A captive adult female bottlenose dolphin presented with stillbirth. The placenta appeared oedematous. No other gross lesions were evident in the placenta or the stillborn calf. Histopathology revealed mild multifocal placentitis and foetal encephalitis. Brucella sp. was isolated from lung, liver, spleen and kidney. Sequence and phylogenetic analysis demonstrated this organism to be most similar to Brucella ceti sequence type (ST) 27. Brucella sp. DNA was detected in formalin-fixed paraffin-embedded placenta and brain by real-time PCR using primers targeting the IS711 gene. Immunohistochemical staining revealed Brucella sp. antigen in placental inflammation. This is the first report of isolation of Brucella sp. from a marine mammal in the Southern Hemisphere and the first report of marine Brucella-associated disease in Australia.

There are limited techniques available to assess the health of sea turtles as physical examination has little correlation to clinical findings, and blood reference intervals are broad and provide limited prognostic significance. Advances in the portability of ultrasound machines allow echocardiography to be increasingly used in the health assessments of wild animals. This study performed blood analysis and echocardiograms on 11 green sea turtles upon admission to a rehabilitation clinic and six animals before release. Significant differences were seen between groups, with admission animals having significantly smaller diameters of the cavum arteriosum at systole and diastole, smaller E-waves and an increased fractional shortening. Pre-release animals displayed significant increases in the maximum blood velocities of both the pulmonary artery and the left aorta. Significant negative correlations were seen between fractional shortening and uric acid and between the velocity time integral of the pulmonary artery and urea. The pulmonary artery velocity time integral was also significantly correlated to the E wave. Furthermore, there was asynchrony between the cavum arteriosum and the cavum pulmonale and the detection of a parasitic granuloma in the ventricular outflow tract of one animal. Overall, the results suggest that cardiac function in stranded green sea turtles is significantly impaired and that echocardiography has applications in the health assessments of green sea turtles.

This study investigated the influence of exercise and fasting state on haematologic and biochemical parameters in juvenile green turtles (Chelonia mydas). Animals were divided into two groups; one group was fasted for 72 h and one group was fed 1 h prior to exercise. Exercise was induced by repeated righting reflexes and blood values were measured prior to and post-exercise. Prior to exercise, fasted animals showed significantly decreased levels of urea, pH, PVCO2 and HCO3- and significant increases in Cl- and PVO2 compared to fed animals and fasted animals had significantly poorer exercise performance. Following exercise both fasted and fed animals had significant increases in Na+, K+, Cl-, PVCO2, PVO2, urea and lactate and significant decreases in pH and HCO3-. The magnitude of increase in lactate levels was significantly less in fasted animals. Prior to exercise, a significant correlation was calculated in fasted animals between pH and HCO3-. Following exercise, significant correlations were calculated in fed animals between pH and HCO3-, PVCO2 and lactate, and between pH and HCO3- in fasted animals. These results show that analytical method, fasting state and the physiologic changes induced during the intense exercise can affect haematologic and biochemical analytes and these factors should be considered when interpreting results from health assessment of wild animals.

Blubber and respiratory vapour ('blow') are now commonly used for endocrine studies on cetaceans, primarily because they can be obtained using minimally invasive methods. For many species, these samples have yet to be validated for these purposes. The objective of this study was to examine the performance of blow and blubber hormone monitoring, relative to serum hormone monitoring, for evaluating the reproductive and adrenal condition of captive bottlenose dolphins (Tursiops spp.). Eighteen bottlenose dolphins were sampled five times for serum and blow and twice for blubber throughout a one-year period. Concentrations of progesterone, testosterone, oestradiol and cortisol were measured in each sample type. Hormone levels were examined in relation to dolphin age, sex, reproductive status, season, time of sample collection (morning/afternoon) and collection type (in- or out-of-water sampling). Patterns in hormone levels were similar for serum and blubber. For instance, in both sample types, progesterone levels were significantly higher in pregnant (serum: 34.10 +/- 8.64 ng/mL; blubber: 13.01 +/- 0.72 ng/g) than in non-pregnant females (serum: 0.32 +/- 0.09 ng/mL; blubber: 1.17 +/- 0.10 ng/g). This pattern was not detected in blow, primarily because seawater contamination, nylon sampling materials and variable sample volumes influenced measured concentrations. In addition, the respiratory water content of a blow sample is known to affect measured hormone levels. Two methods were trialled to control for variability in sample volumes and dilution: (1) normalising blow hormone concentrations relative to urea nitrogen levels (a potential endogenous standard), and (2) measuring the relative proportions (i.e. ratios) of blow hormones. These correction measures had little influence on blow hormone results. Further refinement of blow hormone monitoring methods is required before they can be used for reproductive or adrenal assessments of bottlenose dolphins. Blubber, on the other hand, should be a suitable proxy for serum when attempting to classify pregnancy status and male maturity in these species.

The aim of this study was to describe two epizootics of high mortalities from infection with Streptococcus agalactiae, occurring in captive rays held in a marine display aquarium in south-east Queensland, Australia, in 2009 and 2010. Five different species of rays were affected, including mangrove whiprays (Himantura granulata), estuary rays (Dasyatis fluviorum), eastern shovelnose rays (Aptychotrema rostrata), white-spotted eagle rays (Aetobatus narinari) and blue-spotted mask rays (Neotrygon kuhlii). This report describes the history of both epizootics including collection, quarantine and husbandry of rays, the disease epizootics, clinico-pathological features of the disease, antimicrobial therapy, autogenous vaccine production, and laboratory studies including clinical and histopathology, bacteriology, PCR, molecular serotyping and sequencing of the bacterium S. agalactiae.

Sperm DNA fragmentation is one of the major causes of infertility; the sperm chromatin dispersion test (SCDt) evaluates this parameter and offers the advantage of species-specific validated protocol and ease of use under field conditions. The main purpose of this study was to evaluate sperm DNA fragmentation dynamics in both fresh and post-thaw bottlenose dolphin sperm using the SCDt following different cryopreservation protocols to gain new information about the post-thaw differential sperm DNA longevity in this species. Fresh and cryopreserved semen samples from five bottlenose dolphins were examined for sperm DNA fragmentation dynamics using the SCDt (Halomax(®)). Sperm DNA fragmentation was assessed immediately at collection and following cryopreservation (T0) and then after 0.5, 1, 4, 8, 24, 48 and 72 h incubation at 37°C. Serially collected ejaculates from four dolphins were frozen using different cryopreservation protocols in a TES-TRIS-fructose buffer (TTF), an egg-yolk-free vegetable lipid LP1 buffer (LP1) and human sperm preservation medium (HSPM). Fresh ejaculated spermatozoa initially showed low levels of DNA fragmentation for up to 48 h. Lower Sperm DNA fragmentation (SDF) was found in the second fresh ejaculate compared to the first when more than one sample was collected on the same day (p < 0.05); this difference was not apparent in any other seminal characteristic. While there was no difference observed in SDF between fresh and frozen-thawed sperm using the different cryopreservation protocols immediately after thawing (T0), frozen-thawed spermatozoa incubated at 37°C showed an increase in the rate of SDF after 24 h. Sperm frozen in the LP1(℗) buffer had higher levels (p < 0.05) of DNA fragmentation after 24- and 48-h incubation than those frozen in TTF or HSPM. No correlation was found between any seminal characteristic and DNA fragmentation in either fresh and/or frozen-thawed samples.

Over the last two decades, there have been significant advances in the use of assisted reproductive technology for genetic and reproductive management of captive dolphin populations, including evaluation of sperm DNA quality. This study validated a customized sperm chromatin dispersion test (SCDt) for the bottlenose dolphin (Tursiops truncatus) as a means of assessing sperm DNA damage both in the field and in the laboratory. After performing the SCDt, two different sperm morphotypes were identified: (i) sperm with fragmented DNA showed large haloes of dispersed DNA fragments emerging from a compact sperm nucleoid core and (ii) sperm containing non-fragmented DNA displayed small compact haloes surrounded by a dense core of non-dispersed DNA and protein complex. Estimates of sperm DNA fragmentation by means of SCDt were directly comparable to results obtained following a two-tailed comet assay and showed a significant degree of correlation (r = 0.961; p < 0.001). This investigation also revealed that the SCDt, with minor modifications to the standard protocol, can be successfully conducted in the field using a LED florescence microscopy obtaining a high correlation (r = 0.993; p = 0.01) between the data obtained in the laboratory and in the field.

There has been significant development over the last 20 years to improve genetic management of the captive bottlenose dolphin (Tursiops truncatus) by means of genome resource banking and assisted reproduction. Although standard semen parameters have been analysed in some detail, very little is known about sperm DNA fragmentation (SDF) in this species. The aim of this study was to develop a sperm chromatin dispersion test (SCDt) for the bottlenose dolphin to establish the baseline level of SDF immediately after ejaculation and cryopreservation and to determine the dynamic loss of sperm DNA quality after ex vivo handing and incubation in conditions that mimic the female reproductive tract. Semen from 8 bottlenose dolphins was collected by manual stimulation. Initial validation of the SCDt was conducted by means of in situ nick translation and neutral comet assay using a proven fertile male. To investigate the dynamic loss of sperm chromatin (rate of sDF loss), thawed sperm samples were incubated at 37.9°C for up to 48 h, and aliquots of spermatozoa were assessed after 1, 4, 8, 24, and 48 h. Dolphin sperm nuclei with fragmented DNA exhibited large halos of dispersed DNA fragments, whereas those with nonfragmented DNA displayed small halos of nondispersed DNA. A high correlation (r2 = 0.82; P ≤ 0.01) was found between the respective assessments of the SCDt and the neutral comet assay. All nucleoids resulting in a large halo of dispersed chromatin were intensely positive to in situ nick translation. The level of sDF fragmentation observed immediately after ejaculation in fresh and frozen samples was relatively low (1–5%). After comparing different ejaculates of the same individual, differences were found. Chromatin stability was high during the first 48 h of ejaculation or post-thawing and incubation. Evaluation of the sDF dynamics of fresh and frozen–thawed spermatozoa revealed no significant increase in the baseline level of sDF or in the relative increase of DNA damage after 48 h of incubation. Our data suggest that cryopreservation does not induce a dramatic increase in sperm chromatin damage. Interestingly, sperm samples derived from aged animals resulted in an increased rate of DNA loss, which was observed after 60 min post-incubation.

Ninety-three giant Queensland grouper, Epinephelus lanceolatus (Bloch), were found dead in Queensland, Australia, from 2007 to 2011. Most dead fish occurred in northern Queensland, with a peak of mortalities in Cairns in June 2008. In 2009, sick wild fish including giant sea catfish, Arius thalassinus (Ruppell), and javelin grunter, Pomadasys kaakan (Cuvier), also occurred in Cairns. In 2009 and 2010, two disease epizootics involving wild stingrays occurred at Sea World marine aquarium. Necropsy, histopathology, bacteriology and PCR determined that the cause of deaths of 12 giant Queensland grouper, three wild fish, six estuary rays, Dasyatis fluviorum (Ogilby), one mangrove whipray, Himantura granulata (Macleay), and one eastern shovelnose ray, Aptychotrema rostrata (Shaw), was Streptococcus agalactiae septicaemia. Biochemical testing of 34 S.agalactiae isolates from giant Queensland grouper, wild fish and stingrays showed all had identical biochemical profiles. The 16S rRNA gene sequences of isolates confirmed all isolates were S.agalactiae; genotyping of selected S.agalactiae isolates showed the isolates from giant Queensland grouper were serotype Ib, whereas isolates from wild fish and stingrays closely resembled serotype II. This is the first report of S.agalactiae from wild giant Queensland grouper and other wild tropical fish and stingray species in Queensland, Australia.

Despite its vulnerable conservation status there is scant information on male Greater Bilby Macrotis lagotis reproduction. Observations of gross anatomy revealed a testis to body weight ratio of 0.08 - 0.17% (n = 4), the presence of a carrot shaped prostate with an oblique coronal segmentation of ventral and dorsal orientated prostatic tissue, an elongated membranous urethra, two bulbourethral glands and a bifurcated urethra in the glans penis (n = I).The testis contained a high proportion of Leydig tissue (37.5 ± 2.7%) and a seminiferous epithelial cell cycle (8 stages identified) with a predominance of pre-meiotic stages (61.4%) and Sertoli cells with unusually large nuclei. A GnRH stimulation test conducted on four different intact sexually mature Bilbies using 2μg Buserelin resulted in maximal plasma androgen secretion 30 to 60 mins after injection. While steps of M. lagotis spermiogenesis were similar to those described in peramelid marsupials, the morphology of the Bilby spermatozoan at spermiation,was radically different to that of the bandicoot sperm cell. Similar to the bandicoot, M. lagotis spermatozoa in the corpus epididymidis dislocated their neck insertion from the primary implantation fossa, so that by the time the sperm cell had reached the cauda epididymidis, the acetabulum had migrated cranially Into a secondary implantation fossa and the nucleus had become streamlined with respect to the flagellum.This study reports the first description of large crystalloid inclusions in the principal cells of the caput epididymidal epithelium, the significance of which remains unknown. Male Bilby reproduction reported in this study supports the present taxonomic position of the Thylacomyidae.

The aim of this study was to determine the relative cryopreservation success of koala and wombat spermatozoa and to investigate reasons for their respective post-thaw survival by examining the sperm’s response to a range of osmotic media and determining the presence and distribution of F-actin. An hypothesis was proposed that F-actin may be imparting a degree of structural inflexibility to the koala sperm plasma membrane; hence, exposure of spermatozoa to cytochalasin D (5μM), a F-actin depolymerisation agent, should result in increased plasticisation of the membrane and greater tolerance of cell volume changes that typically occur during cryopreservation. In experiment 1, koala (n=4) and wombat (n=4) spermatozoa packaged in 0.25mL straws were cryopreserved using two freezing rates (fast—3cm above liquid N2 interface; slow—6°C/min in a freezing chamber) and two glycerol concentrations (8 and 14% v/v) in a tris–citrate glucose buffer with 15% (v/v) egg yolk. Wombat spermatozoa showed better (P<0.01) post-thaw survival (% motile, % intact plasma membranes, % decondensed sperm heads) than koala spermatozoa. When exposed to media of varying osmolality, koala spermatozoa were less tolerant (% intact plasma membrane) of hyper-osmotic conditions (920 and 1410mOsmol/kg) than wombat spermatozoa. F-actin was localised using a monoclonal antibody but only found in the wombat sperm head. When koala and wombat spermatozoa were exposed to media of varying osmolality, cytochalasin D had no beneficial effect on sperm survival (% intact plasma membranes). This study has demonstrated that wombat spermatozoa are highly tolerant of cryopreservation when compared to koala sperm but that spermatozoa from both species show greatest post-thaw survival when frozen slowly in 14% glycerol. Koala sperm are also particularly susceptible to hyper-osmotic environments but lack of detectable F-actin in the koala spermatozoan suggests that poor cryopreservation success in this species is unlikely to be associated with F-actin induced plasma membrane inflexibility.

Seven captive male African wild dogs (Lycaon pictus) weighing 25-32 kg each, were anesthetized by i.m. injection via hand syringe with a combination of 1.5 mg/kg ketamine, 40 mu g/kg medetomidine, and 0.05 mg/kg atropine. Following endotracheal intubation, each animal was connected to a bain closed-circuit system that delivered 1.5% isoflurane and 2 L/min oxygen. Atipamezole (0.1 mg/kg i.v.; 0.1 mg/kg i.m.) was given at the end of each procedure (60 min following injection of medetomidine/ketamine/atropine). Time to sternal recumbency was 5-8 min. Times to standing after atipamezole administration were 8-20 min. This anesthetic regimen was repeated on three separate occasions (September 2000, February 2002, and October 2002) on all males to perform electroejaculation procedures. Each procedure was < 80 min from injection to standing. Dogs showed excellent muscle relaxation during the procedures. Arterial blood samples were collected at 10-min intervals for blood gases in one procedure (September 2000). Separate venous samples were taken from each dog during each procedure for hematology and biochemistry. These values were within the normal range for this species. Arterial hemoglobin oxygen saturation (SpO2) and heart rate (HR) were monitored continuously in addition to other anesthesia monitoring procedures (body temperature, respiratory rate [RR], capillary refill time, blink response, pupil position, deep pain perception reflex). All dogs maintained relatively stable SpO2 profiles during monitoring, with a mean (+/- SD) SpO2 of 92% +/- 5.4%. All other physiological variables (HR, RR, body temperature, blood pressure) were within normal limits. Following each procedure, normal behavior was noted in all dogs. All the dogs were reunited into the pack at completion of their anesthetic procedures. An injectable medetomidine-ketamine-atropine combination with maintenance by gaseous isoflurane and oxygen provides an inexpensive, reliable anesthetic for captive African wild dogs.

The aim of the present study was to compare cryopreservation, osmotic tolerance and glycerol toxicity between mature and immature epididymal kangaroo spermatozoa to investigate whether the lack of cryopreservation success of cauda epididymidal spermatozoa may be related to the increased complexity of the sperm ultrastructure acquired during epididymal transit. Caput and cauda epididymidal spermatozoa were recovered from red-necked wallabies (RNW; Macropus rufogriseus) and eastern grey kangaroos (EGK; M. giganteus). In Experiment 1, caput and cauda epididymidal spermatozoa were frozen and thawed using a standard cryopreservation procedure in Tris-citrate buffer with or without 20% glycerol. Although cryopreservation of caput epididymidal spermatozoa resulted in a significant increase in sperm plasma membrane damage, they were more tolerant of the procedure than spermatozoa recovered from the cauda epididymidis (P < 0.05). In Experiment 2, caput and cauda epididymidal EGK spermatozoa were diluted into phosphate-buffered saline media of varying osmolarity and their osmotic tolerance determined. Plasma membranes of caput epididymidal spermatozoa were more tolerant of hypo-osmotic media than were cauda epididymidal spermatozoa (P < 0.05). In Experiment 3, caput and cauda epididymidal RNW spermatozoa were incubated in Tris-citrate buffer with and without 20% glycerol at 35 and 4 degrees C to examine the cytotoxic effects of glycerol. At both temperatures, caput epididymidal spermatozoa showed less plasma membrane damage compared with cauda epididymidal spermatozoa when exposed to 20% glycerol (P < 0.05). These experiments clearly indicate that epididymal maturation of kangaroo spermatozoa results in a decreased ability to withstand the physiological stresses associated with cryopreservation.

This study examined the hypothesis that filamentous actin associated with the complex cytoskeleton of the kangaroo sperm head and tail may be contributing to lack of plasma membrane plasticity and a consequent loss of membrane integrity during cryopreservation. In the first study, the distribution of G and F actin within Eastern Grey Kangaroo (EGK, Macropus giganteus) cauda epididymidal spermatozoa was successfully detected using DNAse-FITC and a monoclonal F-actin antibody (ab205, Abcam), respectively. G-actin staining was most intense in the acrosome but was also observed with less intensity over the nucleus and mid-piece. F-actin was located in the sperm nucleus but was not discernable in the acrosome or sperm tail. To investigate whether cytochalasin D (a known F-actin depolymerising agent) was capable of improving the osmotic tolerance of EGK cauda epididymal spermatozoa, sperm were incubated in hypo-osmotic media (61 and 104 mOsm) containing a range of cytochalasin D concentrations (0-200 microM). Cytochalasin D had no beneficial effect on plasma membrane integrity of sperm incubated in hypo-osmotic media. However, when EGK cauda epididymidal sperm were incubated in isosmotic media, there was a progressive loss of sperm motility with increasing cytochalasin D concentration. The results of this study indicated that the F-actin distribution in cauda epididymidal spermatozoa of the EGK was surprisingly different from that of the Tammar Wallaby (M. eugenii) and that cytochalasin-D does not appear to improve the tolerance of EGK cauda epididymidal sperm to osmotically induced injury.

Implementation of assisted breeding in the captive African wild dog is restricted by a current lack of knowledge on their reproductive physiology and the apparent difficulty of effectively manipulating the complex social dynamic of the pack in order to conduct reproductive procedures. In this study, we describe protocols for the safe and repeated capture and restraint of the African wild dog (n = 7) as well as techniques for assessment of male reproductive function, semen collection and preservation. In a serendipitous finding, captive African wild dogs appeared to display significant seasonal change in male reproduction. Testicular volume and tone, spermatorrhea and the ability to collect semen by electroejaculation all increased significantly during late summer and then subsequently declined by early spring. While there were no detectable seasonal changes in testosterone concentration in the population as whole, the alpha-dominant male in both years of the study, had a highly elevated testosterone concentration compared to subordinate males. Semen collection by electroejaculation during the late summer was most effective in peri-pubertal males (15 months) when all seven electroejaculates were of adequate quality for cryopreservation. In the second breeding season (27 months), there were numerous changes in the pack hierarchy and electroejaculation was not as successful (3/7). The characteristics of electroejaculated semen collected in the breeding season are described for seven animals including the first descriptions and incidence of sperm abnormalities in the species. Semen (n = 7) was frozen using a Tris–citrate fructose buffer and final egg yolk and glycerol concentration of 20% and 4%, respectively. Sperm were loaded into 0.25 mL straws, frozen in liquid nitrogen vapor and then thawed at 37 °C. Initial post-thaw survival of spermatozoa was encouraging (% motile: 31.8 ± 5.8%; rate: 2.8 ± 0.3; % intact plasma membranes: 33.4 ± 5.3% and the % of damaged acrosomes: 4.4 ± 1.5%) but following 2 h incubation at 37 °C, post-thaw survival declined markedly.

Despite their abundance in the wild, common wombats (Vombatus ursinus) do not breed regularly in captivity, such that there is little published about their captive reproductive management.As part of the development of an artificial insemination program in this species and in an attempt to establish important baseline reproductive information, 4 captive male common wombats at Western Plains Zoo (Dubbo, NSW) were systematically examined over a 12-month period in order to assess whether male reproductive function was seasonally dependent.The reproductive parameters investigated included, peripheral plasma testosterone secretion, testis volume and quality of semen (% motility, % live and % normal) collected by electro-ejaculation.An attempt was also made to determine the extent of seasonal change in male reproductive function of common wombats in wild populations.Plasma samples and reproductive tracts were collected from 12 wild male common wombats from the Kangaroo Valley district (NSW) in June (n = 7) and November (n = 5); plasma testosterone secretion, epididymidal sperm characteristics (% motile, % live and % normal) and measurements of testis, prostate and bulbourethral glands were compared.Our results indicated that male common wombats in captivity at Western Plains Zoo did not appear to be seasonal breeders in terms of testosterone secretion or electroejaculate quality (% motile, % live and % normal); there was, however, a significant increase in testis size between the months of June to October (F = 4.04; P = 0.05).Lack of distinct seasonal variation in male reproduction was also evident in wild common wombats in Kangaroo Valley.There was no significant difference (P > 0.05) between wild wombats collected in June or November with respect to plasma testosterone concentration, epididymidal sperm quality or testis, prostate and bulbourethral gland dimensions.10.

The effects of exposure of red blood cells (RBC) of three species of marsupial to a mercury-containing sulfhydryl-modifying reagent, p-chloromercuribenzene sulfonate (PCMBS), on the water diffusional permeability (P d ) of their membranes were monitored by using an Mn2+-doping 1H nuclear magnetic resonance (NMR) technique at 400 MHz. For koala (Phascolarctos cinereus), RBC the maximal inhibition was reached at 37 °C in 60 min with 1 mmol.l–1 PCMBS or in 15–30 min with 2 mmol. l–1 PCMBS. In contrast, in the case of red kangaroo (Macropus rufus) or swamp wallaby (Wallabia bicolor) RBC, maximal inhibition required an incubation of 90 min at 37 °C with 2 mmol.l–1 PCMBS. For the RBC of all three species the value of maximal inhibition was very high, being 50–70% when measured at 25 °C, 60–80% at 30 °C and 60–70% at 37 °C. The lowest values of P d appeared to be around 2×10–3–3×10–3 cm.s–1 in the temperature range of 25–37 °C. The mean value of the activation energy of water diffusion (E a,d ) was ~20–25 kJ.mol–1 for control and ~40 kJ.mol–1 for PCMBS-inhibited RBCs. These results show that marsupial RBC have a basal permeability to water similar to that previously reported for human RBC, but a higher value of the PCMBS-inhibitable water permeability. This indicates that the higher water permeability of marsupial RBC compared with human RBC is associated with a higher fraction of protein-mediated water permeability.

Australian Veterinary JournalVolume 80, Issue 1-2 p. 94-95 Birth of a Banteng (Bos javanicus) calf at Western Plains Zoo after fixed time artificial insemination SD JOHNSTON, SD JOHNSTON School of Animal Studies, The University of Queensland, Gatton, Queensland 4343Search for more papers by this authorMR MCGOWAN, MR MCGOWAN Royal Veterinary College, Hertfordshire, AL9 7TA, United Kingdom.Search for more papers by this authorD BLYDE, D BLYDE Western Plains Zoo, Dubbo, New South Wales 2830Search for more papers by this author SD JOHNSTON, SD JOHNSTON School of Animal Studies, The University of Queensland, Gatton, Queensland 4343Search for more papers by this authorMR MCGOWAN, MR MCGOWAN Royal Veterinary College, Hertfordshire, AL9 7TA, United Kingdom.Search for more papers by this authorD BLYDE, D BLYDE Western Plains Zoo, Dubbo, New South Wales 2830Search for more papers by this author First published: 09 October 2008 https://doi.org/10.1111/j.1751-0813.2002.tb12061.xCitations: 8AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat No abstract is available for this article. References 1 Solti L, Crichton EG, Loskutoff NM, Cseh S. Economical and ecological importance of indigenous livestock and the application of assisted reproduction to their preservation. Theriogenology 2000; 53: 149–162. 2 Asa CS, Read B, Houston EW, et al. Serum oestradiol and progesterone concentrations during the ovulatory cycle and pregnancy in banteng cattle (Bos javanicus). Theriogenology 1993; 39: 1367–1376. 3 Asa CS, Jenness B. Cryopreservation of semen from banteng (Bos javan-icus). Proceedings of the Wild Cattle Symposium, Omaha, Nebraska, 1991; 17–26. 4 Asa CS. Synchronization of ovulation in banteng (Bos javanicus) with PGF2alpha. Proceedings of the Society of Theriogenology , 1991; 351–353. 5 Barnes FL, Balke JME, Eyestone WH, First NL, Read BR. In vitro fertilisation of banteng (Bos javanicus): A case report. Theriogenology 1988; 29: 215. 6 Barth AD. Evaluation of frozen bovine semen by the veterinary practitioner. Proceedings of Bovine Short Course. American College of Theriogenologists and Society for Theriogenology, Hastings, New England, 1995; 105–110. 7 Hafez ESE. Preservation and cryopreservation of gametes and embryos.In: ESE Hafez, B Hafez Reproduction in Farm Animals. 7th edn.Lippincott Williams and Wilkins, Sydney , 2000: 431–442. Citing Literature Volume80, Issue1-2January 2002Pages 94-95 ReferencesRelatedInformation