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Concept: Indian Rhinoceros

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Reproductive tract tumours, specifically leiomyoma, are commonly found in female rhinoceroses. Similar to humans, tumour growth in rhinoceroses is thought to be sex hormone dependent. Tumours can form and expand from the onset of ovarian activity at puberty until the cessation of sex-steroid influences at senescence. Extensive tumour growth results in infertility. The aim of this study was to down regulate reproductive function of tumour-diseased and infertile females to stop further tumour growth using a Gonadotropin releasing factor (GnRF) vaccine. Four infertile southern white (Ceratotherium simum simum) and three Greater one-horned rhinoceroses (rhinoceros unicornis) with active ovaries and 2.7 ± 0.9 and 14.0 ± 1.5 reproductive tract tumours respectively were vaccinated against GnRF (Improvac®, Zoetis, Germany) at 0, 4 and 16 weeks and re-boostered every 6-8 months thereafter. After GnRF vaccination ovarian and luteal activity was suppressed in all treated females. Three months after vaccination the size of the ovaries, the number of follicles and the size of the largest follicle were significantly reduced (P<0.03). Reproductive tract tumours decreased significantly in diameter (Greater-one horned rhino: P<0.0001; white rhino: P<0.01), presumably as a result of reduced sex-steroid influence. The calculated tumour volumes were reduced by 50.8 ± 10.9% in Greater one-horned and 48.6 ± 12.9% in white rhinoceroses. In conclusion, GnRF vaccine effectively down regulated reproductive function and decreased the size of reproductive tract tumours in female rhinoceros. Our work is the first to use down regulation of reproductive function as a symptomatic treatment against benign reproductive tumour disease in a wildlife species. Nonetheless, full reversibility and rhinoceros fertility following GnRF vaccination warrants further evaluation.

Concepts: Reproductive system, Rhinoceros, White Rhinoceros, Black Rhinoceros, International Rhino Foundation, Rhinoceroses, Northern White Rhinoceros, Indian Rhinoceros

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The parathyroid gland was first identified in the Indian rhinoceros in 1849 by Sir Richard Owen. We performed a necropsy in an Indian rhinoceros, recapitulating Owen’s dissection and display what appear to be the initial identification of the recurrent laryngeal nerve in situ and the anatomy and histology of the largest rhinoceros parathyroid glands yet identified.

Concepts: Calcitonin, Parathyroid hormone, Thyroid, Parathyroid gland, Glands, Rhinoceros, Indian Rhinoceros

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A novel strain, YIM 100770(T), was isolated from Rhinoceros unicornis faeces collected from Yunnan Wild Animal Park, China. The taxonomic status was determined based on the physiological, biochemical and phylogenetic characteristics. Strain YIM 100770(T) was observed to be rod-shaped, non-motile, Gram-stain negative and aerobic. The G+C content of the genomic DNA was determined to be 68.5 mol%. The cells of strain YIM 100770(T) contain ubiquinone Q-10 as the respiratory quinone. The major fatty acids (>1%) were identified as Summed feature 8 (C18:1 ω7c and/or C18:1 ω6c; 78.1%), Summed feature 4 (iso-C17:1-I and/or anteiso-C17:1-B; 12.9%), C19:0 cyclo ω8c (2.8%), C16:0 (2.2%) and C18:0 (2.2%). Comparison of 16S rRNA gene sequences revealed the strain show high similarities with the members of the genera Psychroglaciecola (94.5%), Methylobacterium (90.5-94.1%) and Microvirga (92.0-93.3%) in the family Methylobacteriaceae. In addition, the strain also showed high similarities with the members of the genera Chelatococcus (93.7-94.0%) and Pseudochelatococcus (93.1-93.7%) in the family Beijerinckiacea, and the genus Bosea (93.1-93.8%) in the family Bradyrhizobiaceae. The phylogenetic analysis, combined with the chemical characteristics, suggest that the strain represents a novel genus in the order Rhizobiales of the class Alphaproteobacteria, for which the name Enterovirga rhinocerotis gen. nov., sp. nov. is proposed. The type strain of E. rhinocerotis is YIM 100770(T) (=DSM 25903(T) = CCTCC AB 2012048(T)).

Concepts: Fatty acid, Biology, Organism, Species, Ribosomal RNA, 16S ribosomal RNA, Rhizobiales, Indian Rhinoceros

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Seventy-eight soil samples were collected from the various locations in the vicinity of Kaziranga National Park (Assam), India, during April to October 2009 and screened for the presence of keratinophilic fungi using the hair baiting techniques for isolation. Thirty-nine isolates were recovered and identified by recognition of their macro- and micromorphological features. Their identification was also confirmed by the BLAST search of sequences of the ITS1-5.8S-ITS2 rDNA region against the NCBI/GenBank data and compared with deposited sequences for identification purpose. Eleven species related to seven genera were recorded viz. Aphanoascus durus (1.28%), Arthroderma tuberculatum (3.84%), Arthroderma corniculatum (1.28%), Chrysosporium indicum (16.66%), C. tropicum (3.84%), Ctenomyces serratus (5.12%), Keratinophyton punsolae (1.28%), Microsporum appendiculatum (1.28%), Microsporum gypseum complex (11.53%), Trichophyton mentagrophytes (11.28%) and T. terrestre (2.56%).

Concepts: Species, Assam, Microsporum, BLAST, Arthrodermataceae, Kaziranga National Park, Indian Rhinoceros, Tourism in Assam

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The objective of this study was to design an artificial insemination (AI) protocol using cryopreserved spermatozoa to obtain pregnancies in captive Indian rhinoceroses (Rhinoceros unicornis). Four methods developed varied by timing and approach, as follows; Method 1: females (n=2) were inseminated pre- and post-ovulation under general anesthesia, Method 2: females (n=2) were inseminated pre-ovulation without anesthetic via endoscopy, Method 3: females (n=1) were inseminated pre-ovulation without anesthetic via manual insertion of an insemination catheter, Method 4: females (n=2) were inseminated same as Method 3 with the addition of standing sedation. Semen deposition site varied as a result of changes in AI technology and experience. All females conceived following intrauterine AI using three methods. Four pregnancies (n=3 females) produced via Method 3 and 4 resulted in term births (n=2 male calves, n=2 female calves) at 481.8±12.8days post-AI. Unfortunately, two early pregnancy losses were documented in a fourth female conceiving via Method 2. Pregnancy rates were 0%, 22%, 17%, and 50% for Method 1-4, respectively. Method 3 and 4 rates improved to 29% and 67%, respectively when accounting for AI’s conducted only on ovulatory estrous cycles. Spermatozoa (n=5 males) were cryopreserved 0.3-9.3 y prior to successful AI procedures. The lowest dose of frozen-thawed sperm resulting in conception was 500×10(6) motile sperm. Mean time from AI to ovulation in conceptive and non-conceptive cycles was 26±11.8h and 66±80.7h, respectively.

Concepts: Male, Reproduction, Female, Sperm, Spermatozoon, Menstrual cycle, Rhinoceros, Indian Rhinoceros

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Preventing obesity in zoo animals is increasingly recognized as an important husbandry objective. To achieve this goal, body condition scoring (BCS) systems are available for an ever-increasing number of species. Here, we present a BCS for the greater one-horned rhinoceros (Rhinoceros unicornis) based on an evaluation (on a scale from 1 to 5) of seven different body regions, and report resulting scores for 62 animals from 27 facilities, based on digital photographs. In animals above 4 years of age, this BCS correlated with the body mass:shoulder height ratio. Although differences between the sexes for individual regions were noted (with consistently higher scores in males for the neck and shoulder and in parous females for the abdomen), the average BCS of all regions did not differ significantly between males (4.3 ± 0.4) and females (4.1 ± 0.5). Linking the BCS to results of a questionnaire survey and studbook information, there were no differences in BCS between animals with and without foot problems or between parous and non-parous females. In a very limited sample of 11 females, those eight that had been diagnosed with leiomyoma in a previous study had a higher BCS (range 3.9-4.9) than the three that had been diagnosed as leiomyoma-free (range 3.5-3.7). The BCS was correlated to the amount of food offered as estimated from the questionnaire. Adjusting the amounts and the nutritional quality of the diet components is an evident measure to maintain animals at a target BCS (suggested as 3-3.5). Zoo Biol. XX:XX-XX, 2016. © 2016 Wiley Periodicals, Inc.

Concepts: Rhinoceros, Indian Rhinoceros, Dürer's Rhinoceros

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Circulating concentrations of fat-soluble vitamins A, D, and E, minerals, fatty acids, and lipids were quantified in five captive greater one-horned rhinoceroses (Rhinoceros unicornis) throughout two time periods, during which two diets were offered. Animals were fed mixed-grass hay and concentrate pellets while managed in barns for winter housing (April sampling, winter diet). During the spring and summer, animals were fed the same amount of concentrate pellet but had free access to North American browse and grasses instead of dried forage (November sampling, summer diet). Levels of 25-hydroxyvitamin D and α-tocopherol were statistically higher in summer diet samples than in winter diet samples. Retinol was not statistically different between seasons, and β-carotene concentrations were undetectable at both time periods. Cholesterol, triglycerides, and non-esterified fatty acids were all significantly elevated following access to unlimited fresh forages in summer. Serum electrolytes were not different between the two time periods but differences in circulating minerals were noted (cobalt, inorganic iodine, and magnesium elevated in winter diet samples; selenium and zinc elevated in summer diet). Access to non-native fresh green forages resulted in improvement of several nutritional parameters in greater one-horned rhinoceroses, implying a benefit when fresh browse and access to grass is provided. Zoo Biol. XX:XX-XX, 2014. © 2014 Wiley Periodicals, Inc.

Concepts: Nutrition, Fatty acid, Triglyceride, Fat, Lipid, Rhinoceros, Indian Rhinoceros, Dürer's Rhinoceros

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In Indian rhinoceros, extensive leiomyoma, a benign smooth muscle tumour, was sporadically diagnosed post mortem and commonly thought of as contributing factor for reduced fecundity of this species in captivity. However, to date, the prevalence of reproductive tract tumours and their relevance for fecundity are unknown. Our analysis of the international studbook now reveals that females cease reproducing at the age of 18.1±1.2 years; equivalent to a reproductive lifespan of just 9.5±1.3 years. This short reproductive life is in sharp contrast to their longevity in captivity of over 40 years. Here we show, after examining 42% of the captive female population, that age-related genital tract tumours are highly prevalent in this endangered species. Growth and development of these tumours was found to be age-related, starting from the age of 10 years. All females older than 12 years had developed genital tumours, just 7-9 years past maturity. Tumour sizes ranged from 1.5-10 cm. With age, tumours became more numerous, sometimes merging into one large diffuse tumour mass. These tumours, primarily vaginal and cervical, presumably cause widespread young-age infertility by the age of 18 years. In few cases, tumour necrosis suggested possible malignancy of tumours. Possible consequences of such genital tract tumour infestation are hindered intromission, pain during mating, hampered sperm passage, risk of ascending infection during pregnancy, dystocia, or chronic vaginal bleeding. In humans, leiomyoma affect up to 80% of pre-menopause women. While a leading cause for infertility, pregnancy is known to reduce the risk of tumour development. However, different from human, surgical intervention is not a viable treatment option in rhinoceroses. Thus, in analogy to humans, we suggest early onset and seamless consecutive pregnancies to help reduce prevalence of this disease, better maintain a self-sustained captive population and improve animal welfare.

Concepts: Human, Male, Reproduction, Female, Uterus, Reproductive system, Fertility, Indian Rhinoceros

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Because the digenetic trematode fauna of Nepal is poorly known, we began to search for schistosomes in and around Chitwan National Park (CNP) of southern Nepal. Both domestic and wild Indian elephants (Elephus maximus) are present, and we found one of two dung samples from wild elephants and 1 of 22 (4.5%) dung samples from domestic elephants to be positive for schistosome eggs. The morphology of the eggs and both cox1 and 28S sequences derived from the eggs/miracidia were consistent with Bivitellobilharzia nairi, reported here for the first time from Nepal. Also, 7 of 14 faecal samples from the Asian or greater one-horned rhinoceros (Rhinoceros unicornis) contained viable eggs indistinguishable from those of B. nairi. This identification was confirmed by comparison with both cox1 and 28S sequences from B. nairi eggs/miracidia derived from Nepalese and Sri Lankan elephants. This represents the first sequence-verified identification of a schistosome from any species of rhinoceros, and the first verified occurrence of a representative of Bivitellobilharzia (a genus of ‘elephant schistosomes’) in mammals other than elephants. Our work suggests that elephants and rhinos share B. nairi in CNP, even though these two members of the ‘charismatic megafauna’ belong to unrelated mammalian families. Their shared life style of extensive contact with freshwater habitats likely plays a role, although the snail intermediate host and mode of definitive host infection for B. nairi have yet to be documented. This report also supports Bivitellobilharzia as a monophyletic group and its status as a distinct genus within Schistosomatidae.

Concepts: Schistosoma, Digenea, Mammal, Elephant, Rhinoceros, Schistosomatidae, Indian Rhinoceros, Chitwan National Park

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Equine herpesvirus type 1 (EHV-1) was detected in an Indian rhinoceros (Rhinoceros unicornis), which was euthanized because of severe neurological disease. Encephalitis was suspected and EHV-1 DNA was detected in brain, lung, and spleen tissues. The viral IR6 protein was detected in lung tissues by Western blot analysis. Phylogenetic analyses of EHV-1 sequences amplified from various tissues was nearly identical to one recently described that resulted in both non-fatal and fatal encephalitis in polar bears. This represents transmission of EHV-1 to a species that is not naturally sympatric with the natural host of the virus and broadens the host range to Asian non-equid perissodactyls.

Concepts: Molecular biology, Virus, Western blot, Mammal, Horse, Rhinoceros, Host, Indian Rhinoceros