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.     

A review of drugs and techniques used for sedation and anaesthesia in captive rhinoceros species

Captive rhinoceros species are most frequently sedated and/or anaesthetised with the potent opioid, etorphine hydrochloride in combination with an alpha-2 adrenoreceptor agonist or the butyrophenone, azaperone. Carfentanil citrate based combinations have also been used to a lesser extent. In recent years butorphanol tartrate based combinations have been used with good success to induce neuroleptanalgesia. Sedation and anaesthesia are complicated by the large size of all rhinoceros species and their sensitivity to potent opioids. Potential complications include respiratory depression, hypoxaemia, hypertension, pulmonary shunting and ventilation/perfusion mismatch. The pharmacology of the principal drugs used for sedating/anaesthetising rhinoceros is reviewed. Techniques for sedating/anaesthetising the various species and potential complications associated with chemical restraint are discussed.     

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.     

Utilization of macro-nutrients and minerals in captive greater one-horned rhinoceros fed season-specific diets

This experiment was conducted to study nutrient and mineral utilization in greater one-horned rhinoceros fed season-specific diets. Nine adult greater one-horned rhinoceros (774–2407 kg BW) of Sanjay Gandhi Biological Park, Patna, Bihar, India, were used in this experiment. Three digestion trials of 60-d duration were conducted per animal, out of which 55 days was adaptation period and 5 days was collection period. The amount of concentrate was kept constant during all the trials. Green fodder sources were sugarcane, berseem (Trifolium alexndrinum) and chari (Sorghum bicolor) during trials I, II and III respectively. Green fodders were offered 20% in excess of previous day's intake in the afternoon. Apparent digestibility of dry matter and crude fibre was higher (p < .01) in group II as compared to other groups. Apparent digestibility of crude protein was lowest (p < .01) in group I, followed by group III, and the highest value was observed in group II. Apparent digestibility of Ca and P was lower in greater one-horned rhinoceros fed sugarcane and sorghum as green fodder source. In spite of this, sugarcane and sorghum-based diets were able to meet the requirement of Ca, P, Fe and Mn. Berseem supplied Ca in excess of requirement. All the diets were deficient in Zn. Hence, a suitable supplement of Zn should be added to the conventional zoo diet of greater one-horned rhinoceros.     

不同山羊草细胞质效应的比较研究

以 2 4种不同山羊草细胞质的稳定同核异质杂种为材料 ,进行了植株生长特性、病害抗性、抗低温能力、光合特性、主要农艺性状、自交育性和种子蛋白质含量等方面的比较研究。同时 ,还分析了异源细胞质效应的遗传特点 ,指出了生产上有应用价值的细胞质材料     

Studies on digestive physiology and feed digestibilities in captive Indian rhinoceros (Rhinoceros unicornis)

We performed intake, digestibility and ingesta passage studies in 11 Indian rhinoceroses (Rhinoceros unicornis) from four zoological institutions, using total faecal collection for the quantification of faecal output. The regularly fed zoo ration of roughage and concentrates (ration RC) and a roughage-only ration (ration R) were used; the roughage source differed between the facilities and comprised grass hay, grass silage, straw and lucerne hay. Dry matter intake ranged between 0.8 and 1.3% of body weight on ration RC and 0.5-1.2% on ration R. Digestibility coefficients achieved were similar to those reported for horses on diets of comparable composition. Endogenous losses as determined by linear regression analysis were within the range reported for horses. Measurements of faecal volatile fatty acids, faecal lactate and faecal pH also showed similarity to similar measurements in horses. The mean retention times of fluids (Co-EDTA) and particles (Cr-mordanted fibre <2 mm) in the whole gastrointestinal tract averaged 42 and 61 h, respectively, and were the longest ever recorded in a monogastric ungulate with this marker system. The results suggest that the horse is a useful model animal for designing diets for Indian rhinoceroses. Why digestive parameters are similar between these species in spite of enormous differences in body weight and retention times remains to be answered.     

Salmonella surveillance in a herd of asymptomatic captive black rhinoceros (Diceros bicornis) using fecal culture and PCR.

Feces were collected from captive black rhinoceros (Diceros bicornis minor) housed at Disney's Animal Kingdom to examine the frequency of Salmonella spp. shedding in asymptomatic animals using enrichment culture and broth culture- polymerase-chain-reaction (PCR) for detection. Three samples per animal were collected during the first week of each month between February 2001 and December 2003. During the study period, six different individual animals from one herd participated in the study, including two growing calves. A total of 550 cultures, using duplicate samples at two different laboratories, and 464 PCR tests were performed. When culture and PCR results were compared by the same laboratory, similar herd prevalence was found (2.4% positive cultures compared with 2.6% positive PCR tests). However, even though tests were performed on replicate samples, not every sample that was positive by culture was positive by PCR and vice versa. These results suggest that using multiple diagnostic methods and increasing the number of samples submitted may increase the likelihood of finding an asymptomatic Salmonella shedder. Although all of the rhinos shared the same environment throughout the study period, only four out of the six animals tested shed Salmonella spp. even though a minimum of 37 fecal samples were taken from each of the negative animals. Although this study followed a small number of rhinoceros, it suggests that asymptomatic shedding probably occurs more frequently in captive black rhinoceros than was previously believed. The prevalence appears to be similar to that reported for domestic ungulates.     

不同动物来源转移因子对肉鸭T细胞转化增殖影响的比较研究

为了解不同动物来源转移因子(TF)对T细胞的作用,本研究应用不同动物来源TF进行肉鸭T细胞体外转化增殖(MTT法)和T细胞体内转化增殖实验(α-乙酸萘酯酶染色方法).实验结果进行t检验分析,各实验组TF与对照组数据进行比较极显著差异,p<0.01;乌骨鸡来源TF与其他实验组数据进行比较极显著差异,p<0.01:T细胞体内转化增殖实验结果表明TF口服作用24 h内极显著提高T细胞转化增殖效果,而且240 h内T细胞的增殖效果与对照组相比仍然保持极显著差异.实验结果表明不同动物来源TF对樱桃谷商品肉鸭T细胞的转化增殖效果均有显著的影响,其中乌骨鸡来源TF效果最佳.     

Pasteurella multocida antigen-induced in vitro lymphocyte immunostimulation, using whole blood from cattle and turkeys.

A whole blood lymphocyte stimulation assay to study cell-mediated immune responses in bovine pasteurellosis was developed. Peripheral blood lymphocytes from cattle artificially immunised with three Asiatic haemorrhagic septicaemia strains of Pasteurella multocida exhibited higher stimulation indices when incubated with antigen preparations from homologous strains than with the heterologous shipping fever strain. Lymphocytes from cattle immunised with the shipping fever strain of P multocida exhibited a higher stimulation index when incubated with an antigen preparation from the homolgous strain than with antigen preparations from heterologous haemorrhagic septicaemia strains. These results suggest that immunogenic differences exist between the haemorrhagic septicaemia strains and the shipping fever strain of P multocida. An assay using turkey whole blood lymphocytes was also developed. The use of small amounts of whole blood, microtitre plates, either 125I iododeoxyuridine or 3H-thymidine as the labelling agent, and a multiple cell-culture harvester makes the method simple, rapid and suitable for the study of immune competence and cell-mediated immune responses in turkeys on a flock basis.     

Comparative electrocardiography in four species of macaws (genera Anodorhynchus and Ara)

Electrocardiograms (ECGs) were recorded during isoflurane anaesthesia from 52 macaws of four species of the genera Anodorhynchus and Ara in order to establish electrocardiographic reference values. The birds examined were clinically healthy macaws of the following species: hyacinth macaw (HM; Anodorhynchus hyacinthinus, n = 14); green-winged macaw (GWM; Ara chloroptera, n = 11); blue-throated macaw (BTM; Ara glaucogularis, n = 15); and red-fronted macaw (RFM; Ara rubrogenys, n = 12). All ECGs were recorded using a paper speed of 50 mm/s and a calibration of 10 mm = 1 mV. Significant differences were determined between species for the heart rate, duration and amplitude of the P wave, amplitude of the T wave, and amplitude of the QRS complex, specially comparing the RFM to the other macaw species. No significant differences were found between two species of similar body weight: the HM and the GWM.     

Regulation of mitogen- and TCGF-induced lymphocyte blastogenesis by prostaglandins and supernatant from equine embryos and endometrium

Immunosuppressive substances which interfere with lymphocyte blastogenesis are released in vitro by embryos and endometrium from mares in early pregnancy. Immunosuppression was not evident when tissues were cultured in the presence of indomethacin (a prostaglandin-synthesis inhibitor). Various prostaglandins (PGs) were added to equine lymphocytes and lymphocyte proliferation was measured after the addition of concanavalin A (Con A) or phytohaemagglutinin A (PHA). PGE2 and PGF2 alpha inhibited Con A-induced blastogenesis down to final concentrations of 1.8 x 10(-9) M and 1.3 x 10(-6) M, respectively. Other PGs tested (6-keto-PGF1 alpha and 13,14-dihydro-15-keto-PGF2 alpha) did not affect Con A-induced blastogenesis. PHA-induced blastogenesis was inhibited by concentrations down to 1.8 x 10(-9) M PGE2, 3.3 x 10(-7) M PGF2 alpha and 2.8 x 10(-4) M 6-keto-PGF1 alpha. At all concentrations, 13,14-dihydro-15-keto-PGF2 alpha only slightly reduced PHA-induced blastogenesis. Therefore, PGE2 was the only prostaglandin tested which, at physiological concentrations, significantly inhibited incorporation of [3H] thymidine. The mechanism of PGE2-mediated suppression was studied by adding PGE2 and T cell growth factors (TCGF) to TCGF-responsive lymphocytes. PGE2 reduced the TCGF-mediated blastogenic response in a dose-dependent manner. Furthermore, culture supernatant from embryos and endometrium from 14-day pregnant mares inhibited lymphocyte blastogenesis induced by TCGF. These results show that PGE2 interferes with lymphocyte blastogenesis by TCGF-dependent mechanisms. It is suggested that the PGE2 present in the uterus of the early pregnant mare may be one of the factors involved in immunosuppression at the time of maternal recognition of pregnancy.     

Comparative studies on in vitro mitogen-induced proliferation of peripheral blood lymphocytes in dog and breeding fox.

In vitro blastogenesis of dog and fox lymphocytes was compared by a microculture technique. The highest 3H-thymidine incorporation in cultures of dog lymphocytes was observed at day 3, while in those of fox at day 2, incubated either at 37 degrees C or at 39 degrees C. Lymphocytes cultured at 39 degrees C incorporated more tritiated thymidine than did cells cultured at 37 degrees C. The stimulation index (SI) of dog peripheral blood lymphocytes to both mitogens concanavalin A (Con A) and leucoagglutinin (LA) was in a similar range, while pokeweed mitogen (PWM) showed a weaker but significant stimulatory action. The blastogenesis of fox lymphocytes was the greatest in Con A stimulated cultures. The mitogenic potency of LA and PWM was about half of that of Con A, with no essential difference between them. Maximum lymphocyte proliferation of dog and fox was observed when culture media were supplemented with 10% fetal calf serum (FCS).     

四个燕麦品种对锶耐受性的比较研究

为探明燕麦对锶的富集能力和耐受机制,选取了4个燕麦品种(本德、白燕2号、白燕7号和定莜6号),施加不同浓度锶(0、25、100、500和1000 mg/kg)处理30 d,分析燕麦对锶的富集和分配特征、生长指标、丙二醛(MDA)含量以及超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性。结果显示:1)燕麦幼苗的根部和地上生物量都随着土壤中锶浓度的升高呈先增加后降低的趋势;2)各器官的富集浓度随着土壤中锶浓度的升高而增加,并表现为叶>根>茎。转运系数随着土壤中锶浓度的升高而降低,在1000 mg/kg时小于1,而在其他锶处理中均大于1。本德在25 mg/kg时的转运系数最高(2.026);3)高浓度锶增加了燕麦的MDA含量并抑制了POD活性,而SOD活性随着锶浓度增加而升高。结果表明,燕麦的根、茎、叶均能积累锶,叶片的富集能力最强。4个燕麦品种具有锶超富集植物的特征。低浓度锶可以诱导燕麦中抗氧化酶活性的增强,促进其生长;高浓度锶处理中SOD和CAT在清除活性氧中发挥重要作用。综合评价,本德可用于治理锶轻度污染地区,而定莜6号可用于治理锶重度污染地区,为燕麦用于锶污染地区的植物修复提供理论依据。     

Serum lipid concentrations in six canid and four ursid species in four zoos.

Serum lipid levels were measured in healthy captive wild canids and ursids, and the values were compared with previously published data. Serum lipid levels were evaluated in blood samples collected from eight African wild dogs (Lycaon pictus), three arctic foxes (Alopex lagopus), nine gray wolves (Canis lupus), four maned wolves (Chrysocyon brachyurus), two Mexican wolves (Canis lupus baleiyi), nine red wolves (Canis rufus), two brown bears (Ursus arctos), six polar bears (Ursus maritimus), six spectacled bears (Tremarctos ornatus), and five sun bears (Ursus malayanus). Samples were analyzed for total cholesterol, triacylglycerides, high-density lipoprotein-cholesterol, and low-density lipoprotein-cholesterol. Although the results showed a great variation among species, circulating lipids appeared especially high, sometimes extremely so, in the spectacled bears, polar bears, sun bears, and maned wolves compared with all other species sampled. The study provides a substantial basis for comparing lipid levels in presumed healthy animals and indicates a need for controlled study of the effects of diet on circulating lipid levels.     

Oviduct adenocarcinoma in some species of captive snakes

This article reports 5 cases of oviduct adenocarcinoma in adult captive snakes from Smithsonian's National Zoological Park. This neoplasm was found in 1 of each of the following species: emerald tree boa (Corallus caninus), Amazonian tree boa (Corallus enydris enydris), Burmese rock python (Python molurus bivittatus), Northern pine snake (Pituophis melanoleucus melanoleucus), and corn snake (Elaphe guttata). Grossly, tan to red firm masses were found within oviducts in 3 cases. In an additional 2 cases, the primary tumor was detected only histologically. Microscopically, neoplasms were papillary, and often extended transmurally. The neoplastic cells were polygonal and organized in acini or cords, with often abundant fibrovascular stroma. Hemorrhages and necrosis were present in all cases. Inflammation, myxomatous material, desmoplasia, and bacteria were often observed. Histologic evidence of metastasis was present in all cases. Solid metastases were seen in all animals except the Northern pine snake and involved several organs including the liver, lung, and heart. Emboli of neoplastic cells were observed in all animals but the Burmese rock python and corn snake.     

The influence of dietary tannin supplementation on digestive performance in captive black rhinoceros (Diceros bicornis)

Free-ranging browsers such as the black rhinoceros (Diceros bicornis) consume a diet that contains tannins, whereas the diets offered to them in captivity consist mostly of items known to contain hardly any such secondary plant compounds. Tannins could have potentially beneficial effects, including the chelation of dietary iron (iron storage disease is a common problem in black rhinos). Here, we tested the acceptance, and the consequences on digestion variables, of a low-dose tannin supplementation in captive animals. Eight black rhinoceroses from three zoological institutions were used. Faecal output was quantified by total faecal collection. Diets fed were regular zoo diets supplemented with either tannic acid (T, hydrolysable tannin) or quebracho (Q, condensed tannins); overall tannin source intake increased at 5-15 g/kg dry matter (DM) in relation to regular zoo diets. Adaptation periods to the new diets were >2 months. Additional data were taken from one hitherto unpublished study. Data were compared to measurements in the same animals on their regular zoo diets. All animals accepted the new diets without hesitation. There was no influence of tannin supplementation on digestion coefficients of DM and its constituents, or faecal concentrations of short-chain fatty acids or lactate. Water intake did not increase during tannin supplementation. Should the inclusion of dietary tannin sources be an objective in the development of diets for captive rhinoceroses, moderate doses such as used in this study are unlikely to cause relevant depressions of digestive efficiency and will not interfere with bacterial fermentation in a relevant way.     

Quantification of Enterobacteriaceae in faeces of captive black rhinoceros (Diceros bicornis) in relation to dietary tannin supplementation

Free-ranging browsing herbivores ingest a range of secondary plant compounds, such as tannins, with their natural diet. As many of these substances have been shown to have antibacterial properties, it could be speculated that a lack of such compounds in captive zoo diets could favour the growth of potentially pathogenic intestinal bacteria. The effect of a supplementation of a conventional diet (N, consisting mainly of grass hay and/or lucerne hay and pelleted compound feeds) fed to eight captive black rhinoceroses (Diceros bicornis) from three zoological institutions with either tannic acid (T), a source of hydrolysable tannins, or quebracho (Q), a source of condensed tannins, was investigated. The number of faecal colony forming units (CFU) of Enterobactericeae was determined by colony count of dilution series from fresh faeces applied to MacConkey agar plates. Tannins were added to the diets at approximately 5-15 g/kg dry matter, depending on the varying intake of roughage and compound feeds by the animals. There was no difference in the number of CFU between diets N (95.0 x 10(5) +/- 225.3 x 10(5)/g fresh faeces) and T (164.3 x 10(5) +/- 225.1 x 10(5)/g fresh faeces); in contrast, diet Q led to a significant reduction in CFU (4.3 x 10(5) +/- 6.5 x 10(5)/g fresh faeces) compared with the other diets. These findings suggest that condensed tannins could have the potential to reduce the number of potentially pathogenic intestinal bacteria, and that the deliberate inclusion of tannin sources in the diets of captive wild animals should be further investigated. The fact that tannic acid, shown to have antibacterial effects in various in vitro studies, did not have an effect in this study, emphasizes that the relevance of tannin supplementation for intestinal health must be verified in vivo.