Reproductive

Agnew, D.W., L. Munson, and E.C. Ramsay (2004). Cystic endometrial hyperplasia in elephants. Veterinary Pathology 41(2): 179-83.
NAL Call Number: 41.8 P27
Abstract: Most captive female elephants are nulliparous and aged and many have endometrial disease, factors that may hinder fertility. This study characterized the pathologic features and demographic distribution of endometrial lesions from 27 captive Asian (Elephas maximus) and 13 African elephants (Loxodonta africanus), 12- to 57-years of age. The principal lesion was marked cystic and polypoid endometrial hyperplasia (CEH), present in 67% of Asian and 15% of African elephants ranging from 26 to 57 years. The lower prevalence in African elephants likely reflects their younger age range in this study. Fourteen of 15 affected elephants with breeding information were nulliparous. These results suggest that CEH and polyps are common in aged nulliparous elephants, and the severity of these lesions may impair fertility. These findings will be useful in the interpretation of ultrasonographic findings during reproductive examinations of potential breeding cows. Also, breeding programs should focus on younger animals.
Descriptors: zoo animals, endometrial hyperplasia, endometrium pathology, fertility physiology, polyps, endometrial hyperplasia pathology, histological techniques, polyps pathology, species specificity.

Ball, R.L., J.L. Brown, J. Meyer, J. St. Leger, and J.H. Olsen. (2004). Treatment of anestrus due to hyperprolactinemia with cabergoline in a captive Asian elephant (Elephas maximus). Proceedings: American Association of Zoo Veterinarians, American Association of Wildlife Veterinarians, Wildlife Disease Association: Health and Conservation of Captive and Free-Ranging Wildlife. Joint Conference, San Diego, California, American Association of Zoo Veterinarians: p. 363-365. 660 p.
Descriptors: Asian elephant, anestrus, treatment, hyperprolactinemia, Elephas maximus, cabergoline, plasma, serum prolactin levels, hormones, prolactin, ovary.

Belterman, R., T. Dorresteyn and M. van Wees (Editors) (2005). Asian Elephant Studbook Elephas Maximus. Europe Regional EAZA - EEP, Rotterdam Zoo: Rotterdam, 76 p.
Descriptors: Asian elephant, Elephas maximus, Europe, studbook.

Brown, J.L., F. Goritz, N. Pratt Hawkes, R. Hermes, M. Galloway, L.H. Graham, C. Gray, S.L. Walker, A. Gomez, and R. Moreland (2004). Successful artificial insemination of an Asian elephant at the National Zoological Park. Zoo Biology 23(1): 45-63. ISSN: 0733-3188.
NAL Call Number: QL77.5.Z6
Descriptors: Asian elephant, successful artificial insemination, National Zoo, reproduction.

Brown, J.L., D. Olson, M. Keele, and E.W. Freeman (2004). Survey of the reproductive cyclicity status of Asian and African elephants in North America. Zoo Biology 23(4): 309-321. ISSN: 0733-3188.
NAL Call Number: QL77.5.Z6
Descriptors: reproductive cyclicity, status, survey, African elephant, Asian elephant, North America.

Brown, J.L., S.L. Walker, and T. Moeller (2004). Comparative endocrinology of cycling and non-cycling Asian (Elephas maximus) and African (Loxodonta africana) elephants. General and Comparative Endocrinology 136(3): 360-70.
NAL Call Number: 444.8 G28
Abstract: Up to 14% of Asian and 29% of African elephants in captivity are not cycling normally or exhibit irregular cycles based on progestin profiles. To determine if ovarian acyclicity is related to other disruptions in endocrine activity, serum pituitary, thyroid, adrenal, and ovarian hormones in weekly samples collected for 6-25 months were compared between normal cycling (n=22 each species) and non-cycling (n=6 Asian; n=30 African) elephants. A subset of cycling females (n=4 Asian, 7 African) also were blood sampled daily during the follicular phase to characterize the peri-ovulatory period. In normal cycling females, two leutinizing hormone (LH) surges were observed 3 weeks apart during a normal follicular phase, with the second inducing ovulation (ovLH). Serum FSH concentrations were highest at the beginning of the non-luteal phase, declining to nadir concentrations within 4 days of the ovLH surge. FSH remained low until after the ovLH surge and then increased during the luteal phase. A species difference was noted in prolactin secretion. In the African elephant, prolactin was increased during the follicular phase, but in Asian elephants concentrations remained stable throughout the cycle. Patterns of thyroid hormones (thyroid-stimulating hormone, TSH; free and total thyroxine, T4; free and total triiodothyronine, T3) and cortisol secretion were not affected by estrous cycle stage or season in cycling elephants. In non-cycling elephants, there were no fluctuating patterns of LH, FSH, or prolactin secretion. Overall mean concentrations of all hormones were similar to those in cycling animals, with the exception of FSH, prolactin, and estradiol. Mean serum FSH concentrations were lower due to females not exhibiting normal cyclic increases, whereas serum estradiol was higher overall in most acyclic females. Prolactin concentrations were significantly increased in 11 of 30 non-cycling females, all of which were African elephants. In sum, while there were no consistent endocrine anomalies associated with ovarian acyclicity, hyperprolactinemia may be one cause of ovarian dysfunction. The finding of elevated estrogens in some acyclic females also deserves further investigation, especially determining how it relates to reproductive tract pathologies.
Descriptors: physiology, estrous cycle physiology, ovary physiology, blood, estradiol blood, estrous cycle blood, follicle stimulating hormone blood, hydrocortisone blood, luteinizing hormone blood, prolactin blood, seasons, species specificity, thyrotropin blood, thyroxine blood, triiodothyronine blood.

Czekala, N.M., E.A. MacDonald, K. Steinman, S. Walker, N.W.I. Garrigues, D. Olson, and J.L. Brown (2003). Estrogen and LH dynamics during the follicular phase of the estrous cycle in the Asian elephant. Zoo Biology 22(5): 443-454. ISSN: 0733-3188.
NAL Call Number: QL77.5.Z6
Descriptors: Asian elephant, estrogen, LH, dynamics, follicular phase, estrus cycle, reproduction.

Dahl, N.J., D. Olson, D.L. Schmitt, D.R. Blasko, R.S. Kristipati, and J.F. Roser (2004). Development of an enzyme-linked immunosorbent assay (ELISA) for luteinizing hormone (LH) in the elephant (Loxodonta africana and Elephas maximus). Zoo Biology 23(1): 65-78. ISSN: 0733-3188.
NAL Call Number: QL77.5.Z6
Descriptors: African elephant, Asian elephant, ELISA, luteinizing hormone, enzyme-linked immunosorobent assay, LH, development.

Dehnhard, M., J.M. Hatt, K. Eulenberger, A. Ochs, and G. Strauss (2003). Headspace solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) for the determination of 5alpha-androst-2-en-17-one and -17beta-ol in the female Asian elephant: application for reproductive monitoring and prediction of parturition. Journal of Steroid Biochemistry and Molecular Biology 84(2-3): 383-91.
NAL Call Number: QD426.A1J6
Abstract: Asian elephants are not self-sustaining in captivity. The main reasons for this phenomenon are a low birth rate, an aging population, and poor calf-rearing. Therefore, it is essential that reproductive rates had to be improved and there is need for rapid quantitative measures to monitor reproductive functions focussing on estrous detection and the prediction of the period of parturition. The objective of this study was to develop a method which combines headspace solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) for analyses of 5alpha-androst-2-en-17beta-ol and -17-one to prognose estrous and to predict the period of parturition. SPME was carried out with a CTC Combi Pal system.The course of the luteal phase-specific substance 5alpha-androst-2-en-17beta-ol and -17-one followed a cyclic pattern in which the follicular and luteal phases could be clearly distinguished (mean estrous cycle length, 15+/-1.4 weeks). Based on daily urine samples, estrous prognosis might be possibly based on the initial 5alpha-androst-2-en-17beta-o1 increase at the end of the follicular phase. Parturition prognosis was performed in three elephant cows based on the 5alpha-androst-2-en-17beta-o1 drop to baseline levels 5-4 days prior parturition. Experiments revealed that 5alpha-androst-3alpha-ol-17-one and probably 5alpha-androst-3alpha-ol-17beta-ol are generated from sulfate conjugates by a thermal process.
Descriptors: androstane 3,17 diol blood, androsterone blood, chemistry, clinical methods, mass fragmentography methods, parturition blood, chromatography, gas, estrous cycle, pregnancy, animal blood, temperature, time factors.

Freeman, E.W., E. Weiss, and J.L. Brown (2004). Examination of the interrelationships of behavior, dominance status, and ovarian activity in captive Asian and African elephants. Zoo Biology 23(5): 431-448. ISSN: 0733-3188.
NAL Call Number: QL77.5.Z6
Descriptors: ovarian activity, Asian elephants, African elephants, dominance status, behavior, interrelationships.

Graham, L.H., J. Bando, C. Gray, and M.M. Buhr (2004). Liquid storage of Asian elephant (Elephas maximus) sperm at 4 degrees C. Animal Reproduction Science 80(3-4): 329-40.
NAL Call Number: QP251.A5
Abstract: The Asian elephant (Elephas maximus) population in the wild has been in decline for several decades and breeding in captivity has not been self-sustaining. The use of artificial insemination (AI) can help overcome many of the difficulties associated with breeding elephants in captivity; however, the ability to store semen for extended periods of time is critical to the successful application of AI to elephants. The objective of the present study was to assess the effects of four different semen extenders and the presence of egg yolk on the viability and motility of Asian elephant semen stored at 4 degrees C. High quality ejaculates (n=4) were collected from two Asian elephant bulls by rectal massage. Aliquots of each ejaculate were extended in four different diluents (Beltsville thawing solution (BTS); Tris-citric acid (TCA)/fructose-based; Beltsville F5 (BF5); dextrose-supplemented phosphate-buffered saline (PBS)) with or without egg yolk then cooled and stored at 4 degrees C. The percentages of viable (viability) and motile (motility) sperm were evaluated at 8, 24 and 48 h following collection. The addition of egg yolk significantly reduced the percentage loss in viability from initial collection to 48 h compared to extenders without egg yolk (17.0 +/- 8.2 versus 32.6 +/- 8.9 decline in percent viable sperm in the population, respectively; P<0.05). Extender and egg yolk affected (P<0.005) total motility and percent progressively motile sperm at all evaluation times during incubation. TCA + egg yolk maintained higher (P<0.05) levels of progressive motility compared to other extenders supplemented with egg yolk. These results indicate that Asian elephant semen extended in TCA diluent supplemented with egg yolk can maintain at least 50% viability and motility when stored at 4 degrees C for 48 h.
Descriptors: semen preservation, spermatozoa physiology, breeding, buffers, cell survival, cold, egg yolk, ejaculation, artificial insemination, semen preservation methods, sperm motility, tissue and organ harvesting methods.

Graham, L.H., C. Gray, and M.M. Buhr (2004). Influence of cryoprotectant type and concentration on post-thaw characteristics of cryopreserved Asian elephant (Elephas maximus) semen. Biology of Reproduction (Special Issue): 236. ISSN: 0006-3363.
NAL Call Number: QL876.B5
Descriptors: semen, cryopreservation, Asian elephant, post thaw, characteristics, cyroprotectant, influence.

Hama, N., A. Yamada, A. Noda, K. Murata, Y. Shimada, M. Ashida, K.M.Y. Ishikawa, and K. Okuno (2003). Serum hormonal changes in a female Asian elephant (Elephas maximus) with stillbirth. Japanese Journal of Zoo and Wildlife Medicine 8(2): 109-113. ISSN: 1342-6133.
Descriptors: Asian elephant, female, hormonal changes, seasonal, stillbirth, serum, progesterone, estradiol, prolactin, radioimmunoassay, RIA.
Language of Text: Japanese, with English summary.

Kirtland, J. (2003). Dreams and false promises: Asian elephant reproduction in North America. American Zoo and Aquarium Association Annual Conference Proceedings 2003: 49-54.
NAL Call Number: QL76.5.U6A472
Descriptors: Asian elephant, Elephas maximus, breeding programs, North America, reproduction.

Lazar, J., L.E. Rasmussen, D.R. Greenwood, I.S. Bang, and G.D. Prestwich (2004). Elephant albumin: a multipurpose pheromone shuttle. Chemistry and Biology 11(8): 1093-100.
Abstract: (Z)-7-dodecenyl acetate (Z7-12:Ac) is present in the urine of female Asian elephants (Elephas maximus) approaching ovulation and functions as a female-to-male sex pheromone. Here we show that a significant fraction of the pheromone in the urine is bound to a protein, elephant serum albumin (ESA), and provide evidence for key physiological functions of urinary ESA. Our biochemical and behavioral experiments suggest a three-fold role of ESA in pheromone signaling: (1) transporting Z7-12:Ac from serum into urine; (2) extending the presence of the pheromone in the environment without hampering detection; and (3) targeting pheromone delivery to chemosensory organs through localized release of the ligand induced by a pH change. The exploitation of albumin in pheromone transport clearly distinguishes the elephant from other mammals studied, and complements the uniqueness of elephant anatomy, physiology, and behavior.
Descriptors: acetates metabolism, albumins metabolism, pheromones metabolism, acetates urine, albumins chemistry, albumins genetics, biological availability, biological transport, cloning, molecular, hydrogen ion concentration, molecular structure, pheromones urine, substrate specificity, time factors, urine chemistry.

Rasmussen, L., V. Krishnamurthy, and R. Sukumar (2005). Behavioural and chemical confirmation of the preovulatory pheromone, (Z) -7-dodecenyl acetate, in wild Asian elephants: its relationship to musth. Behaviour 142(3): 351-396. ISSN: 0005-7959.
NAL Call Number: 410 B393
Descriptors: Asian elephant, Elephas maximus, age, reproductive behavior, musth, mating strategies, female preovulatory hormone, pheromones, preovulatory urinary hormone, social behavior, India, preovulatory hormone identification, male chemosensory responses.

Rees, P.A. (2004). Some preliminary evidence of the social facilitation of mounting behavior in a juvenile bull Asian elephant (Elephas maximus). Journal of Applied Animal Welfare Science 7(1): 49-58.
NAL Call Number: HV4701.J68
Abstract: This study recorded sexual behavior within a captive herd of 8 Asian elephants for approximately 230 hr on 50 days over a period of 10 months. The study observed a single adult and a single juvenile bull mounting cows more than 160 times. When the juvenile bull was between 4 years, 2 months and 4 years, 8 months old, he exhibited mounting behavior only on days when adult mounting occurred. Adult mounting always occurred first. Beyond the age of 4 years, 8 months, the juvenile bull exhibited spontaneous mounting behavior in the absence of adult mounting. This suggests that mounting behavior may develop because of social facilitation. Determining the significance of the presence of sexually active adults in the normal development of sexual behavior in juveniles will require further studies. Encouraging the establishment of larger captive herds containing adults and calves of both sexes-if their presence is important-would improve the welfare of elephants in zoos and increase their potential conservation value.
Descriptors: sex behavior, social environment, zoo animals.

Sa Ardrit, M., J. Saikhun, N. Thongtip, M. Damyang, S. Mahasawangkul, T. Angkawanish, S. Jansittiwate, T. Faisaikarm, Y. Kitiyanant, K. Pavasuthipaisit, and A. Pinyopummin (2006). Ultrastructural alterations of frozen-thawed Asian elephant (Elephas maximus) spermatozoa. International Journal of Andrology 29(2): 346-52.
NAL Call Number: QP251.I55
Abstract: Intact plasma and acrosome membranes and functional mitochondria following cryopreservation are important attributes for the fertilizing ability of spermatozoa. In the present study, functional and ultrastructural changes of Asian elephant spermatozoa after cryopreservation either in TEST + glycerol or HEPT + dimethyl sulphoxide (DMSO) were evaluated by fluorescent techniques and electron microscopy. Sperm frozen in TEST + glycerol had higher proportion of sperm with intact plasma (49.1 +/- 9.2% vs. 30.9 +/- 3.9%) and acrosomal (53.7 +/- 4.9% vs. 35.8 +/- 6.1%) membranes, as well as active mitochondria (57.0 +/- 7.2% vs. 42.0 +/- 5.0%) than those cryopreserved in HEPT + DMSO. The results obtained from electron microscopy were similar to those obtained by fluorescence microscopy. The percentage of normal spermatozoa was higher when spermatozoa were frozen in TEST + glycerol than those frozen in HEPT + DMSO (31.8 +/- 5.6 vs. 28.5 +/- 6.4). The ultrastructural alterations revealed by transmission electron microscopy could be classified as (i) distension of plasma membrane, while the acrosome was swollen; (ii) disruption or loss of plasma membrane, while acrosome was swollen with distended outer acrosomal membrane; (iii) disruption or loss of plasma and outer acrosomal membrane with leakage of acrosome content; (iv) extensive vesiculation of plasma and outer acrosomal membrane and leakage of acrosome content; (v) a complete loss of both plasma membrane and outer acrosomal membrane; and (vi) swelling of mitochondria. These findings suggest that the freezing and thawing procedure caused structural damage to elephant spermatozoa, especially in the plasma membrane, acrosome and mitochondria. Fluorescence and electron microscopic evaluations are potentially a powerful tool in the analysis of elephant spermatozoa after freezing and thawing.
Descriptors: Asian elephant, spermatozoa, frozen, thawed, ultrastructural, alterations, cryopreservation, fertilizing ability.

Saragusty, J., T.B. Hildebrandt, Y. Natan, R. Hermes, S. Yavin, F. Goeritz, and A. Arav (2005). Effect of egg-phosphatidylcholine on the chilling sensitivity and lipid phase transition of Asian elephant (Elephas maximus) spermatozoa. Zoo Biology 24(3): 233-245. ISSN: 0733-3188.
NAL Call Number: QL77.5.Z6
Descriptors: semen extenders, cooling, liposomes artificial, cryopreservation, cryoprotectants, spermatazoa.

Slade, B.E., B. Schulte, and L.E.L. Rasmussen (2003). Oestrous state dynamics in chemical communication by captive female Asian elephants. Animal Behaviour 65(4): 813-819. ISSN: 0003-3472.
NAL Call Number: Film S-1802
Descriptors: reproductive status, urine, estrus, female, Asian elephants, chemical communication, social group dynamics, chemosensory responses.

Teng MingSheng, Yang XiaoLi, and Wu DengHu (2003). Characteristics of reproductive biology of Asian elephants. Chinese Journal of Zoology 38(6): 86-90. ISSN: 0250-3263.
NAL Call Number: QL1.T8
Descriptors: reproductive biology, characteristics, Asian elephant, mating, estrus, parturition, pregnancy, reproduction, Elephas maximus, fetuses, gestation.
Language of Text: Chinese, with English summary.

Thongtip, N., M. Damyang, S. Mahasawangkul, A. Kongsila, T. Angkawanich, S. Jansittiwate, C. Thitaram, and P. Phongsopawijit. (2003). Frozen semen artificial insemination in Asian elephants (Elephas maximus) using endoscope and ultrasound guide . Proceedings of 41st Kasetsart University Annual Conference, Subject: Animals and Veterinary Medicine, Bangkok, Thailand: Kasetsart University, Bangkok, Thailand, p. 652-657.
Descriptors: Asian elephant, frozen semen, artificial insemination, endoscopy, cervix, thawed semen, ultrasonography, Elephas maximus.
Language of Text: Thai, with English summary.

Thongtip, N., J. Saikhun, M. Damyang, S. Mahasawangkul, P. Suthunmapinata, M. Yindee, A. Kongsila, T. Angkawanish, S. Jansittiwate, and W. Wongkalasin (2004). Evaluation of post-thaw Asian elephant (Elephas maximus) spermatozoa using flow cytometry: the effects of extender and cryoprotectant. Theriogenology 62(3-4): 748-760. ISSN: 0093-691X.
NAL Call Number: QP251.A1T5
Abstract: Although the development of semen cryopreservation in the African elephants (Loxodonta africana) has been accomplished, effective procedures for cryopreservation of Asian elephant (Elephas maximus) spermatozoa have not been established. In the present study, we investigate the freezing methods for conservation of Asian elephant spermatozoa under field conditions and identify the most suitable freezing protocols which provide acceptable post-thaw semen quality. Semen was collected from two Asian elephant bulls (EM1 and EM2, 10 ejaculates from each bull) by manual manipulation and were assessed for volume, pH, sperm cell concentration, and progressive motility. Eight out of 20 ejaculates were of acceptable quality (progressive motility greater than or equal to 60%), and were used for cryopreservation studies. Semen were frozen in TEST+glycerol, TEST+DMSO, HEPT+glycerol, or HEPT+DMSO. The post-thaw progressive sperm motilities were assessed, and sperm cells were stained with PI and FITC-PNA for membrane and acrosomal integrity assessment using flow cytometry. Post-thaw progressive motility of spermatozoa (EM1: 42.0½4.3%; EM2: 26.0½17.3%) and the percentage of membrane and acrosome intact spermatozoa (EM1: 55.5½8.1%; EM2: 46.3½6.4%) cryopreserved in TEST+glycerol were significantly higher than (P<0.05) those frozen in the other medium investigated choices for cryopreservation of Asian elephant spermatozoa. The data support the use of TEST+glycerol as an acceptable cryopreservation media of Asian elephant semen for the establishment of sperm banks.
Descriptors: Elephas maximus, spermatozoa, cryopreservation, semen extenders, cryoprotectants, flow cytometry, freezing, thawing, sperm motility, glycerol, dimethyl sulfoxide, plasma membrane, membrane permeability, acrosome reaction, male fertility, membrane integrity.

Vandebona, H., N.C.W. Goonesekere, W.D. Ratnasooriya, J. Alahakoon, and M.B. Gunasekera (2005). Using dna fingerprinting to establish paternity of Asian elephants, Elephas maximus, born at Pinnawela Elephant Orphanage, Sri Lanka. Annales Academiae Regiae Scientiarum Upsaliensis 39: 214-221. ISSN: 0504-0736.
Descriptors: Asian elephants, paternity identification, DNA fingerprinting, blood samples, genetic diversity, breeding programs, estrus, mating, Sri Lanka.