Greater One-horned Rhinoceros Research Articles

Detection and characterization of vector-borne parasites and Wolbachia endosymbionts in greater one-horned rhinoceros (Rhinoceros unicornis) in Nepal

Vector-borne parasite infections affect both domestic and wild animals. They are often asymptomatic but can result in fatal outcomes under natural and human-induced stressors. Given the limited availability of molecular data on vector-borne parasites in Rhinoceros unicornis (greater one-horned rhinoceros), this study employed molecular tools to detect and characterize the vector-borne parasites in rescued rhinoceros in Chitwan National Park, Nepal. Whole blood samples were collected from thirty-six R. unicornis during rescue and treatment operations. Piroplasmida infections were first screened using nested polymerase chain reaction (PCR) targeting 18S ribosomal RNA gene. Wolbachia was detected by amplifying 16S rRNA gene, while filarial nematodes were detected through amplification of 28S rRNA, COI, myoHC and hsp70 genes. Our results confirmed the presence of Theileria bicornis with a prevalence of 75% (27/36) having two previously unreported haplotypes (H8 and H9). Wolbachia endosymbionts were detected in 25% (9/36) of tested samples and belonged to either supergroup C or F. Filarial nematodes of the genera Mansonella and Onchocerca were also detected. There were no significant association between T. bicornis infections and the age, sex, or location from which the animals were rescued. The high prevalence of Theileria with novel haplotypes along with filarial parasites has important ecological and conservational implications and highlights the need to implement parasite surveillance programs for wildlife in Nepal. Further studies monitoring vector-borne pathogens and interspecies transmission among wild animals, livestock and human are required.

GUT MICROBIOME DIVERSITY OF THREE RHINOCEROS SPECIES IN EUROPEAN ZOOS.

The wild rhinoceros populations have declined drastically in the past decades because the rhinoceros are heavily hunted for their horns. Zoological institutions aim to conserve rhinoceros populations in captivity, but one of the challenges of ex situ conservation is to provide food sources that resemble those available in the wild. Considering that the mammalian gut microbiota is a pivotal player in their host's health, the gut microbiota of rhinoceros may also play a role in the bioavailability of nutrients. Therefore, this study aims to characterize the fecal microbiome composition of grazing white rhinoceros (WR; Ceratotherium simum) and greater one-horned rhinoceros (GOHR; Rhinoceros unicornis) as well as the browsing black rhinoceros (BR; Diceros bicornis) kept in European zoos. Over the course of 1 yr, 166 fecal samples in total were collected from 9 BR (n = 39), 10 GOHR (n = 56), and 14 WR (n = 71) from 23 zoological institutions. The bacterial composition in the samples was determined using 16S rRNA gene Illumina sequencing. The fecal microbiomes of rhinoceros clustered by species, with BR clustering more distantly from GOHR and WR. Furthermore, the data report clustering of rhinoceros microbiota according to individual rhinoceros and institutional origin, showing that zoological institutions play a significant role in shaping the gut microbiome of rhinoceros species. In addition, BR exhibit a relatively higher microbial diversity than GOHR and WR. BR seem more susceptible to microbial gut changes and appear to have a more diverse microbiome composition among individuals than GOHR and WR. These data expand on the role of gut microbes and can provide baseline data for continued efforts in rhinoceros conservation and health status.

Vesicular stomatitis virus in two species of rhinoceros at a California zoological park.

To describe an outbreak of vesicular stomatitis virus (VSV) in southern white rhinoceros (SWR; Ceratotherium simum simum) and greater one-horned rhinoceros (GOHR; Rhinoceros unicornis) at a safari park in San Diego, CA, from May to September 2023. 21 SWR and 5 GOHR in professionally managed care. Rhinoceros of both species presented with a range of clinical signs and severities. Lesion locations were categorized as cutaneous (coronary bands, heels and soles, limbs, ventrum, neck folds, and ears) and mucocutaneous (lips, nostrils, mucous membranes of the oral cavity, and vulva). Clinical signs included lethargy, lameness, difficulty with prehension, hyporexia to anorexia, and hypersalivation. Severely affected rhinoceros had clinical pathology findings consistent with systemic inflammation. Vesicular stomatitis New Jersey virus was confirmed via PCR from swabs of lesions in 10/26 (38%) rhinoceros. Of these 10 confirmed cases, 9 (90%) were SWR and 1 (10%) was a GOHR. A further 6/26 (24%) were considered probable cases, and 10/26 (38%) were considered suspect cases based on clinical signs, but the inability to appropriately sample due to the housing environment precluded confirmation. Histopathology samples from 3 rhinoceros were consistent with VSV, and viral RNA was localized in histologic lesions via RNA in situ hybridization for 1 case. All rhinoceros survived infection despite severe systemic illness in 2 animals. This case series describes the clinical appearance and progression of VSV in 2 rhinoceros species. To the authors' knowledge, this is the first report of VSV in a rhinoceros.

Parturition and Neonatal Parameters of Three Species of Rhinoceros under Managed Care in the United States

Rhinoceros species range from near threatened to critically endangered due to habitat loss and poaching. A sustainable ex situ breeding population is critically important to maintain genetic diversity and help ensure the survival of the species; however, not all populations under human care are self-sustaining. While rhinoceros reproductive physiology and pathology have been well studied, there is still a paucity of information describing the normal parameters of parturition and neonatal landmarks. Using video recordings, medical records, and keeper logs, we reviewed and compared data regarding the parturition of three rhinoceros species (black rhinoceros (BR) (Diceros bicornis), n = 4; greater one-horned rhinoceros (GOHR) (Rhinoceros unicornis), n = 21; and southern white rhinoceros (SWR) (Ceratotherium simum simum), n = 22) managed under human care in the United States. Using equine parameters as a model for comparison, we compiled the following data: the signs of impending parturition, durations of the parturition phases, calving presentation, frequency of dystocia or stillbirth, and time from birth to neonatal landmarks. Data from 47 births, including 26 videos, were examined. The durations of parturition phases I, II, and III had median lengths of 153 min (n = 18), 28 min (n = 21), and 205 min (n = 15), respectively. Anterior presentation of the calf was observed in 59% births, whereas posterior presentation occurred in 41% births. Posterior calving presentation was associated with a longer phase II of parturition (p = 0.04), although more data are needed to determine whether the posterior presentation of the calf carries a higher risk for stillbirth. Most (83%) stillbirths occurred in GOHR, indicating that this species might be at a higher risk for stillbirth compared to SWR (17%) (p = 0.07). The median time from birth to the calf standing was longer in the GOHR (64 min) compared to the SWR (30 min) (p = 0.02). Detailed descriptions of the parturition parameters and neonatal landmarks in rhinoceros will aid facilities with rhinoceros breeding programs to recognize abnormalities in the parturient or post-partum periods and guide indications for veterinary intervention.

Empire, Nature and Agrarian World: A History of Rhino Preservation in the Kaziranga Game Reserve, India (1902–1938)

The greater one-horned rhinoceros or Indian rhinoceros ( Rhinoceros unicornis ) faced extinction in British India at the end of the nineteenth century. In 1908, the Government of Assam established the Kaziranga Game Reserve (KGR, now Kaziranga National Park) to preserve the vanishing rhino. As the twentieth century progressed, creating wilderness – by demonising the presence of the peasants and graziers – became a global panacea for protecting wildlife. Contrary to that belief, this article will show how the rhino population revived amidst human existence dictated by agro-ecological interactions and bureaucratic expediencies. The rhino’s ethology and its place in the imagination of rural people minimised its enemies. Moreover, in fluvial geography that constantly transformed the KGR’s boundaries, peasants and graziers creatively negotiated their usufruct rights and supported rhino preservation. Locating the KGR in the historical analysis of fluvial agro-ecology, this study illuminates how a critical interaction between different actors, i.e. human and non-human and coloniser and colonised, accentuated the cultural and material contestations amidst which the rhino eventually survived.

Comparative analysis of gestation in three rhinoceros species (Diceros bicornis; Ceratotherium simum; Rhinoceros unicornis)

This comparative analysis evaluated endocrine profiles and gestation length data of captive pregnant black rhinoceros (Diceros bicornis), white rhinoceros (Ceratotherium simum), and greater one-horned (GOH) rhinoceros (Rhinoceros unicornis). Hormone profiles were collected over three decades as part of pregnancy diagnoses. After the third month of gestation, the luteo-placental shift in progesterone production in pregnant rhinoceroses causes a significant increase in the concentration of faecal progesterone metabolites. We defined a laboratory-specific value of 1000 ng/g faeces as a threshold for incipient feto-placental progesterone production. Using this value allowed a comparison between species and revealed significant individual differences within a species. The mean ± SEM gestation days for reaching the 1000 ng/g faeces threshold were 89.5 ± 2.9 (range 56–138 days; n = 39) in black, 96.0 ± 2.6 (58–138; n = 39) in white, and 117.8 ± 5.3 (74–173; n = 19) in GOH rhinoceroses. For the calculations of gestation length, we complemented our results from three decades of reproductive monitoring with data from the literature, resulting in about 70 values for each species. Gestation length in the black, the white and the GOH rhinoceros was 460.6 ± 1.5 (range: 436 – 486), 503.8 ± 1.3 (range: 480 – 525) and 480.5 ± 1.1 (range: 453 – 505) days, respectively. Daylight length significantly affected gestation length, while the sex of offspring had no effect. On average, pregnancies with parturitions in spring and summer were one week shorter than those in autumn and winter. Although rhinoceroses are non-seasonal breeders, most parturitions in captivity occur in autumn and winter. We also analysed preconception endocrine profiles in the white rhinoceros. Conceptions in this species occurred after oestrous cycles of approximately 35 days (n = 18), 70 days (n = 3), 15 days (n = 1), after periods of ovarian inactivity (n = 5), and during a foal heat within one month after stillbirth parturition (n = 1). In conclusion, this study provides a comprehensive overview of gestational parameters in three rhinoceros species.

Empire, Nature and Agrarian World: A History of Rhino Preservation in the Kaziranga Game Reserve, India (1902–1938)

The greater one-horned rhinoceros or Indian rhinoceros ( Rhinoceros unicornis ) faced extinction in British India at the end of the nineteenth century. In 1908, the Government of Assam established the Kaziranga Game Reserve (KGR, now Kaziranga National Park) to preserve the vanishing rhino. As the twentieth century progressed, creating wilderness – by demonising the presence of the peasants and graziers – became a global panacea for protecting wildlife. Contrary to that belief, this article will show how the rhino population revived amidst human existence dictated by agro-ecological interactions and bureaucratic expediencies. The rhino’s ethology and its place in the imagination of rural people minimised its enemies. Moreover, in fluvial geography that constantly transformed the KGR’s boundaries, peasants and graziers creatively negotiated their usufruct rights and supported rhino preservation. Locating the KGR in the historical analysis of fluvial agro-ecology, this study illuminates how a critical interaction between different actors, i.e. human and non-human and coloniser and colonised, accentuated the cultural and material contestations amidst which the rhino eventually survived.

Reduced genetic diversity in greater one-horned rhinoceros in Chitwan National Park A new challenge and opportunity for rhino conservation in Nepal

Understanding the genetic diversity of a species is vital to improve the effectiveness of conservation management interventions. The greater one-horned rhinoceros (Rhinoceros unicornis) is one of the most iconic megaherbivores in South Asia but is classified as Vulnerable by the IUCN. The species is now broadly confined to two isolated populations in Chitwan National Park (CNP), Nepal and Northeast India, which are both constrained by poaching, and the loss or degradation of habitat. Since 2016, rhino mortalities have increased in CNP for unknown reasons. To assess the impact of any current or previous mortality events, we investigated the genetic diversity in the current CNP rhino population. We collected skin samples from 67 dead rhinos found in CNP and its buffer zone between 2012 and 2019. Fragments 428 bp of mitochondrial DNA D-loop were amplified from extracted DNA using PCR and sequenced to compare with complementary sequences derived from a previous study of CNP rhinos conducted during 1986-1987. A total of six haplotypes were detected in the older sample set with a haplotype diversity (Hd) of 0.38012 compared to only four haplotypes and a Hd of 0.3374 in our cohort. This decline in haplotype diversity was unexpected as the CNP rhino population has increased by 69% over the previous 27 years. In addition, we found the haplotypes were not equally distributed across CNP, with a greater variation detected in the eastern sector (4 haplotypes) compared to the west (1 haplotype). We recommend that CNP management authorities consider translocations within the park boundaries to promote breeding between the eastern and western subpopulations to disseminate and maintain haplotype diversity throughout the population.

Rhinoceros serum labile plasma iron and associated redox potential: interspecific variation, sex bias and iron overload disorder disconnect.

A consequence of the poaching crisis is that managed rhinoceros populations are increasingly important for species conservation. However, black rhinoceroses (BR; Diceros bicornis) and Sumatran rhinoceroses (SR; Dicerorhinus Sumatrensis) in human care often store excessive iron in organ tissues, a condition termed iron overload disorder (IOD). IOD research is impeded by the challenge of accurately monitoring body iron load in living rhinoceroses. The goals of this study were to (i) determine if labile plasma iron (LPI) is an accurate IOD biomarker and (ii) identify factors associated with iron-independent serum oxidative reduction potential (ORP). Serum (106 samples) from SRs (n = 8), BRs (n = 28), white rhinoceros (n = 24) and greater one-horned rhinoceros (GOH; n = 16) was analysed for LPI. Samples from all four species tested positive for LPI, and a higher proportion of GOH rhinoceros samples were LPI positive compared with those of the other three species (P < 0.05). In SRs, the only LPI-positive samples were those from individuals clinically ill with IOD, but samples from outwardly healthy individuals of the other three species were LPI positive. Serum ORP was lower in SRs compared with that in the other three species (P < 0.001), and iron chelation only reduced ORP in the GOH species (P < 0.01; ~5%). Serum ORP sex bias was revealed in three species with males exhibiting higher ORP than females (P < 0.001), the exception being the SR in which ORP was low for both sexes. ORP was not associated with age or serum iron concentrations (P ≥ 0.05), but was positively correlated with ferritin (P < 0.01). The disconnect between LPI and IOD was unanticipated, and LPI cannot be recommended as a biomarker of advanced rhino IOD. However, data provide valuable insight into the complex puzzle of rhinoceros IOD.

Gut microbiome architecture of wild greater one-horned rhinoceros: a vulnerable species from Kaziranga National Park, India

Rhinoceros unicornis, also known as the greater one-horned rhinoceros (GoHR), is a vulnerable wildlife species found in the Indian subcontinent with an estimated global population of 3582, of which an estimated 2995 resides in India. The Kaziranga National Park of Assam is the home to ~80.56% of the GoH population in India. Recent advances in genetics and microbial studies underscored the importance of gut microbial symbiosis as a crucial factor for host metabolic health and environmental interaction, particularly for higher mammals. Alteration of the normal microbiome can also be an indicator of chronic disease and infection. Freshly voided dung samples from nine dung heaps of free ranging or wild GoH rhinoceros were collected from Kaziranga National Park for mapping the gut microbial architecture through 16S-metagenomic approach. In our sample, the GoH gut harbours 168.8±12.55 (SE) bacteria-specific OTUs belonging to 21 phyla of which the gram-negative Proteobacteria is the most abundant phyla. Other abundant phylas found in the GoH gut are Firmicutes and Bacteroidetes. Although the GoH rhinoceros gut can utilize fibrous plant by microbial fermentation, the aerobic, nonfermenting Acinetobacter (20.7%), Stenotrophomonas (17.8%) and Brevundimonas (9.1%) constitute about 50% of all identified genus. Functional prediction of the GoH microbiome reveals that>50% of the bacteria present are involved in metabolism followed by cellular processes and information processing. A significant proportion (>1%) are associated with different diseases. In summary, our study characterized bacterial communities of nine wild GoH to identify some unique features and its implication in disease and survival of GoH.

Feasibility of reintroducing grassland megaherbivores, the greater one-horned rhinoceros, and swamp buffalo within their historic global range

Reintroduction of endangered species is an effective and increasingly important conservation strategy once threats have been addressed. The greater one-horned rhinoceros and swamp buffalo have declined through historic hunting and habitat loss. We identify and evaluate available habitat across their historic range (India, Nepal, and Bhutan) for reintroducing viable populations. We used Species Distribution Models in Maxent to identify potential habitats and evaluated model-identified sites through field visits, interviews of wildlife managers, literature, and population-habitat viability analysis. We prioritize sites based on size, quality, protection, management effectiveness, biotic pressures, and potential of conflict with communities. Our results suggest that populations greater than 50 for rhinoceros and 100 for buffalo were less susceptible to extinction, and could withstand some poaching, especially if supplemented or managed as a metapopulation. We note some reluctance by managers to reintroduce rhinoceros due to high costs associated with subsequent protection. Our analysis subsequently prioritised Corbett and Valmiki, for rhino reintroduction and transboundary complexes of Chitwan-Parsa-Valmiki and Dudhwa-Pilibhit-Shuklaphanta-Bardia for buffalo reintroductions. Establishing new safety-nets and supplementing existing populations of these megaherbivores would ensure their continued survival and harness their beneficial effect on ecosystems and conspecifics like pygmy hog, hispid hare, swamp deer, hog deer, and Bengal florican.

Climate change vulnerability of Asia’s most iconic megaherbivore: greater one-horned rhinoceros (Rhinoceros unicornis)

Climate change is an emerging threat for biodiversity conservation. It has already started impacting species assemblages and ecosystem dynamics. The greater one-horned rhinoceros (Rhinoceros unicornis) is an iconic and globally threatened megaherbivore. Once widespread across the northern part of the Indian subcontinent, there were fewer than 500 rhinoceros during the early 1960s, confined to isolated patches of suitable habitats in the southern part of Nepal and northern foothills of India, including Brahmaputra floodplains. Following both governments’ successful conservation strategies, the species has been recovering, and its global population at present is over 3500. However, the likely impacts of climate change has not been adequately incorporated into conservation plans for the species and may challenge this success. In this study, we developed a set of 21 vulnerability indicators and assessed the vulnerability of rhinoceros to climate change in Nepal through a review of literature, site observations of prime rhinoceros habitat, key informant interviews, a two-day stakeholders’ consultation workshop, and expert elucidation. Our findings suggest that rhinoceros in Nepal is likely to be ‘moderately vulnerable’ to the impacts of climate change, mainly due to (1) the likelihood of invasive plant species and severe floods in its prime habitat ‘Chitwan National Park’, and (2) fragmented habitat, small population size, droughts and forest fires in Bardia and Shuklaphanta National Parks. We further identified and recommended adaptation measures intended to enhance the resilience of rhinoceros to these likely threats.

Risks from disease caused by Mycobacterium orygis as a consequence of Greater one-horned Rhinoceros (Rhinoceros unicornis) translocation in Nepal.

The greater one-horned rhinoceros (Rhinoceros unicornis) is listed as vulnerable by the IUCN Red List. Mycobacterium orygis-associated disease was identified in a single greater one-horned rhino in Chitwan National Park in February 2015 prior to a planned translocation of five greater one-horned rhinoceros from Chitwan National Park to Bardia National Park for conservation purposes. This paper describes a qualitative disease risk analysis conducted retrospectively post-translocation for Mycobacterium orygis and this translocation, with the aim to improve the understanding of disease threats to the conservation of greater one-horned rhino. The disease risk analysis method used was devised by Sainsbury & Vaughan-Higgins (Conservation Biology, 26, 2017, 442) with modifications by Bobadilla Suarez et al (EcoHealth, 14, 2017, 1) and Rideout et al (EcoHealth, 14, 2017, 42) and included the use of a scenario tree and an analysis of uncertainty as recommended by Murray et al. (Handbook on import risk analysis for animals and animal products. Volume 1. Introduction and qualitative risk analysis, 2004), and the first time this combination of methods has been used to assess the risk from disease in a conservation translocation. The scenario tree and analysis of uncertainty increased the clarity and transparency of the analysis. Rideout et al.'s (EcoHealth, 14, 2017, 42) criteria were used to assess the source hazard and may be useful in comparative assessment of source hazards for future conservation translocations. The likelihood of release into the destination site of Mycobacterium orygis as a source hazard was estimated as of low risk, the risk of exposure of populations at the destination was of high risk and the likelihood of biological and environmental consequences was low. Overall, the risk from disease associated with Mycobacterium orygis as a result of this translocation was found to be low. Recommendations on disease risk management strategies could be improved with a better understanding of the epidemiology including the presence/absence of Mycobacterium orygis in greater one-horned rhino to develop effective disease risk management strategies.

Reduced Gut Microbiome Diversity and Metabolome Differences in Rhinoceros Species at Risk for Iron Overload Disorder.

Iron overload disorder (IOD) affects many wildlife species cared for ex situ. Two of the four rhinoceros species in human care, Sumatran rhinoceros (Dicerorhinus sumatrensis) and black rhinoceros (Diceros bicornis), are susceptible, whereas the other two, white rhinoceros (Ceratotherium simum) and greater one-horned (GOH) rhinoceros (Rhinoceros unicornis), are relatively resistant to IOD. Complex interrelationships exist between mammalian hosts, their indigenous gut microbiota, metabolome, physical condition, and iron availability. The goal of this study was to gain insight into these relationships within the family Rhinocerotidae. Specific objectives were to (1) characterize the gut microbiome and metabolome of four rhinoceros species; (2) compare the microbiome and metabolome of IOD-susceptible and IOD-resistant rhinoceros species; and (3) identify variation in the microbiome and metabolome associated with compromised health or disease in IOD-susceptible rhinoceroses. Fecal samples were collected from 31 rhinoceroses (Sumatran rhinoceros, n = 3; black rhinoceros, n = 6; GOH rhinoceros, n = 9; white rhinoceros, n = 13) located at five facilities, and matched fecal aliquots were processed for microbiome and metabolome analyses using 16S rRNA gene sequencing and nuclear magnetic resonance spectroscopy, respectively. Despite the phylogenetic disparity and dissimilar zoo diets of the hosts, the structure of the fecal microbiota of the two IOD-susceptible rhinoceros species were more closely related to each other than to those of the two IOD-resistant species (Bray–Curtis dissimilarity; IOD-susceptible vs. IOD-resistant p-value < 0.001). In addition, IOD-susceptible rhinoceroses exhibited less microbial diversity than their IOD-resistant relatives (Shannon diversity; p-value < 0.001) which could have health implications. Of note, the black rhinoceros was distinct among the four rhinoceros species with the most divergent fecal metabolome; interestingly, it contained higher concentrations of short chain fatty acids. Neither age nor sex were associated with differences in microbial community composition (p = 0.253 and 0.488, respectively) or fecal metabolomic profile (p = 0.634 and 0.332, respectively). Differences in the distal gut microbiomes between IOD-resistant and IOD-susceptible rhinoceroses support hypotheses that gut microbes play a role in host iron acquisition, and further studies and experiments to test these hypotheses are warranted.

Human-Rhino Conflict: Local People’s Adaptation to Impacts of Rhino

The greater one-horned rhinoceros, a protected species, significant to growing eco-tourism industry and conservation efforts in Nepal poses a number of challenges to the local people residing in the vicinity of the Chitwan National Park (CNP). The article is based on a study of local people’s perceptions on impacts of rhino and various coping and adaptation measures being adopted by them. The study also aims to explore ways to enhance adaptation by securing local people’s participation in rhino conservation. Review of secondary literature, surveys, Focus Group Discussions (FGDs), and Likert scale analysis were carried out for the purpose of the study. Five per cent of the households (50) in Bachauli Village Development Committee that hosts Bagmara Buffer Zone Community Forest User Group of the CNP were randomly selected. The study found that despite 80 per cent of the respondents incurred damage in some form from rhinos, 40 per cent of them exhibited a positive perception towards rhino conservation. Among several coping and adaptive measures to damages, a combination of electric/solar fencing and trench is found to be most effective. Applied research through collaborations between social scientists and ecologists can help to address the gaps in the understanding of different elements of human-wildlife conflict.

189 Isolation and purification of rhinoceros and horse spermatogonial stem cells

The northern white rhinoceros (NWR; Ceratotherium simum cottoni) is critically endangered. Testicular tissue (TT) of an NWR that died at 46 years was cryopreserved and stored. Spermatogonial stem cells (SSC) have been isolated from TT and, following transplantation or in vitro culture, differentiated into mature spermatozoa. These SSC have been phenotypically characterised by the expression of markers specific for SSC, and in vitro culture methods optimized to isolate, purify, and enrich populations of SSC from mixed germ cells. The reproductive age may affect the percent of SSC in TT, affecting recovery from older animals. We previously identified in TT of NWR the surface markers GFRa1 and GPR125, both of which were expressed at various stages of spermatogenesis. However, characterisation, isolation, and purification of rhinoceros SSC have not been studied. Because of the limited availability of rhinoceros TT, we used the horse as a model for rhinoceros (both species are in the same order: Perissodactyla). In this study, we (1) identified and compared the expression of markers specific for SSC (GFRa1, GPR125, PLZF) and pluripotent markers (SSEA-1, SSEA-4, OCT-4) in rhinoceros and horse mixed germ cells; (2) evaluated whether rhinoceros and horse SSC could be purified and enriched by sequential culture with collagen (2 days) and laminin (3 weeks); and (3) determined whether age may affect the percentages of SSC by comparing the expression of SSC and pluripotent markers in horse mixed germ cells at different reproductive stages: pubertal (PU=1-1.5 years) v. post-pubertal (PP=2-3 years) v. adult (AD=&amp;gt;5 years). Mixed germ cells were isolated from testes of one male of each of 3 rhinoceros species: NWR, southern white rhinoceros (SWR; Ceratotherium simum simum, 49 years), and greater one-horned rhinoceros (GOHR; Rhinoceros unicornis, 5 years), and horses at different reproductive stages (PU=2, PP=2, AD=7). The SWR and GOHR testes were refrigerated overnight, whereas NWR mixed germ cells were isolated from frozen-thawed TT. Flow cytometry analysis showed expression of SSC and pluripotent markers but not for transcription factor PLZF. The marker expression was similarly distributed between the 3 species of rhinoceros, with a larger portion of cells positive for SSEA-4 (mean%±s.e.m.: 6.3±1.1%) and smaller proportion for GDNFa1 (0.4±0.2%), GRP125 (0.1±0.1%), OCT-4 (0.4±0.2%), and SSEA-1 (0.4±0.2%; P&amp;lt;0.05). Similarly, horse expressed the markers GDNFa1 (0.9±0.3%), GRP125 (0.9±0.4%), OCT-4 (1.7±0.5%), and SSEA-1 (0.12±0.1%), but the abundance of SSEA-4 (1.8±0.8%) was less than that of rhinoceros (P&amp;lt;0.05). In rhinoceros, sequential culture enhanced the numbers of cells expressing all markers compared with that before culture, whereas in horses, the increase was observed only for cells expressing GDNFa1, GPR125, and SSEA-1. Reproductive stage did not affect the percentages of horse cells expressing germ cell and pluripotent markers. Overall, these results showed that rhinoceros SSC can be isolated from TT and expressed the same SSC markers as horses, and that differential culture enriched a population of SSC.

The Return of the Unicorns: The Natural History and Conservation of the Greater One-horned Rhinoceros

The Return of the Unicorns: The Natural History and Conservation of the Greater One-horned Rhinoceros

Coprophilous fungi from dung of the Greater One-Horned Rhino in Kaziranga National Park, India and its implication to paleoherbivory and paleoecology

AbstractFungal spores, especially those of coprophilous fungi, are present in dung middens ofRhinoceros unicornis(greater one-horned rhinoceros) in both forest and grassland areas of the Kaziranga National Park, India. The presence of coprophilous fungi on rhino dung, chieflySporormiella,Saccobolus,Ascodesmis,Cercophora, andSordaria, is documented for the first time. TheSporormiella–Ascodesmis–Saccobolusassemblage is abundant and characterizes the rhino dung in forest and grassland areas. The presence of coprophilous fungi spores allows for an examination of the relationship between rhinoceros ecology and the flora and other fauna in the region. The overall dataset is useful in interpreting the present and past distribution of rhino and other associated animals based on the relative abundance of different types of coprophilous fungi spores and their relationship to paleoherbivory and paleoecology in India and adjoining areas.

Global lessons from successful rhinoceros conservation in Nepal.

Global populations of rhinoceros have declined alarmingly, from about 500,000 at the beginning of the 20th century to 29,000 in 2016, largely due to an escalation of poaching for rhinoceros horn (Traffic 2016; Biggs et al. 2013). The current global rhino population is comprised of three Asian Species and two African species, the latter located in South Africa, Kenya, Tanzania, Namibia and Zimbabwe,. In Africa, the Southern white rhinoceros population is estimated at 20,700; and there are estimated to be around 4,885 black rhinoceros. The greater one-horned rhinoceros, found in Nepal and India, has a population of approximately 3,555. The other Asian rhino species are confined to Indonesia and have much lower numbers; there are fewer than 100 Sumatran rhinos and only 58–61 Javan rhinos. The number of African rhino killed by poachers in the last ten years is estimated at 5,957 (Traffic 2016; Emslie et al. 2013; Poaching fact2016), about 1,338 of these were taken in 2015, a year in which the highest number of rhino were taken since the late 1980s (Traffic 2016; Gaworecki 2016; Figure 1). At current poaching rates, Africa’s rhino populations may be extinct within 20 years (Di Minin et al. 2015). The Sumatran and Javan rhino populations continue to decline due to habitat destruction, poaching and inbreeding (Save the Rhino, 2016b) pushing them to the verge of extinction.

Ovarian down Regulation by GnRF Vaccination Decreases Reproductive Tract Tumour Size in Female White and Greater One-Horned Rhinoceroses.

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.