Cutaneous hemangioma in a giant panda (Ailuropoda melanoleuca).

An adult male giant panda (Ailuropoda melanoleuca) was presented with a cutaneous mass ventral to the eye. The animal was anesthetized and the mass was surgically excised. Histopathologic examination determined that the mass was a benign cavernous hemangioma, the first reported case in a giant panda.     

Dietary shifts affect the gastrointestinal microflora of the giant panda (Ailuropoda melanoleuca)

Giant pandas exhibit seasonal changes in bamboo plant part preference. The influences on the gastrointestinal tracts (GIT) microbial populations were evaluated during a 14‐month period for a pair of adult male and female giant pandas housed at the Memphis Zoo using traditional culturing methods to enumerate eight bacterial groups (total anaerobes, total aerobes (TAR), streptococci (STR), total enterics, Escherichia coli, Bacteroides spp., lactobacilli and Clostridium spp.). Both the male and female pandas altered bamboo consumption behaviours, with a sharp decrease in leaf preference in April 2010 and returning to high levels of leaf preference from June to October, corresponding to significant shifts in the densities of TAR, STR, and lactobacilli and Bacteroides spp. These findings indicate seasonal changes in food preference affect the assemblages of microbial populations within the GIT of the giant panda and contribute to a better understanding of the importance of bamboo in this species’ foraging strategy.     

A trial of intrauterine insemination using a fiberscope in the giant panda (Ailuropoda melanoleuca)

An intrauterine insemination technique using a fiberscope was investigated in the giant panda. A septal wall was present about 5 cm from the vulva, the external urethral orifice was present in the ventral region, and the entrance (pseudocervix) to the vagina was present in the dorsal region. The uterovaginal region protruded in the dorsal region about 15 cm from the pseudocervix. The external uterine orifice was present in the uterovaginal region, revealing that intrauterine insemination can be easily performed. This technique may greatly contribute to artificial reproduction of the giant panda.     

Post mortem computed tomography as a complementary tool for diagnosing cholangiohepatitis in a giant panda (Ailuropoda melanoleuca)

A geriatric female giant panda developed grave signs of illness and was diagnosed with suspected hepatobiliary tract obstruction or other severe hepatic disease such as advanced cholangiohepatitis. The giant panda was euthanized and post mortem computed tomography was performed prior to necropsy. Common bile duct obstruction at the major duodenal papilla by a mineral attenuating calculus causing dilatation of common bile and gallbladder with concurrent multiple areas of liver abscess were detected by postmortem computed tomography. These were confirmed with gross necropsy. This is the first case report of common bile duct obstruction by mineral calculus with concurrent severe cholangiohepatitis in a giant panda.     

Serologic response to a canarypox-vectored canine distemper virus vaccine in the giant panda (Ailuropoda melanoleuca).

The giant panda (Ailuropoda melanoleuca) is known to be susceptible to natural infection with canine distemper virus (CDV). Vaccination of giant pandas with conventional modified live CDV vaccines has been avoided due to the numerous carnivore species known to have become infected with CDV after vaccination. Serum-neutralizing antibodies to CDV were measured after s.c. and i.m. annual vaccination with a canarypox-vectored recombinant CDV vaccine in an adult male and female giant panda over the period of 2 yr. The vaccine proved to be safe, and serum-neutralizing antibody titers interpreted as protective against CDV disease were measured in each animal.     

Adjuvant effects of recombinant giant panda (Ailuropoda melanoleuca) IL-18 on the canine distemper disease vaccine in mice

Canine distemper virus (CDV) is a morbillivirus known to cause morbidity and mortality in a broad range of animals. Giant pandas (Ailuropoda melanoleuca), especially captive ones, are susceptible to natural infection with CDV. Interleukin-18 (IL-18) is a powerful adjuvant molecule that can enhance the development of antigen-specific immunity and vaccine efficacy. In this study, a giant panda IL-18 gene eukaryotic expression plasmid (pcAmIL-18) was constructed. Female BALB/c mice were muscularly inoculated with the plasmids pcAmIL-18, pcDNA3.1 and PBS, respectively. They were subsequently injected with an attenuated CDV vaccine for dogs, and the induced humoral and cellular responses were evaluated. The results showed that pcAmIL-18 remarkably improved the level of specific antibody, IFN-γ and IL-2 in mice sera, the T lymphocyte proliferation index and the percentage of CD4⁺ and CD8⁺ cells. These data indicated that pcAmIL-18 is a potential adjuvant that promotes specific immunity.     

Isolation and identification of a canine coronavirus strain from giant pandas (Ailuropoda melanoleuca)

Two giant pandas (Ailuropoda melanoleuca) died of unknown causes in a Chinese zoo. The clinical disease profile suggested that the pandas may have suffered a viral infection. Therefore, a series of detection including virus isolation, electron microscopy, cytobiological assay, serum neutralization and RT-PCR were used to identify the virus. It was determined that the isolated virus was a canine coronavirus (CCV), on the basis of coronavirus, neutralization by canine anti-CCV serum, and 84.3% to 100% amino acid sequence similarity with CCV. The results suggest that the affected pandas had been infected with CCV.     

Comparative anatomy of the radial sesamoid bone in the polar bear (Ursus maritimus), the brown bear (Ursus arctos) and the giant panda (Ailuropoda melanoleuca).

Since we have clarified the manipulation mechanism using the radial sesamoid (RS) in the giant panda (Ailuropoda melanoleuca), our aim in this study is to examine the position, shape and function of the RS morphologically, and to observe the attachment to the RS of the M. abductor pollicis longus and the M. opponens pollicis in the other Ursidae species. So, we focused on the carpus and manus of the polar bear (Ursus maritimus) and the brown bear (Ursus arctos) in this study. The RS was tightly articulated to the radial carpal, and could not adduct-abduct independently of the radial carpal. The M. abductor pollicis longus tendon and the M. opponens pollicis belly were attached to the RS, independently. In the polar bear, the deep concave and the flat surface were confirmed in attachment area for these two muscles. The morphological relationship between the RS and the M. abductor pollicis longus and the M. opponens pollicis in the two species of bears were essentially consistent with that in the giant panda. It also demonstrated that the manipulation mechanism of the giant panda has been completely based on the functional relationship between the small RS, and the M. abductor pollicis longus and the M. opponens pollicis in Ursidae species.     

Serosurvey of ex situ giant pandas (Ailuropoda melanoleuca) and red pandas (Ailurus fulgens) in China with implications for species conservation.

Conservation strategies for the giant panda (Ailuropoda melanoleuca) include the development of a self-sustaining ex situ population. This study examined the potential significance of infectious pathogens in giant pandas ex situ. Serologic antibody titers against canine distemper virus (CDV), canine parvovirus (CPV), canine adenovirus (CAV), canine coronavirus (CCV), canine herpesvirus, canine parainfluenza virus (CPIV), Toxoplasma gondii, Neospora caninum, and Leptospira interrogans were measured in 44 samples taken from 19 giant pandas between 1998 and 2003 at the Chengdu Research Base of Giant Panda Breeding in Sichuan, China. Seroassays also included samples obtained in 2003 from eight red pandas (Ailurus fulgens) housed at the same institution. All individuals had been vaccinated with a Chinese canine vaccine that included modified live CDV, CPV, CAV, CCV, and CPIV. Positive antibody titers were found only against CDV, CPV, and T. gondii. Sera were negative for antibodies against the other six pathogens. Results indicate that the quality of the vaccine may not be reliable and that it should not be considered protective or safe in giant pandas and red pandas. Positive antibody titers against T. gondii were found in seven of the 19 giant pandas. The clinical, subclinical, or epidemiologic significance of infection with these pathogens via natural exposure or from modified live vaccines in giant pandas is unknown. Research in this area is imperative to sustaining a viable population of giant pandas and other endangered species.     

圈养亚成体大熊猫(Ailuropoda melanoleuca)春、夏季行为节律的研究

2010年2～6月,应用瞬时扫描取样法对上海动物园10只圈养亚成体大熊猫的行为分配和节律进行研究.研究结果表明,圈养亚成体大熊猫的行为组成以取食和休息为主.春季,亚成体大熊猫取食行为频次高于夏季,而休息行为频次则低于夏季,春夏两季的行为总体分配间没有显著差异；从行为节律来看,圈养亚成体大熊猫的行为均呈现非正态特征.走动行为主要集中于7:00～8:00和13:00 ～15:00,攀爬行为主要出现于12:00之后,玩耍行为以13:00～14:00为主；亚成体大熊猫的部分行为之间存在相关性,活动行为与取食行为呈正相关,与休息行为呈负相关.     

CASE REPORT: Medical and surgical management of severe inflammation of the nictitating membrane in a Giant Panda (Ailuropoda melanoleuca)

A 10‐year‐old male giant panda (Ailuropoda melanoleuca) presented for severe, acute swelling and protrusion of the right nictitating membrane, unresponsive to topical therapy. Excisional biopsy of the nictitating membrane and its associated lacrimal gland was elected due to necrosis and friability of the tissue. Histopathology revealed suppurative, necrotizing conjunctivitis and dacryoadenitis. Culture grew Stenotrophomonas maltophila and Enterococcus spp with extensive antibiotic resistance. Treatment with topical and systemic antibiotics based on sensitivity results was initiated. All treatments were well tolerated. Healing was uncomplicated with no recurrence of the lesion and no clinical evidence of keratoconjunctivitis sicca. Ophthalmic exams and Schirmer tear tests performed opportunistically during postoperative anesthetic procedures were unremarkable, confirming that excision of the nictitating membrane had not caused clinically significant detrimental effects. The etiology of this lesion remains undetermined, but trauma is suspected. To the authors’ knowledge, this is the first report of nictitating membrane pathology and excision in the giant panda.     

Application of an enzyme-immunoassay (EIA) for rapid screening of 5 alpha-pregnane-3,20-dione (DHP) in blood plasma of the Asian elephant, Elephas maximus.

Populations of African (Loxodonta Africana) and Asian Elephants (Elephas maximus) in zoos and safari parks are at risk due to their low reproductive success. To extend the limited knowledge of their reproductive physiology, the development of easy and practical methods for the analysis of the relevant reproductive hormones is essential to support e.g. assisted reproduction. For the measurement of 5 alpha-pregnane-3,20-dione (DHP), the predominant ovarian gestagen in both species, an enzyme-immunoassay (EIA) based on commercial reagents was applied. Advantages of this EIA are the small volume of plasma needed for evaluation (5 microliters) and the possibility of direct processing without an extraction stage. The lower limit of detection was 0.16 ng/ml, the mean recovery was 101% and the mean coefficients of variation were 7.3% (intra-assay) and 9.9% (inter-assay). In Asian elephants, DHP levels reached 15 ng/ml during the luteal phase and up to 21 ng/ml during pregnancy. Estrous cycle lengths based on the lowest DHP concentrations varied from 12 to 20 weeks (mean 15.4 +/- 2.3). In two Asian elephant cows a calf was still-borne. Thereafter, the animals reassumed ovarian activity after approximately 8 and 13 weeks, respectively. In one animal estradiol implants for hormonal contraception caused a down regulation of the ovarian function as demonstrated by an irregular pattern of DHP secretion over a period of 48 weeks. We propose the direct DHP-EIA as a suitable method for reproductive monitoring in elephants, as it can be easily established in laboratories.     

Development and evaluation of a DAS-ELISA for rapid detection of avian influenza viruses

Rapid detection of avian influenza virus (AIV) infection is critical for control of avian influenza (AI) and for reducing the risk of pandemic human influenza. A double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was developed for this purpose. The method employed a monoclonal antibody (MAb) as the capture antibody and rabbit polyclonal IgG labeled with horseradish peroxidase as the detector antibody, and both antibodies were against type-specific influenza A nucleoprotein (NP). The DAS-ELISA could detect minimally 2.5 ng of influenza viral protein in virus preparations treated with Triton X-100, which is equvilent to 2.5 x 10(2) EID50 virus particles. This DAS-ELISA could detect all 15n AIV subtypes (H1-H15) and did not cross react with other avian pathogens tested. The DAS-ELISA were directly compared with virus isolation (VI) in embryonated chicken eggs, the current standard of influenza virus detection, for 805 chicken samples. The DAS-ELISA results correlated with VI results for 98.6% of these samples, indicating a sensitivity of 97.4% and specificity of 100%. The method was further tested with H5N1 and H9N2 AIV experimentally infected chickens, ducks, and pigeons, as well as field samples obtained from central China in 2005. The DAS-ELISA method has demonstrated application potential as an AIV screening tool and as a supplement for virus isolation in Asia.     

LUX^TM荧光PCR快速检测沙门氏菌的研究

本研究针对沙门氏菌invA基因的保守序列,设计特异的LUX^TM荧光标记引物,通过优化反应奈件和参数,建立可快速检测沙门氏菌的LUX^TM荧光PCR检测方法。结果显示,该方法高度敏感,其对纯菌的检测低限达到10^2cfu／mL,经6h增菌培养后检测,对样品液的检测低限达到1cfu／mL;特异性强,测试的全部13株沙门氏菌标准和参考菌株均呈阳性反应,测试的全部27株非目标菌均呈阴性反应;重复性好,定量检测批内和批间的变异系数均小于2％。应用本方法检测食品样品240份,结果检出阳性4份,与TaqMan荧光PCR和SN标准检测结果完全一致。本方法可在8h内完成对样品中沙门氏菌的检测,其检测的快速性、敏感性和特异性与TaqMan荧光PCR技术相当,且检测成本较低。     

氯霉素残留检测阻断ELISA试剂盒的研制及性能测定

在建立氯霉素单克隆抗体（CAP mAb）杂交瘤细胞株和阻断ELISA方法的基础上．研制出CAP残留快速检测试剂盒（CAP-kit），并对其性能进行了测定。结果表明，CAP-kit的标准曲线呈典型的S型，符合4参数logit曲线拟合，相关系数R^2=0．9953，检测范围为1．0μg/L～128．0μg/L，灵敏度为0．85μg／L，半数抑制浓度（IC50）为7．54μg/L，检测限为1．0μg/L；牛奶样、猪肉样的平均添加回收率为88．3％、82．5％，平均批内和批间变异系数均〈15％；CAP-kit与氯霉素琥珀酸钠的交叉反应（CR％）为150％，与其它酰胺醇类和抗菌素药物无交叉反应；试剂盒在4℃可保存6个月。     

Accuracy of rectal palpation and of a rapid milk progesterone enzyme-immunoassay for determining the presence of a functional corpus luteum in subestrous dairy cows

The accuracy of rectal palpation and of a rapid milk progesterone enzymeimmunoassay (EIA) for determining the presence of a functional corpus luteum (CL) in subestrous dairy cows was investigated, using the results of a radioimmunoassay (RIA) for progesterone in skimmed milk as the “gold standard”. The ovaries and uterus of each of 359 subestrous cows from 32 dairy herds were palpated per rectum, the presence or absence of a functional CL was predicted, and a milk sample was collected for analysis by both the rapid EIA and the RIA.

The nine clinicians participating in the study had a combined sensitivity of 82.6% and specificity of 52.6%, compared to the EIA which had a sensitivity of 90.3% and a specificity of 84.2% for predicting the presence of functional luteal tissue.

It was concluded that the technique of rectal palpation was inaccurate at assessing the functional (progesterone-secreting) status of ovarian structures. In addition, a qualitative EIA was as sensitive and more specific than rectal palpation in predicting the presence of a functional CL in the subestrous cow.

     

Development and evaluation of a model for prediction of fecal and urinary nitrogen excretions in cattle

The reduction of nitrogen (N) excretion in animal production is crucial in intensive farming systems particularly in the developed countries. In this study, a model to predict N excretion in cattle was developed based on existing feeding standards and evaluated using independent N balance experiments for Holstein steers and lactating cows and Japanese Black (JB) steers. Although model predictions for fecal and urinary N excretions appeared to be close to observed values in plot figures, statistical analysis showed that the model tended to over-predict both fecal and urinary N excretions, especially in Holstein lactating cows. This was because body weight changes of cows during lactation period were not considered in the model due to the lack of information (i.e., body weight gain or loss) available in the experimental data for evaluation. There were large mean bias and small line bias for urinary N prediction, but reverse results were obtained for fecal N prediction. The largest mean square prediction errors for both N excretions were due to random variation in all cases. When all data were pooled (combined), the accuracy for predictions for fecal N excretion was considerably high (r² = 0.94), indicating that the model may predict fecal N excretion beyond breeds, sexes and physiological states (growing and lactating). More information and accumulated data will be required to predict urinary N excretion under a wide range of genotype and environmental situation.     

Development and clinical evaluation of a rapid diagnostic kit for feline leukemia virus infection

Feline leukemia virus (FeLV) causes a range of neoplastic and degenerative diseases in cats. To obtain a more sensitive and convenient diagnosis of the disease, we prepared monoclonal antibodies specific for the FeLV p27 to develop a rapid diagnostic test with enhanced sensitivity and specificity. Among these antibodies, we identified two clones (hybridomas 8F8B5 and 8G7D1) that specifically bound to FeLV and were very suitable for a diagnostic kit. The affinity constants for 8F8B5 and 8G7D1 were 0.35 × 10⁹ and 0.86 × 10⁹, respectively. To investigate the diagnostic abilities of the rapid kit using these antibodies, we performed several clinical studies. Assessment of analytical sensitivity revealed that the detection threshold of the rapid diagnostic test was 2 ng/mL for recombinant p27 and 12.5 × 10⁴ IU/mL for FeLV. When evaluating 252 cat sera samples, the kit was found to have a kappa value of 0.88 compared to polymerase chain reaction (PCR), indicating a significant correlation between data from the rapid diagnostic test and PCR. Sensitivity and specificity of the kit were 95.2% (20/21) and 98.5% (257/261), respectively. Our results demonstrated that the rapid diagnostic test would be a suitable diagnostic tool for the rapid detection of FeLV infection in cats.