Conservation genetics of the black rhinoceros (Diceros bicornis)

We analysed genetic variation in mitochondrial DNA (mtDNA) control region in the Diceros bicornis minor and D. b. michaelianimals at the Western Plains Zoo, which form part of an international ex-situ breeding program. Six of the nine D. b. minor animals were wild-caught from Zimbabwe during the 1990s, and our study revealed five distinct mtDNA haplotypes, and a haplotype diversity of 0.86 in the colony. Phylogenetic relationships between mtDNA haplotypes analyzed using the neighbor joining method reveal that for the small sample available, D. b. minor and D. b. michaeli are reciprocally monophyletic and represent separate ancestral lineages. Nucleotide divergence between the black and and white rhinoceros(Ceratotherium simum) was 14.0%, and nucleotide divergencebetween the D. b. minor and the D. b. michaeli subspecies was 2.6%. This suggests a divergence time for the two blackrhinoceros subspecies of between 0.93 MY and 1.3 MY.     

Evaluation of habitat use and ecological carrying capacity for the reintroduced Eastern black rhinoceros (Diceros bicornis michaeli) in Ruma National Park,Kenya

We carried out a postrelease evaluation to determine predictors of habitat use and carrying capacity for the black rhinoceros (Diceros bicornis michaeli), which are critical for monitoring how the Ruma National Park sub-population may contribute to Kenya's meta-population strategy. We determined whether level of elevation, rockiness, shade, distance to fence, roads, and human settlements predict habitat use, differences in habitat and diet preference between female and male black rhinoceros, and the ecological carrying capacity (CC) of black rhinoceros in the park. We used standard ecological methods to collect data on predictors of habitat use, habitat preference and to estimate CC. Results show, first, that none of the environmental and anthropogenic factors evaluated predicted habitat use by black rhinoceros in the park. Second, although there was no significant difference in habitat preference between the sexes (U = 16.50, p = 0.306), there was a 60% difference in Jaccard's dissimilarity in diet selection between the sexes. Third, the park can support 65 black rhinoceros. Altogether, the findings suggest that the park has potential to support other sub-populations in Kenya. We recommend that future similar studies should incorporate population viability analysis and a community-based approach to forecast the species health and extinction risk.     

Characteristics of black rhinoceros (Diceros bicornis) bedding sites

Black rhino numbers have decreased greatly since the early 1970s, primarily as a result of poaching. A recent strategy to protect rhinos in Kenya has been to establish fenced sanctuaries. This has increased the rhino population and that of other species, but problems have emerged because of limited dispersal and rising animal densities. Unfortunately, how rhino utilize habitat, especially areas called bedding sites, is not well understood. These areas provide shade and may be a critical component of rhino habitat. We measured habitat variables at bedding sites located in bedding plots and compared them with control plots at Sweetwaters Reserve, Kenya. Euclea divinorum was the most common tree in the bedding site comprising 64.3% of the vegetation. Elephant dung was significantly more likely to be found in bedding plots than in control plots which suggests that elephants and rhinos use overlapping habitats. Elephants may be causing damage to the tree species that are important for bedding sites. Resource competition between large herbivores in small reserves is likely to negatively affect the tree species. Black rhino habitat, particularly bedding sites, may be at risk and rhino numbers may decrease.     

Gamete rescue in the African black rhinoceros (Diceros bicornis)

Mortality rates are high among captive African black rhinoceroses (Diceros bicornis), due to increased susceptibility to disease. The ability to rescue genetic material from individuals that die unexpectedly represents a practical approach to assist ex situ conservation efforts. The objectives of the present study were to attempt postmortem oocyte recovery from ovaries of African black rhinoceroses (N = 6) and to test the efficacy of equine protocols for rhinoceros oocyte IVM and IVF using cryopreserved rhinoceros sperm. The interval from ovary removal to oocyte recovery was 25.3 ± 13.9 h (mean ± SD). Ovaries were transported at 4 °C or 22 °C and effects of temperature on postmortem oocyte competence was evaluated. Numbers of oocytes collected per female averaged 15.8 ± 6.9. In total, 95 oocytes were recovered. Of these, 85 were inseminated using homologous sperm and 10 were inseminated using heterologous sperm. Overall, substantial numbers of viable oocytes were retrieved from African black rhinoceros ovaries 1 to 2 days postmortem from ovaries stored at ambient temperature. A proportion of these oocytes matured and underwent penetration and fertilization by heterologous or homologous frozen-thawed rhinoceros sperm. The reproductive competence of postmortem oocytes was further demonstrated by development of a single two-cell embryo. Despite the need for further refinements, gamete rescue in the rhinoceros has promise for producing rhinoceros embryos, as well as testing sperm functions in vitro.     

Genetic structure of the black rhinoceros (Diceros bicornis) in south-eastern Africa

Despite an on-going struggle to conserve the endangered black rhinoceros (Diceros bicornis) since the 1980s, huge capital investment and several genetic surveys, the level of genetic structure and connectivity among populations in southern Africa is not well understood. Here, we undertake a major population genetic study of black rhinoceros in the Zimbabwe Lowveld, an area inhabited by over half of that country’s original Zambezi descendants plus one large population sourced from the relict KwaZulu stock of South Africa. Using nuclear microsatellite and mitochondrial DNA data, we found much higher levels of genetic diversity in the indigenous Zimbabwean populations, where observed multilocus heterozygosity was 0.54 versus 0.40 in KwaZulu, and maternal haplotype diversity was 0.77 versus 0.03. We show, for the first time, that both gene pools can be differentiated from each other on the basis of nuclear markers. This, along with the discovery of recent gene flow between all Lowveld populations, suggests that Zimbabwean and South African gene pools were prehistorically connected.     

Effects of browse availability and quality on black rhino (Diceros bicornis michaeliGroves 1967) diet in Nairobi National Park,Kenya

Investigations were conducted to study the effects of browse availability and quality on black rhinoceros (Diceros bicornis michaeliGroves 1967) diet in Nairobi National Park, Kenya, between September 1994 and February 1995. Line intercept method was used to assess availability of browse. The ratios of browsed crowns to the total crown were applied in the estimate of browse use. Proximate analysis and extraction methods were used, respectively, to appraise browse nutritional quality and phytochemical status of twelve species. Thirty‐four plant species were identified as potential black rhino browse, whose availability differed significantly (P < 0.001). Lippia javanica (20.581 ± 3.101, n = 544) and Lannea cornuta (0.963 ± 0.529, n = 544) were the most and least available species, respectively. Only 32 species, however, were utilized, as L. cornuta and Rhynchosia hirta did not show any level of rhino feeding activity. Browse use was found to vary significantly (P < 0.001) during the study period. Grewia similis was most (20.564 ± 2.764, n = 544) and Commelina africana least (0.974 ± 0.974, n = 544) exploited, respectively. Differences in total alkaloids and phenols, crude fibre, crude protein and total ash contents among species investigated were highly significant (P < 0.001). Ether extract values were similar (P= 0.0933) across plant taxa. The study shows that the black rhino is a selective browser feeding on plants with low phenol and alkaloid contents and high fibre values. Of the nutritionally similar species, browsing was driven by availability. These findings suggest that a key habitat factor important in black rhinoceros conservation include diverse plant species with low levels of phenols and alkaloids.     

Pregnancy diagnosis in the black rhinoceros (Diceros bicornis) by salivary hormone analysis

Saliva samples collected from 12 black rhinoceroses (Diceros bicornis) were analyzed for 20 alpha-hydroxyprogesterone-like and total estrogen immunoreactivity. Five nonpregnant, two conception, and nine pregnancy intervals were monitored. HPLC co-chromatography of immunoreactive free steroid components in saliva indicated that one of three immunoreactive progestin components eluted with 20 alpha-hydroxypreg-4-en-3-one, whereas the single major estrogen peak eluted with estradiol. Direct radioimmunoassay measurements of ether extracts for these two reproductive hormones in saliva provided accurate and consistent profiles in which pregnancy was detected 13 months prior to parturition, and parturition was predicted by approximately 1 month. Measurement of 20 alpha-hydroxypreg-4-en-3-one was more useful in pregnancy diagnosis, and estradiol measurement was useful as an indicator of impending parturition. © 1996 Wiley-Liss, Inc.     

Mortality and female fecundity in an expanding black rhinoceros (Diceros bicornis minor) population

Identifying factors that affect demographic parameters and how those factors act is vital for understanding population dynamics, especially of endangered species. Moreover, specific ideas in the population dynamics of large herbivores underpin the management of the critically endangered black rhinoceros (Diceros bicornis). We studied an expanding black rhinoceros population since its establishment in 1986 in the Great Fish River Reserve, South Africa, through 2008 when managed removals interrupted natural dynamics. During the study, only 13 animals died, of which nine were subadults. In a linear modelling context, we used information-theoretic methods to evaluate the influence of independent variables expected to affect demographic parameters. For females, age at first reproduction (AFR) increased with abundance, but there was no effect of abundance on adult fecundity as measured by inter-birth intervals (IBIs). We evaluated these results in the theoretical context of population dynamics of large herbivores, in particular, Eberhardt’s proposal of a specific sequence in which demographic parameters first respond to increasing density. Our observations are consistent with Eberhardt’s prediction that immature individuals are impacted before adults, but the relative timing and magnitude of density effects on immature individuals was unclear. Rainfall did not influence AFR or IBIs. Maternal age influenced IBIs but much of the observed variation in IBIs was not accounted for by structural variation. Studies of populations more nearly approaching a stable age distribution and carrying capacity are needed to resolve remaining uncertainties and ambiguities in the life history of the black rhinoceros in particular and megaherbivores in general.     

Serum alpha-tocopherol, all-trans retinol, total lipids and cholesterol in the black rhinoceros (Diceros bicornis)

1. Mean concentration of serum alpha-tocopherol (Vitamin E) in 28 free-living black rhinoceroses sedated during translocation in Zimbabwe was 1.92 (SD, 0.43) mg/l. 2. Alpha-tocopherol was not detectable (less than 0.15 mg/l) in five captive black rhinoceroses held at London Zoo. 3. Circulating levels of all-trans retinol (Vitamin A) were not different between the two groups. 4. The low level of alpha-tocopherol in captive rhinoceroses suggests a risk of acute haemolytic anaemia.     

Predicting the habitat usage of African black rhinoceros (Diceros bicornis) using random forest models

Species distribution models are often used in ecology to ascertain relationships between environmental variables and species presence. Modelling to understand this relationship can be used to aid conservation management strategies. In this paper, we applied the random forest classification method to predict habitat used by black rhino for browsing. The random forest model was created using detailed habitat data collected from Ol Pejeta Conservancy in Kenya. Variables from plots where rhino had been present were compared to those not used by rhino. Independent data were used to test the predictive accuracy of the rules generated. The model performed well with the independent test data, correctly classifying 69% of the sampling plots where black rhino were present. Important habitat features that affected rhino presence were browse availability and density of vegetation, with Vachellia drepanolobium (formerly Acacia) and Euclea divinorum being important components. The analysis also highlighted areas of potential high browse pressure, which should be the focus of continued monitoring and management.     

Serum and fecal steroid analysis of ovulation,pregnancy, and parturition in the black rhinoceros (Diceros bicornis)

Studies were conducted to determine: (1) if fecal hormone metabolite concentrations correlated with serum estrogen and progesterone concentrations, follicular activity and reproductive behavior in the black rhinoceros (Diceros bicornis) and (2) if threshold values of respective fecal metabolite concentrations correlated with pregnancy. Blood and fecal samples were collected, in conjunction with transrectal ultrasound and behavior observations, for an 18-month period from one black rhinoceros female. Subsequently, serial fecal samples were collected from 13 females in 10 zoos. Quantitative analysis of serum progesterone (P₄) and estradiol (E₂) was performed by radioimmunoassay (RIA): analysis of fecal estrogen metabolites (E) and fecal progesterone metabolites (P) were performed by enzyme immunoassay (EIA). Serum P₂ concentrations identified two luteal phase patterns and two nadirs which corresponded with behavioral estrus. Fecal E patterns indicated a sharp peak which corresponded with breeding. concentrations of fecal P illustrated identifiable nadirs and several peaks which corresponded to serum P₄ nadirs and luteal phases. Serum P₄ concentrations were not different between the luteal phase and pregnancy. Fecal P concentrations started to rise above luteal phase concentrations approximately 150 days postbreeding and remained elevated until immediately before parturition. Serum E₂ and fecal E concentrations rose and subsequently declined after parturition. In the fecal samples from seven pregnant females, fecal P concentrations were similarly elevated compared to six nonpregnant females. Results indicated that fecal steroid metabolites accurately reflected serum steroid hormone concentrations and that the measurement of P and E concentrations permitted the characterization of the estrous cycle, the diagnosis of pregnancy, and the onset of parturition. Zoo Biol 16:121–132, 1997. © 1997 Wiley-Liss, Inc.     

Management parameters affecting the reproductive potential of captive,female black rhinoceros,Diceros bicornis

With deterioration of the wild population over the last two decades, captive reproduction of black rhinoceros has become a high priority for zoological gardens. Several reproductive parameters of female black rhinoceros were analyzed with data from the international studbook, and compared to data from field studies. These analyses yielded comparisons for ages of females at first calving, length of birth intervals, and span of reproductive life. The implications for rhino productivity are discussed, and some suggestions for increasing productivity are presented. © 1992 Wiley-Liss, Inc.