Alkaline phosphatase as an indicator of true ejaculation in the rhinoceros

The objective was to determine if seminal alkaline phosphatase (ALP) can serve as an indicator of true ejaculation in the rhinoceros. Concentrations of ALP activity were determined in seminal fractions collected from African black rhinos (Diceros bicornis), an African white rhino (Ceratotherium simum), and an Indian rhino (Rhinoceros unicornis) during electroejaculation. In addition, seminal fractions collected during penile massage of a Sumatran rhino (Dicerorhinus sumatrensis) were assessed. Correlations between ALP activity and sperm concentration, fraction pH, and fraction osmolality were evaluated in the Indian rhino and black rhino. Concentrations of ALP activity in rhino ejaculate fractions ranged from < 5 to 11,780 U/L and were positively correlated (P < 0.05) with sperm concentration for both Indian rhino (r = 0.995) and black rhino (r = 0.697), but did not exhibit a strong correlation with either pH or osmolality (P > 0.05). Data were insufficient for establishing meaningful correlation coefficients in the Sumatran rhino and white rhino, but preliminary results were in accordance with findings in the Indian rhino and black rhino. We concluded that ALP was present in rhinoceros semen, likely originated from the epididymides and/or testes, and could serve as a useful tool for assessing the production of ejaculatory versus pre-ejaculatory fluid in the rhinoceros.     

Alkaline phosphatase as an indicator of true ejaculation in the rhinoceros

The objective was to determine if seminal alkaline phosphatase (ALP) can serve as an indicator of true ejaculation in the rhinoceros. Concentrations of ALP activity were determined in seminal fractions collected from African black rhinos (Diceros bicornis), an African white rhino (Ceratotherium simum), and an Indian rhino (Rhinoceros unicornis) during electroejaculation. In addition, seminal fractions collected during penile massage of a Sumatran rhino (Dicerorhinus sumatrensis) were assessed. Correlations between ALP activity and sperm concentration, fraction pH, and fraction osmolality were evaluated in the Indian rhino and black rhino. Concentrations of ALP activity in rhino ejaculate fractions ranged from < 5 to 11,780 U/L and were positively correlated (P < 0.05) with sperm concentration for both Indian rhino (r = 0.995) and black rhino (r = 0.697), but did not exhibit a strong correlation with either pH or osmolality (P > 0.05). Data were insufficient for establishing meaningful correlation coefficients in the Sumatran rhino and white rhino, but preliminary results were in accordance with findings in the Indian rhino and black rhino. We concluded that ALP was present in rhinoceros semen, likely originated from the epididymides and/or testes, and could serve as a useful tool for assessing the production of ejaculatory versus pre-ejaculatory fluid in the rhinoceros.     

Mineral concentrations in serum/plasma and liver tissue of captive and free‐ranging Rhinoceros species

Mineral implications in health issues of captive rhinos have received much attention lately. This study was undertaken to establish reference values for the mineral status of rhinos. Serum/plasma samples of free‐ranging black (Diceros bicornis) and white (Ceratotherium simum) rhinos and of captive black, white, Indian (Rhinoceros unicornis), and Sumatran (Dicerorhinus sumatrensis) rhinos, as well as liver tissue samples of captive black, white, and Indian rhinos were analyzed for mineral content. Circulating mineral levels of free‐ranging animals were subject to variation according to region. In free‐ranging animals, high molybdenum (Mo) values compared to horse normals were striking. Captive animals displayed even higher circulating Mo concentrations. The significance of iron (Fe) overload in captive specimens of the browsing rhinos (black and Sumatran) was confirmed. Hepatic Fe levels increased in blacks with age. Although this Fe overload is suspected to be linked with diets, the data indicate that this is not due solely to an excessive dietary Fe supply. Whereas the grazing species (white and Indian) had high liver copper (Cu) levels, the browsing species had low to marginal liver Cu concentrations. Liver concentrations of K, Mg, Co, and Mo increased with age in captive black rhinos. Additional findings include high circulating Se levels in all rhino species. Future research should be directed at investigating factors leading to high Fe levels, and at investigating Cu metabolism in captive rhinoceros species. Zoo Biol 24:51–72, 2005. © 2005 Wiley‐Liss, Inc.     

Haltung und Zucht des Spitzmaulnashorns (Diceros bicornis) im Zoologischen Garten Berlin

The first Black rhino was kept in Berlin Zoo in 1870. After second world war the keeping of Diceros bicornis started in 1954. Between 1981 and 2006 16 births occurred in Berlin Zoo, of which 14 calfs were raised.     

Rhinoceros accounting in Kruger National Park,South Africa

Trends in rhinoceroses (rhinos) in Kruger National Park (Kruger) is of key concern. Poaching drives trends in the Park. Reconciling annual population estimates with yearly reports of poached carcasses fuels public critique. We account for trends in rhinos by extracting time series of estimates. Progressively modelling influences of management introductions and removals, effects of environmental variation and rhino density, direct impacts of poaching, consequences of imperfect carcass detection, and indirect impacts of deaths of dependent calves form the basis of accounting for rhinos. Models that considered all these influences explained 93% of white and 83% of black rhino population trends. In addition, the models predicted 2,515 white and 225 black rhinos, similar to estimates of 2,607 (95% CI: 2,475–2,752) and 202 (95% CI: 172–237) during 2020 respectively. The best model, however, predicts slow recovery with a white rhino population equivalent to pre-poaching achieved between 2030 and 2040. For black rhinos, recovery to pre-poaching population size would be between 2040 and 2050. The poaching onslaught in Kruger disrupted eruptive white rhino dynamics and prevented black rhinos from transitioning into eruptive dynamics. Authorities require innovative approaches within and beyond Kruger to help re-ignite rhino conservation.     

Feed intake,diet utilization,and composition of browses consumed by the Sumatran rhino (Dicerorhinus sumatrensis) in a North American zoo

The Sumatran rhino (Dicerorhinus sumatrensis), presently indigenous to the Malay peninsula and the islands of Sumatra and Borneo, is faced with extinction. It is estimated that fewer than 400 individuals still exist in the wild. In response, concerted efforts have been made to maintain and breed this species in captivity, and proper diet and nutrition underlie reproduction. The present feeding trial characterizes a diet fed to three Sumatran rhinos at the Cincinnati Zoo and Botanical Garden. The diet consists of mixed alfalfa/orchard grass hay, a variety of browses, grain pellets, produce (apples and bananas), and a vitamin E supplement. All three rhinos maintained body weight (614–761 kg), regular patterns of bowel movements, and fecal consistency and composition throughout the three 5‐day trials. Dry matter intake (DMI) ranged from 1.40 to 2.49% of body mass. Browse contributed by far the majority of nutrients to the diets in this study, from 62 to 83% of DMI. The nutrient composition of six species of subtropically grown browses (two from differing locations, for a total of eight browses characterized) showed that leaves did not differ from twigs in water or lignin content, but leaves contained higher concentrations of protein (both crude and bound) and ash, as well as lower fiber fractions. Of the macrominerals analyzed, only phosphorus differed between leaves and twigs, with twigs containing significantly more. No differences were seen between leaves and twigs in iron, molybdenum, or zinc content, but leaves contained lower concentrations of copper and higher concentrations of manganese and selenium compared with twigs from the same plants. Dry matter digestibility averaged approximately 50%. The only overt imbalances detected when comparing rhino diets with nutrient recommendations for domestic equids were excess calcium relative to phosphorus intake and low copper intake. These data, combined with information on native browse composition, intake, and digestibility, should greatly assist in providing suitable diets for this highly endangered species. Zoo Biol 19:169–180, 2000. © 2000 Wiley‐Liss, Inc.     

Early Pleistocene Dicerorhinus sumatrensis remains from the Liucheng Gigantopithecus Cave, Guangxi, China

The Liucheng Gigantopithecus Cave is very famous for its rich mammalian fauna and Gigantopithecus fossils. The Gigantopithecus Cave Fauna has been regarded as the typical Early Pleistocene fauna in South China. The majority of the fossils unearthed has been studied and published during the past decades. The only group remaining unpublished is the rhinoceros, which is studied in this paper. The rhino materials available today are only mandibles and lower cheek teeth. The dimensions of the tooth rows and the isolated teeth fall well within the range of the recent Dicerorhinus sumatrensis. Therefore, the rhino fossils from the Liucheng Gigantopithecus Cave can be referred to this living species. The reduced lower incisors and the U-shaped lingual contour of the mandible also support this taxonomic determination. The rhino materials from the Liucheng Gigantopithecus Cave represent the earliest known record of this species and the smallest Pleistocene rhinocerotid in China.     

EQUIPMENT REVIEW

ABSTRACT

The Northern White Rhinoceros Ceratotherium simum cottoni is a subspecies of the White Rhino that is almost extinct in the wild. We studied the last reproducing herd kept in Zoo Dvur Kralove to describe its vocal repertoire. The calls produced by eight individuals were recorded and analysed as concerned both sound properties and behavioural contexts in which they were emitted. We distinguished 11 calls belonging to four categories: (1) tonal harmonic sounds, (2) puffing sounds, (3) growling sounds, and (4) repetitive sounds. We found an apparent similarity between acoustic parameters of homologous calls recorded in both white rhino subspecies. We further confirmed that the repertoire of white rhino calls is much larger than that reported in other rhino species. We tentatively interpret this finding as an adaptation to increased sociality. Four calls reach the infrasound range; nevertheless, they are probably not used for communication over distance. This is obvious in the case of the grouch call, which contains the highest infrasound component. There are, however, other candidates for such a communication function: the repetitive pant sound, which is not known in other rhino species. We hypothesise that the repetitions may enhance their audibility in the typical open habitats of white rhinos.     

Anthropogenic Influences on Conservation Values of White Rhinoceros

White rhinoceros (rhinos) is a keystone conservation species and also provides revenue for protection agencies. Restoring or mimicking the outcomes of impeded ecological processes allows reconciliation of biodiversity and financial objectives. We evaluate the consequences of white rhino management removal, and in recent times, poaching, on population persistence, regional conservation outcomes and opportunities for revenue generation. In Kruger National Park, white rhinos increased from 1998 to 2008. Since then the population may vary non-directionally. In 2010, we estimated 10,621 (95% CI: 8,767–12,682) white rhinos using three different population estimation methods. The desired management effect of a varying population was detectable after 2008. Age and sex structures in sink areas (focal rhino capture areas) were different from elsewhere. This comes from relatively more sub-adults being removed by managers than what the standing age distribution defined. Poachers in turn focused on more adults in 2011. Although the effect of poaching was not detectable at the population level given the confidence intervals of estimates, managers accommodated expected poaching annually and adapted management removals. The present poaching trend predicts that 432 white rhinos may be poached in Kruger during 2012. The white rhino management model mimicking outcomes of impeded ecological processes predicts 397 rhino management removals are required. At present poachers may be doing “management removals,” but conservationists have no opportunity left to contribute to regional rhino conservation strategies or generate revenue through white rhino sales. In addition, continued trends in poaching predict detectable white rhino declines in Kruger National Park by 2016. Our results suggest that conservationists need innovative approaches that reduce financial incentives to curb the threats that poaching poses to several conservation values of natural resources such as white rhinos.     

Social and spatial relationships in captive southern white rhinoceros (Ceratotherium simum simum)

Although critical to the conservation of white rhinoceros, captive breeding has proven challenging because of the poor and irregular reproductive health of many captive rhinos, and social interactions may play a significant role. This research investigated the social and spatial relationships of two captive groups of southern white rhinoceros (Ceratotherium simum simum) by examining the frequency of companion changes, the number of space maintenance vocalizations made per hour by each reproductively mature female, and dominant/subordinate interactions. The observed captive rhinos did not change their companionships during the study. They exhibited space maintenance vocalizations and display greater than once per hour, particularly when feeding. Females housed with four calves on 0.033 km² exhibited space maintenance vocalizations more frequently (X±SE = 6.19±0.199/hr) than females housed with one calf and more space (0.06 km², X±SE = 0.55±0.182/hr) and females housed without calves and more space (0.65 km², X±SE = 1.90±0.086/hr). Wider separation of food piles and of females with young calves is suggested to reduce the interpreted spatial stress. The presence of a large number of rhinos in restricted captive space resulted in the formation of herds with dominance hierarchies that were enforced during competition for food and shade. The most subordinate rhino in each of the herds exhibited unusual behaviors such as dung‐kicking and nonestrus urine squirting, and neither has ever reproduced. Suppression of subordinate rhinos might lead to social stress that could negatively impact reproductive success. Zoo Biol 26:487–502, 2007. © 2007 Wiley‐Liss, Inc.     

Methods of semen collection in an ambulatory greater one-horned rhinoceros (rhinoceros unicornis)

Illegal poaching and reduced habitats have led to the endangered status of rhinoceroses. Collection of semen for preservation and future artificial insemination would therefore increase the reproductive potential of the rhinoceros. Although various collection methods have been attemped with individual rhinos, comparison between methods on a single animal have not been reported. This report details the application of six semen collection techniques on an unsedated and unrestrained greater one-horned rhino. The methods included different types of penile and/or rectal stimulation. Artificial vaginas and an inflatible probe for electroejaculation were specially constructed for semen collection. Of the various methods employed, penile massage consistently resulted in sperm-poor seminal fluid, but when preceded by either rectal massage or electroejaculation, seminal fluid with high sperm concentration was obtained.     

Reserve Size,Conspecific Density,and Translocation Success for Black Rhinoceros

Abstract: Fighting and accidental injury commonly cause black rhinoceros (rhino; Diceros bicornis) death after release. Smaller reserves and higher conspecific density after release (release density) might increase a rhino's encounter rate with hazards like fenced boundaries and conspecifics. We conducted a science-by-management experiment on the influence of reserve size and release density on rates of movement, association, and injury and death amongst 39 black rhinos during the first 100 days after their release into 4 Namibian and 8 South African reserves ranging in size from 670 ha to 45,000 ha. Association rates were negatively related to reserve size and positively correlated with release density. There was also a negative relationship between the proportion of the reserve traversed by individual rhinos and reserve size. In reserves ≥18,000 ha association rates were consistently zero but became elevated in reserves ≤11,500 ha and at release densities ≤9 km²/rhino. Daily displacement did not increase with increasing reserve size >8,500 ha but in smaller reserves daily displacements indicated higher encounter rates by released rhinos with fenced boundaries. Three rhinos received fight-related injuries requiring intervention and 2 of 4 deaths were fight-related. All injuries and 3 deaths occurred in reserves ≤11,500 ha. Model selection based on Akaike's second-order Information Criterion indicated that the parameter release density alone best explained mortality risk. Traditionally considered risk factors, rhino sex, age, and presence of resident conspecifics, were superseded by the risk posed by releases into smaller reserves. Reserves ≤11,500 ha and release densities ≤9 km²/rhino pose an increasing risk to rhino survivorship and so larger reserves and lower densities than these should be favored as release sites.     

Characterization of microsatellite loci in the black rhinoceros (Diceros bicornis) and white rhinoceros (Ceratotherium simum): their use for cross-species amplification and differentiation between the two species

As the population sizes of the black and white rhinoceroses continues to decline, more efforts are needed in multiple areas to help with the conservation efforts. One area being explored is the use of genetic diversity information to aid conservation decisions. In this study, we designed 21 microsatellite primers for white and black rhinoceroses, 16 and 17 of which amplified bands in the white and black rhinoceros, respectively. Out of these primers all 16 were polymorphic in the white rhinoceros and 12 of the 17 were polymorphic in the black rhinoceros. The mean number of alleles was 3.31 and 2.12, the expected heterozygosities were 0.420 and 0.372, and the observed heterozygosities were 0.436 and 0.322 for the white and black rhinoceroses, respectively. Seven of the primers produced different allele sizes and variations that distinguished between black and white rhinoceroses. Further genetic analyses with larger wild population sample sizes and markers are recommended to obtain a better understanding of the genetic structure of the black and white rhinoceros populations in order to be useful in the conservation efforts of these critically endangered species. A. Kilbourn—In memoriam.     

Reproductive behaviour of captive Sumatran rhinoceros (Dicerorhinus sumatrensis)

The Sumatran rhinoceros (Dicerorhinus sumatrensis) is on the verge of extinction in Malaysia. At the Sumatran rhinoceros Conservation Centre in Sungai Dusun, the reproductive behaviour of two female and two male rhinoceroses were studied for 8-10 months during attempts to breed them in captivity. Due to the paucity of scientific information on the reproductive biology of the Sumatran rhinoceros, this study was conducted to obtain information on the reproductive behaviour of this species. The male rhino was introduced to a female rhino in the morning for 1-2 h daily in order to observe for behavioural oestrus. Observations were made on the signs of oestrus and mating behaviour. Oestrus was determined by receptivity towards the male and lasted about 24 h. Common signs of oestrus were an increase in frequency of urine spraying, tail raising or swinging, anogenital and other contacts. Although the males exhibited mounting, the inability of the male to achieve intromission was poor. The study demonstrated that the pattern of courtship and copulation of the captive Sumatran rhinos were comparable with those of other rhino species, reported previously by other scientists and flehmen reflex was also exhibited by the male Sumatran rhinos. In a captive breeding programme, it is recommended that only an oestral female is introduced into a male enclosure due to the male solitary behaviour and to avoid serious injuries inflicted onto the females.     

A potential link between lateral semicircular canal orientation,head posture,and dietary habits in extant rhinos (Perissodactyla,Rhinocerotidae)

Extant rhinoceroses share the characteristic nasal horn, although the number and size of horns varies among the five species. Although all species are herbivores, their dietary preferences, occipital shapes, and common head postures vary. Traditionally, to predict the “usual” head posture (the most used head posture of animals during normal unstressed activities, i.e., standing) of rhinos, the occipital shape was used. While a backward inclined occiput implies a downward hanging head (often found in grazers), a forward inclined occiput is related to the horizontal head posture in browsing rhinos. In this study, the lateral semicircular canal (LSC) of the bony labyrinth was virtually reconstructed from µCT‐images in order to investigate a possible link between LSC orientation and head posture in extant rhinoceroses. The usual head posture was formerly reconstructed for several non‐rhinoceros taxa with the assumption that the LSC of the inner ear is held horizontal (parallel to the ground) during normal activity of the living animal. The current analysis of the LSC orientation resulted in a downward inclined usual head posture for the grazing white rhinoceros and a nearly horizontal head posture in the browsing Javan rhinoceros. The other three browsing or mixed feeding species show subhorizontal (closer to horizontal than a downgrade inclination) head postures. The results show that anatomical and behavioral aspects, like occipital shape, presence and size of horns/tusk‐like lower incisors, as well as feeding and feeding height preferences influence the usual head posture. Because quantitative behavioral data are lacking for the usual head postures of the extant rhinos, the here described relationship between the LSC orientation and the resulting head posture linked to feeding preferences gives new insights. The results show, that the inner ear provides additional information to interpret usual head postures linked to feeding preferences that can easily be adapted to fossil rhinoceroses.     

Genetic diversity, phylogeny and conservation of the Javan rhinoceros (Rhinoceros sondaicus)

With a total population of less than 60 individuals limited to two locations, the Javan rhinoceros is perhaps the most endangered large mammal on earth. Although species specific information is crucial to its conservation, its precarious status, habitat inaccessibility, and behavioral adaptations pose major obstacles to its study. Here we report on the first genetic analysis of the two extant populations, in Ujung Kulon, Indonesia, and Cat Tien, Vietnam, and discuss their conservation. As its critically endangered status precluded invasive sampling, we extracted DNA from dung, amplifying and sequencing segments of the mtDNA 12S rRNA gene and the non-coding D-loop. Divergence between Javan rhinos from Ujung Kulon and Cat Tien was similar to that between recognized subspecies of African rhinos, and exceeded that between Sumatran rhinos. The Ujung Kulon and Cat Tien populations represent separate Evolutionary Significant Units, advocating independent management. However, given the precariousness of the Cat Tien population, demographic considerations may override genetic issues in the short term. Genetic diversity of Javan rhinos was low and population expansion in the immediate future will be critical for its survival.     

Energy and mineral nutrition and water intake in the captive Indian rhinoceros (Rhinoceros unicornis)

In the captive Indian rhinoceros (Rhinoceros unicornis), two disease complexes with a high incidence—chronic foot problems and uterine leiomyomas—may be linked to excess body weight (BW). In this study, intake and digestion trials were conducted (by means of 7‐day weigh‐backs, and 5‐day total fecal collections, respectively) with 11 Indian rhinoceroses at four zoological institutions in Europe and the United States to quantify energy and mineral nutrition on conventional or roughage‐only diets. Diets comprising a variety of forages (grass hay only, a combination of grass hay and grass silage, straw, or a mixture of grass and legume hay) were offered as the roughage source, along with various concentrates, produce, and supplements. Water intake was quantified, and urine samples were obtained opportunistically. The animals consumed 0.5–1.1% of their BW in dry matter (DM) daily, with calculated digestible energy (DE, in megajoules MJ) values ranging from 0.27 to 0.99 MJ DE/kg BW^0.75/day compared to an estimated requirement of 0.49–0.66 MJ DE/kg BW^0.75/day. Seven of 11 rhinos (64%) fed restricted levels of concentrate plus forage consumed DE in excess of this estimate. Even on roughage‐only diets, some individuals consumed energy well above the presumed metabolic requirements. Hence, restriction of both concentrates and roughage may be important for weight management in this species. Water intake ranged from 30 to 49 mL/kg BW daily (3.4–5.2 L/kg ingested DM), similar to values that have been reported for domestic equids. Excretion amounts and patterns also resembled those found in horses. Endogenous fecal losses measured for Ca, P, Cu, Fe, and Zn indicate that the maintenance requirements of these minerals should be met in Indian rhinoceroses by diets that meet recommendations for domestic horses. It is particularly important to evaluate dietary adequacy in mineral nutrition in this species in concert with the need for restricted energy intake, especially with regard to the hypothetical involvement of a low Zn supply in chronic foot problems. Zoo Biol 24:1–14, 2005. © 2005 Wiley‐Liss, Inc.     

First successful artificial insemination with frozen-thawed semen in rhinoceros

The first successful artificial insemination (AI) in a rhinoceros was reported in 2007 using fresh semen. Following that success, we decided to evaluate the possibility of using frozen-thawed semen for artificial insemination. Semen, collected from a 35-36 year old Southern white rhinoceros (Ceratotherium simum simum) in the UK was frozen using the directional freezing technique. This frozen semen was used in two intrauterine AI attempts on a 30 years old female rhinoceros in Hungary. The first attempt, conducted 30 days postpartum with an insemination dose of ∼135 × 10⁶ motile cells, failed. The second attempt, conducted two estrus cycles later with an insemination dose of ∼500 × 10⁶ motile cells, resulted in pregnancy and the birth of a healthy offspring. This represents the first successful AI using frozen-thawed semen in a rhinoceros, putting it among very few wildlife species in which AI with frozen-thawed semen resulted in a live birth. The incorporation of AI with frozen-thawed semen into the assisted reproduction toolbox opens the way to preserve and transport semen between distant individuals in captivity or between wild and captive populations, without the need to transport stressed or potentially disease carrying animals. In addition, cryopreserved spermatozoa, in combination with AI, are useful methods to extend the reproductive lifespan of individuals beyond their biological lifespan and an important tool for managing genetic diversity in these endangered mammals.     

Breeding fireflies at Tama Zoo: An ecological approach

Techniques to breed the Genji firefly, Luciola cruciata, a lampyrid (firefly) native to Japan, we developed at Tama Zoo, Tokyo. In order to determine captive husbandry procedures, the staff made close observations on the life cycle of this species in nature. It took more than a decade to establish the breathing program in an outdoor water channel at the Tama Zoo. Patterned after a freshwater ecosystem, the channel system enables the insect to undergo the simulated “food-chains” cycle for completing the metamorphic process. Captive-reared larvae, also known as “glowworms”, and knowledge on husbandry have been utilized to reintroduce the species in its former natural ranges.