Use of the brucellosis card test for screening cattle in Saskatchewan.

One group of 28,714 bovine sera were tested by both the brucellosis tube serum agglutination test and the brucellosis card test. The tube serum agglutination test confirmed 99.8% of the negative brucellosis card test results. The brucellosis card test identified 63% of the tube serum agglutination test reactors. In a second group of 496 sera reacting to either the tube serum agglutination test, complement fixation test, plate serum agglutination test or acid antigen serum agglutination test the brucellosis card test identified 99.1% of the complement fixation test positive sera and 91.3% of the sera reacting to any of the other serological tests. The brucellosis card test showed satisfactory agreement with both the complement fixation test and tube serum agglutination test. It appears to be a useful screening test in operations involving large numbers of animals since under these conditions the reactors can be quickly identified and isolated.     

Study on Sterility Test Method of Ivermectin Microemulsion

The assay was aimed to study the sterility test and validation test of ivermectin microemulsion preparation and establish a sterility test method for ivermectin microemulsion preparation.The test method was carried out according to the method in volumeⅠ, Chinese Veterinary Pharmacopoeia Edition 2005.By choosing positive control bacteria and defining washing volumes in sterility test, the membrane-filter method was used to test the quantity of 10 bottles of test samples, and the sterility test was established.The result of method validation test showed that the test and all of positive control bacteria and microorganism growth after each filter being washed with 400 mL 0.1% peptone solution.It illustrated that the samples had no antimicrobial activity under the sample quantity and test condition.This method was available for sterility test of ivermectin microemulsion preparation.Using this method to test three lots test samples, the results showed that the positive control bacteria grew well within 24 h.The negative control bacteria and three lots test samples were sterile.It indicated that sterility test results met the requirements.     

伊维菌素微乳无菌检查方法学的研究

本试验旨在对伊维菌素微乳制剂进行无菌检查方法学验证和无菌检查试验,确认本试验所用的方法适用于该制剂的无菌检查。按《中国兽药典》2005版一部(附录118)所载"无菌检查法"项下进行试验,通过对阳性对照菌、不同量冲洗液等条件的选择,采用薄膜过滤法对10瓶供试品(每种试验菌的样品量)进行检测,建立了无菌检查方法。经方法验证,用400mL 0.1%蛋白胨水溶液冲洗后,含供试品容器中的7个阳性菌试验组与阳性菌对照组相比均生长良好,说明供试品的该检验量在该检验条件下无抑菌作用或其抑菌作用可以忽略不计,可以用该方法进行供试品的无菌检查。对3批供试品进行无菌检查,阳性对照菌均在24h内生长良好,阴性对照均澄清,无菌生长,3批供试品均澄清,无菌生长,无菌检查试验结果符合规定。     

猪轮状病毒阳性血清(G5型)制备及初步鉴定

采用常见猪病毒抗体阴性仔猪制备了猪轮状病毒G5型阳性血清,经冻干鉴定,其性状、无菌检验、特异性检验、剩余水分测定、真空度测定结果均符合要求,效价高达1∶11482,为我国猪轮状病毒活疫苗或其联苗成分的外源病毒检验、鉴别检验提供阳性血清,以满足疫苗检验需求。     

Validation of the indirect fluorescent antibody and the complement fixation tests for the diagnosis of Theileria equi

The indirect fluorescent antibody (IFA) test for Theileria equi was evaluated to assess test's suitability for the serological diagnosis of equine piroplasmosis, to provide performance parameters for the purpose of test validation, and to compare it with the complement fixation (CF) test. Using a protocol that included Evan's blue, the specificity of the IFA test was estimated at 99.0% for T. equi by the classical method of analysis, and 96.6% by the Bayesian method. The use of Evan's blue in the test protocol increased test specificity and contributed to an excellent test agreement between two collaborating laboratories (kappa = 0.96). Using Bayesian analysis, the sensitivity estimate for the IFA test was 89.2%. The CF test sensitivity and specificity estimates for T. equi were 63.1 and 96.4%, respectively, as determined by Bayesian analysis. The IFA test was more sensitive than the CF test but the specificity estimates were similar.     

Comparison of a modified counterimmunoelectrophoresis test and a microimmunodiffusion test for detection of pseudorabies virus antibodies in porcine sera.

The correlation of a modified counterimmunoelectrophoresis (CIE) test and a microimmunodiffusion test for detecting pseudorabies virus antibodies in porcine sera was investigated, using as reference a standard virus neutralization test. The counterimmunoelectrophoresis test exhibited a sensitivity comparable to the microimmunodiffusion test but was not as sensitive as the virus neutralization test. The best feature of the modified counterimmunoelectrophoresis test is that it is a rapid test. It provides an alternative to currently used diagnostic tests for detection of pseudorabies virus antibodies in sera from field reared and experimentally reared swine exposed to pseudorabies virus.     

An improved immunodiffusion test for antibodies to bovine leukemia virus antigens

A new immunodiffusion (ID) test for antibodies to bovine leukemia virus (BLV) antigens was established. This test, refered to as the tannic acid enhanced, indirect, double immunodiffusion (IID-T) test, includes the following steps: (a) double micro-immunodiffusion using diluted references reagents, (b) treatment of the gel plate with antiglobulin serum, (c) treatment of the gel plate with 1% tannic acid. The IID-T test has proved to be eight times more sensitive than the double immunodiffusion test (ID) commonly used for anti-BLV. Diagnostic efficiency at individual levels of the IID-T test for anti-gpBLV is higher than this parameter of the ID test for anti-gpBLV and radioimmunoassay (RIA) for anti-p24BLV. The IID-T test is simple, reproducible, and more economic than the ID test in the amount of the reference reagents required. The IID-T test is more reliable than the ID test especially when weak, low titer sera are tested. Thus, the IID-T test seems to be suitable for large scale serodiagnosis of BLV infection in cattle.     

Performance of 2 microtiter canine Coombs' tests

BACKGROUND: The Coombs' test, which detects immunoglobulin or complement on RBC surfaces, has long been the standard for laboratory confirmation of immune-mediated hemolytic anemia (IMHA), a common cause of hemolytic anemia in the dog. This test, however, suffers from relatively low sensitivity. Optimization of test sensitivity would lead to fewer discrepancies between laboratory results and clinical diagnoses, and in some cases institution of appropriate therapy in a timely manner. OBJECTIVES: The objectives of this study were to 1) characterize the sensitivity and specificity of 2 canine Coombs' tests for the detection of IMHA, 2) document the efficacy of using multiple antisera dilutions beyond what is directed by manufacturers, and 3) evaluate the necessity of monovalent antisera in the test protocol. METHODS: Sixty-five canine whole-blood samples submitted for Coombs' testing were evaluated. Patients were classified as IMHA positive or negative based on a set of predetermined criteria. IMHA classification was compared to Coombs' test results from 2 different Coombs' tests adapted to a microtiter-plate format. One Coombs' test (VMRD Coombs' test) utilized a single polyvalent antiserum (VMRD, Inc, Pullman, WA, USA), while a second Coombs' test (University of Minnesota [U of MN] Coombs' test) used both polyvalent and monovalent antisera. RESULTS: Sensitivity and specificity were 61% and 100% for the VMRD Coombs' test, and 82% and 95% for the U of MN Coombs' test. The use of multiple antisera dilutions resulted in 6 additional Coombs' positive test results. All positive Coombs' test results were positive by polyvalent antisera. CONCLUSIONS: When used in a microtiter-plate format, the U of MN Coombs' test was a more sensitive test for the detection of IMHA in canine patients when compared to the VMRD Coombs' test. The use of multiple antisera dilutions increased test sensitivity. Sensitivity, however, was not increased by the use of monovalent antisera in the Coombs' test protocol.     

Accuracy of an indirect fluorescent-antibody test and of a complement-fixation test for the diagnosis of Babesia caballi in field samples from horses

We evaluated the indirect fluorescent-antibody (IFA) test and complement-fixation (CF) test for diagnosis of equine piroplasmosis in the absence of a gold standard. Using Evan's blue, we estimated the specificity of the IFA test on a parasite-free, field horse population to be 98% (95% confidence interval=97, 99). We observed an excellent test agreement (kappa=0.83) between two collaborating laboratories when the IFA test was performed on identical samples from an endemic area. Using Bayesian analysis with informative prior probability distributions, we estimated the sensitivity of the IFA test to be 92% (95% probability interval, PI=81, 98), and specificity to be 95% (95% PI=88, 99). The CF test sensitivity and specificity estimates were 28% (95% PI=15, 47) and 99% (95% PI=96, 100), respectively. We found the IFA to be superior to the CF test, and the inclusion of Evan's blue in test protocol improved the performance of the IFA test. We conclude that the IFA test for Babesia caballi is a sensitive and specific test for the diagnosis of equine piroplasmosis.     

低频电子耳标协议一致性测试系统研究

电子耳标的低频一致性测试是在测试步骤、测试装置、测试环境的限定下,对协议符合性进行的测试。本文选定改进的激励/响应测试模式,硬件层选取合适的模块化硬件,软件层包括RFID协议仿真软件及RFID协议一致性测试软件。根据电感耦合原理计算线圈电感,进而设计天线相关参数。最终完成低频电子耳标协议一致性测试系统的搭建,测试过程符合ISO24631.1的要求。     

An evaluation of the delayed-type hypersensitivity test for diagnosing brucellosis in individual cattle: a field study

A field study was conducted to evaluate the delayed-type hypersensitivity (DTH) test in diagnosing brucellosis in cattle, in particular the diagnosis of infection in individual cows. A total of 93 cows that were negative, suspect, or positive to the serum agglutination test (SAT), complement fixation test (CFT), or the milk ring test (MRT) were subjected to the DTH test. The cows were then slaughtered and the supramammary lymph nodes were collected for bacteriologic examination. In 989 cows the DTH test, MRT and serologic tests were negative. When the DTH test results were compared with bacteriologic results, 12 of the 93 cows with CFT titres greater than 1:200 tested negative in the DTH test while bacteriologic results were positive. The sensitivity of the DTH test (calculated on the remaining 81 cows) was 100%; the specificity was 83%. The sensitivity of the DTH test (calculated on 93 cows) was 81%; the specificity was 83%. The sensitivity and specificity of the DTH test correlated well with those of the CFT (86-83%). We conclude that the DTH test is very sensitive, and specific enough to diagnose brucellosis in individual cows. The DTH test should be used in combination with serologic tests in the diagnosis of brucellosis.     

Establishment of a potency test by ELISA for a rabies vaccine for animal use in Japan

The ELISA we developed was able to determine the antigen content and was suitable for a potency test, and we described a relative potency assay method which determines the potency of test vaccines by comparing the ELISA value of a test vaccine to that of a reference vaccine. In the present study, we standardized the reference vaccine used for determining the potencies of test vaccines, and established a potency test by ELISA. We evaluated the proposed reference vaccine by the neutralizing antibody responses in dogs after vaccination, by the challenge protection test in guinea pigs (GP potency test), which is the earlier official potency test used in Japan, and by the NIH potency test, which is widely used throughout the world. The results showed that a 4-fold dilution of the proposed reference vaccine induced sufficient immunity in dogs. A 3-fold dilution of the proposed reference vaccine passed the GP potency test. The international units (IU) calibrated by the NIH potency test were 3.7 IU/dose. From the results and the WHO recommendation that veterinary rabies vaccines should have a potency of at least 1.0 IU/dose, we determined to dilute the proposed reference vaccine by 3 fold and regarded it as the reference vaccine. Finally, we confirmed that there is a good agreement between the results of the potency test by ELISA and the results of the GP potency test. The establishment of the potency test by ELISA has made it possible to monitor the potency in the production process and has contributed to the stable production of the vaccine.     

猪链球菌2型分离株的生物学特性试验

对从济南某猪场急性死亡的病例中分离得到的猪链球菌2型病原菌,进行生化试验、药敏试验及致病性试验研究。生化试验表明其各项指标均符合链球菌的标准,药敏试验结果表明该菌对多种抗菌素均有耐药性,对复方新诺明、氨苄西林、恩诺沙星等高度敏感,动物致病性试验表明能100%致死小白鼠。     

Potency test for inactivated rabies vaccine (author's transl)

Vaccination schedules and a challenge carried out in laboratory animals are necessary to test the Rabies vaccines for potency.Two main techniques, described by W.H.O., are compared:—The Habel test is a system involving a constant vaccine and a variable virus.—The N.I.H. test is the inverse system with a few major differences during the immunization phase.Although the Habel test is easier to implement, it is quite impossible to make a rigourous statistical interpretation of its results.Although it is more difficult to implement the N.I.H. test as it requires a reference vaccine and a titrated viral suspension, the statistical interpretation is simple.For a same vaccine, the variation amplitude is 1 to 250 for the Habel test and 1 to 7,6 for the N.I.H. test. The N.I.H. test is preferred to the Habel test     

Diagnosis of tuberculosis due to Mycobacterium bovis in New Zealand red deer (Cervus elaphus) using a composite blood test and antibody assays

A blood test for tuberculosis in deer was developed as an ancillary test to clarify the status of skin test-positive deer, with non-specific sensitisation following exposure to saprophytic mycobacteria. The blood test incorporates the measurement of the relative humoral and cellular immunological responses to Mycobacterium bovis and M. avium antigens to provide a composite test with high levels of sensitivity (>95%) and specificity (>98%). The specificity of the test has allowed it to be used in parallel with the skin test to salvage thousands of tuberculosis-free deer with non-specific skin test-positive reactions, while its high sensitivity has consistently identified M. bovis-specific reactivity in tuberculous skin test-positive animals. The rules for establishing the diagnostic parameters for the cellular and antibody assays were developed by retrospective analysis of the laboratory results using blood samples from many thousand tuberculous or disease-free deer. The sensitivity of the blood test was tested in this study using 150 animals with tuberculosis diagnosed by the isolation of M. bovis. It had sensitivity values of 95.7-95.9% in herds with a low (<2.0%) or a high (>30.0%) incidence of tuberculosis. The test had a specificity of 98.0% when tested on 218 disease-free animals, 118 of which were skin test-positive. An antibody test was developed to diagnose M. bovis in skin test-negative anergic deer from tuberculosis infected herds. When this test was used with deer blood taken 10 days after reading the skin test, it had a sensitivity of 85.3% for 102 M. bovis-positive deer. When used in combination with skin test, the antibody test complemented the skin test to raise the sensitivity of the combined tests to 95.0%, when antibody-positive or skin test-positive tests were used to diagnose tuberculosis. The specificity of the antibody test was 100% when used to evaluate 218 disease-free deer from non-infected herds.     

牛型结核菌素皮内变态反应试验和γ-干扰素ELISA试验检测奶牛结核病的比较研究

本研究采用PPD皮内变态反应试验和γ-干扰素ELISA试验,对甘肃省3个地区的1585头奶牛进行结核病检测.结果表明,PPD皮内变态反应共检出7份阳性样品;经γ-干扰素ELISA检测2份为阳性、其余5份为假阳性或禽型阳性,假阳性或禽型阳性样品再经细菌分离鉴定表现为阴性;PPD皮内变态反应检出的21份疑似样品再经γ-干扰素ELISA检测,表现为禽型阳性、假阳性或阴性;PPD皮内变态反应阴性样品经γ-干扰素ELISA试验检测,结果为阴性或禽型阳性.在检测奶牛结核病时,PPD皮内变态反应试验特异性较差,γ-干扰素ELISA试验结果与牛结核分枝杆菌细菌分离鉴定结果一致,而且该技术敏感性、特异性和鉴别假阳性均优于PPD皮内变态反应试验.     

Comparison of results of three commercial heartworm antigen test kits in dogs with low heartworm burdens

OBJECTIVE: To compare results of 3 commercial heartworm antigen test kits performed on serum samples from dogs infected with low numbers of adult female heartworms. DESIGN: Blinded laboratory evaluation. Sample Population-Serum samples from dogs (n = 208) proven at necropsy to be infected with 1 to 4 adult female heartworms and from dogs (32) without heartworms. PROCEDURE: Samples were sequentially tested with each test kit, following the manufacturers' instructions, by licensed veterinary technicians in private practice who were not aware of infection status of the dogs. The order of test kit evaluations was randomly chosen. For each test kit, sensitivity, specificity, accuracy, positive predictive value, and negative predictive value were evaluated. RESULTS: All tests yielded some false-negative results, and there were significant differences among tests in regard to ability to detect low heartworm burdens. Sensitivity of the test kits ranged from 78 to 84%. For all test kits, sensitivity increased as number of female heartworms increased. All 3 test kits had high specificity (97%). CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that sensitivity of the 3 commercially available heartworm antigen test kits ranged from 78 to 84% when used to test serum samples from dogs with low heartworm burdens, and that sensitivity varied among test kits. For all 3 test kits, specificity was 97%. All 3 test kits yielded false-positive and false-negative results for some dogs with low heartworm burdens.     

An evaluation of the gamma interferon test for detecting bovine tuberculosis in cattle 8 to 28 days after tuberculin skin testing

The gamma interferon (IFN -gamma) test was evaluated for its ability to diagnose bovine tuberculosis in cattle that 8 to 28 days previously had a positive caudal fold skin test. The sensitivity of the test was determined in a group of 163 Mycobacterium bovis -infected cattle from 21 herds. The specificity was estimated in a group of 213 cattle which had reacted to a caudal fold test, but were from 82 herds that had no evidence of infection with M bovis. The sensitivity and specificity of the IFN -gamma test was 85 and 93 per cent respectively. No significant differences in the sensitivity and specificity of the test were observed between blood samples that were cultured on the day of collection and those cultured the day after collection. These findings support the use of the IFN -gamma test as a practical serial test that can be used to complement the caudal fold skin test.     

紫花苜蓿种子活力测定方法的比较

采用标准发芽法、胚根突出法、低温发芽法、电导率法对紫花苜蓿(Medicago sativa)10个种子样品的种子活力进行了测定,结果表明,基于标准发芽法的发芽率和活力指数,可将10个种子样品的种子质量分为3级;基于胚根突出法中胚根突出数可将其分为6级,计数胚根突出数的最适时间为24和28h;基于低温发芽法的活力指数和发芽势可将其分为6级;而电导率法可将其分为7级。因而,电导率法作为紫花苜蓿种子活力评价指标最为敏感,其次为胚根突出法和低温发芽法,标准发芽法最不敏感。     

Individual behavioral characteristics of pigs and their impact on production

Two studies were carried out with pigs to determine the relationship between back test results and production parameters and between back test results and other factors. In the first study, 823 piglets were tested with the back test at 10 and 17 days of age. Production parameters such as average daily weight gain and lean meat percentage were determined. In the second study, the back test was performed on 566 piglets at 3, 10, and 17 days of age. The number of escape attempts in the back test (back test score) of the mother was known for 364 piglets. Parameters concerning the health of the sow and piglets were recorded, as well as the sow's reaction to piglet removal for testing. The relationships between production parameters and back test scores of the animals were calculated, as well as the influence of birth weight, sex (all males were castrated), parents, and health parameters on back test scores. Back test scores were fairly consistent over successive tests for each piglet. Males had higher back test scores than females, and piglets from sows with low back test scores also had low scores. Finally, a higher back test score was correlated with a higher lean meat percentage and a better carcass grading at slaughter; no relation with daily weight gain was found. It is concluded that there are individual differences in the way pigs cope with a stressful situation, as measured with the back test, and that this coping behaviour is consistent. A positive correlation exists between back test scores at a young age and lean meat percentage at slaughter. The response to stress, and hence back test scores, is assumed to be inheritable.