granules of pancreatic beta-cells. Its imbalance basically influence beta cell function and may contribute to development of glucose intolerance.

Antioxidant Effect

In diabetes mitochondria produce more reactive free oxigen radicals which have direct (modification of insulin signaling) and indirect (diabetes-induced complications) effects [9,10]. This is one of the hypotheses why use of antioxidants in diabetes is recommended. Zinc exert antioxidant effect in various targets and normalize the in diabetes low levels of glutathion peroxidase, restore catalase and superoxide dismutase activity and activates antioxidant metallothionein sysnthesis [11].

Interactions among the Trace Elements

Copper administration in rats impairs body gain, diminishes the se-zinc and tissue zinc levels and worsened clinical chemical blood parameters like alkaline phophatase, lactic dehydrogenase and amylase in diabetes as well as in non-diabetes animals. Addition of high dose of zinc to food ameliorates the referred parameters [12]. High doses of zinc hinders absorption of copper. Presence of moderate amounts of iron and iron-fortificated food does not influence significantly the Zn-absorption. In contrast, zinc supplementation inprove iron uptake in the diabetic intestine [13]. Zn inhibit N-methyl-D-aspartase (NMDA) receptors therefore Zn deficit results in increased intracellular Ca-levels [14]. These and other examples demonstrate that the abovementioned bivalent cations are normally in balance and supplementation of the trace elements should be administered simultaneously.

2. Zinc Supplementation

The most natural way of zinc intake is food consumption. In general, fruits and vegetables are poor in zinc except potatoes and in less extent green beans and kale. Concentrated dairy products like cheese contains more Zn and one serving of nuts (cashew, almonds, pine nut) or seeds (especially hemp, flax or pumpkin-seed) is also containing the estimated average requirement of this cation. Best sources from this aspect are red meat (beef, in lower degree lamb, pork) and shellfish (oysters are paticularly high in Zn but, crab, shrimps, mussels are also very rich in zinc). Zinc content of some groups of food is presented in Table 1. Recommended Daily Allowance (US Institute of Medicine) for zinc is between 8 – 11 mg/day. It seems in case of zinc deficiency restoration of body zinc content by food intake is difficult.

Table 1. Zinc content of selected foods.

Food

Cereals (ready-to-eat)

Shellfish, mussels

Beef, meat

pumpkin seed

Lamb, chicken

Cheese

Fruits, fruit coctail

Milk, dairy

Mushrooms

Mg Zn/100g food

10–30

50–70

6–12

10

2–4

3–6

0,5–7

0,3–0,5

0,6–1,2

Various attempts have been made to improve Zn-content of staple foods. It seems high Zn-content of soil or Zn-spaying of plants during the growth (biofortification of food) may help in zinc-deficiency on population level.

Zinc absorption takes place mainly in the proximal jejunum and distal duodenum. Absorption from food ranges from 11 to 22% but absorbed proportion depends on the body Zn-content and the form of zinc as well as on the constituents of the specific food (enhancers and inhibitors) [15]. The high protein-content may rise the Zn-absorption and high Cu++ hampers its absorption. These conditions may explain why zinc absorption from the US diet is estimated around 30% [16]. In contrast, high phytate content of food (phytate: zinc molar ratio >ca.14) can inhibit zinc intake [17]. The list of high phytate-containing foods are presented in Table 2. The aggregation of zinc deficiency in low-income developing countries supposedly due to regular consumption of high phytate-containing plant-based food [18]. Various pure chemical supplements, like zinc oxide, zinc sulfate, zinc citrate, zinc gluconate, demonstrated an absorption of 49,9 – 61,3% in human [19]. Constant cytosolic Zn concentration is maintained by SLC 30 and SLC39 gene families controlled zinc transporters , named ZnT-s with 10 members and ZIP-s with 14 members, respectively [20]. Zinc transporter gene SLC 30A1 plays a direct role in inorganic zinc (eg. ZnCl, ZnSO4) absorption. The absorption from Zn-gluconate is better than that from ZnO (Cmax: 18,3% but not Tmax) and also AUC was better by 8,1% [21]. The uptake of organic Zn-compounds (eg. Zn-glycine and Zn-methionine) is less dependent on this transporter-system [22].

Table 2. Phytates in foods.

Food

Phytate content (mg/100g)

kidney beans

610–2.380

peas

510–1.770

lentils

270–1510

mais

720–2.200

wheat (whole)

390–1.350

rice

60–1.080

peanuts

170–4.470

almonds

650–9.420

walnuts

200–6.690

Dietary supplements usually contain supraphysiologic doses (20–40 mg or more) of zinc in inorganic (oxide compound or chloride/sulphate salt), or recently rather in organic (eg. Zn-gluconate) form. Or just endogenous vehicles transport Zn better to the target, like in case of Zn-carnosine. Carnosine is a small, water-soluble endogenous peptide physiologically present in skeletal muscle, brain and nearly every organ. It is prone to form chelates of metals, inclusive Zn [23].

3. Clinical Experiences

Serum zinc is supposed to be low in diabetes patients [24, 25]. Moreover, the total blood zinc concentration gradually sink in T2DM patients in relation to the duration of diabetes [26]. However Skalnaya reported elevated se zinc level in type 2 diabetes patients in a cohort of 413 postmenopausal women [27]. Zinc concentration in cellular level needs a tight control in order to keep healthy functions. In diabetes even a sin healtly subjects, the Zn-transporters and metallothioneins basicly influence the cellular zinc homeostasis [28]. In a group of pre-diabetes youngsters (between 10–17years of age) se-zinc levels positively correlated with elevated protein intake and negatively correlated with higher carbohydrate intake [29]. The onset of diabetes however is not linked to the zinc levels as describes Park and coworkers based on the results of assesment of toenail-concentration of 3.960 american young population in a periode of 23 years [30].

Meta-analysis of Jayawardena demonstrate beneficial effet of zinc supplementation on glycaemic control [31].The systematic literature survey of de Carvallho demonstrated that plasma zinc levels negatively correlated with glycated hemoglobin percentage (HbA1c) and zinc supplementation tendentiously has ameliorated glycemic control of type 2 diabetes patients [25]. According to the available data zinc is beneficial only for patients having low serum-zinc levels [32]. From the prevention aspect Cochrane evaluation of 2015 conclude to neutral regarding zinc supplementation and insulin resistance (HOMA_IR) as well as major complications like cholesterol and triglyceride levels [33]. The cardiovascular complications are, according to the systematic review of Chu, also not clearly correlat with the zinc status of the T2DM patients [34]. The microvascular complications natheless seems to be Zn-concentration dependent [35]. This is partly due to the antioxidant effect of Zn as demonstrated Cruz in his summary [36].

Discussion

Scientific literature of zinc is rich enough however there are few evidences from clinical point of view, ie. randomized controlled studies supporting confident knowledge about the use of zinc supplementation in diabetes mellitus are scarce. Physiological zinc levels in somatic cells are very low but extremly constant. Therefore the presence of stored Zn and the concerted regulation seems to be essential. If zinc levels in blood were low, symptoms appear slowly and signs usually are aspecific, in this situation supplementation is recommended. In diabetes plasma zinc concentration should be kept within the normal range otherwise insulin-resistance as well as diabetes complications worsen. In case of hypozincemia zinc supplementation improves glycemic control. In prediabetes, when zinc levels are still normal or near normal, supplemental administration of zinc has no beneficial effect. Collectively: if diabetic patient had low zinc concentration, permanent zinc supplementation is needed and should be administered in supraphysiological doses. Contemporary formulations, especially nano-formulations are effective in normalization of zinc homeostasis and dysregulations of diabetes originate in hypozincemia can be corrected by Zn supplementation.

References

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Analysing Three Decades of Land cover Change in Chilika Lake Ecosystem

DOI: 10.31038/ESCC.2019115

Abstract

To understand how an area changed over time, change detection analysis using multi-decadal satellite remote sensing data can be quite useful. Out study here uses 30m Landsat data acquired at 30 years apart from 1987 to 2016 collected during the same time of the year (December) to understand the landscape dynamics in the Chilika Lake and neighbouring areas. ISODATA unsupervised classification technique is applied in ArcGIS 10.4.1 for our analysis. The Landsat images of our study area were categorized into bare ground, grass, shrubs, forests and surface water cover types. Our analysis showed that during the last thirty years, bare ground, grass and surface water cover decreased by 139 sqkm, 115 sqkm and 139 sqkm respectively while shrub and forest cover types increased by 180 sqkm and 217 sqkm respectively. The results from our study provides a baseline understanding of the changes happening in this key coastal ecosystem during the last three decades and thus provides information towards developing understanding of long-term coastal ecosystem change in the study area.

Keywords

Decadal Change, Change Detection, Ecosystem Change, Coastal Ecosystems, Landsat

Introduction

Land use and land-cover (LULC) change is in the mainstream of studying Global Change impacts. Land use and land cover are two separate terminologies often used interchangeably [1]. The changes in the physical characteristics of the earth’s surface e.g. deforestation, afforestation, distribution of water bodies, soil and types of vegetation as well as anthropogenic changes such as proliferation of manmade structures are captured in the term land use. The land use and land cover changes of a region is characteristic of the human use of the land and can also be representative of the economic activities. The patterns of these changes are normally a combination of natural and socio-economic drivers over time. Therefore, developing an understanding of the land use and land-cover changes of a region is important to develop management schemes to meet increasing stresses on the natural resources of a region.

Over the years, Land-change science established itself as the foundational ground for studying global environment change and overall sustainability of an ecosystem [2]. This field helps to better understand the human and environment dynamics leading to changes land uses and land covers. This takes care of the change happening in terms of their type, magnitude and location as well. This requires the integration of social, natural, and geographical information sciences. LULC is considered as one of the major concerns in global environmental change and hence overall sustainability of an ecosystem. The LULC driven by rapid urbanization and increasing economic activities put a lot of pressure on natural resources. This is especially true for a rapidly developing country such as India. The environmental degradation associated with LULC change has shown to impact various ecosystem good and services. Various studies have demonstrated that conversion of various land cover types to agriculture and urban landscapes have negative impacts on nutrient cycling, erosion control and climate regulation and water availability and soil fertility [3–7]. The direct consequences of these changes degrade various ecosystem goods and services provided to the human beings through various ecosystem functions. Thus it is important to understand the consequences of the LULC change and hence the overall impact it can have on a whole ecosystem using satellite based measurements over decades.

Remote sensing and GIS has taken a significant role in developing the LULC change science [8–10]. While remote sensing has made possible to study the changes of large areas with precision and over decade’s time-scale, GIS provided a platform for data analysis and updating of mapping products. With emergence of high resolution and long-term satellite programs such as Landsat combined with advanced GIS software, the LULC science has been able to provide routine services for monitoring and modelling of land use / cover patterns. The Landsat series of satellite data archive since 70s has provided one of the strongest resources for LULC science. The freely made available Landsat archive represents a huge information source for studying the changes and developing monitoring capabilities in our manmade and physical environments [11].

Coastal ecosystems are key ecosystems considering their importance in providing goods and services. In addition to the aesthetic and recreational value provided by the coastal ecosystems, economic benefits provided by them in supporting human livelihood via food and materials, nutrient cycling, waste processing and other essential goods and services are quite important. Millennium Ecosystem report of 2005 identifies coastal ecosystems as one of the most productive yet highly vulnerable ecosystems of the world. Coastal areas are vulnerable to climate change impacts due to factors such as: sea level rise, changes in storm surge and precipitation, increased coastal water temperature and ocean acidification. Additionally, factors such as increasing human habitats and technological advances also put additional pressures on coastal ecosystems and thus contribute to the exploitation of coastal resources. In recent times, there are evidences of dramatic declines in various types of coastal ecosystems such as coral reef, mangroves, estuaries, marshes, dunes, deltas, seagrass beds and kelp forests. In this study, an attempt has been made to develop a baseline understanding of changes happening in the Chilika and its neighbouring areas using long-term satellite data records.

Study Area

Our Study area is Chilika Lagoon and its neighbouring areas. Chilika Lagoon is the largest brackish water wetland of India and a Ramsar site. The highly productive eco-system, and its rich fishery resources provides livelihood for more than 0.2 million people who live in and around the lagoon. The lagoon was encountering serious ecological as well as anthropogenic problems leading to change in its ecological characters for which it was included in the Montreux record in 1993 by Ramsar Bureau. This serious threat to the lagoon eco-system had also adversely affected the biodiversity and livelihood of local communities. To restore this unique ecosystem the Government of Orissa created Chilika Development Authority. Rising to the occasion Chilika Development Authority initiated the restoration of the lagoon with ecosystem approach and active community participation. Our interest in the particular area is mainly due its dynamic ecological nature as a coastal area which also provides essential goods and services for a sizeable population and thus providing services for regional economy.

Methods

Two cloud-free Landsat scenes at 30m spatial resolution were selected in December, 1987 (Landsat-TM) and December, 2016 (Landsat-8) for the land use / cover analysis. The data was downloaded from the USGS Earthexplorer site (https://earthexplorer.usgs.gov). The datasets were pre-processed in open source software QGIS using Semi-Automatic Classification Plugin for QGIS (Ref). First, the raw DN data for individual bands were converted to radiance values and TOA reflectance. Then the TOA reflectance was converted to surface reflectance using the DOS1 correction. An area of interest (AOI) was created for the Chilika Lake and the adjoining areas and both the scenes were subset to the AOI. Following this, NDVI values were calculated using NIR and Red bands for respective sensors for both the dates. The Landsat data provided by USGS are already georegistered and orthorectified and hence these steps were not performed. As we calculated NDVI i.e. a standard spectral index, inter-sensor calibration was also not needed.

For our classification, we wanted to have a simple classification scheme for a few classes to have an understanding of how the landscape changed over three decades. Iso Cluster Unsupervised Classification analysis of the Spatial Analyst Extension in ESRI ArcGIS 10.4.1 software was used to classify the two NDVI images into five classes each. The number of classes that we wanted to have for our study was determined based on our estimation of broad land-use classes for the area after reviewing the NRSC LULC map. The Iso Cluster Unsupervised Classification analysis combines the functionalities of the Iso Cluster and Maximum Likelihood Classification tools and outputs to a classified raster. The algorithm automatically finds the clusters in an image and outputs a classified image and an optional signature file. We identified our broad classes as Bare ground, Grass, Shrub, Forrest and Water.

To perform change detection analysis for land use / cover change in the study area, a post classification detection analysis was employed. First a pixel-based comparison was used to identify change information for each of the classes and thus interpret changes using “-from, -to” information from 1987 to 2016. Then each of the raster images for each of the classes were converted to vector files and total areas for each of the classes was calculated for each of the years i.e. 1987 and 2016. This information was used to calculate gain or loss of the total area of each of the classes over three decades time period. These data were compiled and presented in table forms.

Results

The results from our analysis are shown below in Figs 2 – 8. Figure-1 Shows the study area and Table-1 summarize the results from the change detection analysis. The following paragraphs provide a brief account of the results obtained.

Table 1. Summary of Landcover change during 1987–2016

Land-cover Type

Area in 1987 (Sq. Km)

Area in 2016 (Sq. Km)

Area Change (Sq. Km)

% of total area in 1987

% of total area in 2016

% change

Bare ground

1893

1754

-139

30.9

28.6

-7.3

Grass

1302

1187

-115

21.2

19.4

-8.8

Shrub

866

1046

180

14.1

17.1

20.7

Forrest

1070

1287

217

17.4

21.0

20.2

Water

999

860

-139

16.3

14.0

-13.9

Our initial visual assessment of the study area using natural colour RGB images of Landsat images in December showed visible changes which is shown in Figure 1. Particularly, the northern sector of the lake showed some distinct visible changes in vegetation as marked with dotted circles in the figure. This was also visible from the LULC analysis as shown in Figure 2. Figures 3 – 7 are created by overlaying spatial distribution of the particular land cover type for 1987 and 2016 for all the (a) figures and overlaying the 2016 on top of 1987 for all the (b) figures. The purpose of these figures is to show the dynamics of each of the landcover types and how they lost and/or gained in spatial extension from 1987 to 2016. Figure 3 shows the increase in bare ground class from 1987 to 2016 (Fig 3a) and decrease in bare ground spatial class from 1987 to 2016 (Fig 3b). This reveals that bare ground coverage decreased by 7.3% of the total from 1987 to 2016 which is equal to an area of 139 Sq. Km.

ESCC 2019-105 - Santonu India_F1

Figure 1. The figure shows natural colour RGB images of Landsat in December of 1987, 1999, 2006 and 2013 for a subset of the study area. The image depicts some visible changes in vegetation change. The type of vegetation change was identified in consultation with the Chilika Development Authority and also in a recent field visit to the study area.

ESCC 2019-105 - Santonu India_F2

Figure 2. The figure shows the five classes in December, 1987 and December, 2016. The legend inside each figure marks the classes with respective colours.

ESCC 2019-105 - Santonu India_F3

Figure 3. Dynamics of bare ground change between 1987 and 2016. The lightly shaded areas in Fig 3a shows the bare ground extent gained in 2016 compared to 1987 while the dark shaded areas in Fig 3b shows the bare ground extent lost in 2016 compared to 1987. Overall, the bare ground coverage decreased by 7.3% of the total from 1987 to 2016 with an area of 139 Sq. Km.

ESCC 2019-105 - Santonu India_F4

Figure 4. Dynamics of grass change between 1987 and 2016. The dark shaded areas in Fig 4a shows the grass extent gained in 2016 compared to 1987 while the light shaded areas in Fig 4b shows the grass extent lost in 2016 compared to 1987. Overall, the grass coverage decreased by 8.8% of the total from 1987 to 2016 with an area of 115 Sq. Km.

ESCC 2019-105 - Santonu India_F5

Figure 5. Dynamics of shrub change between 1987 and 2016. The lightly shaded areas in Fig 5a shows the shrub extent gained in 2016 compared to 1987 while the dark shaded areas in Fig 5b shows the shrub extent lost in 2016 compared to 1987. Overall, the shrub coverage was increased by 20.7% of the total from 1987 to 2016 with an area of 180 Sq. Km.

ESCC 2019-105 - Santonu India_F6

Figure 6. Dynamics of forest change between 1987 and 2016. The dark shaded areas in Fig 6a shows the forest extent gained in 2016 compared to 1987 while the light shaded areas in Fig 3b shows the forest extent lost in 2016 compared to 1987. Overall, the forest coverage increased by 20.2% of the total from 1987 to 2016 with an area of 217 Sq. Km.

ESCC 2019-105 - Santonu India_F7

Figure 7. Dynamics of Surface Water change between 1987 and 2016. The red shaded areas in Fig 7a shows the surface water extent lost in 2016 compared to 1987 while the blue shaded areas in Fig 7b shows the surface water extent lost in 2016 compared to 1987. Overall, the surface water extent coverage decreased by 13.9% of the total from 1987 to 2016 with an area of 139 Sq. Km.

Similarly, figure 4 shows the change of cover type grass from 1987 to 2016. The figure depicts that this cover type decreased by 8.8% of the total from 1987 to 2016 with an area of 115 Sq. Km. For land cover type shrub shown in figure 5, the cover type has seen an overall increase by 20.7% of the total from 1987 to 2016 which equals to an area of 180 Sq. Km. Figure 6 depicts the change dynamics of the land cover type forest i.e. dense vegetation from 1987 to 2016. The figure reveals that this land cover type increased by 20.2% of the total from 1987 to 2016 with an area of 217 Sq. Km. The surface water land cover type change increased decreased by 13.9% of the total from 1987 to 2016 which equals an area of 139 Sq. Km.

Table 1 summarizes the analysis for the land use / cover type carried out on the Landsat data for December 1987 and December 2016. The total area in 1987 and 2016 and how much they changed are shown in sq.Km. Each of the land cover types in percent of the total area in 1987 and 2016 and how much they changed from 1987 to 2016 as a percentage are also shown in positive and negative changes.

Discussions and Conclusion

This study conducted near Chilika and its adjoining areas provided us the first glimpse of land cover change over three decades. The basic land cover types we chose helps us to have an baseline understanding of the changes happening in the area which is important while trying to develop long-term studies of change detection in finer spatial and temporal resolution. This study also shows us how we can exploit the long-term Landsat archive for ecosystem studies at a reasonably fine spatial resolution which is otherwise not available from any of the other satellites data archive. The results from our study reveal that bare ground, grass and water land cover types decreased from 1987 to 2016 while shrub and forest types increased from 1987 to 2016. We did our analysis on December Landsat data mainly due to better coverage because of less cloudy conditions. The timing of the season also has an impact on the land cover types we chose. For example, the bare  ground included areas that are normally left barren after rice cultivation in December months in addition to open soils. Therefore, decreasing of bare ground from 1987 to 2016 probably means that less areas being cultivated in 2016 compared to 1987. The decrease of grass land cover types from 1987 to 2016 might mean that more naturally vegetated areas are being exploited for built-up areas. The shrub land cover type included shrubby vegetation which also included some of the weed infestation in Chilika area such as Phragmites Karka. Chilika Development Authority recognizes this as a major problem in Chilika area and also states the proliferation of this vegetation around Chilika over the last few decades. So, increase of shrub from 1987 to 2016 by about 180 sq.km probably captures the proliferation of this particular weed infestation problem that is happening in Chilika which is an interesting result. The forest land cover type included any dense vegetation including actual forests. The increase of forest type from 1987 to 2016 is mostly because of replantation that is happening in the area which was confirmed by the Chilika Development Authority. The decrease of surface water in the area by 139 sq km also complements the finding of increase of shrubs in the area which is actually proliferation of weeds in the chilika area. The decrease in surface water probably also depicts the degradation of coastal wetlands in the areas over three decades duration. Degradation of wetlands is a recognised problem in India and elsewhere and hence our finding provides us important baseline information for developing further studies. This finding needs further investigation with finer spatial and temporal resolution data and ground validation.

Acknowledgements

We would like to acknowledge support from the NRSC-NICES program to conduct this study as part of the Ecosystem Change and Climate Change research theme. We would also like to thank Chilika Development Authority, Govt. of Odisha for helpful feedback during our studies and also to interpret our results.

References

  1. Dimyati MUH, Mizuno K, Kobayashi S, Kitamura T (1996) An analysis of land use/cover change in Indonesia. International Journal of Remote Sensing 17: 931–944.
  2. Turner BL, Lambin EF, Reenberg A (2007) The emergence of land change science for global environmental change and sustainability. Proceedings of the National Academy of Sciences 104: 20666–20671.
  3. Ahearn DS, Sheibley RW, Dahlgren RA, Anderson M, Johnson J et al (2005) Land use and land cover influence on water quality in the last free-flowing river draining the western Sierra Nevada, California. Journal of Hydrology 313: 234–247.
  4. Leh M, Bajwa S, Chaubey I (2013) Impact of land use change on erosion risk: an integrated remote sensing, geographic information system and modeling methodology. Land Degradation & Development 24: 409–421.
  5. Li S, Gu S, Liu W, Han H, Zhang Q (2008) Water quality in relation to land use and land cover in the upper Han River Basin, China. Catena 75: 216–222.
  6. Xiong X, Grunwald S, Myers DB, Ross CW, Harris WG et al (2014) Interaction effects of climate and land use/land cover change on soil organic carbon sequestration. Science of the Total Environment 493: 974–982.
  7. Gallo KP, Easterling DR, Peterson TC (1996) The influence of land use/land cover on climatological values of the diurnal temperature range. Journal of climate 9: 2941–2944.
  8. Dewan AM, Yamaguchi Y (2009) Land use and land cover change in Greater Dhaka, Bangladesh: Using remote sensing to promote sustainable urbanization. Applied Geography 29: 390–401.
  9. Basnyat P, Teeter LD, Lockaby BG, Flynn KM (2000) The use of remote sensing and GIS in watershed level analyses of non-point source pollution problems. Forest Ecology and Management 128: 65–73.
  10. Gupta M, Srivastava PK (2010) Integrating GIS and remote sensing for identification of groundwater potential zones in the hilly terrain of Pavagarh, Gujarat, India. Water International 35: 233–245.
  11. Chander G, Markham BL, Helder DL (2009) Summary of current radiometric calibration coefficients for Landsat MSS, TM, ETM+, and EO-1 ALI sensors. Remote sensing of environment 113: 893–903.

Understanding Ecosystem-Based Adaptation to Climate in Kenya’s Mt Elgon Forest Ecosystem: Definitions, Opportunities and Constraints

DOI: 10.31038/ESCC.2019114

Abstract

A number of approaches have been employed across the world to address adaptation to climate change impacts. The role of ecosystems in adaptation to climate change impacts has been recognized at the international level more so upon the realization that conservation, sustainable management and the restoration of ecosystems can help people adapt to the impacts of climate change. This concept of using ecosystems for climate change adaptation otherwise known as Ecosystem based Adaptation (EbA) utilizes the premise that healthy, well managed ecosystems have climate change mitigation potential. The approach is gaining increasing attention as it is accessible to the rural poor in developing countries due to its cost-effectiveness and due to the fact that it uses infrastructure that is already established by nature. Research has it that the Mount Elgon ecosystem in the south-rift part of Kenya has EbA characteristics which can offer longer term solutions to adaptation to climate change impacts while providing a range of other benefits in terms of ecosystem goods and services. This paper seeks to profile Mt. Elgon ecosystem’s natural infrastructure in improving resilience of the forest adjacent community to the impacts of climate change. This was achieved by carrying out, a descriptive survey that involved 405 household and 51 civil servant and civil society respondent drawn from Saboti, Kiminini, Endebess, and Kwanza sub- counties, Transnzoia County in Kenya. Results show that residents of the study area grow maize (90.6%) being their staple food as compared with other crops such as beans (3.7%), vegetables (2.7%) and millet (1.5%).The household incomes centre around crop farming (47.4%) followed by formal employment (21.5%),family business (12.9%), casual employment in the agricultural sector (10.9%), while other sources accounted for 0.5%. This specialty economy exposes residents to the effects of climate.   A major conclusion in this study is that beneficiary decentralized governance systems must seize opportunities presented by the Mt Elgon ecosystem to develop initiatives that improve the resilience of ecosystems and people to climate change impacts.

Keywords

Ecosystem based adaptation, opportunities, challenges, Mt. Elgon

1. Introduction

Ecosystem based adaptation is a nature-based approach, has the potential to increase adaptive capacity and social and ecological resilience to climate change in both developed and developing countries [1]. It provides a cost effective, economically beneficial, as well as longer term solutions, with a range of co-benefits in terms of the goods and services provided by ecosystems

Mount Elgon Forest Ecosystem is one of Kenya’s five major water towers and the second highest mountain in the country. It doubles up as an important biodiversity hotspot of global significance, supporting several endemic plant and animal species (CIFOR, 2017). This gazetted montane forest reserve was recognized as a Biosphere Reserve by UNESCO in 2003 due to its significance as a water tower and biodiversity reservoir [2]. The key values of Mt Elgon are presented in the form of natural heritage, biodiversity, water catchments, agricultural base, and tourism that support a poor human population in its landscape. This ecosystem is characterized by peasant farmers whose population density average at 600 people per km2 near forest. This community depends on the forest for most of their subsistence needs. The bimodal pattern of rainfall with annual rainfall of 1,400 – 1,800 mm comes in March to May and September to November. The dry seasons run from June to August and from December to March. The reliable climatic conditions, coupled with other ecological services of this ecosystem supports the adjoining human population of about 2 million people, a majority of whose livelihoods and economic activities depend solely on the goods and services that they derive from this forest ecosystem.

As a key afro-montane ecosystem, the effectiveness of management policies and institutional arrangement has direct impacts on the livelihoods of the surrounding community and other support sectors across large watersheds in Kenya and Uganda (CIFOR, 2017).  Improved understanding of the ecosystem health of this ecosystem, biodiversity status and its contribution to provision of ecosystem goods and services is a key step in developing a cost effective and economically viable policy strategy to increase community resilience to climate change impacts. In view of the foregoing, this paper seeks to profile Mt. Elgon ecosystem’s natural infrastructure in improving resilience of the forest adjacent community to the impacts of climate change.

1.1 Study area

The study area covers four sub-counties whose residents directly or indirectly interact and depend on the Mt. Elgon forest for their livelihoods (Figure 1).

ESCC 2019-104 - Jusper M. Omwenga USA_F1

Figure 1. Study area.

(Source: Moi University Geography Department GIS Lab, 2013.)

This landscape which consist of a forest reserve and a national park extend and border with the local communities who live adjacent to the forest and depend on its forest for their livelihood. The forest provides most of the goods and services, which form the basis of their subsistence. The rivers and many rivulets which emanate from this forest have for a long time influenced greatly the livelihoods of the immediate and downstream communities.

The rich agro-ecosystem that is supported by this afro-montane forest has recently experienced the impacts of climate change. This has manifested itself through increasing mean annual temperatures and shifting of precipitation means of the crucial agricultural calendar (figures 2 & 3)

ESCC 2019-104 - Jusper M. Omwenga USA_F2

Figure 2. Mean annual temperature changes in the Mt. Elgon ecosystem.

ESCC 2019-104 - Jusper M. Omwenga USA_F3

Figure 3. Rainfall variability in MAM and OND over the years (2009 -2018).

1.2 Methodology

A descriptive survey was used to collect data from fro 405 households who are   residents of Saboti, Kiminini, Endebess, and Kwanza sub- counties (table 1).

Table 1. Sample size of households.

Sub-county

No of households

Number in sample

Kwanza

139,708

ESCC 2019-104 - Jusper M. Omwenga USA_F5

Endebess

53,811

ESCC 2019-104 - Jusper M. Omwenga USA_F6

Saboti

141,575

ESCC 2019-104 - Jusper M. Omwenga USA_F7

Kiminini

58,437

ESCC 2019-104 - Jusper M. Omwenga USA_F8

Total

393,531

405

Another set of 51 responds drawn from the civil servants and civil society organizations was selected using purposive sampling. The selection was based on their knowledge on Mt.  Elgon ecosystem and climate change adaptation matters. The civil servant respondents were drawn from the County Government of Transnzoia, the national government ministries, departments and agencies. Civil society respondents were drawn from the private sector, local NGOs and CBOs. Information obtained from this category of respondents was correlated with existing secondary information.

2. Results and Discussion

2.1 Opportunities for Ecosystem based management in Mt. Elgon ecosystem

2.1.1. Ecosystem goods and services in enhancing resilience to climate change

There is a wide range of ecosystem goods and services that are obtained from the Mt. Elgon forest ecosystem (table 2). The community regards these as important in both directly and indirectly supporting their livelihoods.

Table 2. Goods and services obtained from the Mt. Elgon forest ecosystem.

HOUSEHOLD RESPONSES (N=405)

GOVERNMENT/CIVIL SOCIETY RESPONSE (N=41)

Goods/service

Trend

Freq

Percent

Goods/service

Trend

Freq

Percent

Food

Increase

111

27.4

Food

Increase

5

12.2

Decrease

289

71.4

Decrease

36

87.8

Fuel wood

Increasing

100

24.7

Fuel wood

Increasing

7

17.1

Decreasing

304

75.1

Decreasing

34

82.9

Fresh water

Increasing

78

19.3

Fresh water

Increasing

7

17.1

Decreasing

321

79.3

Decreasing

34

82.9

Medicinal plants

Increasing

65

16.0

Medicinal plants

Increasing

7

17.1

Decreasing

339

83.7

Decreasing

34

82.9

Air quality

Increasing

81

20.0

Air quality

Increasing

8

19.5

Decreasing

321

79.3

Decreasing

33

80.5

Natural hazard regulation

Increasing

142

35.1

Natural hazard regulation

Increasing

16

39.0

Decreasing

259

64.0

Decreasing

25

61.0

Water flow regulation

Increasing

127

31.4

Water flow regulation

Increasing

8

19.5

Decreasing

275

67.9

Decreasing

33

80.5

Cultural and spiritual

Increasing

99

24.4

Cultural and spiritual

Increasing

6

14.6

Decreasing

304

75.1

Decreasing

35

85.4

Biodiversity regulation

Increasing

118

29.1

Biodiversity regulation

Increasing

7

17.1

Decreasing

284

70.1

Decreasing

34

82.9

Explanation

Goods such as food, fiber, fuel wood, freshwater and medicinal plants are obtained from this forest ecosystem besides offering diverse ecosystem services. Generally, the benefits obtained from Mt. Elgon ecosystem can be grouped into four main categories. These are provisioning, regulating, supporting, cultural, and recreational services. They are all relevant in contributing to the reduction of vulnerability of the community to the effects of climate change.

A healthy, fully functioning Mt. Elgon ecosystem can enhance the provision of these much needed ecosystem goods and services. Further, a healthy ecosystem can be more resilient to stressors and thus better able to support adaptation to climate change impacts. Further, a healthy ecosystem implies a greater element of flexibility in adaptation response options. Strengthening and protecting this ecosystem is a sustainable investment that ensures an array of environmental, social and financial benefits especially under adverse climatic situations.

2.1.2 Eba in improving food security in Mt. Elgon Ecosystem

Table 3 shows the main crops grown by most households in the study area. They include maize (90.6%), beans (3.7%), vegetables (2.7%), millet (1.5%) and sugarcane (1.5%). The physiographic and edaphic factors are suitable for growing other crops such as millet, sunflower and sorghum (GoK, 2013) [2] but this opportunity has not been exploited.

Table 3. Main crops grown by respondents.

Crop type

Frequency

Percent

Valid Percent

Cumulative Percent

Beans

15

3.7

3.7

3.7

Vegetable

11

2.7

2.7

6.4

Maize

367

90.6

90.6

97.0

Millet

6

1.5

1.5

98.5

Sugarcane

6

1.5

1.5

100.0

Total

405

100.0

100.0

Source: (Author, 2015)

Maize farming is widely adopted in the study area with the size of land owned not being an obstacle to people growing maize (α = 0.05, p = 0.207). It is the most preferred irrespective of the size of land owned by a household (table 4).

Table 4. Relationship between sizes of land owned by respondent and the main crop grown.

Size of Land Owned by Respondent

Main crop

Beans

Vegetable

Maize

Millet

Sugarcane

Total

< 0.5 Acre

3

4

126

2

3

0.5 – 1 Acre

6

4

47

0

0

2 – 4Acres

5

2

106

4

3

5 – 10 Acres

1

1

55

0

0

> 10 Acres

0

0

15

0

0

None

0

0

18

0

0

Total

15

11

367

6

6

Source: (Author, 2018)

Explanation

Owing to the food preferences, maize is a staple food in both the study area and in Kenya as a nation. The adequacy of this preferred crop determines how food secure the Mt. Elgon community is at any one given moment. In the event of a short supply occasioned by the effects of climate change, most households will have limited choices of other food stuffs and will be considered to be food insecure. This causes maize to be widely grown as compared to other crops. Its production is relies mainly on rain-fed agriculture, which is in turn influenced by the prevailing climatic conditions in the Mt. Elgon forest ecosystem. This climatic fluctuations that are characteristic of the area exposes Maize production to climate related uncertainties and may in turn affect the production pattern of this staple food. The major climatic uncertainties include precipitation variability, seasonal temperature change, extreme weather events such as; drought, floods, emergence of new crop pests and diseases, which may in turn increase the community’s vulnerability to climate change impacts.

Herrero [4] observes that overreliance on one staple food crop by a society may expose them to the impacts of climate change. This is especially so when the production systems rely on the prevailing climatic patterns of the area. The uncertainties associated with natural climatic patterns may strongly affect the stability of food supplies.  This results to multiple effects that include the reduction of the society’s ability to access food at affordable prices besides leading to critical effects on food security. Seizing ecological opportunities that come with the Mt. Elgon’s ecosystem can diversify crop farming and thus broaden the variety of agricultural activities in the area.  These favorable conditions can positively be used to increase resilience to impacts of climate change.

2.1.3 EbA in increasing income diversity in the Mt. Elgon ecosystem

Crop farming (47.4%) is the main source of family income followed by formal employment (21.5%). Other income sources include family business (12.9%), casual employment in the agricultural sector (10.9%), while other sources accounted for 0.5% (table 5).

Table 5. Common source of household income.

Family business

Common income Household income source

Total

Crop farming

Livestock farming

Formal employment

Casual employment

Other

Age of the Respondent

Below 18

0

6

0

0

0

0

6

18-25

7

17

3

7

7

0

41

26-32

7

28

2

14

10

0

61

33-38

24

49

4

26

14

1

118

39 and Above

14

92

19

40

13

1

179

Total

52

192

28

87

44

2

405

Source: (Author, 2018)

Explanation

Agriculture, being the main economic activity has a strong ripple effect on the vibrancy of other sectors in the same region. This sector emerged as the single largest source of income supporting a majority of homesteads. It was established that even those in formal employment and are with a relatively stable income are actually working in agriculture-allied institutions. For instance, the Kenya Seed Company, Agricultural Development Corporation (ADC), Educational facilities, and flower farms in the study area have a workforce that receives a relatively regular income to supports their livelihoods. All these employees are categorized as working in the agriculture sector and its value chain facilities. This implies that the agricultural share of total labor force and associated income is big and largely influences the vulnerability to climate change narrative in the study area. This is largely due to the fact that agricultural activities practiced by the community in the area heavily rely on the natural climatic patterns.

Indeed the findings point out to lack of diversification in income sources in the study area. This narrow range of income streams is an important socio-economic exposure to the effects of climate change. Diversified income sources can help in cushioning households against the negative effects of climate change by providing alternatives should another fail. Herrero et al [4] observes that over-reliance on limited economic livelihood options is a strong predisposing factor to vulnerability to the effects of climate.   Broadening of income-generating opportunities by vulnerable groups especially with the imminent threats of climate change and its impacts is urgent.  This then calls for the need to avoid the overreliance on climate dependent agriculture and its income sources. Adger [5] too observes that dependency on income from agriculture is an important aspect of vulnerability and is caused by reliance on a narrow range of limited resources. Such dependency may often lead to social and economic stresses. He further points out that there are links between poverty and lack of diversification of livelihood activities by the farmers and thus leading to enhanced poverty.

The Mt. Elgon ecosystem which has diverse ecological niches can wistfully be utilized to broaden the economic base of the residents. This call on enhanced investment in research directed at birthing economic ventures that site match the different ecological niches presented by this ecosystem. Eco-tourism and commoditization guided by sustainable exploitation of some renewable natural resources such as water may provide additional avenues of income generation. Restoration of degraded ecosystems is an important aspect of EbA because it provides a mechanism for carbon sequestration and hence climate change mitigation, sources of employment and enhancement of resources to support livelihoods [6,7].

Payment for ecosystem services also known as payments for environmental services or benefits are other possible income streams in the Mt. Elgon ecosystem. It implies that incentives can be offered to farmers, landowners and natural resource conservancies in exchange for managing their land to provide some sort of ecological service [8]. The programs are voluntary and mutually beneficial contracts between the riparian consumers of environmental services and the suppliers of these services.

Challenges to Ecosystem based adaptation

3.1. Biodiversity loss

The Mt. Elgon ecosystem has lately suffered from a shrinking biodiversity resource base, a situation that has been attributed to over-abstraction of some species. Diminished biodiversity is a pointer of poor ecosystem health and consequently, reduced ability to buffer the community against the effects of climate changes. A publication by IUCN [2] points out that there has been a marked reduction in forest cover due to clearing of land for agricultural production (figure 4).

ESCC 2019-104 - Jusper M. Omwenga USA_F4

Figure 4. Land use and land cover changes in Mt. Elgon ecosystem (1973–2013)

Source: ACCCESS/IUCN, 2014

The use of land and natural resources in and around the Mt Elgon ecosystem has resulted to significant alteration of ecosystem structure, function and processes, including connectivity within and between ecosystems, a case that has also been observed by Western [9]. Ongugo (op cit) [10] observes that water streams emanating from the Mt.  Elgon have in the recent past reduced significantly in terms of volumes and annual discharge due to anthropogenic factors. The soils too have lost their fertility due to poor soil management practices. Deterioration of these resources has in turn impacted negatively on the community livelihood resources, whose existence is closely linked to the ecosystem health. Modification of the Mt. Elgon ecosystem may reduce its health, productivity and resilience, and must be managed to ensure sustainable supply of ecosystem goods and services.

Over-abstraction of the ecosystem goods in the Mt. Elgon has been linked to people viewing the resources as free goods that are open to exploitation by all. IPCC (op cit) observes that, because many of the ecosystem goods and services have always been freely available, with no markets and no prices, their true long-term value is not included in society’s economic estimates. It calls for “hercynian” decision to value these goods and services in monetary terms. Further, promoting better governance, and strengthening the rules that help to protect this ecosystem is encouraged in order to enhance the ecosystem health, which culminates in enhanced EbA services.

3.2. Uncoordinated management of the ecosystem

Cumulatively, 77% of household are of the opinion that there is uncoordinated approach towards managing the ecosystem resources. This has led to the degradation of the individual natural resources and consequently affecting their livelihood sources (table 6).

Table 6. Respondent views on coordination the management of ecosystem resources.

There is poor coordination in the management of  natural resources

Frequency

Percent

Valid Percent

Cumulative Percent

Strongly Agree

160

39.5

39.5

39.5

Agree

152

37.5

37.5

77.0

Don’t know

35

8.6

8.6

85.7

Disagree

46

11.4

11.4

97.0

Strongly disagree

12

3.0

3.0

100.0

Total

405

100.0

100.0

Source: (Author, 2018)

Responses indicate that 32% of households have not witnessed nor do they have knowledge these collaborative meetings while 26.7% have seen these meetings being held yearly (table 7).

Table 7. Household view on inter-sectoral ecosystem management meetings.

Frequency of inter-sectoral meetings

Frequency

Percent

Valid Percent

Cumulative Percent

Weekly

6

1.5

1.5

1.5

Monthly

43

10.6

10.6

12.1

Bi-monthly

11

2.7

2.7

14.8

Quarterly

107

26.4

26.4

41.2

Yearly

108

26.7

26.7

67.9

Never

130

32.1

32.1

100.0

Total

405

100.0

100.0

 Source: (Author, 2015)

A major pre-requisite for effective utilization of ecosystem-based adaptation to climate change is the presence of inter-agency collaborative management of the natural environment. Mt. Elgon ecosystem is not a homogenous landscape. It is made up of at least four discrete eco-climatic zones that support different plant and animal communities [11]. The situation calls for a management arrangement that reflect the ecological diversity and inter-connectedness of ecosystem processes. Contrary to this expectation, ecosystem restoration activities geared towards addressing ecosystem restoration, resource management and conservation in the Mt. Elgon ecosystem often take a sectoral approach. This is often associated with to an old age common practice in government in which departments are used to working sectorally while enjoying the benefits of selective mandate. In this kind of setup, the departments lobby and direct much effort to oppose any move towards holism and collaborative management. Under this arrangement, ecosystem restoration activities and natural resource management are often characterized by sector fragmentation with a number of government departments managing specific resources which focuses on specific uses.

There are consequences of fragmentation that may affect the ecosystem health of Mt. Elgon and by extension EbA. Mhlanga et al [12] observes that sectoral fragmentation of conservation efforts in a homogenous ecosystem can lead to some negative outcomes such as; lack of continuity caused by almost constant programmatic and structural change, loss of public confidence in both the processes of governance and government; and the rapid shift of natural resource management policies to community-based programs without adequate funding and other support. Mt. Elgon ecosystem risks the same consequences should this challenge of sectoral fragmentation fail to be addressed.

3.3. Duplication of conservation efforts

Fragmented management of strategic activities by different actors in one ecosystem is contributing significantly to ecosystem degradation. This poses a threat of interfering with institutional frameworks.  There are possibilities of weak enforcement of polices and legislation at all levels of governance, right from the ecosystem to the national level. Morrison et al [13] notes that governments regularly restructure departments in the natural resource management and environmental arena, sometimes for the purpose of improved policy integration, but most frequently to meet ministerial and bureaucratic aspirations. He further observes that fragmentation of policies and their implementation seriously diminishes the overall effectiveness of natural resource management programs.

A major recommendation given in his study is the adoption of an integrated ecosystem management system which requires active but sustained involvement of all resource users and stakeholders on how the available financial and human resources are allocated and utilized. Conflicts mitigation is crucial too in order to encourage long-term supply of ecosystem goods and services for improved livelihoods.

Diversity of statutory instruments in the Mt. Elgon can make the management of an ecosystem’s resources very effective if properly coordinated. Such coordination can create synergies between and among various government departments which is necessary for sustainable production and utilization of ecosystem goods and services hence cushioning against the effects of climate change.

3.4. Natural resource governance challenges

Findings show that 73.2% of government and civil society respondents report that there exist inter-sectoral policy inconsistencies (table 8) and incoherencies as regards the management of Mt. Elgon ecosystem natural resources whereas a paltry 26.8% have not noticed any policy inconsistencies.

Table 8. Presence of inter-agency policy inconsistencies.

Presence of inter-sectoral conflicts

Frequency

Percent

Valid Percent

Cumulative Percent

Yes

30

73.2

73.2

73.2

No

11

26.8

26.8

100.0

Total

41

100.0

100.0

Mt. Elgon Ecosystem offers a range of benefits and opportunities for local and national economic development, improved livelihoods and provision of environmental goods and services. This call for a good environmental policy that ensure that there is harmonized and integrated approach towards the management of natural resources to ensure that there is sustainable provision of goods and services to beneficiaries.  In this manner there will be reduced vulnerability to effects of climate change by ecosystem goods and service dependent communities. Management of renewable natural resources in Mt. Elgon as separate entities becomes difficult because of the complexity of their interlinked social and ecological components.

The observed policy inconsistencies in managing natural resources found in the same ecosystem is a major factor that could determine the health of this ecosystem and its sustainable supply of livelihood goods and services. A good ecosystem plan and management policy should aim at maintaining an ecosystem in a healthy, productive and resilient condition so that it can meet human needs into the future [14]. The plan should embrace an all-inclusive approach to management that considers the entire ecosystem, including the various stakeholders.

Co-management, where management responsibility is shared between government and resource-users, may improve the suitability and perceived legitimacy of management rules when there is policy coherence. However, despite the potential for such arrangements to improve the resilience of natural resource systems, how co-management works in the face of the glaring policy inconsistencies remains poorly understood [14].

4. Conclusion and Recommendations

Conclusion

The study area is characterized by a specialty economy dominated by maize farming. The fact that it is basically supported by rain-fed agriculture, this specialty agricultural practice exposes the community to climate change impacts. Since the prevailing climate still favors agriculture, residents should be encouraged to adopt mixed farming which caters for both crop and livestock farming. This, coupled with complete value chains, will lead to diversified income sources.

For effective utilization of EbA, the decentralized governance system must make deliberate effort to enhance the ecosystem health of Mt Elgon through sustainable management, conservation and restoration of natural and agro-ecosystems, taking into account anticipated climate change impact trends to reduce the vulnerability and improve the resilience of ecosystems and people to climate change impacts.

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  10. Ongugo PO, Langat D, Oeba VO, Kimondo JM, Owuor B, et al (2014). A review of Kenya’s national policies relevant to climate change adaptation and mitigation: Insights from Mount Elgon. Working Paper 155, Bogor, Indonesia: CIFOR.
  11. KEFRI (2018) Kenya Forestry Research Institute (KEFRI).
  12. Mhlanga L, Nyikahadzoi K, Haller T  (2014) Fragmentation of Natural Resources Management: Experiences from Lake Kariba Volume 2 of Defragmenting African resource management 2: LIT Verlag Münster.
  13. Morrison TH, Mcdonald GT, Lane MB (2004) Integrating Natural Resource Management for Better Environmental Outcomes, Australian Geographer 35: 243–258.
  14. Plummer R, Fitzgibbon J (2004) Co-management of Natural Resources: A Proposed Framework. Environmental Management 33: 876–885.

Ecotoxicological Genetics: from Mussel Watch to Crop Watch

DOI: 10.31038/ESCC.2019113

Commentary

A good researcher usually specializes and enjoys a specific area of study. For example, the researcher is passionate to spend and devotes most of his/her time in studies like biomonitoring, ecology, ecotoxicology, genetics and plant crop improvements. The aim of this paper is to review Ecotoxicological Genetic (EG) study by using marine mussel Perna viridis under the Mussel Watch program and to discuss the potential EG o be applied to crop plants as Crop Watch.

From worldwide scenario, the pace of knowledge increment on EG studies highly indicates that this area is not a new knowledge since it can be easily found in the literature (see Nevo et al., 1986 [1]). However, in Malaysia, the first (perhaps) paper on such topic was published by Yap et al. [2], which will be further discussed in the following paragraph.

Anthropogenic activities have created significant impacts on chemical levels in the coastal environment [3], including inorganic and organic (persistent and emerging) chemicals. The evaluation of EG study in Malaysia is based on the marine mussel P. viridis as a model in this review paper.

Firstly, the most detailed ecotoxicological and biomonitoring study on P. viridis was that reported by Yap et al. [4] which reported on four heavy metals. As found in the literature, heavy metal pollution in Malaysia is increasingly reported in the literature since early 2000 see Yap et al., [5,6]. Application of the biomonitoring data of heavy metals in the marine mussels from Malaysia has been assessed for human health risks [7].

Secondly, the first genetic structures of P. viridis were investigated by Yap et al. [8] by using electrophoretic allozyme study. The genetics differentiation/composition of P. viridis is heavily dependent on free-swimming larvae along the west coast of Peninsular Malaysia especially in Malacca (Al-Barwani et al., 2007) [9]besides the physical barrier (the Johore Causeway [10] and heavy metal contamination in the east part of the Causeway [11,12]. These causative agents should merits more studies in future.

Yap and Tan [13] has made a comprehensive review on the EG studies, based on P. viridis, in Malaysia. This emerging research perspective, perhaps, has started with the use of allozyme polymorphism of P. viridis in relation to heavy metal stress [14]. Preliminary studies on the EG studies on biomonitors in Malaysia have been focused on green-lipped mussels [2,15,16], horseshoe crabs [17] and guppy fish [18].

Perhaps, the earliest EG study was that on the heavy metal stress on allozyme polymorphisms in Malaysia by Yap et al. [19]. They found a positive relationship between allozyme polymorphisms and heavy metal levels in P. viridis sampled from contaminated and uncontaminated coastal waters. Following that, Yap and Tan [15] reported changes in the enzymes GOT, EST and ME in direct connection to Zn stress. This was explained by a lower rate of filtration in the gills and a decreased value of condition index. The above laboratory experiment was conducted by using P. viridis as a test organism. The significance finding of Yap and Tan [15] supported the previous study by Yap et al. [20] that allozymes of P. viridis could be influenced by heavy metal stress based field collected samples. Yap et al. [16] reported significant (P< 0.01) relationships between heavy metal levels and RAPD primers in the byssus and soft tissues of P. viridis. This implied that correlation analysis between a specific primer of RAPD marker and a particular metal could be employed for the identification of metal pollution in the mussels.

Following the above EG studies by using P. viridis as a model, heavy allozyme polymorphisms and heavy metal levels were investigated in female guppy populations collected from two contrasting sites, namely polluted and unpolluted ecosystems [17]. They reported that the levels of Fe and Cu were significantly (P< 0.05) higher in guppy population sampled from polluted drainage than those from unpolluted ecosystem. This finding was largely supported by the significantly (P< 0.05) higher levels of Fe and Cu in the surface sediments, showing contamination by Fe and Cu in the polluted drainage. Based on allozyme study, they found that the banding pattern of the unpolluted wild guppy population with monomorphic alleles which were similar and comparable to unpolluted domesticated guppy population bought from a pet shop. This confirmed that LDH in the guppy can be used as a good biomarker of Fe and Cu contamination. Almost similar ecotoxicological genetic approach was applied to horseshoe crab populations in Malaysia by Yap et al. [18]. They sampled populations of horseshoe crab (Carcinoscorpius rotundicauda) from contaminated and uncontaminated coastal areas of Peninsular Malaysia.

Future ecotoxicological genetic studies should focus on crop plants since they are the major food sources to the ever increasing world populations nowadays. Food crop such as oil palm is an important focus. This is due to the fact that oil palm (Elaeis guineensis Jacq.) has arisen as a key economic crop nourishing the world population nowadays [21]. For example, ecotoxicological monitoring study has been conducted in the oil palm by Yap et al. [22] while genetic studies on the oil palm by Wahid et al. [23] for the high-quality planting material through genetic improvements. However, the above two studies were conducted separately and interpretations were made based on ecotoxicology and genetics, respectively. Future studies should merge the two areas as EG study to make the our understanding in a more holistic ecologically and genetically. Other ecotoxicological monitoring study in crops such as papaya and bananas have been published by Yap et al. [24] and Yap et al. [25], respectively. However, the genetic studies on the above crops are lacking in the literature.

Therefore, Crop Watch is a new research approach incorporating ecotoxicology and genetics. This EG studies hold a great potential research in the future, not only in academia but also commercial industries.

In conclusion, the above literature review indicated Crop Watch by means of EG studies is a potential (although not a new) research area. Considering the importance and combination of knowledge on ecology, ecotoxicology and genetics would help to monitor the growth and yield besides human health risk assessment of the crop better. This Crop Watch approach is expected to continue in future, especially in Malaysia and other Asian countries.

References

  1. Nevo E, Noy R, Lavie B, Beiles A, Muchtar S (1986) Genetic diversity dan resistance to marine pollution. Biol J Linn Soc 29: 139–144.
  2. Yap CK, Ismail A, Tan SG and Rahim Ismail A (2004) Assessment of different soft tissues of the green-lipped mussel Perna viridis (Linnaeus) as biomonitoring agents of Pb: Field and laboratory studies. Wat AirSoil Pollut 153: 253–268.
  3. Amin B, Ismail A, Arshad A, Yap CK and Kamarudin MS (2009) Anthropogenic impacts on heavy metal concentrations in the coastal sediments of Dumai, Indonesia. Environ Monitor Assess 148: 291–305.
  4. Yap CK, Ismail A and Tan SG (2003) Background concentrations of Cd, Cu, Pb and Zn in the green-lipped mussel Perna viridis (Linnaeus) from Peninsular Malaysia. Mar Pollut Bull 46: 1043–1048.
  5. Yap CK, Ismail A, Din Mohd A, Said ZB Tan S. and Siraj SS (2005) Heavy metal (Cd, Cu, Pb dan Zn) concentrations in the green-lipped mussel Perna viridis (L.) from artificial substrates at aquacultured farm of Sebatu. Malays Fisheries J 4: 81–87.
  6. Yap CK, Yeow KL, Edward FB and Tan SG (2009) Revealing copper contamination at the penang industrial area by using Malaysian Mussel Watch Approach. Asian J Microbiol Biotechnol Environ Sci 11: 683–689.
  7. Yap CK, Cheng WH, Karami A and Ismail A (2016) Health risk assessments of heavy metal exposure via consumption of marine mussels collected from anthropogenic sites. Sci Tot Environ 553: 285–296.
  8. Yap CK, Tan SG, Ismail A and Omar H (2002) Genetic variation of green-lipped mussel Perna viridis (Linnaeus) from the west coast of Peninsular Malaysia. Zool Stud 41: 376–387.
  9. Al-Barwani SM, Arshad A, Nurul Amin SM, Japar SB, Siraj SS, Yap CK (2007) Population dynamics of the green mussel Perna viridis from the high spat-fall coastal water of Malacca, Peninsular Malaysia. Fish Res 84: 147–152.
  10. Yap CK, Cheng WH, Ong CC and Tan SG (2013) Heavy metal contamination and physical barrier are main causal agents for the genetic differentiation of Perna viridis populations in peninsular Malaysia. Sains Malays 42: 1557–1564.
  11. Yap CK, Mohd Nasir S, Edward FB and Tan SG (2012a) Anthropogenic inputs of heavy metals in the east part of the Johore Straits as revealed by their concentrations in the different soft tissues of Perna viridis (L.). Pertanika J Trop Agric Sci 35: 827–834.
  12. Yap CK, Shahbazi A and Zakaria MP (2012b) Concentrations of heavy metals (Cu, Cd, Zn and Ni) and PAHs in Perna viridis collected from seaport and non-seaport waters in the Straits of Johore. Bull Environ Contam Toxicol 89: 1205–1210.
  13. Yap CK and Tan SG (2011) Ecotoxicological genetic studies on the green-lipped mussel Perna viridis in Malaysia. In: L.E. McGevin, ed., Mussels: Anatomy, habitat and environmental impact. Nova Science Publishers USA 221–244.
  14. Tan SG, Yap CK (2006) Biochemical and molecular indicators in aquatic ecosystems: Current status and further applications in Malaysia. Aquat Ecosyst Health Manage 9: 227–236.
  15. Yap CK, Tan SG (2007) Changes of allozymes (GOT, EST and ME) of Perna viridis subjected to zinc stress: A laboratory study. J Appl Sci 7: 3111–3114.
  16. Yap CK, Chua BH, Teh CH, Tan SG, Ismail A (2007) Primers of RAPD markers and heavy metal concentrations in Perna viridis (L.), collected from metal-contaminated and uncontaminated coastal waters: Are they correlated with each other? Russian J. Genetics 43: 544–550.
  17. Yap CK, Chong CM, Tan SG (2011a) Lactate dehydrogenase in the guppy fish (Poecilia reticulata) as a biomarker of heavy-metal pollution in freshwater ecosystems. J. Sust. Manage. 6(2): 240–246.
  18. Yap CK, Chong CM, Tan SG (2011b) Allozyme polymorphism in the horseshoe crabs Carcinoscorpius rotundicauda collected from polluted intertidal area in Peninsular. Malaysia. Environ Monitor Assess 174: 389–400.
  19. Yap CK, Ismail A, Tan SG, Rahim Ismail A (2004b) The impact of anthropogenic activities on heavy metal (Cd, Cu, Pb and Zn) pollution: Comparison of the metal levels in green-lipped mussel Perna viridis (Linnaeus) and in the sediment from a high activity site at Kg. Pasir Puteh and a relatively low activity site at Pasir Panjang. Pertanika J Trop Agric Sci 27: 73–78.
  20. Yap CK, Tan SG, Ismail A, Omar H (2004c) Allozyme polymorphisms and heavy metal levels in the green-lipped mussel Perna viridis (Linnaeus) collected from contaminated and uncontaminated sites in Malaysia. Environ Int 30: 39–46.
  21. Kushairi A, Soh KL, Azaman I, Elina H, Meilina-Ong A, et al.(2018) Oil palm economic performance in Malaysia and R & D progress in 2017. J Oil Palm Res 30: 163–195.
  22. Yap CK, Nur Aishah H, Cheng WH, Zakaria MP, Al-Shami SA (2019a) Bioaccumulation of Cu and Pb in the different parts of oil palm (Elaeis guineensis) in comparison to their habitat topsoils. In: Soil Pollution: Sources, Management Strategies and Health Effects, Editor: Chee Kong Yap, Nova Science Publishers, New York, USA.
  23. Wahid MB, Nor S, Abdullah A, Henson IE (2005) Oil Palm – achievements and Potential. Plant Prod Sci 8: 1–13.
  24. Yap CK, Nur Sakinah MJ, Cheng WH, Omar H, Nulit R, et al (2019b) Distribution of Ni and its human health risk assessment in the Carica papaya from Peninsular Malaysia. In: Soil Pollution: Sources, Management Strategies and Health Effects, Editor: Chee Kong Yap, Nova Science Publishers, New York, USA.
  25. Yap CK, Ahmad Zulfadhli T, Cheng WH, Sharifinia M, Ye F, et al (2019c) Health risks of heavy metals via consumption of Musa paradisiaca and the ecological risk assessment of heavy metals in the habitat topsoils. In: Soil Pollution: Sources, Management Strategies and Health Effects, Editor: Chee Kong Yap, Nova Science Publishers, New York, USA.

Public Participation in the Malaysian Environmental Impact Assessment

DOI: 10.31038/ESCC.2019112

Abstract

The environmental impact assessment in Malaysia can generally be categorized as preliminary and detailed. Public participation in preliminary EIA is confined to the review of environmental impact statements by technical committees. Public participation in detailed EIA is more extensive involving inputs of ad-hoc panel members during scoping, public display and comments of EIS as well as review of the EIS by the panel members. Consultation with affected communities is conducted by consultants during EIS preparation though the process is not spelled in the legislation. At only the initial level of involvement in the spectrum of public participation, there is still much room for improvement in engaging the public in the Malaysia EIA. Public participation in the Malaysian EIA could be extended beyond the EIS review stage by engaging the public in scoping and assessment and even during development of policies and plans. Public participation in Malaysia requires further facilitation to improve accessibility to the necessary information for effective provision of comments.

Keywords

Public participation, consultation, EIA, EIS, Environment, Malaysia

Introduction

Environmental impact assessment (EIA) is a process to systematically identify, predict, evaluate and mitigate impacts of development proposals to facilitate decision-making by relevant authorities on the worthiness of the proposals. The impacts evaluated consist mainly of the biological, physical and social aspects [1]. The EIA has its origin in the United States (US) with the enactment of the National Environment Policy Act (NEPA) in 1970. The act was developed in response to mounting public awareness for environmental protection stemming from increasing pollution across the US due to industrialization and urbanization [2]. The Santa Barbara oil spill in 1969 and construction of the Interstate Highway System resulting in extensive losses of ecosystems both pushed for the subsequent passing of NEPA [3]. Since then, other countries began to model their environmental laws after NEPA and to date, there are more than 100 countries in the list [4].

Only 4 years later, the Environmental Quality Act 1974 was enacted in Malaysia requiring development proposals with significant environmental impacts to have EIA conducted under Section 34A [5]. However, it was until 1987 that the Environmental Quality (Prescribed Activities) (Environmental Impact Assessment) Order was made and the order came into effect on the 1st April 1988. While prescribed activities had not been well defined under Section 34A of the Environmental Quality Act 1974, thus limiting the ability of the Act to dictate EIA for certain proposals, it became clear in 1988 that the prescribed activities listed in the Order would be subject to EIA [6]. The order was replaced by the Environmental Quality (Prescribed Activities) (Environmental Impact Assessment) Order 2015 [7]. The major difference of the two Orders, other than a revision of the prescribed activities, is that the 2015 version specifies the prescribed activities whose environmental impact statements (EIS) require public display and comments. Both the Orders do not illustrate the EIA process and the process is not easily accessible on the official portal of the Malaysian Department of Environment [6, 7].

EIA Process in Malaysia and Public Participation

There are two types of EIA, namely the preliminary and detailed EIAs. The types of EIA have not been stated in the Orders but have been mentioned in the guide for investors published by DOE [6-8]. Prescribed activities listed in the First Schedule of the Environmental Quality (Prescribed Activities) (Environmental Impact Assessment) Order 2015 do require to have the EIS public displayed and commented, and are generally subject to the preliminary EIA. However, those listed in the Second Schedule of the Order prompting public display and comments of the EIS are put through the detailed process [7, 8]. Prescribed activities in the Second Schedule are deemed to have larger impacts than those in the First Schedule due to comparatively larger scale of the activities. Both the preliminary and detailed EIAs undergo the typical EIA stages of screening, scoping, EIS preparation and review, decision-making and follow-up but the stages differ in levels of details and activities [9]. A comparison of both the processes is shown in Table 1.

Table 1. Comparison of Preliminary and Detailed EIAs in Malaysia.

EIA Stage

Preliminary EIA

Detailed EIA

Screening

Prescribed activities in the First Schedule of the EIA Order 2015

Prescribed activities in the Second Schedule of the EIA Order 2015

Scoping

Terms of Reference (TOR) is submitted to the state DOE office.

The scope of EIS is confirmed through issuance of a formal letter by the office.

TOR is submitted to the national DOE headquarters.

DOE calls for ad-hoc panel meeting for the TOR at the headquarters. The ad-hoc panel comprises government officers, academics of universities and representatives of non-governmental organizations (NGOs).

If additional scope is required, the TOR is then revised and resubmitted.

EIS Preparation

Preparation of preliminary EIS based on the scope stated in the letter issued by the state DOE.

No public display and comment of EIS is required.

Preparation of detailed EIS based on the scope in the final TOR submitted.

Copies of EIS are displayed at locations specified by the DOE, including the state DOE offices, the headquarters, universities and public libraries for public comments.

EIS review

EIS is distributed to technical committee members for review.

The technical committee usually comprises government officers.

Technical committee meeting is held at the state DOE office for evaluation of whether the EIS meets the legal requirement and addresses all relevant impacts satisfactorily.

EIS is distributed to ad-hoc panel members for review.

Ad-hoc panel meeting is held at the DOE headquarters for evaluation of whether the EIS meets the legal requirement and addresses all relevant impacts satisfactorily.

Decision-making

The state DOE director approves or rejects the EIS or requires provision of additional information before approval.

If approval is granted, it comes with a set of approval conditions.

The Director General of DOE approves or rejects the EIS or requires provision of additional information before approval.

If approval is granted, it comes with a set of approval conditions.

Follow-up

Post-EIA monitoring which involves submission of quarterly environmental monitoring reports to the DOE.

Post-EIA monitoring which involves submission of quarterly environmental monitoring reports to the DOE.

From Table 1, it appears that only detailed EIA involves public participation. To understand the extent of public participation in Malaysia, a further probe of the definition of public participation and the definition of public is necessary. The IAIA defines public participation as ‘involvement of individuals and groups that are positively or negatively affected by, or that are interested in, a proposed project, program, plan or policy that is subject to decision-making process” [10]. The undertakings of public participation implies the democratic approach of a country [11]. For instance, a deliberative approach provides limited means for the undertaking of public participation in comparison to the collaborative approach which upholds inclusiveness, openness and consideration of multiple perspectives for effective planning and decision-making [12]. Lawrence perceived EIA as a form of social learning in the quest for sustainable development during which all stakeholders have the opportunity to enhance their knowledge [13].

Participation is fundamentally different from consultation in the sense that participation involves active engagement in decision-making while consultation is confined to the request of information and inputs from intended parties [14].Putting participation on a scale, consultation can be visualized as a lower level of participation providing minimum opportunity for the public to be involved in decision-making [15]. The International Association for Public Participation (IAP2) divides public participation into 5 levels starting with informing at the bottom which revolves around provision of unbiased information to the public so that they can understand the issues at hand. Consultation is at the next level up. Involvement sits higher that consultation and focuses on engagement with the public to ensure public needs and concerns are continuously gathered and considered. Next on the spectrum is collaboration which forges a partnership with the public in identification of alternatives and solutions. The highest level is empowerment which grants the decision-making power to the public [15]. The EPA’s spectrum of public involvement aligns with that of IAP2 in Figure 1.

ESCC 2019-102 - Daniel Tang Kuok Ho Malaysia_F1

Figure 1. Spectrum of Public Involvement [16].

The Level of Public Participation in the Malaysian EIA

Referring again to Table 1, elements of public participation can now be identified in both the preliminary and detailed EIA processes. In preliminary EIA, it is confined to the review stage involving stakeholders in the technical committee who comprise largely of government officers. In detailed EIA, public participation is garnered via public display and comments of the EIS at designated locations as well as the review of the TOR and the EIS by ad-hoc panel members consisting of government officers, academics of universities and representatives of NGOs.

There is minimal involvement of other members of the public particularly those affected by the proposals in preliminary EIA as reflected by the process in Table 1. In practice, consultants appointed to prepare the EIS may conduct social survey to gather opinions of the communities around the project area but such practice widely varies as it is not required in preliminary EIA, hence a lack of model to be adopted [8, 17]. For detailed EIA, there is a higher extent of public participation starting from the review and comments of the TOR by the ad-hoc panel members to the review of the EIS by the members. Public participation has also been extended to the public display of EIS at designated locations for comments and announcement of the display via DOE’s website and newspapers. This is akin to the level of involvement of the public participation spectrum proposed by IAP2 and the level of recommendation in the EPA’s spectrum. Public display of EIS invites comments not only from those affected by the proposal but other members of the public whom are concerned about the proposal [17].

At this point, it becomes clear that public participation in Malaysia is often confined to particular stages of EIA for instance, the EIS preparation and review stages of preliminary EIA if social survey is conducted during impact assessment. Based on Wood’s model of EIA [9], public participation should permeate every stage of EIA starting from consideration of alternatives to monitoring action impacts. Nonetheless, different countries may incorporate public participation to varying extents at different stages of the EIA. Taking the Western Australia for example, public comments are invited during screening on the need for assessment and the level of assessment of a proposal. This extends to the scoping stage with publication of the Environmental Scoping Document, equivalent to the TOR in Malaysia on the website of the environmental authority for public comments. The EIS is also published for comments and the final decision on EIS approval can be appealed [18]. In New Zealand, while there is no public scoping, public participation is facilitated via public hearing and review of EIS. For public hearing to be held, it must either be requested by members of the public who have provided comment on a proposal or come under the decision of the ministry or environmental agency. Nonetheless, public participation in New Zealand is not confined to EIA [19]. The Resource Management Act 1991promotes public participation via open standing at the stage of national policies and plans establishment and application by members of the public to Environmental Court for enforcement order in pursuit of sustainable management. At regional level, consultation with Tangata Whenua i.e. the local Maori people is conducted for development of regional and district plans [20].

In Malaysia, based on the author’s experience in environmental consultation, public meetings are conducted for detailed EIA involving the communities at the vicinity of the proposed development sites and such meetings are usually conducted once or twice to gather their inputs and perceptions of the proposals. Certain concerns are addressed by the proponents during the meetings but follow-up of the inputs, comments and perceptions via a feedback mechanism is lacking. Such approach is at best a form of consultation. The affected communities have very little influence over the alternatives of the proposal unlike in the Western Australia where public comments are invited from the screening stage of EIA and in New Zealand, where public participation comes even earlier at the strategic assessment stage [20, 21]. The public display and comments of EIS at designated locations can be perceived as involvement or recommendation level engaging the general public and not just the affected communities in providing concerns. The practice, however, is not well-facilitated. The availability of EIS at specific locations limits the accessibility of the public to the information necessary to provide their comments.  The public can also purchase the EIS from the appointed EIA consultant at a cost which is often quite prohibitive [22]. This forms a stark difference to Western Australia where information related to the EIA ranging from scoping documents, EIS to assessment of the authority and final approval are posted on the website of the environmental authority [21]. In New Zealand, the EIS are also readily available online for public comments and only commercially confidential information is withheld [23]. A search on the World Wide Web reveals very few complete EIS of proposals in Malaysia and in most instances, only the executive summaries are made available.

The EIA process may differ in states in Malaysia having their own environmental legislation and it appears that public participation could elude the state’s EIA legislation [24]. Other than engagement of government officers and NGOs in the review of EIS, public participation could be missing from the EIA process and the access of EIS could be made tedious, thus further hampering public engagement in EIA.

Conclusion

Public participation has been incorporated into the EIA processes in Malaysia but it is still far from maturity. The most common form of public participation is consultation involving representatives of government departments and NGOs in the review of EIS. For detailed EIA, the extent of public participation is increased with public display of EIS for comments and the review of TOR by ad-hoc panel members as well as their inputs during scoping meeting. However, general members of the public have not been actively engaged in the EIA processes. It can be argued that the knowledge level of the Malaysian public members in providing constructive comments is still relatively low [17]. However, in a democratic system, it is the right of the public to have access to the necessary information to provide their thoughts and feedback, and such right should be respected in decision-making. Besides, public participation in EIA should be viewed as a form of social learning through which higher level of public knowledge in this respect can be achieved. It is therefore recommended that public participation in Malaysia should be extended beyond the EIS review stage and should be facilitated by making necessary information available on an official online portal.

References

  1. MacKinnon AJ, Duinker PN, Walker TR (2018) The Application of Science in Environmental Impact Assessment. United Kingdom: Routledge 2018.
  2. Rychlak RJ, Case DW (2010) Environmental Law: Oceana›s Legal Almanac Series. New York: Oxford University Press 2010: 111–120.
  3. Mohl RA (2007) Stop the road: Freeway revolts in American cities. J Urban Hist 2007 July 01.
  4. Eccleston CH (2008) NEPA and Environmental Planning: Tools, Techniques, and Approaches for Practitioners. Fluoride. US: CRC Press 2008.
  5. https://www.doe.gov.my/portalv1/wp-content/uploads/2015/01/Environmental_Quality_Act_1974_-_ACT_127.pdf
  6. http://www.doe.gov.my/eia/wp-content/uploads/2012/03/Environmental-Quality-Prescribed-Activities.pdf
  7. https://www.doe.gov.my/portalv1/wp-content/uploads/2015/01/Perintah-Kualiti-Alam-Sekeliling-Aktiviti-Yang-Ditetapkan-Eia-2015.pdf
  8. Environmental requirements: A guide for investor. Putrajaya, Malaysia: Department of Environment 2010.
  9. Wood C (2003) Environmental Impact Assessment: A Comparative Review 2nd ed. Harlow, UK: Prentice Hall 2003.
  10. Andre P, Enserink B, Connor D, Croal P (2006) Public Participation International Best Practice Principles. Special Publication Series No. 4. USA: International Association for Impact Assessment 2006.
  11. Carpenter J, Brownill S (2008) Approaches to democratic involvement: widening community engagement in the English Planning System. Planning Theory & Practice 2008.
  12. Tewdwr-Jones M,  Allmendinger P (2002) Conclusion: communicative planning, collaborative planning and the post-positivist planning theory landscape. In P Allmendinger & M Tewdwr-Jones (eds). Planning Futures: New Directions for Planning Theory. Oxon: Routledge 2002: 206-216.
  13. Lawrence DP (2003) Environmental Impact Assessment Practical Solutions to Recurrent Problems. New Jersey: John Wiley & Sons 2003.
  14. Petts J (1999) Public participation and environmental impact assessment. In J Petts (ed). Handbook of Environmental Impact Assessment. V.1, Environmental Impact Assessment: Process, Methods and Potential Oxford, UK: Blackwell Science. 1999: 145-177.
  15. https://cdn.ymaws.com/www.iap2.org/resource/resmgr/foundations_course/IAP2_P2_Spectrum_FINAL.pdf
  16. https://www.epa.gov/sites/production/files/2015-09/documents/spectrum508.pdf
  17. Marzuki A (2009) A review on public participation in environment impact assessment in Malaysia. Theoretical and Empirical Researches in Urban Management. 2009.
  18. http://www.epa.wa.gov.au/step-step-through-proposal-assessment-process
  19. ELAW. New Zealand. 2017. Available: https://www.elaw.org/eialaw/new-zealand
  20. Ministry for the Environment. An everyday guide to the RMA – Applying for a resource consent. Wellington. New Zealand: Ministry for the Environment 2015.
  21. EPA. EPA assessment reports. 2017. Available: http://www.epa.wa.gov.au/epa-assessment-reports
  22. https://www.thestar.com.my/news/nation/2017/12/26/myhsrs-eia-report-ready-for-public-viewing/
  23. https://www.epa.govt.nz/public-consultations/
  24. NREB (2019) The Natural Resources and Environment Ordinance (Prescribed Activities) Order 1994. 2019.

Comparative Treatment between Sitagliptin vs. Metformin, Alone or In Combination, In Patients with Polycystic Ovarian Syndrome. A Clinical Entity with a High Risk of Developing Diabetes Mellitus and Gestational Diabetes

DOI: 10.31038/EDMJ.2019341

Summary

Objective: To determine the efficacy of sitagliptin alone or in combination with metformin in women with polycystic ovary in terms of ovarian cyclicity, fertility and cardio metabolic profile compared to metformin alone.

Rationale: Polycystic Ovarian Syndrome (PCOS) affects a percentage of 5–10% of women of reproductive age worldwide and has a prevalence of 6.6% (95% CI 2.3 to 10.9%). Mexican women and the most common cause of infertility in developed countries.

It has been observed that treatment with insulin-sensitizing drugs (metformin and pioglitazone) improve menstrual cyclicity and fertility in the metabolic profile with polycystic ovary patients. Incretins and DPP-4 inhibitors have been shown to improve the activity of pancreatic β cell would increase weight loss by its anorectic effect and the existence of adequate weight control and improvement of fertility.

The previous tests have compared the effect of exenatide and alone or in combination with metformin in the treatment of PCOS, in this article we are going to compare sitagliptin and metformin alone or in combination.

Study design: Blind, controlled and randomized clinical trial.

Patients: Women between 18 – 40 years of age, with a BMI> 20 and with diagnosis of PCOS with the Rotterdam criteria.

Results: In the normalized menstruation index it was found that there was a statistically significant intragroup increase in each of the treatments. The sitagliptin had a higher percentage of change with 127%, followed by that of metformin with 87.5% and then the COMBO with 60%. No statistically significant differences were found between treatment groups.

Conclusion: The therapeutic effect of Sitagliptin was observed in patients with PCOS comparable to metformin and the combination of metformin-sitagliptin is more effective in terms of ovulation than the other two treatments alone

Keywords

Polycystic Ovarian Syndrome, Diabetes Mellitus, Prediabetes, Insulin Resistance

Background

Polycystic Ovarian Syndrome is a syndrome of ovarian dysfunction whose main characteristics are hyperandrogenism, hyperandrogenemia and the presence of polycystic ovaries. This syndrome affects a percentage of between 5 to 10% of women of reproductive age [1]; however, in Mexican-American women a prevalence of 12.8% has been reported. In 2010, Moran et al. They conducted a prospective cross-sectional study in 150 Mexican women to determine the prevalence of PCOS in this population. By Rotterdam criteria, a prevalence of 6.6% was found (95% CI: 2.3–10.9%) [2–4].

Its etiology remains unknown and is the most common cause of infertility in developed countries [1]. Polycystic Ovarian Syndrome is associated with important metabolic alterations. The prevalence of Diabetes Mellitus 2 is 10 times higher in women with PCOS than among women without this entity. An alteration in glucose tolerance or the development of Diabetes Mellitus 2 is found in 30 to 50% of women over 30 years of age obese with PCOS, so screening for glucose intolerance has been recommended in women with PCOS [3]. The prevalence of metabolic syndrome is 2 to 3 times higher among women with PCOS than among women without this entity and 20% of women with PCOS less than 20 years of age have metabolic syndrome [1]. There is also a significant risk among patients with PCOS of developing Gestational Diabetes [5].

A significant number of patients with PCOS are overweight and many are obese; however, obesity is not considered as a cause for the development of this syndrome [6].

Regarding Pathophysiology in studies, it is suggested that teak cells in women with Polycystic Ovarian Syndrome are more efficient in the conversion of androgenic precursors to testosterone than teak cells in normal women. The concentration of LH has a relative increase over FSH and the ovaries preferentially synthesize androgens. An increase in the frequency of the pulses of the Gonadotropin Releasing Hormone (GnRH) was observed. The increase in the frequency of GnRH pulses favors transcription of the Beta subunit of LH over the Beta subunit of FSH [5].

The role of insulin in the pathophysiology of PCOS is very important because it acts in synergy with LH to increase the synthesis of androgens in the cells of teak and the ovaries of women with PCOS seem to have greater sensitivity to the effect of insulin, perhaps hypersensitivity to it, even when the classic white organs of insulin, such as muscle and fat, show resistance to its action [6–8].

Insulin prevents ovulation both by direct affection of follicular development and by the indirect increase of intra-ovarian androgen levels or alteration of gonadotropin secretion. A decrease in circulating insulin levels results in an increase in the frequency of ovulation or menstruation, reduction of testosterone concentrations or both [1].

Metformin is the most widely used biguanide for the treatment of type 2 diabetes mellitus worldwide. Its most important action is the inhibition of hepatic glucose production but also increases the sensitivity of peripheral tissues to insulin. Increased insulin sensitivity, which contributes to the efficacy of metformin in the treatment of diabetes, has also been found in non-diabetic women with polycystic ovarian syndrome [1].

In women with PCOS, long-term treatment with metformin can increase ovulation, improve menstrual cyclicity and reduce androgen levels; the use of metformin can even improve hirsutism. However, it has not shown any risk modification to develop DM2 [1,9].

The results of a randomized clinical study reported in 1998, that pretreatment with metformin, compared with placebo, increased the incidence of ovulation after a subsequent treatment with clomiphene. The meta-analysis of Lord and cols in 2002 included data from 13 trials and 543 women with PCOS and concluded that metformin is effective and increases the frequency of ovulation (odds ratio, 3.88, 95% confidence interval, 2.25 to 6.69) [9,10].

The signals derived from the intestine and stimulated by the intake of oral nutrients have an important role in the release of insulin. Studies suggest that glucagon-like peptide (GLP-1) and Glucose-Dependent Insulinotropic Polypeptide (GIP) represent the dominant peptides in most intestinal insulin-stimulating hormones. GIP and GLP-1 are members of the glucagon peptide superfamily and share amino acids [11].

Incretins increase insulin secretion in a glucose-dependent manner by activation of other specific β-cell receptors [11]. An intracerebroventricular injection of GLP-1, or GLP-1 receptor agonists, produces a reduction in food intake that is associated with weight loss in some but not in all studies [11].

There are other actions of GLP-1 on the β cell independent of the acute stimulation of insulin secretion. GLP-1R agonists (GLP-1 receptor) also promote insulin biosynthesis, proliferation of β-cells and stimulate exocrine or precursor cells towards greater differentiation to the β cell phenotype. The increase in the volume of the β cell dependent on GLP-1 receptors has been demonstrated in several experiments in animals. The expansion of the β cell after the administration of GLP-1R receptor agonists prevents or delays the incidence of Diabetes Mellitus in mice [12].

GLP-1 also activates anti-apoptotic pathways, leading to a reduction in β cell death. Studies in mice have shown a reduction in the activation of caspase 3. The antiapoptotic action of GLP-1R agonists is probably directed to the reduction of peroxide-induced apoptosis of Min6 cells [12].

Giovani Paacini et al. They carried out a study whose objective was the characterization of the secretion of GIP and GLP-1 after a load of 75 g of glucose in women with PCOS without glucose intolerance compared with healthy women. The concentrations of GLP1 were the same in the women with PCOS with respect to the control women in the initial phase of the tolerance curve until 60 minutes and were significantly lower in women with PCOS at 180 minutes of the curve [11].

In a study conducted by Pontikis et al. In 20 women with PCOS who underwent a glucose tolerance curve and isoglycemic test after a night of fasting in a two-week interval, they measured levels of insulin, glucose, C-peptide, GIP and GLP-1. Obese women with PCOS were found to have low levels of GIP concentrations in response to the glucose tolerance curve compared to the control group. Age, sensitivity to insulin (QUICKI), SHBG, and basal GIP did not differ between the control group and patients with PCOS. However, baseline GLP-1 was significantly lower in obese women with PCOS compared to both control groups (p 0.023) and in thin women (p <0.02). The group with PCOS showed a decrease in GIP levels after the glucose load compared to the control group [13,14].

A novel drug, exenatide, is an incretin mimetic that simulates the glucorregulatory properties of GLP-1 [12].
Exenatide therapy often results in a loss of weight which can result in a decrease in insulin resistance. The optimal treatment of PCOS should not only improve anovulation but should also reduce comorbidities such as obesity, insulin resistance and DM2, which are linked to this syndrome [11].

Exenatide, which is an analogue of incretin glp-1, apparently has beneficial effects on the β cell mass when given in pharmacological doses to rodents. The effect of DPP4 inhibitors on the mass of the β cell is less clear. In mice in which diabetes was induced and treated with sitagliptin, it was observed that this drug preserved the β cells of apoptosis but there was no increase in the β cell mass [6].

A study conducted by Elkind-Hirsch K. et al. In patients with Polycystic Ovarian Syndrome, overweight and with insulin resistance evaluated the treatment with exenatide and metformin in terms of menstrual cyclicity, hormonal parameters, metabolic profile and inflammatory markers. We included 60 overweight women (BMI> 27) and oligo-ovulation with PCOS, between 18 and 40 years of age [15].

The results of the study showed a statistically significant increase in menstrual frequency in all treatment groups (p 0,001). More regular menses were reported with combination therapy compared with single-drug therapy (p 0.018). Compared with the baseline, ovulation periods improved in all groups, with a significantly higher proportion with the combined therapy (p 0.01) [15].

The weight decreased significantly from the first to the last visit in all groups (p 0.001). The reduction in body weight was associated with an increase in menstrual frequency significantly (p <.006) [15].

HOMA-IR decreased significantly with all treatments (p 0.043). Similarly, insulin sensitivity, determined by IS OGTT, improved significantly with treatment (p <0.002). The improvement in sensitivity was significantly higher with combination therapy than with treatment with exenatide alone (p <0.02) but not compared with metformin (p <0.085) [16].

The most frequent adverse effects were gastrointestinal to medium to moderate, nausea was the most frequent adverse effect and was greater during combination therapy [15].

Sitagliptin is a molecule that belongs to the family of selective inhibitors of the enzyme Dipeptidyl Peptidase 4 (DDP-4) that normally degrades the endogenous incretins GIP and GLP-1 [16].

In humans, it has been observed that a daily dose of sitagliptin for 10 days resulted in a nearly double increase in GLP-1 after meals.

A study conducted by Kazutaka Aoki et al. It evaluated the effect of miglitol, sitagliptin and its combination on plasma concentrations of glucose, insulin and incretins in non-diabetic men. The results showed that insulin sensitivity among the group taking sitagliptin significantly improved, endogenous GIP and GLp1 concentrations increased and a statistically significant increase in pancreatic insulin secretion was observed [16].

A systematic review and meta-analysis of drugs belonging to DDP-4 showed that there is no risk of gastrointestinal adverse effects but there was an increased risk of urinary tract infections, headache and especially nasopharyngitis [16].

It has been observed that treatment with insulin-sensitizing drugs (metformin and pioglitazone) improves menstrual cyclicity, fertility and the metabolic profile in patients with polycystic ovary [17]. However, they have no effect on the activity of the beta cell and therefore on the progression to DM2 or Gestational Diabetes [18,19]. Incretins and DPP-4 inhibitors have been shown to improve the activity of the pancreatic β-cell, inhibit apoptosis, in addition to promoting weight loss due to its anorexigenic effect, thus providing an adequate control of weight and an improvement in fertility [17]. In addition, there was a deficit in the secretion and concentrations of GIP and GLP-1 in women with PCOS [13,14]. In a previous pilot study conducted by Paredes Palma JC et al. The statistically significant effect of sitagliptin on ovarian cyclicity was observed, increasing the normalized rate of menstruation by 60% and observing ovulation in terms of comparable progesterone secretion in women who were treated with Metformin [20]. This study carried out an extended study with a larger number of patients with PCOS to compare the use of Sitagliptin Vs Metformin Vs Metformin + Sitagliptin in patients with PCOS with a greater number of patients.

Hypothesis

Treatment with sitagliptin alone or in combination with metformin in women with polycystic ovary syndrome will be more efficient in terms of ovarian cyclicity, fertility and cardiometabolic profile compared to metformin alone.

Study Design

Blinding controlled and randomized clinical trial.

Objectives

  • Evaluate the change in menstrual frequency with the use of sitagliptin and metformin, alone and in combination, in obese and nonobese women with polycystic ovarian syndrome and assess the effect on the hormonal, metabolic and inflammatory profile.

Primary Objective

  • To evaluate the changes in the menstrual pattern of patients with thin and obese SOP with the use of sitagliptin and metformin, alone and in combination.

Secondary Objectives

  • Evaluate changes in anthropometry (absolute weight, BMI, waist circumference, hip waist index).
  • Evaluate changes in insulin sensitivity and secretion.
  • Evaluate changes in the concentration of reproductive hormones (FSH, LH, PRL, testosterone, androstenedione, DHEA, DHEAS, 17 OHP4, TSH).
  • Evaluate changes in ovulation rhythm (progesterone in luteal phase).
  • Evaluate changes in the lipid profile (total cholesterol, HDL, LDL, VLDL, LDL, non-HDL cholesterol, triglycerides).
  • Evaluate changes in inflammatory markers (C-reactive protein, VSG and adiponectin, IL6, SHBG).

Inclusion Criteria

  • Age between18–40 years
  • BMI> 20
  • SOP diagnosis by Rotterdam criteria

Exclusion Criteria

  • Women diagnosed with diabetes mellitus.
  • Smokers
  • Hormonal use in the 6 months prior to entering the study.
  • Drugs that affect intestinal motility.
  • Consumption of lipid-lowering drugs.
  • Drugs that reduce weight in the last 3 months.
  • Metformin intake in the last 6 months.
  • No history of assisted fertilization treatment in the previous 6 months.

Elimination Criteria

Informed consent letter is not signed.

  • There is no attachment to treatment.
  • Do not attend scheduled appointments of pancreatic insulin secretion [16].

Description of the Experimental Maneuver

The participating patients were summoned every Friday from 8 am to 2 pm. The reason for the study, its advantages and disadvantages, was amply explained and the signing of an informed consent was submitted for consideration. To the patients who accepted to enter, clinical evaluation was applied (determination of menstrual pattern and application of the Ferriman Gallwey scale to determine the degree of hyperandrogenism), transvaginal USG and hormonal quantification were requested (LH, FSH, testosterone, androstenedione, dehydroepiandrosterone, prolactin, cortisol, ACTH, TSH, T4, T3) in order to identify patients who meet the Rotterdam criteria and exclude other diseases with a clinical picture similar to PCOS. Patients who were identified with PCOS were cited in the follicular phase of the menstrual cycle (from the 1st to the 5th day of menstruation), special mention are those patients who undergo amenorrhea who were cited from the 1st to the 5th day of bleeding after the application of 5 mg daily of medroxyprogesterone, at this time randomization was done to assign them to one of three groups:

  • Group 1, Metformin with an initial dose of 425 mg VO before breakfast and before dinner until reaching a dose of 850 mg. every 12 hours.
  • Group 2, Sitagliptin 100 mg vo. every 24 hours.
  • Group 3, Sitagliptin plus Metformin in the doses described above.

Before the administration of the first dose, they were programmed for a glucose tolerance curve of 5 hours with 75 g. of glucose. In the first sample, 20 ml were obtained to quantify: lipid profile (total cholesterol, HDL, LDL, VLDL, LDL, non-HDL cholesterol triglycerides) and inflammation markers (C reactive protein, VSG and adiponectin, IL6, SHBG) taking counts the following times 0, 30, 60, 120, 180, 240 and 300 minutes. At each time 3 ml were taken to quantify glucose and insulin.

The same was done 24 weeks after the completion of the treatment according to the assigned group with only 24 hrs of suspension of the assigned medication. An individual with a normal menstrual pattern was considered if she presented 5 menses in 24 weeks of intervention with medications (Menstruation Normalization Index, INM) (FIGURE 1).

Statistical Analysis

It was performed for quantitative variables, average and standard deviation. For proportional variables, proportions were calculated. The quantitative variables were compared with a paired student’s T. For qualitative variables, chi square test was performed. A p less than 0.05 will be considered as statistical significance.

Results

Thirty patients who were diagnosed with PCOS according to the Rotterdam criteria were included in this study, other causes of hyperandrogenism were ruled out. The patients presented clinical or biochemical signs of hyperandrogenism and 28 (93.3%) had ultrasound images compatible with polycystic ovaries. The age range was between 18 and 37 years old. Ten and nine patients (63.3%) presented menstrual alterations, 8 patients (26.6) presented opsomenorrhea and 11 (36.6%) amenorrhea. Regarding weight, 7 patients (2.3%) presented obesity grade I, 2 patients (0.6%) obesity grade II, 5 (1.6%) patients presented obesity grade III. Considering that in a time of 6 months it is normal to present 5 menstrual cycles, an index was created to normalize the number of menses per group according to their frequency. (Table 1).

TABLE 1. CLINICAL CHARACTERISTICS BY TREATMENT GROUP.

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In the hormonal characteristics of the study patients it was observed that 12 patients (4%) presented the characteristic dissociation of LH and FSH and 2 patients (0.6%) presented testosterone concentrations compatible with androgen-producing ovarian tumor, which was discarded by ultrasonography, It is also to be noted that all the patients had a normal TSH concentration except two to which they were given the treatment and still continued with criteria to establish the diagnosis of PCOS; and all patients had a concentration of 17OHP4 (17 hydroxyprogesterone) below 4 ng / mL which is the cut-off point for suspecting 21-hydroxylase deficiency. (TABLE 2)

TABLE 2. BASAL HORMONAL CHARASTERISCTIC BY TREATMENT GROUP.

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In the metabolic characteristics of the patients under study, it should be mentioned that 21 patients (7%) presented hypercholesterolemia, 15 patients (5%) presented hypertriglyceridemia, 20 patients (6.6%) presented LDL hypercholesterolemia, 8 patients (2.6%) presented abnormally low concentrations of HDL cholesterol.

On the other hand 5 patients (1.6%) had fasting altered glycemia, as well as 20 patients (6.6%) had basal hyperinsulinemia and 7 patients (2.3%) had insulin resistance and 28 patients (9.33%) disinsulinism.
Regarding the glycosylated hemoglobin in the 30 patients (100%), the value was normal, so the diagnosis of Diabetes Mellitus was ruled out by this criterion. (TABLE 3)

TABLE 3. BASAL METABOLIC CHARACTERISTICS BY TREATMENT GROUP.

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Ten patients were located in the group treated with metformin, ten patients in the group treated with sitagliptin and ten patients in the group treated with metformin + sitagliptin. The hormonal, anthropometric and metabolic characteristics between the groups were homogeneous at the beginning of the study except that in the combo group (MET + SITA) pretreatment vs the Sitagliptin group pre-treatment, initiation with greater insulin resistance (P <0.05) as well as the average basal insulin (p <0.05) but with a homogeneous index of menstruations in the three groups (P> 0.05), a condition that must be taken into account when analyzing the results for this group. (TABLE 4) (GRAPH 1)(GRAPH 2)

TABLE 4. TEST OF HOMEGENEITY OF THE TREATMENT GROUPS BEFORE THE CLINICAL ASSAY.

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GRAPH 1. BASAL INSULINE PRE Y POST TREATMENT

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GRAPH 2. BASAL GLUCOSE PRE AND POST TREATMENT

Although the patients were given counseling for the aspects of nutrition and exercise; only the recommendations of the international guidelines were proposed as part of the SOP management and it was sought that they had an adequate attachment, all the patients in each group reported having adhered to these recommendations in a percentage greater than 90%, so the effect of the intragroup and intergroup absolute weight loss and in the BMI, represents the effect of the medication in the corresponding group.
In all treatment groups there was a decrease in weight associated with the use of the medication, (p <0.05), with the percentage of change (5.8%) being higher for the metformin group and lower for the sitagliptin group (2.5%). The percentage of change for the COMBO group was 3.45%. There were significant differences in the intra-group reduction (p <0.05) in all the groups and in the comparison between groups a greater effect of the metformin group Vs sitagliptin (p <0.05) and in Metformin Vs Combo (p <0.05) was observed without having Differences between the sitagliptin group and the COMBO
(GRAPH 3)

In terms of BMI in all groups there was a statistically significant reduction intragroups (p <0.05) not so between groups (p> 0.05) after 6 months of treatment (GRAPH 4)

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GRAPH 3. WEGHT PRE AND POST TREATMENT

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GRAPH 4. MASS BODY INDEX BMI PRE AND POST TREATMENT

A reduction in the statistically significant intra-group abdominal circumference was observed in that of metformin and COMBO (P <0.05), but not in that of sitagliptin and the metformin group showed a greater difference in the reduction of statistically significant BMIs Vs the group of sitagliptin and COMBO .
In the case of the normalized menstruation index, it was found that there was a statistically significant intragroup increase in each of the treatments. The group with the highest percentage of change was that of sitagliptin with 127%, followed by metformin with 87.5% and then COMBO with 60%. When comparing the effect of the treatment after 6 months, no statistically significant differences were found between Metformin Vs Sitagliptin, nor between Metformin vs. Combo. But the effect of the COMBO Vs Sitagliptin group was greater. (Student T test p 0.05) (GRAPH 5)

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GRAPH 5. MENSTRUAL NORMALIZED INDEX (MNI) PRE AND POST TREATMENT

An increase in the number of ovulations was observed in all the groups reflected in progesterone concentrations as the treatment time was completed (P <0.05). It should be noted that none of the groups had statistically significant differences in the mean progesterone concentrations at entry into the study. No statistically significant differences were observed when comparing after 6 months of treatment between Metformin Vs Sitagliptin. But if greater effect between the group COMBO Vs Metformin and Vs Sitagliptin respectively. (GRAPH 6) (GRAPH 7)

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GRAPH 6. PROGESTERONE LUTEAL PHASE (OVULATION) PRE Y POST TREATMENT

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GRAPH 7. PROGESTERONE LUTEAL PHASE MEAN FROM 21 AND 24 DAYS (OVULATION) MONTHLY FOLLOW UP

Regarding the effect of the drugs in each treatment group in terms of the HOMA pre Vs after treatment, in all the groups a tendency of decrease of the resistance was observed only reaching the statistically significant difference in the COMBO group, there were no statistically significant differences compare the effect on insulin resistance between the treatment groups (GRAPH 8). In the secretion of insulin it was observed that both in the group of metformin and sitagliptin there was a decrease in the average concentrations of insulin, especially in the middle and final part of the secretion curve. However, a greater delta of change was observed in the sitagliptin group, especially in the 2nd, 3rd and 4th hours of secretion. (7, 9 and 9 μU / ml of insulin respectively). Compared to 4.6, 0 and 6 μU / ml of insulin at the same times for metformin. In the case of the COMBO group, a reduction in secretion was observed at all times of the curve with a delta change of 6, 19, 0.8 and 6 μU / ml of insulin in each of the hours of determination. The statistical tests for the group of Metformin and Sitagliptin showed no differences in the intragroup change (paired T> 0.05) as well as between each secretion moment between groups (ANOVA p> 0.05). (GRAPH 9) In the case of the COMBO group I present statistically significant differences intragroup but not when compared with the other two treatments. (GRAPH 1)

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GRAPH 8. HOMA PRE AND POS TREATMENT

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GRAPH 9. INSULIN INTEGRATED SECRETION IN 5 HOURS PRE AND AFTER TEST

In lipids there was no effect of any of the treatments only in the case of metformin in terms of triglycerides if there was a statistically significant reduction (paired T-test p <0.05) with a delta of change of 24.04gr / dl decrease (16 %). An increase in HDL cholesterol was also observed in this group with a change delta of 6.38 (14.9%) (Paired T test p <0.05) (GRAPH 10–13)

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GRAPH 10. CHOLESTEROL PRE AND POST TREATMENT

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GRAPH 11. TRIGLYCERIDES PRE Y POST TREATMENT

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GRAPH 12. TOTAL CHOLESTEROL MONTHLY FOLLOW UP

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GRAPH 13. CHOLESTEROL HDL MONTHLY FOLLOW UP

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GRAPH 14. CHOLESTEROL LDL MONTHLY FOLLOW UP

Discussion

The group of patients with PCOS had homogeneous characteristics and no statistically significant differences. In the case of the COMBO group at the beginning, significantly greater resistance to insulin was found than the Sitagliptin group, part of the effect that was observed 6 months after the start of the treatment superior to sitagliptin could have been due to this fact. The INM Menstruation Normalized Index did not show statistically significant differences between the groups at the beginning of the treatment. It is noteworthy that within the groups of sitagliptin vs metoformine the biochemical, clinical and hormonal characteristics did not show statistically significant differences and the effect on increased frequency of menses is comparable to that observed in the metformin group, considering that the basal rate of menstruation was greater in the sitagliptin group. Although the trend suggests that sitagliptin would have a greater effect on the increase in the frequency of their menses, no significant differences were found after 6 months. This stands out from the pilot study carried out by Paredes et al., Where the group with the highest delta change was metformin and this study coincides with the fact that no statistically significant differences were found after 6 months of treatment. These changes may be due to the greater number of samples in the present work. In the case of the COMBO group, the effect on the frequency of menses also reached group significance and after 6 months had a greater effect than sitagliptin, unlike those found in the pilot study, the latter can also be explained by the greater amount of patients included in the sample of this study. In the case of the Elkind-Hirsch K. study, in which metformin vs exenetide vs COMBO was compared, an increase in the frequency of menses was observed in all groups, however between groups there were also significant differences for weight loss. that the effect of exenetide on the frequency of menses can be due to this phenomenon [16].

In the case of our study, statistically significant differences in weight were observed before and after any treatment group, so we observed in terms of increased frequency of menses and ovulation could be attributed to this fact, however the percentage delta of change in minor weight of the group of sitagliptin Vs Metformin and the fact that after 6 months of treatment there was greater statistically significant weight loss in the group of Metformin Vs Sitagliptin and Vs COMBO and before the results of finding greater effect of sitagliptin in the INM and in terms of ovulation leads us to think that in the case of sitagliptin the effect in the treatment should be explained by other phenomena besides weight loss, at a certain moment we would have to carry out a study evaluate the presence of receptors and their mechanisms of action of sitagliptin within the ovary or within the reg ulation of the pituitary-ovary axis. This also highlights what was found in the pilot study where none of the treatment groups presented differences in weight, supporting the previous statement of the need to study other possible sites of action of sitagliptin in the reproductive regulation of women with PCOS. . It is noteworthy that although the COMBO group was less efficient in terms of increasing the frequency of menses, its effect on ovulation was greater than in other treatment groups, which could suggest that it is due to the additive effect that sitagliptin provides to the treatment. . In the Paredes et al pilot study, the findings were consistent with the above. In terms of insulin secretion, it should be noted that although sitagliptin is not considered an insulin sensitizer, it has been observed that it decreases insulin plasma concentrations, perhaps by influencing a better functioning of the pancreatic beta cell, in our studies showed a significant decrease in insulin secretion, even with delta change greater than in the metformin group in the first part of the secretion curve, but without reaching differences statistically between both groups, these findings are consistent with the observed in the weight loss in the treatment groups as well as with the behavior of the HOMA, where the intra-group trend was observed in all the treatments, the tendency to reduction after 6 months of treatment and with greater effect in the COMBO group, however, there is to consider that this group started with a higher degree of insulin resistance compared to other groups, so its greatest effect can be explained by this cause. This has also been observed in a study conducted by Kazaka Aoki et al [15]. In the case of the lipid profile, the only treatment that showed a pleiotropic effect for triglyceride reduction and an increase in statistically significant HDL cholesterol was Metformin, an effect that seems independent of weight reduction and insulin resistance. In many other studies, there has also been no effect of sitagliptin and even metformin on lipid metabolism [9,8,10,11,15], even in the pilot study conducted by Paredes et al [20].

Conclusion

Sitagliptin improves ovarian cyclicity and ovulation in women with PCOS in comparable terms with respect to metformin and the metformin sitagliptin combination. The combination of sitagliptin metformin is more effective in terms of ovulation than the other two treatments alone. However, sitagliptin showed that it can influence the reproductive and intra-ovarian aspect. Similarly, it showed that it can improve the metabolism of insulin in patients with PCOS, so it would be interesting to show if it could be a treatment that not only improves the clinical, metabolic and reproductive conditions of patients with PCOS, but also prevents the development of Diabetes. Mellitus, a highly frequent consequence of these patients. The weight did not change the results of the findings.

References

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Nucleoprotein Assembly Mediated Transfection

DOI: 10.31038/NAMS.2019213

Abstract

Ordered DNA methyltransferases covalently bound to Y-junction DNA were spontaneously taken up mammalian cells in culture, where a nuclear localization signal carried by the bound methyltransferases caused the fluorescently labeled nucleoprotein complex to decorate the nucleus of several mammalian cell lines. We report proof of concept experiments demonstrating that the Y-junction nucleoprotein assembly can be modified to carry gene expression cassettes that permit spontaneous transfection and expression of a desired protein in mammalian cells.

Introduction

Ordered nucleoprotein assemblies have been developed for a variety of applications in Nanofabrication [1]. Many protein bearing nanoparticles have been shown to target mammalian cells where they can be selectively internalized [2,3].   Bacterial DNA methyltransferases covalently bound to Y-junction DNA have also been shown to be spontaneously taken up by mammalian cells in culture [4].  Further, those experiments showed that the nuclear localization signal carried by the bound methyltransferases caused the fluorescently labeled nucleoprotein complex to decorate the nucleus of several mammalian cell lines. Here we report proof of concept experiments using Green Fluorescent Protein (GFP) expression to demonstrate that the Y-junction nucleoprotein assembly can be modified to carry gene expression cassettes that permit spontaneous transfection and expression of a desired protein in mammalian cells.

Materials and Methods

Cell Culture conditions, EcoRII DNA methyltransferase purification, flurocytidine containing DNA Y-junction synthesis and construction, as well as the construction of the thioredoxin-linked nanoparticle have all been described in detail [4–6].  The Y-Junction was modified for expression cassette delivery as described in figure 1.

NAMS 2019-104 - Steven Smith USA_F1

Figure 1. The NP-Trx2-GFP-DNA was assembled from the component parts.

Synthesis of the component parts. 1.  The EcoRII Methyltransferase fusion protein was cloned, expressed and purified from E. coli.   2. The DNA scaffold is composed of three DNA strands with a Cy 5 tag added during the synthesis. The sequences for two M.EcoRII binding sites and one Tsp RI Ligation site are designed into the sequences. 3.  The GFP DNA was generated by PCR, which will include the GFP open reading frame and the eukaryotic promoter sequence.  A Tsp RI sequence was included in the N-terminal region of the GFP DNA.   B.  The assembly consists of three steps.  First the three strands of the DNA scaffold were annealed. Then the GFP DNA was ligated to the DNA scaffold.  Finally the EcoRII Methyltransferase fusion proteins were coupled to the EcoRII Methyltransferase binding sites.

Transfection Conditions: Comparison of NP-Trx2-GFP -DNA Mediated GFP Delivery with Lipid Mediated Transfection.

We compared the effectiveness of delivering NP-Trx-GFP-DNA to DU145 cells with a traditional transfection reagent (e.g. Lipofectamine™ (Invitrogen)). Cells were plated at 5,000 cells per plate (Costar) before delivery without antibiotics and grown to 50–70% confluence. The media was removed from each well and the cells were washed with PBS.  Then 1.0 µM NP-Trx2-GFP-DNA was added to the cells and incubated for multiple time points at 37oC in 5% CO2. The transfected cells were observed by fluorescent microscopy and fluorimetry for the frequency of green fluorescent protein expression in viable cells per input viable cell. The emission wavelength of Cy5.5 at 707nm, which is on the DNA Y-Junction (Figure1), did not interfere with the appearance of fluorescence from GFP at 509nm.  Following incubation, the cells were washed with PBS and growth media was added and the cells were incubated to permit growth.  The cells were monitored using an inverted fluorescent microscope over the next 10 days for internalization at 707nm and for the appearance of GFP fluorescence at 509nm indicating successful expression of GFP.

Results and Discussion

As noted above the trisubstituted nanoparticle was found to spontaneously enter mammalian cell lines of several types (Figure 2).  Among those, the DU145 cell line, which showed significant uptake in the nucleus and HK293, which showed weak uptake were selected for further work in this proof of concept report.

NAMS 2019-104 - Steven Smith USA_F2

Figure 2. Fluorescent images of cell lines exposed to the NP-Trx3.

We observed differential binding of the NP-Trx3 prepared as described in [4] with different cell lines.  Fluorescence was observed in DU145 (Panel A), LNCaP (Panel B), MCF-7 (Panel C), HK293 (Panel D) but not MCF7cells exposed to Y-Junction DNA without the targeting ligand (Eco RII DNA methyltransferase) (Panel E), Primary Prostate Epithelial Cells (PrEC) (Panel F), PC-3 (Panel G) and COS7.

Final assembly of the carrier nanoparticle (NP-Trx2-GFP-DNA) was carried out as described in materials and methods. Each stage in the process was tested using microfluidics mobility shift analysis [7]. The results of a typical mobility shift analysis are depicted in Figure 3.Transfection efficiency was monitored by GFP fluorescence after transfection.  Nuclear fluorescence indicating GFP expression was observed with DU145 cells using Y-GFP DNA and Lipofectamine mediated transfection or with purified NP-Trx2-Y-GFP-DNA without Lipofectamine mediation.  The mixture of unreacted components of the nanoparticle (Y-DNA, GFP-DNA, and EcoRII-Trx) did not yield GFP expression (Figure 4).  Unpurified Np-Trxn-Y-GFP-DNA was found to exhibit GFP expression without Lipofectamine mediation suggesting that the nanopaticle might support transfection with a single EcoRII-Trx molecule acting as targeting ligand.  HK293 Cells were also found to support NP-Trx2-Y-GFP-DNA without Lipofectamine mediation although with lower efficiency than DU145 cells.

NAMS 2019-104 - Steven Smith USA_F3

Figure 3. Mobility Shift analysis of the Steps in Carrier Nanoparticle Assembly.

Y DNA: Purified Y-Junction. GFP DNA: Purified TspR1 cleaved GFP expression cassette. Ligated Y-GFP DNA: Purified Y-DNA GFP DNA ligation product. NP-Trxn-Y-GFP-DNA: Unpurified EcoRII-Txn + Y-GFP reaction products, showing DNA carrying one or two covalently bound EcoRII-Txn molecules and two covalently bound and one non-covalently bound EcoRII-Txn molecules.

NAMS 2019-104 - Steven Smith USA_F4

Figure 4. GFP Expression after transfection.

A: Lack of GFP expression after exposing DU145 cells to the mixture unreacted components of the nanoparticle (Y-DNA, GFP-DNA, and EcoRII-Trx). B: GFP expression after Lipofectamine mediated transfection of DU145 cells with 300nM Y-GFP-DNA.  C: GFP expression after exposing DU145 cells to 300nM purified NP-Trx2-Y-GFP-DNA alone.

Expression was seen to be transient with the number of fluorescent nuclei diminishing steadily over a period of ten days, suggesting that the DNA from the nanoparticle did not become integrated. Transfection efficiency was comparable to that observed with Lipofectamine mediation transfection, and depended on the cell line used for transfection.

For DU145 cells 38% transfection was observed with Nanoparticle mediated or Lipofectamine mediated transfection assuming the input transfectant was not replicated. However, the efficiency was adequate for most purposes and avoids the undesirable cellular stress of Lipofectamine exposure [8]. We also noted that purification of the NP-Trx2-Y-GFP-DNA DNA was unnecessary since simply allowing the reaction between the Y-GFP-DNA and EcoRII-Trx to occur in 30 min incubation at 37oC resulted in comparable transfection levels. We were not concerned that uncoupled EcoRII activity might alter gene expression by methylating non CG methylation sites since previous work with this enzyme showed that this does not occur [9].  Further, given the ability of the of the nanoparticle to spontaneously enter the cytoplasm of certain cells (e.g. LNCaP see Figure 2B) the system can be modified to carry siRNA by repurposing the truncated Y-Junction so as to permit hybridization of a double stranded siRNA molecule to the nucleoprotein complex, thereby avoiding the deleterious effects of the Lipofectamine transfection generally employed in siRNA transfection [8].

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