Effects of body weight and season on serum lipid concentrations in sloth bears (Melursus ursinus ursinus)

Serum lipid levels were measured in 66 healthy sloth bears (Melursus ursinus ursinus) living under semicaptive conditions with access to natural food resources in the Bannerghatta Biological Park (Karnataka, India), a portion of their native habitat range in the Indian peninsula. Total cholesterol, triglycerides, high-density lipoprotein (HDL) cholesterol, and low-density lipoprotein cholesterol levels were analyzed. The effects of age, body weight, and season on these lipid parameters were statistically evaluated. There were no correlations between age and any of the serum lipid parameters analyzed. Positive correlations of body weight to both triglyceride and HDL cholesterol levels in these bears were identified. In addition, seasonal trends in physiological serum lipid values, potentially due to variations in the sloth bear diet, were identified. Serum triglyceride levels were higher during postmonsoon season and cholesterol levels were higher during winter compared to other seasons. Serum lipid values obtained from sloth bears in this study were also compared to previously published data on other members of the family Ursidae. This is the first report of serum lipid values as a reference for sloth bears. These values can be used as sensitive predictors of overall health and nutritional status to aid in the captive management and feeding of these bears.     

Pandemic (H1N1) 2009 influenza A virus infection associated with respiratory signs in sloth bears (Melursus ursinus)

In 2009, a pandemic influenza A virus (pH1N1) spread globally in humans and infected a broad range of captive animals with close human contact. In February 2014, a pH1N1 virus was isolated from a sloth bear with respiratory signs at a US zoo, demonstrating that recurring epidemics present an ongoing threat to animals, including threatened species. This is the first report of pH1N1 infection in sloth bears. To understand the sloth bear virus within the global context of pH1N1, phylogenetic trees were inferred including full‐length sequences from available non‐human, non‐swine hosts, representing four families in the order Carnivora and one order of birds. A combination of phylogenetic and epidemiological evidence strongly suggests the sloth bear was infected with a human‐origin pH1N1 virus, supporting the implementation of biosecurity measures to protect human and animal health.     

Molecular prevalence and characterization of Hepatozoon ursi infection in Indian sloth bears (Melursus ursinus)

Hepatozoon species are parasites that infect a wide variety of domestic and wild animals. The objective of the study was to detect the occurrence of Hepatozoon ursi in Indian sloth bears and to characterize the parasite based on phylogenetic analysis of the partial 18S rRNA gene sequence. Hepatozoon infection could be detected in 38 (70%) out of fifty-four blood samples of Indian sloth bears (captive and wild), suggestive of high prevalence of Hepatozoon infection in Indian sloth bears. Sequencing of partial 18S rRNA gene of the positive samples and BLAST analysis indicated that the nearest phylogenetic neighbour was H. ursi with which they exhibited 99-100% similarity. Additionally, Hepatozoon sp. isolated from wild sloth bears of India were identical to those in captive sloth bears and phylogenetically related to H. ursi reported from Japanese black bears from Japan. To our knowledge, this is the first report on the molecular characterization of H. ursi infection in Indian sloth bears.     

Evaluation and comparison of native and recombinant LipL21 protein-based ELISAs for diagnosis of bovine leptospirosis

A 21-kDa leptospiral lipoprotein (LipL21) was evaluated for its diagnostic potential to detect bovine leptospirosis by ELISA. Both native LipL21 (nLipL21) and recombinant LipL21 (rLipL21) proteins were tested and compared regarding diagnostic efficiency, and no statistically significant difference was observed. The sensitivity of rLipL21 ELISA for 62 microscopic agglutination test (MAT) positive sera was 100% and the specificity with 378 MAT negative sera was 97.09%. Thus, rLipL21 protein-based ELISA could be used as an alternative to MAT for the diagnosis of bovine leptospirosis.     

Serodiagnosis of bovine leptospirosis by IgG-Enzyme-Linked Immunosorbent Assay and Latex Agglutination Test

The efficacy of a recombinant leptospiral outer membrane protein LipL41 as an antigen for conducting IgG-Enzyme linked immunosorbent assay (ELISA) and latex agglutination test (LAT) for serodiagnosis of bovine leptospirosis was evaluated. The recombinant LipL41 antigen developed and used for detecting the antibodies was specific in detection of the pathogenic serovars of Leptospira, as the expression of the LipL41 antigen is restricted only to pathogenic leptospires. A total of 430 bovine serum samples were subjected to IgG-ELISA and LAT, and the sensitivity and specificity were assessed in comparison with microscopic agglutination test (MAT). The sensitivity and specificity of IgG-ELISA and LAT were 86.84% and 93.16%, and 95.42% and 98.33% respectively. Both the tests are found to be sensitive, specific and concurred with the standard MAT. The study concluded that the rLipL41 protein could be used as a potential diagnostic antigen in different assay formats for bovine leptospirosis.     

2010 ACVIM small animal consensus statement on leptospirosis: diagnosis, epidemiology, treatment, and prevention

This report offers a consensus opinion on the diagnosis, epidemiology, treatment, and prevention of leptospirosis in dogs, an important zoonosis. Clinical signs of leptospirosis in dogs relate to development of renal disease, hepatic disease, uveitis, and pulmonary hemorrhage. Disease may follow periods of high rainfall, and can occur in dogs roaming in proximity to water sources, farm animals, or wildlife, or dogs residing in suburban environments. Diagnosis is based on acute and convalescent phase antibody titers by the microscopic agglutination test (MAT), with or without use of polymerase chain reaction assays. There is considerable interlaboratory variation in MAT results, and the MAT does not accurately predict the infecting serogroup. The recommended treatment for optimal clearance of the organism from renal tubules is doxycycline, 5 mg/kg p.o. q12h, for 14 days. Annual vaccination can prevent leptospirosis caused by serovars included in the vaccine and is recommended for dogs at risk of infection.     

Use of machine-learning algorithms to aid in the early detection of leptospirosis in dogs

Leptospirosis is a life-threatening, zoonotic disease with various clinical presentations, including renal injury, hepatic injury, pancreatitis, and pulmonary hemorrhage. With prompt recognition of the disease and treatment, 90% of infected dogs have a positive outcome. Therefore, rapid, early diagnosis of leptospirosis is crucial. Testing for Leptospira-specific serum antibodies using the microscopic agglutination test (MAT) lacks sensitivity early in the disease process, and diagnosis can take >2 wk because of the need to demonstrate a rise in titer. We applied machine-learning algorithms to clinical variables from the first day of hospitalization to create machine-learning prediction models (MLMs). The models incorporated patient signalment, clinicopathologic data (CBC, serum chemistry profile, and urinalysis = blood work [BW] model), with or without a MAT titer obtained at patient intake (=BW + MAT model). The models were trained with data from 91 dogs with confirmed leptospirosis and 322 dogs without leptospirosis. Once trained, the models were tested with a cohort of dogs not included in the model training (9 leptospirosis-positive and 44 leptospirosis-negative dogs), and performance was assessed. Both models predicted leptospirosis in the test set with 100% sensitivity (95% CI: 70.1–100%). Specificity was 90.9% (95% CI: 78.8–96.4%) and 93.2% (95% CI: 81.8–97.7%) for the BW and BW + MAT models, respectively. Our MLMs outperformed traditional acute serologic screening and can provide accurate early screening for the probable diagnosis of leptospirosis in dogs.     

Sloth bear immobilization with a ketamine-xylazine combination: reversal with yohimbine

Five captive sloth bears (Melursus ursinus) were immobilized with a combination of ketamine (5.80 to 9.75 mg/kg of body weight) and xylazine (1.40 to 2.44 mg/kg), given IM. The youngest bear was immobilized twice, 62 days apart; all other bears were immobilized only once. Induction times were 4 to 25 minutes. After completion of various intended procedures, yohimbine (0.125 mg/kg) was administered IV. Arousal times were 2 to 20 minutes and bears were standing in 17 to 51 minutes. Compared with reported recovery times of 2 to 3 hours for bears immobilized with ketamine-xylazine combinations but not given an antagonist, results of the present study indicated that yohimbine reduces anesthesia recovery times in sloth bears immobilized with ketamine-xylazine combination.     

Evaluation of a recombinant LigB protein of Leptospira interrogans serovar Canicola in an enzyme-linked immunosorbent assay for the serodiagnosis of bovine leptospirosis

A recombinant leptospiral lipoprotein, LigB, was evaluated for use in the diagnosis of bovine leptospirosis by enzyme-linked immunosorbent assay (rLigB IgG ELISA). The standard reference test (Microscopic agglutination test, MAT) of 200 serum samples from cattle suspected of leptospirosis showed that 95 (47.5%) samples had positive agglutination titres, which ranged from 100 to 1600. In rLigB IgG ELISA, 49% of the samples were positive. Sensitivity of IgG ELISA for 95 bovine sera, which had MAT titres of greater than or equal to 100, were 100%. ELISA showed a specificity of 97.1% with 105 bovine sera, which were negative at a 1:50 dilution in MAT for Leptospira interrogans serovars. The results of ELISA and MAT correspond very good. When analytical specificity of IgG ELISA was evaluated using bovine serum samples from animals showing the serum antibodies to other pathogens, no cross-reaction was observed. Thus the recombinant LigB IgG ELISA can be used instead of the MAT as an aid to the diagnosis of bovine leptospirosis.     

猪乙型脑炎乳胶凝集试验与血凝抑制试验方法的比较

用血凝抑制试验 (HI)和乳胶凝集试验 (LAT)两种方法检测乙型脑炎弱毒疫苗免疫猪血清 ,结果均呈阳性反应。用LAT对来自 12个猪场的 94份猪血清进行了乙型脑炎病毒 (JEV)抗体检测 ,并与HI进行了对比 ,两种方法检测结果阳性符合率和总符合率分别为 90 .3% (5 6 /6 2 )和 88.3% (83/94 ) ,两种方法检测结果差异不显著 (p >0 .0 5 )。对来自无乙型脑炎的 12头健康猪血清进行检测 ,两种方法检测结果均为阴性。结果表明 ,用LAT与HI检测乙型脑炎结果符合 ,前者更为简便和实用。     

Prevalence of pathogenic Leptospira spp. in sheep in a sheep-only abattoir in New Zealand

AIM: To determine the prevalence of the two most commonly diagnosed pathogenic Leptospira spp. serovars, Hardjobovis and Pomona, in sheep in a sheep-only abattoir in New Zealand, and to determine the prevalence of kidneys which were leptospire culture-positive collected from sheep seropositive or seronegative to the microscopic agglutination test (MAT). METHODS: A repeated cross-sectional observational study was conducted of serological and kidney culture prevalences of Leptospira borgpetersenii serovar Hardjobovis and Leptospira interrogans serovar Pomona. Lines of sheep and individual sheep were systematically randomly selected at a sheep-only abattoir during 18 May 2004 to November 2004 and 06 December 2004 to 14 June 2005. Additionally, a cross-sectional study examined prevalences in a purposively selected line of sheep from a flock with clinical evidence of an outbreak of leptospirosis. RESULTS: In the study population of 15,855 sheep of which 2,758 were sampled, 5.7 (95% CI=4.9-6.7)% were seropositive to one or both serovars; 44.2 (95% CI=34.6-54.2)% of 95 lines of sheep and 44.9 (95% CI=35.0-55.3)% of 89 farms showed serological evidence of infection. The serological prevalence of serovar Hardjobovis was significantly higher than that of serovar Pomona both at line (33% and 4%, respectively) and individual (5% and 1%, respectively) levels. A low but persistent seroprevalence of Hardjobovis throughout both years suggested low-level endemicity to this serovar, whereas Pomona infections appeared to be sporadic. Leptospires were isolated from kidneys of 8/37 (22%) Hardjobovis- and 1/6 (17%) Pomona-seropositive, and 5/499 (1%) seronegative animals. Of the animals purposively sampled from a farm with a clinical outbreak of leptospirosis, all kidneys from the 13 seropositive animals were culture-positive, indicating a high risk of exposure of meat workers in outbreak situations. Kidneys of MAT-seropositive sheep were 21.7 (95% CI=7.6-61.9) times more likely to test culture-positive than kidneys from animals with negative MAT titres. In general, the results indicated that 13/1,000 sheep slaughtered were potentially shedding leptospires. CONCLUSIONS: The study demonstrated the presence of a definite risk of occupational exposure of meat workers in a sheep-only slaughterhouse to the two most commonly diagnosed pathogenic Leptospira spp. serovars in New Zealand.     

Variability in results of the microscopic agglutination test in dogs with clinical leptospirosis and dogs vaccinated against leptospirosis

Background: The microscopic agglutination test (MAT) is commonly used for the diagnosis of canine leptospirosis. In dogs it is sometimes suggested that the serogroup with the highest MAT titer is the infecting serogroup; however, this is not true in humans with confirmed leptospirosis. We sought to investigate the value of MAT results in predicting the infecting serogroup by comparing results across several laboratories and within individual dogs over time. Objectives: To examine the variability in MAT results across different laboratories in dogs recently vaccinated against leptospirosis, and in dogs with clinical leptospirosis, and to investigate variability over time in MAT results in individual dogs with leptospirosis. Animals: Eighteen dogs from a research colony, 9 of which had been vaccinated against leptospirosis, and 17 dogs clinically diagnosed with leptospirosis. Methods: Serum samples were submitted to up to 5 veterinary diagnostic laboratories for MAT titers from each dog on at least 1 occasion. MAT results also were followed over time in 6 dogs diagnosed with leptospirosis. Results: MAT results were discordant across different laboratories in dogs recently vaccinated against leptospirosis and in dogs with clinical leptospirosis. MAT results varied over time in individual dogs with the disease. Conclusions and Clinical Importance: The MAT is a valuable test for the diagnosis of leptospirosis in dogs, but it is unlikely that test results can be used to predict the infecting serogroup. Laboratories offering the MAT should consider participation in a proficiency scheme.     

乳胶凝集试验与血清中和试验检测猪伪狂犬病血清抗体的 …

应用血清中和试验（ＳＮＴ）和伪狂犬病乳胶凝集试验（ＬＡＴ）诊断试剂盒对两种伪狂犬病是性血清、伪狂犬病病毒（ＰＲＶ）高兔血甭及６０份被检猪血清进行了ＰＲＶ抗体效价测定和相关性分析,两种方法测得的抗体效价之间呈强相关性（ｒ＝０．９６）,且ＬＡＴ效价比ＳＮＴ一般高出一个滴度;能干为自３５个猪场的４１４份猪血清进行了ＰＲＶ抗体检测,并与ＳＮＴ检测结果进行了对比,结果在ＳＮＴ检测为阳笥的１７１份血清中,ＬＡ     

检测猪细小病毒血清抗体乳胶凝集试验方法的建立及初步应用

将猪细小病毒在IBRS-2细胞上同步培养增殖,待出现细胞病变后,反复冻融,收获病毒液,用甲醛灭活。随后用经硫酸胺沉淀、透析后浓缩的细小病毒液进行方阵滴定,选择致敏乳胶的最佳条件,制成细小病毒乳胶凝集试验（LAT）抗原。抗原与细小病毒阳性血清反应出现肉眼可见的凝集颗粒,而与生理盐水、PBS、犊牛血清、猪瘟、衣原体、口蹄疫、伪狂犬病、弓形体及萎缩性鼻炎等阳性血清不出现凝集现象。用所建立的细小病毒乳胶凝集试验（LAT）与血凝抑制试验检测了203份猪血清,其中血凝抑制抗体阳性为161份,阴性为42份,阳性率为79.31％;乳胶凝集抗体阳性为150份,阴性为53份,阳性率为73.89％,两种方法阳性符合率为93.16％,经统计学检验P＞0.05,两者差异不显著。结果表明,乳胶凝集试验可以作为临床上大量血样进行血凝抑制试验前的初筛,具有简便、准确的优点,在检测细小病毒（PPV）抗体上具有较好的应用前景。     

Toxoplasma gondii and Leptospira spp. infection in free-ranging armadillos

Armadillos are primitive mammals used as food, mostly in rural areas. These animals may be sources of toxoplasmosis and leptospirosis infection for humans, but there is little information about their potential risk as reservoirs. In order to determine the prevalence of armadillos infected by Toxoplasma gondii and Leptospira spp., serum samples of 31 nine-banded armadillos (Dasypus novemcinctus), three six-banded armadillos (Euphractus sexcinctus), two naked-tailed armadillos (Cabassous tatouay) and two long-nosed armadillos (D. hybridus), captured in the mid-west region of the state of S?o Paulo, were analyzed for leptospirosis using the Microscopic Agglutination Test (MAT-l), and for toxoplasmosis using the Modified Agglutination Test (MAT-t). Only 4/31 (12.90%) nine-banded armadillos were positive for T. gondii, while 3/31 (9.68%) nine-banded armadillos and 1/3 (33.33%) six-banded armadillos presented antibodies to Leptospira spp., demonstrating the potential risk of T. gondii and Leptospira spp. transmission to humans, mainly due the habit of eating the meat of these animals in rural areas.     

Kidney Injury Molecule-1 in the detection of early kidney injury in dogs with leptospirosis

Renal damage, a common feature in canine leptospirosis, ranges from a subclinical affection to kidney dysfunction and death. Chances of recovery can be improved by early intervention. However, traditional biomarkers (serum urea and creatinine) have limited relevance for precocity. Kidney Injury Molecule-1 (KIM-1) is a transmembrane protein upregulated in early stages of tubular injury. This study evaluated the use of urinary KIM-1 to detect early renal injury in naturally occurring canine leptospirosis. This exploratory research included 30 dogs divided into two groups: (1) dogs with leptospirosis (n = 25) and (2) healthy dogs (n = 5). Leptospira sp. infection was diagnosed through urine PCR and/or direct bacteriologic culture and/or serology (single MAT titters ≥800). Additionally, stage of infection was further characterized in acute and subacute phases based on the onset of clinical symptoms from 3 to 7 days. Urinary KIM-1 (uKIM-1) concentrations were measured in both groups with a commercial canine ELISA kit.uKIM-1 levels were statistically different (P < 0.01) between the studied groups, especially in non-azotemic dogs (P = 0.0042). The biomarker showed 88 % sensibility to diagnosis of kidney injury at> 1.49 ng/mL cut-off. Urine KIM-1 was negatively correlated with urine specific gravity (USG) but accompanied histopathological evidence of renal degeneration, necrosis and regeneration processes, extending information on kidney health. Measurement of KIM-1 in the urine of canine patients was able to detect naturally occurring acute and subacute leptospirosis accompanied by tubular injury in early non-azotemic infections.     

猪繁殖与呼吸综合症血清抗体的检测报告

2001年从四川省6个市7个种猪场采集未经免疫猪繁殖与呼吸综合征（PRRS）疫苗的种猪血清样本598份，用乳胶凝集试验检测PRRS抗体，结果表明，平均阳性率为19．56％，最低的为0，最高者达63％，阳性场占6／7。     

乳胶凝集试验检测鸡传染性鼻炎血清抗体的研究

将Ａ型和Ｃ型鸡副嗜血杆菌培养后,菌落计数,离心计数,离心后用０．０１ＭｐＨ７．４磷酸盐缓冲液洗涤菌体２次。经过对致敏温度、致敏时间、致敏乳胶的菌体浓度及菌体的处理方式等条件的选择,建立了检测鸡传染性鼻炎血清抗体的乳胶凝集试验。与琼脂扩散试验相比,特异性一致,但更加敏感,且操作简便、快速。结果说明,该方法有较好的应用前景,尤其适用于本病的现场快速诊断。     

Serological reactions of leptospirosis-positive (MAR and CFT) bovine sera in ELISA

The collection of test sera for measuring ELISA results was composed of bovine sera with MAT titres of greater than or equal to 1:200 in the leptospirosis MAT and of greater than or equal to 1:5 in the CFT together with sera from a serologically negative and clinically non-suspicious cattle herd. To establish cut-off ODs, the geometric mean net-extinction of the negative serum collection plus 1, 2, and 3 standard deviations were calculated. By comparison of 3 different conjugates from rabbits, it was demonstrated that results from anti-total bovine Ig were superior to anti-IgG and anti-IgM conjugates. Considerations regarding sensitivity and specificity led to the recommendation to use a test serum dilution of 1:160, to apply anti-total bovine Ig conjugates, and to establish the cut-off OD at the geometric mean net-extinction of negative sera plus 3 standard deviations. Under such conditions, agreement between leptospirosis MAT/CFT positivity on the one side and ELISA positivity on the other was reached in 74%. This recommendation is made for cross-sectional studies but not for examinations of clinically suspicious cattle herds.     

Renal and biochemical changes produced in broilers by high-protein, high-calcium, urea-containing, and vitamin-A-deficient diets

Three hundred 18-day-old male chicks (Arbor Acre) were divided into five groups of 60 each and given high-protein (42.28%), high-calcium (3.37%), urea-containing (5%), vitamin-A-deficient, or control diets to study the effect of nutritional imbalances on the development of nephritis and related biochemical changes over 15 weeks. The first four diets increased the levels of glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, uric acid, and nonprotein nitrogen in serum. Blood urea was increased by only the urea diet. Hypoglycemia and a decrease in hepatic glucose-6-phosphatase were also observed in chicks fed the first four diets. The vitamin-A-deficient diet resulted in a depletion of vitamin A in the liver and kidneys. These changes were directly correlated with the prolonged feeding of experimental diets and also with the severity of nephritis and degenerative changes in various organs. It was concluded that increasing the intake of nitrogen or calcium in order to increase production may in fact have the opposite effect, leading to degenerative changes in various tissues and to nephritis.