Prevalence of serum antibodies to Mycobacterium avium subsp. paratuberculosis in cattle in Galicia (northwest Spain)

Prior to establishing a control and prevention program for Johne's disease in cattle in Galicia (northwest Spain), a survey was conducted to estimate the prevalence of the disease. For this survey, 61,069 animals of at least 1-year of age from 2735 randomly selected herds were bled and samples analyzed with a commercial ELISA. The estimated true individual-level prevalences – assuming the manufacturer's reported test sensitivity of 48.5% and specificity of 98.9% – were 3.02% in dairy cattle, 1.03% in beef cattle and 2.83% in animals from farms with both dairy and beef cattle. True herd prevalences (with herds declared positive if one or more animals tested positive) were 10.69% for dairy herds, 0% for beef herds and 2.71% for mixed herds. When herds were declared positive if at least two animals tested positive, true herd prevalences were 14.75% for dairy herds, 1.47% for beef herds and 12.01% for mixed herds. Assuming a higher specificity of 99.4%, true individual-level prevalences increased to 4.03% in dairy herds, 2.07% in beef herds and 3.84% in mixed herds. Herd prevalences were 27.77%/18.79%, 2.78%/2.40% and 5.70%/12.24% (using the one/two-animal cut-offs) in dairy, beef and mixed herds, respectively. In conclusion, these results seem to indicate that a small percentage of cows and a rather high percentage of dairy herds in this region are MAP-seropositive.     

Cut-off points for aggreate herd testing in the presence of disease clustering and correlation of test errors

In order to test if disease is present in a large herd, an investigator will often subject only a small sample of animals to a fallible diagnostic test. The herd is declared positive for disease if the number of test-positive animals is greater than or equal to a previously chosen cut-off value. Such a test, called an aggregate test, has a sensitivity and specificity that depends on the sample size, the cut-off point and the sensitivity and specificity of the individual test. It also depends on the distribution of the disease among the herds being tested and on the fact that factors such as herd-level seropositivity may cause some herds to be more prone to testing errors than others. In this paper, we use the beta-binomial distribution to model all these factors and thereby calculate and tabulate aggregate test sensitivities and specificities under a variety of conditions. Receiver operating characteristic (ROC) curve methodology permits the choice of optimum sample sizes and cut-off values. We also investigate the situation in which an investigator may be willing to miss detecting the disease if the prevalence in the herd is low. A compiled FORTRAN program for the calculation of aggregate test cut-off point properties, including positive and negative predictive values, is available from the authors.     

Analysis of national serological surveys for the documentation of freedom from porcine reproductive and respiratory syndrome in Switzerland

Results of national serological surveys for porcine reproductive and respiratory syndrome (PRRS) conducted in Switzerland in 2001 and 2004 were analyzed. In 2001, 41,124 breeding sows from 2,540 herds out of 6,406 were sampled, and in 2004, 7,498 animals were sampled from 1,074 herds out of 5,320. All serum samples were tested for PRRS using an ELISA developed at the Institute of Virology and Immunoprophylaxis (IVI), Switzerland with a sensitivity (Se) and specificity (Sp) of 94 and 97%, respectively. Positive samples were re-tested with a commercial ELISA (IDEXX) with Se of 100% and Sp of 99%. Samples positive in the second test were confirmed with the fluorescent antibody test (FAT). A stochastic model using data from the main survey conducted in 2001 was done to verify whether the sampling scheme used could detect at least one infected herd with 99% confidence level if the herd designed prevalence was at 0.1 or 0.2%. Additionally, a Bayesian approach was conducted to calculate the post-survey probability of freedom from PRRS using data from the 2001 and 2004 surveys. A Monte Carlo simulation with 5000 iteration was run for each model. Eleven samples in 2001 and six in 2004, all from different farms, could not be conclusively confirmed as negative by the FAT. All other samples were negative. Truly infected animals and herds were not predicted by a stochastic model at the 99% confidence level and 0.1% herd prevalence using data from the 2001 survey. However, it was demonstrated that the prior probability of freedom from PRRS increased from 89.3 to 99.2% after the 2001 survey. Upon completion of the 2004 survey, the probability of freedom from PRRS reached a value of 99.7%. Based on our results, we could conclude that the pig industry in Switzerland is free of PRRS virus with this level of confidence. Restricted import activities over the last decades are a possible explanation for the continuing absence of PRRS-infection in the Swiss swine population. Import requirements defined by the pig industry minimize the risk of introduction of PRRS-infected animals in the future.     

An estimated prevalence of Johne's disease in a subpopulation of Alabama beef cattle.

The objective of this study was to estimate the overall prevalence of animals that were infected with Mycobacterium avium ssp. paratuberculosis in a subpopulation of Alabama beef cattle. This was determined using a commercial enzyme-linked immunosorbent assay (ELISA) for the detection of M. avium ssp. paratuberculosis-specific antibodies in serum. Serum was collected from 79 herds that were participating in the Alabama Brucellosis Certification program. A total of 2,073 beef cattle were randomly tested by selecting 30 animals per herd in herds greater than 30 and selecting all animals in herds 30 and less for testing. It has been estimated that the commercial ELISA test used has a 60% sensitivity and a 97% specificity. Of the 79 herds tested, 29 herds were seronegative, 24 herds had 1-2 positive animals, and 26 herds had 3 or more seropositive animals. The average number of infected animals per positive herd was 3.3. In addition, a calculated minimum of 53.5% of the herds were identified as Johne's positive herds with a 95% confidence level. Of the total number of animals tested, 8.0% (166/2,073) of them were positive by the ELISA. After adjustments for test sensitivity and specificity and the proportion of animals sampled per herd, the true prevalence was calculated to be 8.75%. These data suggest that approximately 50% of the herds are infected with M. avium ssp. Paratuberculosis, and the overall prevalence of infection in Alabama beef cattle is approximately 8%, which correlates with other previously published regional estimates.     

A stochastic-modeling evaluation of the foot-and-mouth-disease survey conducted after the outbreak in Miyazaki, Japan in 2000

When foot-and-mouth-disease (FMD) was identified in Miyazaki prefecture in March 2000, Japan conducted an intensive serological and clinical survey in the areas surrounding the index herd. As a result of the survey during the 21 days of the movement-restriction period, two infected herds were detected and destroyed; there were no other cases in the months that followed. To evaluate the survey used for screening the disease-control area and surveillance area, we estimated the herd-level sensitivity of the survey (HSe) through a spreadsheet model using Monte-Carlo methods. The Reed-Frost model was incorporated to simulate the spread of FMD within an infected herd. In the simulations, 4, 8 and 12 effective-contact scenarios during the 5-day period were examined. The estimated HSes of serological tests (HSeE) were 71.0, 75.3 and 76.3% under the 4, 8 and 12 contact scenarios, respectively. The sensitivity analysis showed that increasing the number of contacts beyond 12 did not improve HSeE, but increasing the number of sampled animals and delaying the dates of sampling did raise HSeEs. Small herd size in the outbreak area (>80% of herds have <20 animals) seems to have helped in maintaining HSeE relatively high, although the serological inspection was carried out before sero-positive animals had a chance to increase in infected herds. The estimated herd-level specificity of serological tests (HSpE) was 98.6%. This HSpE predicted 224 false-positive herds (5th percentile estimate was 200 and 95th percentile was 249), which proved close to the 232 false-positive herds actually observed. The combined-test herd-level sensitivity (serological and clinical inspections combined; CTHSe), averaged 85.5, 87.6 and 88.1% for the 4, 8 and 12 contact scenarios, respectively. Using these CTHSes, the calculated probability that no infected herd was overlooked by the survey was > or =62.5% under the most-conservative, four-contact scenario. The probability that no more than one infected herd was overlooked was > or =89.7%.     

Pooled fecal culture sampling for Mycobacterium avium subsp. paratuberculosis at different herd sizes and prevalence.

A stochastic spreadsheet model was developed to obtain estimates of the costs of whole herd testing on dairy farms for Mycobacterium avium subsp. paratuberculosis (Map) with pooled fecal samples. The optimal pool size was investigated for 2 scenarios, prevalence (a low-prevalence herd [< or = 5%] and a high-prevalence herd [> 5%]) and for different herd sizes (100-, 250-, 500- and 1,000-cow herds). All adult animals in the herd were sampled, and the samples of the individuals were divided into equal sized pools. When a pool tested positive, the manure samples of the animals in the pool were tested individually. The individual samples from a negative pool were assumed negative and not tested individually. Distributions were used to model the uncertainty about the sensitivity of the fecal culture at farm level and Map prevalence. The model randomly allocated a disease status to the cows (not shedding, low Map shedder, moderate Map shedder, and heavy Map shedder) on the basis of the expected prevalence in the herd. Pooling was not efficient in 100-cow and 250-cow herds with low prevalence because the probability to detect a map infection in these herds became poor (53% and 88%) when samples were pooled. When samples were pooled in larger herds, the probability to detect at least 1 (moderate to heavy) shedder was > 90%. The cost reduction as a result of pooling varied from 43% in a 100-cow herd with a high prevalence to 71% in a 1,000-cow herd with a low prevalence. The optimal pool size increased with increasing herd size and varied from 3 for a 500-cow herd with a low prevalence to 5 for a 1,000-cow herd with a high prevalence.     

Prevalence of paratuberculosis (Johne's disease) in the Belgian cattle population

The national bovine paratuberculosis (PTB) seroprevalence (apparent prevalence) in the Belgian cattle population was determined by a serological survey that was conducted from December 1997 to March 1998. In a random sample of herds (N=556, 9.5%), all adult cattle of 24 months of age or older (N=13,317, 0.4%) were tested for the presence of antibodies using a commercially available absorbed ELISA test kit. The PTB median within-herd seroprevalence (proportion of detected animals within the seropositive herds) and the PTB individual-animal seroprevalence (proportion of detected animals) were, respectively, 2.9% (quartiles=1.6-5.6) and 0.87% (95% confidence interval (CI)=0.71-1.03). The PTB herd seroprevalence (proportion of detected herds) was 18% (95% CI=14-21).Assuming a test sensitivity and specificity of 45 and 99% [Sweeney et al., 1995. J. Vet. Diagn. Invest. 7 (4), 488; Sockett et al., 1992. J. Clin. Microbiol. 30 (5), 1134], respectively, the median true within-herd prevalence and the true individual-animal were estimated to be 7 and 2%, respectively. The true herd prevalence of Mycobacterium paratuberculosis infection was first estimated according to currently accepted methodology. This calculation revealed that the specificity of the used test has a dramatic effect on the estimation; assuming a test sensitivity of 45% and a true within-herd prevalence of 7%, the true herd prevalence estimation decreased from 36 to 0.8% if the test specificity decreased from 99. 9 to 99%, respectively. This sensitivity analysis showed that the practical limits of the accuracy of the used screening test jeopardize the estimation of the true herd prevalence within reasonable confidence limits, because the within-herd PTB true prevalence was low.For this reason we augmented the herd specificity for herds with larger adult herd size (>5). This was done by increasing the cut-off number of positive cattle required (>/=2) to classify a herd truly positive and including herds with one positive test result if there was historical evidence of PTB (previous diagnosis and/or clinical signs). This approach resulted in an estimated true herd prevalence of M. paratuberculosis infection of 6%. The true herd prevalence for dairy, mixed and beef herds was, respectively, 10, 11 and 3%.     

A novel method to identify herds with an increased probability of disease introduction due to animal trade

In the design of surveillance, there is often a desire to target high risk herds. Such risk-based approaches result in better allocation of resources and improve the performance of surveillance activities. For many contagious animal diseases, movement of live animals is a main route of transmission, and because of this, herds that purchase many live animals or have a large contact network due to trade can be seen as a high risk stratum of the population. This paper presents a new method to assess herd disease risk in animal movement networks. It is an improvement to current network measures that takes direction, temporal order, and also movement size and probability of disease into account. In the study, the method was used to calculate a probability of disease ratio (PDR) of herds in simulated datasets, and of real herds based on animal movement data from dairy herds included in a bulk milk survey for Coxiella burnetii. Known differences in probability of disease are easily incorporated in the calculations and the PDR was calculated while accounting for regional differences in probability of disease, and also by applying equal probability of disease throughout the population. Each herd's increased probability of disease due to purchase of animals was compared to both the average herd and herds within the same risk stratum. The results show that the PDR is able to capture the different circumstances related to disease prevalence and animal trade contact patterns. Comparison of results based on inclusion or exclusion of differences in risk also highlights how ignoring such differences can influence the ability to correctly identify high risk herds. The method shows a potential to be useful for risk-based surveillance, in the classification of herds in control programmes or to represent influential contacts in risk factor studies.     

A simulated surveillance program for bovine paratuberculosis in dairy herds in Norway

Monte Carlo simulation models were used to evaluate the feasibility and potential results of a proposed national survey of the prevalence of bovine paratuberculosis (PTB) in dairy herds in Norway. The expected herd prevalence was assumed to be 0.2% in the simulations. The low sensitivity of the ELISA test, the assumed low herd prevalence, the typical low within-herd prevalence of PTB and the small herd sizes all present problems in detection of the disease. Simulations with 500, 1000, 2500 and 6000 herds tested were done. Our results suggest that a national survey would not be feasible at present, due to the low probability of detecting infected herds and because of the high number of false-positive reactions that would be expected to occur.     

Effect of a test and control program for bovine Johne's disease in Victorian dairy herds 1992 - 2002

OBJECTIVE: To report on progress in Johne's disease (JD) control in infected dairy herds participating in the Victorian Johne's disease Test and Control Program (TCP). PROCEDURE: Clinical histories and JD testing data recorded by the Department of Natural Resources and Environment (now called Department of Primary Industries) were analysed for 542 dairy herds participating in the TCP. The herds were required to conduct annual herd tests of cattle 2 years old and older with an enzyme linked immunosorbent assay (ELISA), cull the reactors and manage the younger cattle to minimise infection. RESULTS: Testing of over 680,000 animals identified over 10,000 reactors giving an average prevalence of reactors at the first whole-herd test (T1) of 1.78%. There was a relatively rapid increase in the incidence of clinical disease before the TCP started and then it markedly declined. There was a slow and interrupted decline in reactor prevalence, with a marked peak occurring at the fourth herd test (T4). The average age of reactors and clinical cases was 5.7 and 5.9 years, respectively. Of the reactors and clinical cases detected during the TCP, 87% and 95% respectively, were born before the TCP started. Thirty herds completed the program by achieving three successive negative whole herd tests and 91 herds dropped out because of inability to comply with the agreed requirements of the program. There were no home-bred reactors born after the start of the program in 253 (47%) herds and of the 522 herds that were tested more than once, there were 319 (61%) herds in which no home-bred reactors were detected after the first year of testing. The number of ELISA positive animals detected at T1 appeared to be only about 26% of the animals from that round that subsequently became positive or developed clinical disease at later test rounds. CONCLUSION: The TCP caused a marked decline in the number of clinical cases, probably because animals in which clinical disease was imminent were detected by testing and removed. A reduction in prevalence of reactors occurred only when most herd members were born after the TCP started. The sensitivity of the ELISA appears to be low based on the large number of reactors that were negative at T1 but were positive at later tests. Low sensitivity of diagnostic tests and the long incubation period of the disease limits meaningful analysis of the program until it has continued for some years. Measures adopted in the TCP have not broken the cycle of infection in many participating herds. It is unsure if this was because of poor compliance with control recommendations or a poor understanding of methods of transmission by scientists. Eradication is not feasible in the short-term.     

Estimating herd prevalence of bovine brucellosis in 46 USA states using slaughter surveillance

Making valid inferences about herd prevalence from data collected at slaughter is difficult because the observed sample is dependent on the number of animals sampled from each herd, which varies with herd size and culling practices, and the probability of a positive test result, which depends on variable within-herd prevalence levels as well as test sensitivity and specificity. In this study, brucellosis herd prevalence among beef cow-calf operations is estimated from slaughter surveillance data using a method that combines process modeling with Bayesian inference. Inferences are made for two populations; the first population comprises cow-calf beef herds in a typical U.S. state. The second population represents all beef herds in a collection of 46 low-risk states. The Bayesian Monte Carlo method used in this study links process model inputs to observed surveillance results via Bayes Theorem. The surveillance evidence across multiple years is accumulated at a discounted rate based on the probability of introducing new infection into an area. The process model's inputs include herd size, culling rate per herd, within-herd prevalence, serologic test performance, and the probability of successfully investigating positive results. The surveillance results comprise the number of cows and bulls tested at slaughter and the number of affected herds detected each year. The results find at least 95% confidence that brucellosis herd prevalence among beef cow-calf herds is less than 0.014% (3 per 21,500 herds) and 0.00081% (5 per 6,15,770) after 5 years of slaughter surveillance (with no detections of affected herds) in a typical U.S. state and across 46 low-risk U.S. states, respectively. These results were based on conservative modeling assumptions, but sensitivity analysis suggests only slight changes in the results from changing the assumed process model input values. The most influential analytic input was the probability of introducing new infection into a putatively brucellosis-free state or group of states.     

Prevalence and distribution of paratuberculosis (Johne's disease) in cattle herds in Ireland

A simple random survey was conducted in Ireland during 2005 to estimate the ELISA-prevalence of paratuberculosis, commonly called Johne's disease (JD), in the cattle population. Serum samples were collected from all 20,322 females/breeding bulls over 12 months-of-age in 639 herds. All samples were tested using a commercially available absorbed ELISA. The overall prevalence of infected herds, based on the presence of at least one ELISA-positive animal, was 21.4% (95% CI 18.4%-24.9%). Herd prevalence levels amongst dairy herds (mean 31.5%; 95% CI: 24.6%, 39.3%) was higher than among beef herds (mean 17.9%; 95% CI: 14.6%-21.8%). However, the animal level prevalence was similar. The true prevalence among all animals tested, was calculated to be 2.86% (95%CI: 2.76, 2.97) and for animals >= 2 yrs, it was 3.30% (95%CI: 3.17, 3.43). For animals in beef herds, true prevalence was 3.09% (95%CI: 2.93, 3.24), and for those in dairy herds, 2.74% (95%CI: 2.59, 2.90). The majority of herds had only one ELISA-positive infected animal. Only 6.4% (95% CI 4.7%-8.7%) of all herds had more than one ELISA-positive infected animal; 13.3% (CI 8.7%-19.7%) of dairy herds ranging from two to eight ELISA-positive infected animals; and, 3.9% beef herds (CI 2.4%-6.2%) ranging from two to five ELISA-positive infected animals. The true prevalence of herds infected and shedding Mycobacterium avium subspecies paratuberculosis is estimated to be 9.5% for all herd types; 20.6% for dairy herds; and 7.6% for beef herds. If ELISA positive animals <2-years-of-age are excluded, the true herd prevalene reduces to: 9.3% for all herd types; 19.6% for dairy herds; and 6.3% for beef herds based on a test specificity (Sp) of 99.8% and test sensitivity (Se) (i.e., ability to detect culture-positive, infected animals shedding at any level) of 27.8-28.9%.     

Sensitivity of test strategies used in the Voluntary Johne's Disease Herd Status Program for detection of Mycobacterium paratuberculosis infection in dairy cattle herds

OBJECTIVE: To evaluate sensitivities at the herd level of test strategies used in the Voluntary Johne's Disease Herd Status Program (VJDHSP) and alternative test strategies for detecting dairy cattle herds infected with Mycobacterium paratuberculosis. DESIGN: Nonrandom cross-sectional study. SAMPLE POPULATION: 64 dairy herds from Pennsylvania, Minnesota, Colorado, Ohio, and Wisconsin. Fifty-six herds had at least 1 cow shedding M. paratuberculosis in feces; the other 8 herds were free from paratuberculosis. PROCEDURE: For all adult cows in each herd, serum samples were tested for antibodies to M. paratuberculosis with an ELISA, and fecal samples were submitted for bacterial culture for M. paratuberculosis. Sensitivities at the herd level (probability of detecting infected herd) of various testing strategies were then evaluated. RESULTS: Sensitivity at the herd level of the testing strategy used in level 1 of the VJDHSP (use of the ELISA to test samples from 30 cows followed by confirmatory bacterial culture of feces from cows with positive ELISA result) ranged from 33 to 84% for infected herds, depending on percentage of cows in the herd with positive bacterial culture results. If follow-up bacterial culture was not used to confirm positive ELISA results, sensitivity ranged from 70 to 93%, but probability of identifying uninfected herds as infected was 89%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the testing strategy used in the VJDHSP will fail to identify as infected most dairy herds with a low prevalence of paratuberculosis. A higher percentage of infected herds was detected if follow-up bacterial culture was not used, but this test strategy was associated with a high probability of misclassifying uninfected herds.     

Prevalence of antibodies against Chlamydophila abortus and Coxiella burnetii in goat herds in Poland

An epidemiological study was carried out to determine the herd prevalence of Chlamydophila abortus and Coxiella burnetii antibodies in goats covered by a milk recording program in Poland. The survey took place in 2007 and 48 herds located in different parts of the country were involved. A representative sample from each herd was taken by a simple random sampling allowing to detect seropositivity of a herd on a 95% level of confidence. In total 918 goats were tested for specific antibodies against both germs with the use of enzyme-linked immunosorbent assays. In addition, history of reproductive failures was recorded in these herds. The survey revealed that the herd prevalence of C. abortus was 4.2% (2 herds) while no C. burnetii antibodies were found. Abortions were reported to be a problem in 80% of herds while repeating estrus was encountered in 46% of herds. Reproductive failure concerned two seropositive herds as well. Since the germ is present in the population, it has to be taken into consideration in diagnostic process. Nevertheless, the results of the present study indicate that C. abortus infection occurs infrequently in Polish goats. As no antibodies against C. burnetii were detected in the screened sample the risk of goat-to-human transmission of both bacteria in Poland seems to be very low.     

Evaluation of a bulk-milk ELISA test for the classification of herd-level bovine leukemia virus status

The results of a commercial bulk-milk enzyme-linked immunosorbent assay (ELISA) test for herd-level bovine leukemia virus (BLV) status were compared to results obtained from individual agar-gel immunodiffussion (AGID) testing on sampled cattle. A positive herd was defined as a herd having one or more AGID-positive animals. The estimated true herd status was based on the sensitivity and specificity of the AGID test and the number of cattle sampled per herd. Ninety-seven herds were used, with a mean of 13 cows sampled per herd. The AGID test indicated an apparent herd prevalence of 70.1%. After accounting for the number of cows sampled and the sensitivity and specificity of the AGID test, the estimated true herd prevalence of BLV was 52.3%. The ELISA test identified 79.4% of herds as positive for BLV, and had an apparent sensitivity and specificity of 0.97 and 0.62, respectively. However, after accounting for the sensitivity and specificity of the AGID test in individual animals, the specificity of the ELISA test was 0.44. The ELISA test was useful for identifying BLV-negative herds (i.e., ruling out the presence of BLV infection in test negative herds). With the moderately low specificity, herds identified as positive by the ELISA test would require further testing at the individual or herd level to definitively establish their BLV status.     

Spread of pseudorabies virus among breeding swine in quarantined herds.

In theory, pseudorabies virus (PRV) may be eliminated from any size of breeding herd by phased test and removal if replacement gilts are not infected with PRV, culling decisions are partially based on PRV status, and the cull rate is higher than the incidence rate of PRV. Annual cull rates are commonly at least 50%, but little information exists on the incidence of PRV within enzootically infected swine herds. The purpose of this study was to develop a method by which spread of PRV could be detected among breeding swine within enzootically infected herds and to determine the incidence of PRV infection in these herds. Data were collected from 17 herds that were quarantined for PRV and ranged in size from 120 to 1,100 sows. At each herd, within the first 5 days of introduction, a group of approximately 30 replacement gilts was identified, vaccinated with a glycoprotein X-deleted PRV vaccine, and blood sample was collected. The owner of 1 herd had a nonvaccinated breeding herd and elected to leave incoming gilts nonvaccinated. After vaccination, blood samples were collected every 1 to 2 months for an average of 13.6 months. Serum samples from vaccinated gilts were tested for antiglycoprotein X antibodies by a specific differential ELISA. Samples from nonvaccinated gilts were evaluated by serum neutralization test. Product-limit method was used to estimate the probability of not becoming infected with PRV. Spread was detected in 7 of 8 herds that had more than 400 sows and in 2 of 9 herds that had less than 400 sows.(ABSTRACT TRUNCATED AT 250 WORDS)     

A serological survey to determine the prevalence of infection with Treponema hyodysenteriae in Western Australia

A serological survey to detect antibody titres against Treponema hyodysenteriae was conducted on pigs from 106 herds in Western Australia. Titres indicating a positive result in the tests were determined by examining 400 sera from 4 herds known to be free of swine dysentery, and sera from immunised or experimentally infected pigs. Samples of serum from 40 bacon-weight pigs from each of the 106 herds were then collected at 2 abattoirs. Each serum was tested in enzyme-linked immunosorbent assays (ELISA) against the lipopolysaccharide of T hyodysenteriae of serogroups A, B and E, respectively. To assist in evaluating the test, 19 herds were resampled and retested, and faecal samples from 17 herds were cultured for T hyodysenteriae. Thirty-five of the 106 herds (33%) had serological evidence of infection when only one batch of sera from each herd was tested. The ELISA to detect T hyodysenteriae infection in herds using 40 sera was estimated as having a sensitivity of 77.3% and a specificity of 81.8% based on the owners' opinion of their herds disease status. Prevalence of infection within herds ranged from 2.5% to 47.5%, with a mean of 18%.     

Nested PCR on blood and milk for the detection of Mycobacterium avium subsp paratuberculosis DNA in clinical and subclinical bovine paratuberculosis

OBJECTIVE: To determine the potential of PCR on blood and milk to detect cattle infected with Mycobacterium avium subsp paratuberculosis. PROCEDURE: A nested PCR method probing for IS900 was developed and compared to ELISA serology in 11 clinically infected and 46 subclinically infected, lactating Holstein cows from a herd with confirmed paratuberculosis (Johne's disease). RESULTS: When compared to serum ELISA the nested blood- and milk PCRs were equal in identifying DNA from clinically infected animals. The PCR procedures also gave positive DNA results with some subclinically infected animals when these only gave suspicious or negative results in the ELISA test. Most clinically and subclinically infected animals were detected with milk PCR. CONCLUSION: Since there may well be a haematological phase in paratuberculosis, nested PCR testing of blood and milk samples shows potential to detect animals subclinically infected with M a paratuberculosis. More subclinically infected animals need to be tested and confirmed infected before estimates of sensitivity and specificity can be made.     

A Cross-sectional Study of Bovine Tuberculosis in Dairy Farms in Asmara,Eritrea

Bovine tuberculosis in dairy cattle in Asmara, Eritrea, was studied using a cross-sectional study to describe its prevalence and to identify factors associated with it. A total of 72 randomly selected herds were included in the study. The comparative intradermal tuberculin test was used for the diagnosis. Of 1813 individual animals tested, 14.5% were reactors. Thirty herds (41.7%) had at least one reactor but, by defining a reactor herd as any herd with two or more reactors, only 19 (26.4%) herds were classified as reactor herds. Based upon individual animal specificity of 98.5%, the calculated herd specificity was >99%. A multiple logistic model showed that the presence of exotic breeds was associated with a high risk (odds ratio = 5.70; 95% confidence interval 1.13–28.8). An increased risk was also linked to large herds. Keeping the animals always indoors reduced the risk, but could not be fitted to the model owing to empty cells.     

Serological prevalence of Mycoplasma bovis infection in suckling beef cattle in France

The prevalence of Mycoplasma bovis infection in France was assessed by means of a serological survey of suckling beef cattle, using an ELISA. The survey included 824 randomly selected herds in eight French counties and a total of 32,197 animals more than one year old. In each county, the number of herds tested was determined statistically on the basis of the hypothesis that about 40 per cent of herds are infected. The proportion of herds containing at least one infected animal ranged from 28 to 90 per cent depending on the county. Among the 32,197 sera tested, the animal infection rate ranged between 2 per cent and 13 per cent. In infected herds, the average number of positive animals per herd was between 10 and 20 per cent, and the infection was unevenly distributed among the areas tested.