Comparative studies of mitogen- and antigen-induced lymphocyte proliferation in four captive rhinoceros species.

Cellular immune function in four rhinoceros species was evaluated by way of in vitro lymphocyte proliferation responses to mitogenic and antigenic stimuli to establish normative data on white blood cell activity for each species and to identify species-specific differences that might help explain the predisposition of black rhinoceroses (Diceros bicornis) to disease. A cross section of the U.S. rhinoceros population encompassing all four captive species was sampled, including the Sumatran rhinoceros (Dicerorhinus sumatrensis) (n = 3); Indian rhinoceros (Rhinoceros unicornis) (n = 4); African black rhinoceros (n = 16); and African white rhinoceros (Ceratotherium simum) (n = 10). Of the four species evaluated, African black rhinoceroses exhibited the weakest (P < 0.05) lymphocyte proliferative responses to the mitogens: pokeweed (0.1 microg/ml), phytohemagglutinin (0.3 microg/ml), and concanavalin A (5.0 microg/ml). Total cell density at the end of culture was only 70% of that achieved with lymphocytes isolated from African white rhinoceroses, Indian rhinoceroses, and Sumatran rhinoceroses. However, lymphocyte response to bacterial endotoxin lipopolysaccharide was similar (P > 0.05) across species. Antigenic stimulation produced much weaker responses than mitogenic stimulation. No differences (P > 0.05) were observed among rhinoceros species in response to 1 and 10 microg/ml of Leptospira icterohemorrhagiae or Leptospira gryppotyphosa. Lymphocytes from African white rhinoceroses proliferated weakly in the presence of Aspergillus fumigatus filtrate, whereas lymphocytes from the southern black rhinoceros subspecies appeared slightly suppressed in the presence of increasing doses (0.1, 1, and 10 microg/ml) of Aspergillus filtrate. This comparative data set characterizing lymphocyte proliferation in the rhinoceros reveals several differences in immune cell responses among rhinoceros species and provides some evidence that lymphocytes of captive African black rhinoceroses are less vigorous than those of the other rhinoceros species.     

Urinalysis in three species of captive rhinoceros (Rhinoceros unicornis, Dicerorhinus sumatrensis, and Diceros bicornis)

This study reports urinalysis values for three species of captive rhinoceros (Rhinoceros unicornis, Dicerorhinus sumatrensis, and Diceros bicornis) and evaluates individual and species differences. Repeated urinalysis was conducted on 11 individuals to establish normal reference ranges. Although no individual or species differences existed in urinary values for pH, all species differed in specific gravity. Rhinoceros urine demonstrated many physical and chemical properties similar to that of the horse, but reliability of this comparison was limited. Urinary pH in the rhinoceros was within range of that established for the horse and other large herbivores. However, all rhinoceros species exhibited urinary specific gravities below the lower limit of the normal equine reference range. Comparative urinalysis using an outside laboratory source confirmed the results of this study and illustrated the value of conducting in-house analysis. These results are the first data available on reference ranges for urine parameters in the greater one-horned, Sumatran, and African black rhinoceros and provide a useful diagnostic tool for the veterinary care of individuals in captivity.     

Ultrasonographic monitoring of artificially stimulated ejaculation in three rhinoceros species (Ceratotherium simum, Diceros bicornis, Rhinoceros unicornus).

Manual massage of the penis and rectal electroejaculation methods have been minimally effective for collecting semen from the rhinoceros. These two methods for stimulating ejaculation were evaluated by rectal ultrasonography. One individual each of three rhinoceros species (Ceratotherium simum simum, Rhinoceros unicornis and Diceros bicornis) was stimulated by manual massage of the penis and two black rhinoceroses (Diceros bicornis) were electroejaculated. On ultrasonography, neither manual massage nor rectal electroejaculation affected the accessory glands. The pelvic urethra remained empty during manual massage of unconditioned animals; however, this area was filled before commencement of massage of conditioned animals. The pelvic urethra also filled during electroejaculation. Semen accumulated in the pelvic urethra during pauses between electrical stimulations and was moved distally into the penile urethra by rectal and penile massage. The volumes of seminal fluid recovered from electroejaculation in this study exceeded previously reported attempts. This study demonstrated the potential of transrectal imaging for improving the recovery of semen by these methods.     

Hematologic, plasma protein, and biochemical profiles of brown pelicans (Pelecanus occidentalis)

OBJECTIVE: To establish hematologic and biochemical reference values for the brown pelican (Pelecanus occidentalis). ANIMALS: 31 captive, healthy, but permanently disabled pelicans and 35 wild-caught, healthy pelicans from a rehabilitation facility on the east coast of Florida. PROCEDURES: Samples of venous blood were collected from each pelican, and hematologic, plasma biochemical, and electrophoretic protein analyses were performed. Student t-tests were used to compare blood values between captive versus wild-caught, adult male versus adult female, and adult versus juvenile pelicans. RESULTS: Hematologic and electrophoretic values were similar between male and female, adult and juvenile, and captive and wild-caught pelicans. Significant sex differences existed for plasma calcium and triglyceride concentrations. Plasma concentrations of calcium, cholesterol, and CO2 content differed between captive and wild-caught adults. No significant differences were found between wild-caught adult and juvenile pelicans. CONCLUSIONS AND CLINICAL RELEVANCE: Our plasma biochemical results are similar to those of other brown pelicans and confamilial species. Additional studies on seabirds are encouraged, as age, sex, reproductive status, feeding habits, and captivity are important variables for health assessment in this and other aquatic species.     

Radiometric assessment of hexose monophosphate shunt capacity in erythrocytes of rhinoceroses.

OBJECTIVES: To measure metabolic rates of the hexose monophosphate shunt (HMPS) in erythrocytes of rhinoceroses, and to test the hypothesis that low concentrations of endogenous ATP in erythrocytes impair HMPS capacity, thereby increasing susceptibility to oxidant-induced hemolysis. ANIMALS: 13 black and 3 white rhinoceroses, free-ranging in several regions of southern Africa, and 1 Sumatran rhinoceros in US captivity. PROCEDURE: HMPS fluxes were measured in rhinoceros erythrocytes with carbon-labeled glucose in the presence and absence of known HMPS activators. RESULTS: Compared with values for human erythrocytes, mean basal state HMPS fluxes were appreciably lower (22 to 46%) in all 3 rhinoceros species studied. Shunt activators increased HMPS rates approximately 5-fold over basal rates in rhinoceros erythrocytes, compared with increases in humans of 10-fold with ascorbate and 15-fold with methylene blue. Stimulated HMPS rates in human erythrocytes were quantitatively 5- to 10-times greater than those observed in rhinoceros erythrocytes. Overall HMPS catabolic rates were completely independent of intracellular ATP concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: HMPS glycolytic and recycling rates and responses to activators are inherently low in erythrocytes from 3 species of rhinoceros, likely contributing to (but not solely responsible for) the high susceptibility of black rhinoceroses to oxidant-induced hemolysis. Slow erythrocyte HMPS capacities were independent of intracellular ATP concentrations, invalidating a current hypothesis regarding the pathogenesis of hemolytic anemia in captive black rhinoceroses. Limitations in HMPS capacities emphasize the importance of protecting rhinoceroses from exposure to drugs, chemicals, toxins, foodstuffs, and other conditions known to increase production of oxidizing metabolites, reactive oxygen species, and free radicals.     

The Anatomy of the Male Genital System of the Beluga Whale, Delphinapterus leucas, with Special Reference to the Penis

The genital organs of four male adult beluga whales and one newborn animal were dissected and the main characteristics are described. As in other species of cetaceans, the testes and the greatest part of the penis are located inside the abdominal cavity. The penis has a sigmoid flexure and belongs to the fibroelastic type with a thick tunica albuginea and a small amount of vascular spaces in the erectile tissue. The prostate gland, found in other cetaceans, was not seen macroscopically, but only small prostate rudiments could be identified histologically. The os penis and the other accessory glands are absent as in other whales.     

Attempted in vitro maturation and fertilization of postmortem Sumatran rhinoceros (Dicerorhinus sumatrensis) oocytes

A study was conducted opportunistically to evaluate the potential of rescuing immature oocytes from the ovaries of the Sumatran rhinoceros postmortem. Recovered oocytes (n = 30) were placed in maturation culture for 36 hr and inseminated with frozen-thawed homologous spermatozoa. After culture, evaluation of nuclear maturation status revealed that a large number of oocytes were degenerated (n = 21), but nine oocytes were assessed at the germinal vesicle (n = 3), metaphase I (n = 3), and metaphase II (n = 3) stages. Frozen-thawed Sumatran rhinoceros spermatozoa were capable of binding to the zona pellucida of in vitro matured oocytes, but no fertilization or cleavage resulted. In conclusion, relatively large numbers of oocytes can be obtained by ovarian follicular aspiration postmortem in the Sumatran rhinoceros, and some of these oocytes are capable of achieving nuclear maturation in vitro. However, additional studies are required to improve maturation success and achieve fertilization in culture.     

Seminoma in a southern black rhinoceros (Diceros bicornis minor): diagnosis, surgical management and effect on fertility

A testicular mass was identified by ultrasonography performed during a routine reproductive evaluation of an adult male southern black rhinoceros ( Diceros bicornis minor ). Histological examination of a testicular biopsy supported a presumptive diagnosis of testicular neoplasia. Hemi-castration was performed to excise the affected testis and a pathological diagnosis of a seminoma was made. Assessment of semen suggested reduced fertility as a consequence of the neoplastic process, but hemi-castration prevented further growth and metastasis of the tumour and ensured the animal's breeding potential. This is the second documented case of a seminoma in a rhinoceros species and the first case in a black rhinoceros.     

应用B型超声波鉴别马胎儿性别

通过胎儿尾部和脐带之间的生殖器结节定位进行马性别诊断，雄性胎儿准确率97．8％（182／186），雌性胎儿99．4％（160／161）。胎儿性别测定的最适时间为妊娠后59d和68d之间。妊娠中期胎儿性别诊断也非常可靠。超声波性别鉴定准确率达100％。在雄性，从100d到220d妊娠常规获得阴茎或包皮的图像；在雌性，从118d到227d妊娠马观察到胎儿乳腺和乳头。在马繁殖上常规使用的5MHz线性传感器，大多数情况下一直可用到妊娠160d。当胎位太高或其大小超过超声设备监测范围时，须使用3．5MHz探头。若掌握好性别鉴定的最适时间和认识正确的解剖结构，两种方法都是非常精确的。     

Corticosteroid-induced suppression of in vitro lymphocyte proliferation in four captive rhinoceros species.

Captive African black rhinoceroses (Diceros bicornis) are unusually susceptible to several diseases not commonly observed in any of the other three rhinoceros species maintained in captivity. The potential role of corticosteroids (either endogenously produced or exogenously administered) in the development of these sometimes fatal diseases has been questioned. In this study, the suppressive effects of two therapeutic corticosteroids (dexamethasone and hydrocortisone) on in vitro lymphocyte proliferation was examined in four rhinoceros species, including the Sumatran rhinoceros (Dicerorhinus sumatrensis, n = 3), Indian rhinoceros (Rhinoceros unicornis, n = 4), African black rhinoceros (n = 10), and African white rhinoceros (Ceratotherium simum, n = 5). Three blood samples collected from each rhinoceros 1 mo to 1 yr apart provided replicates for the study. Both dexamethasone and hydrocortisone suppressed (P < 0.05) lymphocyte proliferation stimulated by B-cell mitogens (pokeweed and lipopolysaccharide) and T-cell mitogens (phytohemagglutinin and concanavalin A). Suppressive effects of the glucocorticoids differed (P < 0.05) depending on the mitogen used to stimulate the lymphocytes. Overall, dexamethasone was a more potent suppressor of cell proliferation when compared with hydrocortisone (P < 0.05). However, black rhinoceros cell proliferation in response to any of the four mitogens was never completely suppressed, even in cultures containing the highest steroid concentration tested (10(-3) M). The effect of the two corticosteroids differed slightly among the rhinoceros species and subspecies tested, but there was no evidence that eastern or southern black rhinoceros lymphocytes were more sensitive to the suppressive effects of corticosteroids than the other rhinoceros species.     

Reproductive anatomy and physiology of the stallion.

Examination of the stallion's reproductive tract involves assessments of external and internal anatomy. External examinations are performed by visual inspection, palpation, or ultrasonography and include the scrotum, testes, epididymides, penis, and prepuce. Internal examinations may be performed by rectal palpation, transrectal ultrasonography, or endoscopy and include the accessory sex glands, pelvic urethra, and inguinal rings. A fertile stallion must produce, transport, store, and deliver viable spermatozoa to the mare. The physiologic processes involved include neuroendocrine control, spermatogenesis, sperm maturation, semen delivery, and fertilization.     

Standing laparoscopic-guided uterine biopsy in a southern white rhinoceros (Ceratotherium simum simum).

Transrectal ultrasonography of a 35-yr-old captive female southern white rhinoceros (Ceratotherium simum simum) with a history of chronic mucohemorrhagic vulvar discharge revealed right transmural uterine horn enlargement. Abdominal laparoscopic surgery, although extremely difficult because of inadequate instrumentation, permitted uterine visualization and biopsy. Standing anesthesia, incorporating butorphanol and azaperone together with local anesthetic infiltration, facilitated the laparoscopy. A leiomyoma was suspected on the basis of history, physical examination, ultrasonographic appearance, and histopathology. Prior rhinoceros laparoscopies have failed, primarily from limitations imposed by recumbency.     

A comparison of reproductive parameters of female Amur tigers (Panthera tigris altaica) in the wild and captivity

A healthy population of captive Amur tigers might assist recovery of the wild population in Northeast China if individuals were properly prepared and considered suitable for release in the wild. We analyzed the breeding records of 68 female Amur tigers from 1995 to 2010 in the Hengdaohezi Felid Breeding Center of China and compared the reproductive parameters of this population to wild female Amur tigers. We found that the reproductive parameters of the captive population (the age of first parturition, length of gestation and litter survival rate) were not significantly different from those of wild Amur tigers. Differences in birth date and litter size between wild and captive populations may be caused by management protocols for the captive population or insufficient field data from the wild population. Reproductive parameters of females giving birth after losing a litter were similar to parameters of females that did not lose a litter, except for birth date. These results provide no indication of major problems in using captive females for a breeding program for release of cubs into the wild, but additional information is still needed to assess their suitability.     

雄性非洲鸵鸟主要生殖器官的解剖学研究

了解非洲雄鸵鸟主要生殖器官的形态构造，对获取优质多量的精液，改进采精技术和开展鸵鸟的人工授精具有重要意义。本文采用解剖学方法，对雄性鸵鸟主要生殖器官的位置、形态和结构特点进行了系统的研究。结果表明：鸵鸟睾丸呈短棒状，两侧睾丸位置不对称，与其它家禽睾丸的特点有明显的差异；鸵鸟阴茎也很特殊，其阴茎背侧有用于射精的阴茎沟，它不同于哺乳类的尿生殖道，也不同于家禽的螺旋状的排精沟。     

Fungal pneumonia in a captive black rhinoceros

A captive black rhinoceros (Diceros bicornis) with a hoof abscess was treated with long-term antibiotic therapy. After 9 months of treatment, there was rapid deterioration, marked weight loss and reluctance to stand. Profuse, bilateral epistaxis developed accompanied by collapse and the animal was euthanased. Necropsy revealed pulmonary aspergillosis with concurrent Pseudomonas aeruginosa infection. Though a well-recognized disease of black rhinoceros, fungal pneumonia has not been reported in this species in Australia. The cost and efficacy of treatment have been questioned, however, prophylactic antifungal drug administration will be considered in any further cases of chronic, debilitating illness in black rhinoceros at Western Plains Zoo.     

Clinical implications of rhinoceros reproductive tract anatomy and histology.

Reproductive tracts or tissues from five male black rhinoceroses (Diceros bicornis), two male white rhinoceroses (Ceratotherium simum), two male one-horned Asian rhinoceroses (Rhinoceros unicornis), seven female black rhinoceroses, and six female white rhinoceroses from multiple institutions were examined to characterize their anatomy and histology. Some observations and measurements were obtained from in situ tracts of intact animals before or during necropsy. Formalin-fixed tissues were dissected and examined histologically. Retrospective reproductive data from each rhinoceros was obtained from the institutions of origin. Reproductive histology of these species was similar to that of other mammals. Male accessory gland structure varied among species, and the Asian rhinoceros epididymis was more loosely attached and had larger duct diameters than did the epididymides of the African species. Although histology was typically mammalian, rhinoceros reproductive morphology combined characteristics of several different mammals. Defining this unique morphology of rhinoceroses may help in understanding their reproductive physiology and will effect the development of appropriate reproductive techniques.     

Risk factors associated with a skewed natal sex ratio in captive black rhinoceroses (Diceros bicornis) in the United States.

The skew toward males in the sex ratio of calves is considered to be a problem for the captive population of black rhinoceroses (Diceros bicornis) in North America. This study used a retrospective case-control design to determine whether there was a predisposition toward birth of male calves over female calves and to examine risk factors associated with the occurrence of any detected skewed natal sex ratio in captive black rhinoceroses in the United States. The study population included captive female black rhinoceroses housed in the United States that had given birth to at least one calf of known sex. This study confirmed a skewing of the natal sex ratio toward male calves in the captive black rhinoceros population. The skewed ratio was found in calves born to wild-born dams, for which an increased time in captivity, irrespective of age, was associated with an increased likelihood of a male calf. Dams between 12 and 19 yr of age had a decreased likelihood of a male calf. The data also suggested a possible trend for the southern subspecies of black rhinoceros (Diceros bicornis minor) to be more likely to produce male calves than the eastern subspecies (Diceros bicornis michaeli). No associations were found with the sex of offspring in captive-born dams: however, this lack of association could be a result of low power in the study.     

Reproductive cycle observation of the Okinawa rail (Gallirallus okinawae) in the wild

The captive breeding program of the Okinawa rail started in 2008. For successful captive breeding, information related to reproduction, such as age at sexual maturity, testicular cycles and ovulatory cycles, is essential to predict when reproduction is possible and when certain reproductive behaviors are most likely to occur. We made gross and histological observations of the reproductive organs of Okinawa rails to gain understanding of sexual maturity, the testicular cycle and the ovulatory cycle. We found that the weight of the testis was smallest in December and largest in March. Changes in the diameter of the seminiferous tubule showed the same pattern. Mature sperm were observed from March to June. The heaviest ovary was observed in April. A single peak of reproduction, from March to April, was observed in males and females. Our observations suggested that the Okinawa rail is a seasonal breeder. Establishing suitable breeding pairs will be critical to ensure success of the Okinawa rail captive breeding program. Our results suggested that pairing must be started before March. If supportive breeding is used, semen should be collected from March to June and artificial insemination conducted in April.     

Determination of serum biochemical reference intervals for the Iberian lynx (Lynx pardinus)

Biochemical reference intervals were determined for 31 clinically healthy Iberian lynxes (Lynx pardinus) between 1 and 6 years of age. Thirteen of the lynxes were wild-caught and the other 18 were captive-reared animals. The samples were collected between November 2004 and December 2006. The influence of sex (males vs. females), age (juveniles vs. adults) and habitat condition (free-living vs. captive) on the biochemical analytes were evaluated. Serum albumin concentrations were significantly higher in females than in males, while creatine phosphokinase was higher in males. The levels of alkaline phosphatase and lactate dehydrogenase were higher in juvenile lynxes, while gamma glutamyl-transferase and creatinine values were higher in adults. Lynxes captured in the wild had higher concentrations of iron, calcium, alkaline phosphatase and creatinine, but lower aspartate aminotransferase and alanine aminotransferase than lynxes maintained in captivity. The results were generally comparable to commonly reported reference intervals for other lynx species, the domestic cat and other felid species.