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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. [ABSTRACT FROM AUTHOR]

Contents 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. [ABSTRACT FROM AUTHOR]

Contents 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. [ABSTRACT FROM AUTHOR]

Highlights • Steroid hormones are quantifiable in bottlenose dolphin blow and blubber. • Pregnant females had higher blubber progesterone levels than other adult females. • Mature males had higher blubber testosterone levels than juvenile males. • Blow hormone measurements negatively influenced by seawater and sampling materials. Abstract 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. [ABSTRACT FROM AUTHOR]

An abstract of the research paper "Assessment of Fresh and Frozen-Thawed Bottlenose Dolphin (Tursiops Truncatus) Sperm DNA Fragmentation Using the Sperm Chromatin Dispersion Test," by M. J. Sanchez-Calabuig et al. is presented.