Development and evaluation of a rapid absorbed enzyme immunoassay test for the diagnosis of Johne''s disease in cattle

An absorbed enzyme immunoassay (EIA) test for Johne's disease in cattle was developed in which absorption of cross-reacting antibodies occurred as a rapid reaction in solution rather than overnight with whole organisms and a subsequent centrifugation step. Total test time was reduced to less than 2 h with a minimum of manipulations. The test was evaluated in cattle herds from Johne's disease-endemic and Johne's disease-free regions of Australia. Specificity was 99.8%. Calculations of sensitivity were affected by the history of the herd under test. However, the EIA detected in excess of 80% of animals before onset of clinical disease and 65% of faecal shedders were EIA positive on, or before, first detection of Mycobacterium paratuberculosis in their faeces. The test should aid epidemiological studies and be a useful tool in the management and control of Johne's disease.     

Surgical repair of peripheral detachment of the medial meniscus in 34 cattle

A detachment of the periphery of the medial meniscus from the joint capsule and medial collateral ligament in 34 cattle was repaired by securing the meniscus to the joint capsule with vertical mattress sutures during arthrotomy. Of 28 cattle with follow-up reports, 20 had recovered satisfactorily by their owners' evaluations; 17 were either not lame or slightly lame after the operation and three were moderately lame. Three of four cattle without follow-up reports were either not lame or slightly lame when discharged from the hospital.     

Comparison of the complement-fixation and agar gel immunodiffusion tests for diagnosis of subclinical bovine paratuberculosis

The performance of the serum complement fixation (CF) test was compared with that of a serum agar gel immunodiffusion (AGID) test on 74 subclinically infected and 154 uninfected cattle in 6 commercial midwestern dairy herds with Mycobacterium paratuberculosis infection and on 30 cattle in a herd that was free of infection. Infection status of cattle within herds was established by performance of a series of 3 or more fecal cultures and of ileocecal lymph node cultures of culled cattle. In cattle with subclinical infection detected by culturing, the sensitivity estimates of the CF and AGID tests were 10.8% (3.6% SE) and 18.9% (4.5% SE), respectively. In the cattle classified as disease free, the specificity estimates of the CF and AGID tests were 97.4% (1.3% SE) and 99.4% (0.6% SE), respectively. Neither set of estimates was significantly different. Negative test results obtained with the use of either test in apparently normal cattle from suspect herds should be interpreted with caution because both tests suffer from low sensitivities in subclinically infected animals. However, the AGID test may be more useful in regulatory situations in which the CF test is currently used because the AGID test is easier to perform and to interpret.     

Confirmation of vertical transmission of bovine immunodeficiency virus in naturally infected dairy cattle using the polymerase chain reaction.

The purpose of this study was to determine whether bovine immunodeficiency virus (BIV) is vertically transmitted in naturally infected dairy cattle. Twenty-two dam/calf pairs from a Mississippi Agriculture and Forestry Experiment Station dairy were the study group. Blood samples were collected following delivery of calves, the peripheral blood leukocytes were purified from these samples, and the leukocyte DNA was used in polymerase chain reactions targeting the pol gene region of the BIV provirus. Southern blotting and hybridization were used to confirm the BIV specificity of the amplified fragments. BIV provirus was detected in 14 of 22 calves (64%), demonstrating vertical transmission. Eight of the calves were disqualified from the final interpretation of transplacental transfer because they may have nursed their mothers prior to blood collection, allowing the possibility of lactogenic transfer of the virus. Transplacental transmission of BIV was identified in 6 of 22 calves (27%).     

Effect of Weight Gain and Subsequent Weight Loss on Glucose Tolerance and Insulin Response in Healthy Cats

The effects of weight gain and subsequent weight loss on glucose tolerance and insulin response were evaluated in 12 healthy cats. Intravenous glucose tolerance tests (IVGTT) were performed at entry into the study, after a significant gain of body weight induced by feeding palatable commercial cat food ad libitum, after a significant loss of body weight induced by feeding a poorly palatable purified diet to discourage eating and promote fasting, and after recovery from fasting when body weight had returned to pre-study values and cats were eating commercial foods. A complete physical examination with measurement of body weight was performed weekly, a CBC and serum biochemistry panel were evaluated at the time of each IVGTT, and a liver biopsy specimen obtained 2 to 4 days after each IVGTT was evaluated histologically for each cat. Mean serum glucose and insulin concentrations after glucose infusion and total amount of insulin secreted during the second 60 minutes and entire 120 minutes after glucose infusion were significantly (P > .05) increased after weight gain, as compared with baseline. At the end of weight loss, cats had hepatic lipidosis and serum biochemical abnormalities consistent with feline hepatic lipidosis. There was a significant (P > .05) increase in mean serum glucose concentration and t_1/2, and a significant (P > .05) decrease in mean serum insulin concentration and the glucose disappearance coefficient (K) after glucose infusion for measurements obtained after weight loss, compared with those obtained after weight gain and at baseline. Insulin peak response, insulino-genic index, and total amount of insulin secreted during the initial 10 minutes, 20 minutes, and 60 minutes after glucose infusion were decreased markedly (P > .05), compared with measurements obtained after weight gain and at baseline. In addition, the total amount of insulin secreted for 120 minutes after glucose infusion was decreased markedly (P > .05) in measurements obtained after weight loss, compared with those obtained after weight gain. At the end of recovery, all cats were voluntarily consuming food, serum biochemical abnormalities identified after weight loss had resolved, the number and size of lipid vacuoles in hepatocytes had decreased, and results of IVGTT were similar to those obtained at baseline. These findings confirmed the reversibility of obesity-induced insulin resistance in cats, and documented initial deterioration in glucose tolerance and insulin response to glucose infusion when weight loss was caused by severe restriction of caloric intake.     

Treatment of proximal hind-limb lameness in cattle

Lameness of the hind limb (tarsus and proximal) in cattle is considered from the viewpoint of diagnosis and prognosis. Indications and techniques for radiography are discussed, as well as when indicated.     

Serum chemistry alterations in Alaskan sled dogs during five successive days of prolonged endurance exercise

OBJECTIVE: To determine the impact of successive days of endurance exercise on select serum chemistry values in conditioned Alaskan sled dogs. DESIGN: Prospective cohort study. ANIMALS: 10 conditioned Alaskan sled dogs. PROCEDURES: All dogs ran 160 km/d for 5 consecutive days. Serum was obtained prior to exercise and immediately after each exercise run; all samples were obtained before dogs were fed. Serum electrolyte, mineral, protein, total bilirubin, urea nitrogen, creatinine, and cardiac troponin-I concentrations and serum alkaline phosphatase, alanine aminotransferase, creatine kinase, and aspartate aminotransferase activities were measured. Data were analyzed by means of analysis of covariance for a randomized complete block design with dog as a blocking variable, time as a covariate, and distance run as the treatment of interest. Least square mean values were compared with values obtained prior to exercise, and linear and quadratic contrasts were examined. RESULTS: Serum globulin concentration was low prior to exercise (mean +/- SD, 2.2 +/- 0.3g/dL) and progressively decreased as exercise continued. Exercise was associated with increases in serum chloride, urea nitrogen, and cardiac troponin-I concentrations and serum alanine aminotransferase, creatine kinase, and aspartate aminotransferase activities and with progressive decreases in serum potassium, total protein, and albumin concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that multiple successive days of endurance exercise resulted in mild aberrations in serum chemistry variables in conditioned sled dogs. Changes likely reflected the metabolic stresses of prolonged endurance exercise as well as dietary composition. Hypoglobulinemia in resting, conditioned sled dogs may reflect the immunosuppressive or catabolic effects of intense endurance training.     

A direct method for fatty acid methyl ester synthesis: application to wet meat tissues, oils, and feedstuffs

A simplified protocol to obtain fatty acid methyl esters (FAME) directly from fresh tissue, oils, or feedstuffs, without prior organic solvent extraction, is presented. With this protocol, FAME synthesis is conducted in the presence of up to 33% water. Wet tissues, or other samples, are permeabilized and hydrolyzed for 1.5 h at 55 degrees C in 1 N KOH in MeOH containing C13:0 as the internal standard. The KOH is neutralized, and the FFA are methylated by H(2)SO(4) catalysis for 1.5 h at 55 degrees C. Hexane is then added to the reaction tube, which is vortex-mixed and centrifuged. The hexane is pipetted into a gas chromatography vial for subsequent gas chromatography. All reactions are conducted in a single screw-cap Pyrex tube for convenience. The method meets many criteria for fatty acid analysis, including not isomerizing CLA or introducing fatty acid artifacts. It is applicable to fresh, frozen, or lyophilized tissue samples, in addition to oils, waxes, and feedstuffs. The method saves time and effort and is economical when compared with other methods. Its unique performance, including easy sample preparation, is achieved because water is included rather than eliminated in the FAME reaction mixtures.     

中国水稻白叶枯病菌系染色体DNA的RFLP谱型的初析

用２个ＤＮＡ探针ｐＪＥＬ１０１和ｐＢＡＳａｖｒＸａ１０对７８个水稻白叶枯病菌系进行ＲＦＬＰ分型，以分析其群体结构和遗传多样性。分别鉴定出１６种ＲＦＬＰ标记带的谱型。以彼此的带位相似率达８５％为界，可分为簇。参试菌系的群体遗传多样性为０．７７（用ｐＪＥＬ１０１），和０．８３ＲＦＬＰ谱型分析表明：我国多数病原型为杂合组群。白叶枯病菌系的分子表现型的变异，远远大于致病型，两个探针都能有效分析我国菌系的群