This action might not be possible to undo. Are you sure you want to continue?
DETECTION OVARIAN CYCLE OF BEKANTAN (Nasalis larvatus) BASED ON THE PROFILE OF FECAL ESTRADIOL AND PROGESTERONE
Pudji Astuti1), Claude Mona Airin 1), Hera Maheshwari 2), Luthfiralda Sjahfirdi 3)
Department of Physiology, Faculty of Veterinary Medicine, Gadjah Mada University Jl.Fauna 2, Yogyakarta. Indonesia. Email: email@example.com, Phone: 08156851163 2) Department of Physiology, Faculty of Veterinary Medicine, Bogor Agricultural University 3) Department of Biology, Faculty of Mathematics and Science, University of Indonesia, Depok because they have regular cycle whereas 1 animal has an irregular reproductive cycle, so it would be need further observation. There were concluded that fecal samples can be used to determine the endocrine ovarian function and fertility status of Nasalis larvatus. Index Terms: bekantan, estradiol, fecal, ovarian cycle, progesterone I. INTRODUCTION
Abstract--Based on the IUCN, Proboscis monkey (Nasalis larvatus) is one of primate with status endangered species. To support this species conservation programs, it has been conducted the monitoring ovarian cycles using fecal samples. Samples which used in this study were approximately of 210 fecal samples derived from 3 adult females which were kept in Gembira Loka zoo, Yogyakarta, Indonesia. Fecal sampling were withdrawn in the morning between 8-9 am for about 2 months. Subsequently, feces which obtained inserted into the ziplock plastic and stored in the freezer at temperature-20oC until to do the next step such as lyophilization, pulverization and extraction using methanol. Sample then assayed using EIA (Enzyme Immunoassay) method. Length of Estrous cycle was determined by calculating duration of peak estradiol hormone until peak of estradiol in the next cycle. The results showed that fecal estradiol levels were estimated from 5494.00 to 24645.606 pg / g dried weight feces and progesterone were 72.60 - 390.40 ng/g dried weight feces. The average of reproductive cycle is 22.66 days with a range of 22-23 days . Progesterone hormone cycle is shorter than estradiol (21-22 days). Although no further investigation, but this is expected because of differences in time lag between estradiol and progesterone. From the three adult female Bekantan which used in the study, 2 animals were diagnosed as fertile
Severeal years ago, hormone analysis has been done using either plasma or serum, but in the recent time, monitoring steroid fecal metabolites is one important method for determining the profile of the reproductive cycle, especially in wild animals such as bekantan which blood withdrawn is not possible every day. Although his method rather complicated and only inactive metabolites are measurable but this methods are very useful to determine the reproductive cycle of male (Astuti et al., 2006) and female animals such as musth (Ganswindt et al., 2005) and some primates (Mohle et al., 2002); in Hylobates moloch (Astuti et al., 2006; Maheswari et al., 2007); determining chronic stress (Bahr et al., 2000; Astuti et al., 2010); and determine the activity of the adrenal gland; conception, pregnancy (Ziegler and Wittwer., 2005). Some of the benefits were obtained in the use of hormone analysis in noninvasive methods are : 1) sampling does not
112804-7676 IJBAS-IJENS © August 2011 IJENS
METHODOLOGY A. The results show the effect of estradiol and progesterone showed parallel results with the standard.1P 2. fruits such as papaya. ParalelismTest of Estradiol 1. As a validation test. Indonesia. Thus. then each transferred into a plastic tube to the next do lyophilization and pulverization with modified to (Ostner and Heistermann. II. the solution is inserted into the tube polipropilene size 15 ml. fecal should be mixed entirety before placing in plastic tube. and oranges. Estradiol and Progesterone Assay. between the hours of 06. Powder that has been formed is taken 50 mg of dry weight and then extracted with 80% methanol as a solvent of 3 ml.. Proboscis monkey in this study was chosen because bekantan is the endemic species from the island of Borneo. Fecal sampling would be done every morning for about three months. would be done.00 – 07. Animals. Fecal Sampling. Furthermore. of course.6 0. stored in -20oC until Assayed using Enzyme Immuno Assay’s (EIA’s) Estradiol and Progesterone commercial kit product DRG.International Journal of Basic & Applied Sciences IJBAS-IJENS Vol: 11 No: 04 2 require anesthesia in animals. Thus the results obtained would be reflected as actual condition. D. They were given food in the form of a standard diet of vegetables (water spinach and spinach). The purpose of this study were: to determine the metabolite profile of estradiol and period of menstruation.2 1 0. Feed were given 2 times a day namely morning and afternoon. Supernatant was poured into 1. To obtain a high homogenecity. tests on samples could be continued (Figure 1).8 0.29S 2. analyzed using ELISA washer and reader with a wavelength of 450 nm. 3) specifically for urine. Anesthesia for animals cause inconvenience. Indonesia which declared as threatened with extinction by The International Union for Conservation of Nature (IUCN). the samples were placed in ziplock and stored in the freezer-20oC. bananas.5 ml micro tubes.21P 112804-7676 IJBAS-IJENS © August 2011 IJENS IJENS . vortexed for 10 minutes. something that can not be done on animals that aneshtized. The sample were used ± 210 fecal samples derived from 3 adult female bekantan with aged between 5-12 years with a body weight of 7-10 kg. C.4 1.4 0. B.2 0 0 5 10 15 Consent of estradiol stnadart 2. All of samples centrifugated for 10 minutes with speed of 500 g then carried out immediately. Fecal Extraction.30P 2. Animals used in research were adult females bekantan which kept in Gembira Loka zoo. Yogyakarta. 2) incidence because of the stress anesthesia could be reduced so that the stress factor could also be reduced or avoided.00 am. it has been carried out test of parallelism to the hormone estradiol and progesterone using serial dilution. Then. in a single day can be obtained from samples more than once on the same animal. Fecal sample that had been stored in the freezer would be taken. 2003).16S 2. Fecal samples which has been extracted.
measurement of fecal concentration of estradiol is widely used to monitor reproductive cycle..56 6251. RESULTS AND DISCUSSION A.10 20 30 Consent of Progest B1. Sjahfirdi (2006) reported the length of ovarian cycle of Hylobates moloch was found to be 29 until 38 days whereas Maheshwari et al. about 9 days (Martin.9. the length ovarian cycle of Siamang (Symphalangus syndactylus) was 30-33 days (Astuti et al. for animal VI. Based on the presence or absence of menstrual bleeding. (2010) reported the length reproductive cycle of Hylobates moloch using fecal steroid was 21-25 days (Figure 4) ..19 5494. It means that average of the ovarian cycles can be estimated 22. Ovarian cycle are generally vary start from the longest namely squirrel monkey. 2010) whereas using vaginal cytology the length of ovarian cycle of Siamang was 3742 days (Yundiarto et al. 1:4. In this study.10 Figure 1.. In general. 1995).1. Different from two of bekantan. Profile of Estradiol Metabolites.9. (2007). there are some differences in determination the reproductive cycle of primates and other mammals (Bennet et al.00 22 2958. (2009). 1:8. 1:16.94 unknown Although there were a wide variety in estradiol levels. Table 1.9.10 B1. The range of estradiol metabolites from 3 adult female bekantan were vary start from 5494. By using fecal steroid. 1981).606 pg / g dry weight feces (Table 1). In this study the reproductive cycle length determined by the highest concentration of cyclic peak of estradiol to the next highest concentration of estradiol based on Geissmann and Anzenberger.. 1:64. This condition is similar as in the black-handed spider monkeys wich hormone fluctuations were IJENS 112804-7676 IJBAS-IJENS © August 2011 IJENS . 1: 128.66 days with a range (22-23 days). it can be explained that the reproductive cycle of the 3 female bekantan as follows: the ovarian cycle length of bekantan I was 23 days. 2004). the reproductive cycle is used to be the menstrual cycle. peak of estradiol just occurred once. Maheshwari et al..5 0 0 10 standart B1. III. whereas for cattle and other ruminants using estrous cycle.85 11789..77 2146.28. bekantan II was 22 days (Figure 2).94 23 10460..99 4817. 2003) until the range 25 to 84 days for Chimpanzee (Tutin and Mc Ginns.23 251.5 OD 1 0. Concentration of fecal estradiol (pg/G dried weight) Bekantan I Bekantan II Bekantan VI X (average) SD The highest The lowest Length of cycle Days) 9146. 1:32. Paralelism test of estradiol and progesterone with the serial dilution start from 1:2.International Journal of Basic & Applied Sciences IJBAS-IJENS Vol: 11 No: 04 3 Paralelism Test of Progesterone 2 1.74 24645.9.24 2022. It is indicated that the ovarian cycle is irregularly and it can not be further discussed (Figure 3). For primates such as chimpanzees and orang-utans.00 to 24645.06 20173. Comparison of ovarian cycle length in primates yield valuable insight.
2002).9 16. estradiol secretion peak also occurs just before ovulation.9 sampling Figure 2.9 10 12.9 sampling 15000 10000 5000 0 Profile Estradiol of Bekantan VI (pg/dw feces) 1.9 18.9 20.9 8. In the most of primates.9 27. Profile Estradiol of Betina II (pg/dw feces) 30000 20000 10000 0 10. In this study. Such events are often referred to as positive feedback (Brook and Marshall.9 1.9 6. Blanco and Jerrold (2009) reported fecal Estradiol of brown mouse lemur levels increased before or at the day of estrus. it would be expected that high levels of estradiol was needed to surge of LH until ovulation.9 1..9 29.9 7. at least its very important for clueing the length of ovarian cycle. and data IJENS 112804-7676 IJBAS-IJENS © August 2011 IJENS ..International Journal of Basic & Applied Sciences IJBAS-IJENS Vol: 11 No: 04 4 readily apparent in all females.9 10 12. so that the reproductive cycle can not be determined. even sum of animals would be used were very limited. Peak of estradiol just once.9. Profiles of E1C in feces of female Javan Gibbon (Ullah) from Taman Sari zoo (Maheshwari et al.9 28.9 24.9 24.9 22. 2006).9 14. 20000 Profile Estradiol of Bekantan I (pg/dw feces) 10000 0 1.9.9 6.1 sampling Figure 3. Guimaraez (2003) reported that information obtained through non-invasive methods are of great utility to the planning of reproduction programs. but the maximum period between estradiol levels in a cycle can be used as an indicator of female fertility.9 16..9 13.9 22. Profile of estradiol metabolites in Bekantan I and II indicated regularly. Profile of estradiol metabolites in Bekantan VI indicated irregularly. in agreement with results from urinary Estradiol measurements in gray mouse lemurs reported by Perret (2005). the length of cycle are 23 and 22 days.9 16.9 4.9 28.9 22. Figure 4. Progesterone concentrations were higher in old versus young females (Molina et al. 3.9 18.9 14. 3.9 27.9 19. such as the choosing of females with an adequate pattern of cycles to dictate the best moment for an artificial insemination or even optimizing parameters for insemination through natural breeding. Although LH levels as an indicator of ovulation is not detected.9 25. and subsequently decreased.9 8.9 20. 2010).
9 29.9 4.0 300.6 73.9 24.. In animal II.914.9 14. Profile of Progesterone Metabolites.7 220.9 22.9 3.9 prog… Sampling 1.0 400.9 12.9 18.4 Figure 5. 2003).927.10 sampling Profile Progesterone of Bekantan II (ng/dw feces) 112804-7676 IJBAS-IJENS © August 2011 IJENS IJENS . Profile progesterone metabolites of Bekantan.7 31.62 82.8 146.4 150. Maheshwari et al.917. it means that peak of progesterone is later than estradiol. increased hormone estradiol occurred at day 3 and the 26 so that periods of ovarian cycles would be estimated 23 days. 250 200 150 100 50 0 Profile Progesterone of Bekantan I (ng/dw feces) 500.923. In animals I.9 8.9 11.9 6.91 72. This finding is consistent with Blanco et al. Table 2. Concentration of fecal progesterone (ng/G dried weight) Bekantan I Prog/dw Profile Progesterone of Bekantan VI (ng/dw feces) 600.. This fact is supported by the profile of progesterone hormone which has 2 peaks in one cycle of the day of the 8th and the 27th (Figure 5).9 8.92 74.International Journal of Basic & Applied Sciences IJBAS-IJENS Vol: 11 No: 04 5 would be completed with progesterone metabolites.6 390. profile of B.0 1.8 72.9 27.0 200. Besides level of estradiol. (2010) reported the length of ovarian cycle in Javan gibbon is longer than bekantan (Figure 6).9 1. it was only one peak of progesterone. In this study level of fecal progesterone were started from 72.0 100. reproductive cycle can be determined by using profile of progesterone.60 . (2009) which reported in Brown mouse Lemurs fecal progesterone started to rise after estrus in most of the females values were consistently low during ovulation and early pregnancy.9 10.0 200.8 416..0 0.9 20.9 16.0 progesteron … Sampling Bekantan II Bekantan VI X (average) SD The highest The lowest 118.930. low mating concentrations and were attained within 5 days after parturition (Ostner and Heistermann.390.0 0.0 400. peak of progesterone was occurred on day 6 and day 28.4 ng/gr dried weight feces (Table 2). Bekantan I and II has regular cycle whereas bekantan VI has irregularly cycle.920.
. (2000) reported that the time lag for the hormone progesterone squirell and cotton top tamarin monkey respectively 8 and 24 hours while for the hormone estrogen respectively 16 and 48 hours. Diurnal Pattern of Fecal Testosterone and Cortisol Metabolites in the three of Captive-housed Male Javan Gibbons (Hylobates moloch) J. 2005). In Agreement with the first rise of excreted glucocorticoid metabolites in all of monkey which occurred 24 hours poststressor but reached maximum level after 3-4 days. Asmarani Kusumawati. E Hayes. REFFERENCES  Astuti P.66 days with a range of 22-23 days. it seems only one peak of 112804-7676 IJBAS-IJENS © August 2011 IJENS . In addition. Hera Luthfiralda Sjahfirdi. Bahr et al. The last animal is bekantan VI. Detection of Chronic Stress in (Symphalangus syndactylus) Siamang using measurement of fecal Corticoid. the time lags that exist between the stress-induced circulating glucocorticoids and the apparence of their metabolites feces is related to gut passage time (Palme et al. TL Yusuf. Februari 20th. This research is granted by Hibah Kompetensi from The Directorate General of Higher Education.2010. ACKNOWLEDGEMENTS: We would like to thank you very much to The Director of Gembira Loka Zoo. Howler gut passed time range from 23-30 hours to a maximum of 72 hours (Milton. Proc. The 7th National Seminar on Basic Science. In difference case. Claude Maheshwari. This condition is the same as her estradiol profile. Joko Purnomo and Rini for technical assistance. Bpk Machmud Asvan (vet). Bekantan reproductive cycle lengths ranged from 22. Based on the time lags. Estrogen and Progestogen isotopes in feces exhibited an excretion peak at 16 hr post-injection. D Sajuthi 2006. Malang. IV. June:2: 15-18  Astuti P. Maheshwasri L Sjahfirdi. H.. Ibu Fitri (vet) for giving permission to use animals.. (2002) reported the major portion of the urinary metabolites of both hormones estradiol and progesterone in squirrel monkey (Saimiri sciureus) was excreted within 16–24 hr post-injection.. Moorman et al. Pp: 311316 IJENS Figure 6. means indicated an irregular profile. some conclusions can be made as follows: fecal samples can be used as a means to determine endocrine and ovarian function in the female reproductive cycle.International Journal of Basic & Applied Sciences IJBAS-IJENS Vol: 11 No: 04 6 progesterone in day 7 whereas peaks on the day 27 is not clear enough (Figure 5). This fact may explain why estrogen hormone secretion period very close to the secretion of progesterone hormone as in the female reproductive cycle of bekantan I and II. fecal samples can also be used to determine fertility status Proboscis monkey. Yogyakarta Indonesia. Profiles of PdG in feces of female Javan Gibbon (Donna) from Taman Sari zoo (Maheshwari et al.. 2010). CONCLUSION Based on results and discussion. Mona Airin.1984).Hayati Ed.
Sjahfirdi L. Astuti P. 59: 111-135  Milton. Mohle U. Palme R. M. Airin CM. Proc. Jour of Primatol. Rojas-Maya S and Mondragon RC. Heistermann. 2002. Physical. Abstract: 15  Astuti P. 1995. Physiological and Biochemical Zoology: 78:505-514 Guimarães.). Sajuthi D. USA: Academi Press. Physiological. Gen Com Endocrinol 117:427-438 . Bangkok. Hodges K. Hodges JK and Heistermann M. Kusumawati A. Immuno. th International seminar 20 PSSP. Characterization of Urinary and Fecal Metabolites of Testosterone and Their measurement for assesing Gonadal Endocrine Function in Male Nonhuman Primates. Nonhuman Primetes in Biomedical Research: Biology and Management. Hernandez-Lopez L. 2006. Alikodra HS. Hera Maheshwari . Astuti P. Purwantara B. 71:1–8  Bennett BT. Am. Bogor. and a review of ovarian cycles in gibbons (Hylobatidae). In PS Rodman & JGH. 5 – 2009  Ganswindt A. Fecal Steroid Profile of Female Javan Gibbons (Hylobates moloch) Maintained in Pairing-Typed Cage. Hormonal correlates of the ovarian cycle in the yellowcheeked crested gibbon (Nomascus gabriellae). Reprod. J. 2005. MABV. Of the 13th Association of Institutions for Tropical Veterinary Medicine Conference. Heistermann M. 2000._Assessing Reproductive Profiles in Female Brown Mouse Lemurs (Microcebus rufus) From Ranomafana National Park. Sajuthi D. Gen Comp Endocrinol 129:135-145 Molina ALC. The role of foodprocessing factors in primates food choices. Inc. Alikodra HS. 2010. Blackwell Science. Kant (eds.Gibbon Journal Nr. Human Reproduction: a comparative background for medical hypothesis. HAYATI Journal of Biosciences (17) (1): 43-49 Martin RD. Daily Activity of Siamang (Symphalangus syndactylus) in Menoreh Greenland Rescue Center. 1984. Seasonal variations of fecal progesterone and IJENS 112804-7676 IJBAS-IJENS © August 2011 IJENS .2009.2010. Purwantara B. Comparative Aspects of The Metabolism and Excretion of Cortisol in Three Individual Nonhuman Primates. Luthfiralda Sjahfirdi.2009. Abee CR. K. Adaptation for foraging in Nonhuman primates Pp 249-279 Mohle U. 2010. Asmarani Kusumawati. Palme R and Hodges JK. Managing (Symphalangus syndactylus) Siamang in Menoreh Greenland rescue Center: Study of Primates behavior. 2002. Annu Rev Biomed Sci 2003.International Journal of Basic & Applied Sciences IJBAS-IJENS Vol: 11 No: 04 7  Astuti P.  Blanco MB and Jerrold. The Importance of the Application of Noninvasive Techniques in the Study of Wild Animal. 2003. 5: 29-37  Maheshwari H. Yogyakarta. behavioral correlates of musth in captive African Elephants (Loxodonta africana). PP: 101-103  Bahr NI. Widjajakusuma R dan Tolihere M. Henrickson R. Sarmin . Using Fecal Hormone Analysis. hlm 286-297. 5rd Edition. 2003. Southeast Madagascar. and Marshall NJ. Sjahfirdi L. Paez-Ponce DL. Claude Mona Airin. Sjahfirdi L.  Geissmann T and Anzenberger G. Media Konservasi: (XII): 16-21  Maheshwari H. Widjajakusuma R. Essential Endocrinology. Maheshwari H.  Brook CGD. Fecal steroid Profile And Genital Swelling of Female Javan Gibbons (Hylobates moloch Audebert 1797) Maintained in Individual Cage. 2007.
The Gray Mouse Lemur. Heistermann M. Program Studi Biologi. Advanced in Ethology 38: 120 Ziegler T And Wittwer DJ . Fecal Steroid Research in the Field and Laboratory:Improved Methods for Storage. J of Primatol: 67:159– 174 112804-7676 IJBAS-IJENS © August 2011 IJENS IJENS . Universitas Indonesia. Abstrct.International Journal of Basic & Applied Sciences IJBAS-IJENS Vol: 11 No: 04 8 17β-estradiol in captive female blackhanded spider monkeys (Ateles geoffroyi). 2006. New York: 239-264 Yundiarto Y. Transport. Lasley BL. 1981: Chimpnazee reproduction in the wild. 131:274–283. Processing. Endocrine characterization of female reproductive status in wild redfronted lemurs (Eulemur fulvus rufus). Maheshwari H and Sjahfirdi L. Tutin CEG and McGinnis PR. Comparative and biomedical perspectives. and Analysis. 2003. Gen Comp Endocrinol.:1985-1993 Moorman EA.: 57:2: 79-90 Ostner J. CE (ed). Vaginal Cytology and Genital Swelling during menstrual cycle in captive-housed Siamangs (Hylobates syndactylus). 2002.2005. 2004. Theriogenology (66) (8) Pp. Assessment of Reproductive Biology in CaptiveHoused Female Javan Gibbon (Hylobates moloch Audebert 1797): With Special Emphasize On Ovarian Cycle Determination and Daily Activitty Observation. 162-171 Perret M. Setiorini. Sjahfirdi L. Program Pascasarjana. Fakultas Matematika dan Ilmu Pengetahuan Alam. 2005. excretion. Stress hormone in Mammals and birds. Indonesia. and non-invasive measurement in fecal samples. Reproductive biology of the Great Apes. Rettenbacher S. Depok. Blackwell Verlag Berlin. Jawa Barat. Dissertation. Comparatve aspects regarding metabolism. Annals of the New York Academic of Sciences. Excretion and measurement of estradiol and progesterone metabolites in the feces and urine of female squirrel monkeys (Saimiri sciureus). Am J of Primatol. Academic Press. Touma C. 2005. Shideler SE. Relationship Between Urinary Estrogen Levels Before Conception And Sex Ratio At Birth In A Primate. Hum Reprod 20:1504–1510. 1040. In: Graham. Elbahr SM. Palme R. Am. Mendoza SP. Mostl.
This action might not be possible to undo. Are you sure you want to continue?
We've moved you to where you read on your other device.
Get the full title to continue reading from where you left off, or restart the preview.