Research

Clauss, M., Y. Wang, K. Ghebremeskel, C.E. Lendl, and W.J. Streich (2003). Plasma and erythrocyte fatty acids in captive Asian (Elephas maximus) and African (Loxodonta africana) elephants. Veterinary Record 153(2): 54-8.
NAL Call Number: 41.8 V641
Abstract: The fatty acid components of the plasma triglycerides and the phospholipid fractions of the red blood cells of a captive group of two African (Loxodonta africana) and four Asian (Elephas maximus) elephants were investigated. All the animals received the same diet of hay, fruits and vegetables, and concentrates. A comparison with data from free-ranging African elephants or Asian work-camp elephants showed that the captive elephants had lower proportions of polyunsaturated fatty acids (PUFAs), and for several lipid fractions a higher n-6:n-3 ratio, than their counterparts in the wild or under the more natural, in terms of diet, work-camp conditions. The difference in PUFA content was smaller in the African than in the Asian elephants. The captive Asian elephants tended to have lower levels of n-3 and total unsaturated fatty acids in their red blood cells than the captive African elephants.
Descriptors: metabolism, erythrocytes metabolism, unsaturated metabolism fatty acids, phospholipids metabolism, triglycerides metabolism, zoo animals, blood chemical analysis, diet, blood, unsaturated blood fatty acids, triglycerides blood, workload.

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.

Fickel, J., D. Lieckfeldt, L.K. Richman, W.J. Streich, T.B. Hildebrandt, and C. Pitra (2003). Comparison of glycoprotein B (gB) variants of the elephant endotheliotropic herpesvirus (EEHV) isolated from Asian elephants (Elephas maximus). Veterinary Microbiology 91(1): 11-21. ISSN: 0378-1135.
NAL Call Number: SF601.V44
Abstract: The recently described elephant endotheliotropic herpesviruses (EEHV) have been associated with the deaths of numerous captive elephants. A proposed tool for the detection of EEHV infection in elephants is the PCR-based screening for EEHV-DNA in whole blood samples. Unfortunately, this detection method has only been successful in post-mortem analyses or in animals already displaying clinical signs of EEHV disease, thus rendering this method unsuitable for identification of carrier elephants. Here, we focus on glycoprotein B (gB) for serologic assay development, since gB is an envelope protein known to induce a neutralising antibody response in other herpesvirus infections. We sequenced the entire gB gene from five Asian elephants with EEHV, representing four different gB variants. Computer-aided methods were used to predict functionally important regions within EEHVgB. An extra-cytoplasmic region of 153 amino acids was predicted to be under positive selection and may potentially contain antigenic determinants that will be useful for future serologic assay development.
Descriptors: Elephas maximus, viral proteins, glycoproteins, disease transmission, detection, polymerase chain reaction, cytoplasm, amino acid sequences, molecular sequence data.

Greenwood, D.R., D. Comeskey, M.B. Hunt, and L.E. Rasmussen (2005). Chemical communication: chirality in elephant pheromones. Nature 438(7071): 1097-8.
NAL Call Number: 472 N21
Abstract: Musth in male elephants is an annual period of heightened sexual activity and aggression that is linked to physical, sexual and social maturation. It is mediated by the release of chemical signals such as the pheromone frontalin, which exists in two chiral forms (molecular mirror images, or enantiomers). Here we show that enantiomers of frontalin are released by Asian elephants in a specific ratio that depends on the animal's age and stage of musth, and that different responses are elicited in male and female conspecifics when the ratio alters. This precise control of communication by molecular chirality offers insight into societal interactions in elephants, and may be useful in implementing new conservation protocols.
Descriptors: heterocyclic chemistry of bicyclo compounds, heterocyclic pharmacology of bicyclo compounds, physiology, pheromones chemistry, pheromones secretion, drug effects on animal sex behavior, aging physiology, heterocyclic metabolism of bicyclo compounds, pheromones pharmacology, sex behavior, animal physiology, stereoisomerism.

Hildebrandt, T.B., R. Hermes, P. Ratanakorn, W. Rietschel, J. Fickel, R. Frey, G. Wibbelt, C. Reid, and F. Goritz (2005). Ultrasonographic assessment and ultrasound-guided biopsy of the retropharyngeal lymph nodes in Asian elephants (Elephas maximus ). Veterinary Record 157(18): 544-8.
NAL Call Number: 41.8 V641
Abstract: Endotheliotropic herpesvirus causes a fatal disease in young Asian elephants, but there are no methods for identifying latent carriers of the virus. During the postmortem study of one female African elephant and three male and two female Asian elephants, a lymph node located bilaterally caudoventral to the parotid gland, approximately 1.5 to 5 cm below the skin, was identified as suitable for transcutaneous ultrasound-guided biopsy. An ultrasonographic assessment and two biopsies were performed on 39 Asian elephants, and these lymph nodes were classified ultrasonographically as active, inactive or chronically active. The calculated mean (se) volume of 10 active lymph nodes was 17.4 (6.9) cm(3), and that of three chronically active lymph nodes was 10.6 (1.0) cm(3), whereas the mean volume of 17 inactive lymph nodes was 3.1 (0.6) cm(3). The presence of lymph node tissue in samples obtained by ultrasound-guided biopsy from three animals that were maintained under conditions that allowed for additional sampling was confirmed histologically. The dna extracted from the lymphoid tissue and the whole blood of all the elephants was negative for endotheliotropic herpesvirus by PCR.
Descriptors: DNA viral isolation and purification, herpesviridae isolation and purification, herpesviridae infections, lymph nodes pathology, zoo animals, fine needle biopsy methods, disease reservoirs, disease reservoirs virology, Herpesviridae pathogenicity, Herpesviridae infections epidemiology, Herpesviridae infections pathology, lymph nodes ultrasonography, lymph nodes virology, polymerase chain reaction, prevalence, virus latency.

Lamps, L.W., B.R. Smoller, T.E. Goodwin, and L.E.L. Rasmussen (2004). Hormone receptor expression in interdigital glands of the Asian elephant (Elephas maximus). Zoo Biology 23(5): 463-469. ISSN: 0733-3188.
NAL Call Number: QL77.5.Z6
Descriptors: Asian elephant, interdigital glands, hormone receptor, expression, Elephas maximus.

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.E. and D.R. Greenwood (2003). Frontalin: a chemical message of musth in Asian elephants (Elephas maximus). Chemical Senses 28(5): 433-46.
NAL Call Number: QP456.C5
Abstract: Musth is an important male phenomenon affecting many aspects of elephant society including reproduction. During musth, the temporal gland secretions (as well as the urine and breath) of adult male Asian elephants (Elephas maximus) discharge a variety of malodorous compounds together with the bicyclic ketal, frontalin. In contrast, teenage male elephants in musth release a sweet-smelling exudate from their facial temporal gland. We recently demonstrated that the concentration of frontalin becomes increasingly evident as male elephants mature. In the present study, we demonstrate that behaviors exhibited towards frontalin are consistent and dependent on the sex, developmental stage and physiological status of the responding conspecific individual. To examine whether frontalin functions as a chemical signal, perhaps even a pheromone, we bioassayed older and younger adult males, and luteal- and follicular-phase and pregnant females for their chemosensory and behavioral responses to frontalin. Adult males were mostly indifferent to frontalin, whereas subadult males were highly reactive, often exhibiting repulsion or avoidance. Female chemosensory responses to frontalin varied with hormonal state. Females in the luteal phase demonstrated low frequencies of responses, whereas pregnant females responded significantly more frequently, with varied types of responses including those to the palatal pits. Females in the follicular phase were the most responsive and often demonstrated mating-related behaviors subsequent to high chemosensory responses to frontalin. Our evidence strongly suggests that frontalin, a well-studied pheromone in insects, also functions as a pheromone in the Asian elephant: it exhibits all of the determinants that define a pheromone and evidently conveys some of the messages underlying the phenomenon of musth.
Descriptors: bicyclo compounds, heterocyclic pharmacology, physiology, aging physiology, Asia, drug effects on behavior, heterocyclic urine, buffers, urine, pheromones pharmacology, pheromones in urine, drug effects on sex behavior, sex characteristics, smell.

Rezaian, M. and S. Yamashiro (2005). Comparison between elephant and bovine platelet ultra structure. Indian Journal of Animal Sciences 75(3): 267-270. ISSN: 0367-8318.
NAL Call Number: 41.8 IN22
Descriptors: Asian elephant, bovine platelet, ultra structure, elephant platelets, comparison, discoid shape.

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.

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.

Vidya, T. and R. Sukumar (2005). Amplification success and feasibility of using microsatellite loci amplified from dung to population genetic studies of the Asian elephant (Elephas maximus). Current Science (Bangalore) 88(3): 489-492. ISSN: 0011-3891.
NAL Call Number: 475 SCI23
Descriptors: Asian elephant, Elephas maximus, population genetics, nucleic acids, feces, molecular genetics, microsatellite DNA, India, fecal microsatellite DNA samples, dung samples, population genetics analysis, evaluation.