The estimated prevalence of Johne's disease infected sheep flocks in Australia

OBJECTIVE: To estimate the likely geographical distribution and flock-prevalence of ovine Johne's disease (OJD) in Australia. DESIGN: A cross-sectional study design was used. PROCEDURE: The results of abattoir surveillance for OJD carried out during 2000 were analysed to estimate the prevalence of infected flocks in three regions of New South Wales and in other States. A Bayesian approach was used to adjust apparent prevalence estimates for the assumed flock-sensitivity and flock-specificity of abattoir surveillance, and to allow for uncertainty about the true values of these measures. RESULTS: The 95% probability limits for flock-prevalence at 31 December 2000 were 0.04%-1.5%, 8%-15% and 29%-39% for low, moderate and high prevalence regions of New South Wales respectively. The other States generally had an upper 97.5% probability limit of about 1% or less. Based on these estimates about 6 to 10% of flocks in New South Wales and 2.4 to 4.4% of flocks Australia-wide are likely to be infected. CONCLUSION: This study suggests that OJD has a highly clustered distribution in Australia, and provides estimates of the prevalence of infected flocks by State or region. Based on this analysis there were probably between 2000 and 3700 infected flocks in Australia at 31 December 2000, with more than 80% of these in a relatively small geographic area of central and southern New South Wales. Some States, such as Queensland and Western Australia, may have a prevalence equal or close to 0%, however the technique used was unable to demonstrate the absence of infection in these States with the intensity of surveillance undertaken to date.     

Evaluation of an absorbed ELISA and an agar-gel immuno-diffusion test for ovine paratuberculosis in sheep in Australia

The sensitivities and specificities of an absorbed enzyme-linked immunosorbent assay (ELISA) and an agar-gel immuno-diffusion (AGID) test for the detection of Johne’s disease in sheep were estimated using data from six known infected and 12 assumed uninfected sheep flocks. Sensitivities were estimated for all histologically positive sheep, as well as by histological lesion score, lesion type (paucibacillary or multibacillary) and sheep body-condition score, with ELISA sensitivities estimated at 95 and 99% specificity. Logistic-regression analysis was used to test for significant effects of lesion score and condition score, with flock included in the model as a random effect.

Estimated specificities were 95% (95% CI: 93.4, 95.6%) and 99% (98.4, 99.4%) for ELISA cut-point ratios of 2.4 and 3.6, respectively, and 100% (99.7, 100.0%) for the AGID. Estimated sensitivities for all infected sheep were 41.5% (35.0, 48.3%), 21.9% (16.6, 27.9%) and 24.6% (19.1, 30.7%) for ELISA cut-point ratios of 2.4 and 3.6 and for AGID, respectively. Sensitivities of all tests and cut-points varied significantly between flocks and between categories of lesion score and condition score. Sensitivity ranged from 25 to 73, 10 to 47 and 9.2 to 63% between flocks, for the ELISA with cut-points of 2.4 and 3.6, and for the AGID, respectively. Sensitivity was highest in thin sheep and in sheep with multibacillary lesions. The effects of lesion type and condition score on test sensitivity were significant in the logistic regressions for the AGID and ELISA at both cut-points and the flock effect was significant for the AGID but not for the ELISA at either cut-point.     

Ovine brucellosis in alberta

Two parallel surveys of rams from Alberta sheep flocks were conducted to determine the presence of infection with Brucella ovis. In a retrospective study over a period of 24 months, using complement fixation test, 12 flocks out of 142 tested were considered infected. In another 17-month survey of slaughter rams by serology and culture methods 11 flocks out of 124 were found to be infected. The overall prevalence of ovine brucellosis was 8.6% of the flocks tested which represented 12.5% of the estimated sheep flocks in Alberta. Up to 67% of rams in infected flocks reacted to complement fixation test.

The complement fixation test was evaluated for its efficiency in the diagnosis of ovine brucellosis and compared with a limited number of an enzyme-linked immunosorbent assay (ELISA) results and clinical criteria. The complement fixation test as well as ELISA identified all culture positive rams. Both serological tests appeared satisfactory for the diagnosis of B. ovis epididymitis when the results could be interpreted in the light of flock history and clinical findings.

     

Ovine Johne's disease in Australia--the first 20 years

OBJECTIVE: To review the history of ovine Johne's disease in Australia. PROCEDURE: Relevant publications and reports were identified and reviewed to document the spread of ovine Johne's disease (OJD) from 1980 until the end of 2000, as well as the response of industry and government to the spread of this disease. RESULTS: OJD was first diagnosed in the central tablelands region of New South Wales in 1980. Since then it has spread, either from the initial focus or through separate introductions so that by December 2000 a total of 823 infected flocks had been identified. Cases have been confirmed in New South Wales, Victoria, the Australian Capital Territory, on Flinders Island in Tasmania, on Kangaroo Island in South Australia and in Western Australia. In early 1999, agreement was reached to fund and implement a 6-year, $40 million National OJD Control and Evaluation Program (NOJDP). This program is jointly funded by the sheep industries (national and state), and Commonwealth and State governments, and is managed by Animal Health Australia. CONCLUSION: A national program is now in place to support the control of OJD and research to determine the feasibility and cost-effectiveness of eradication. The development of new diagnostic techniques, such as abattoir surveillance and pooled faecal culture, provide opportunities to refine surveillance strategies and to define better the distribution and prevalence of this disease, as required by the national program. Effective control measures, combined with quality surveillance data, will enable informed decision making for the future national management of OJD.     

Sensitivity and specificity of the agar-gel-immunodiffusion test, ELISA and the skin test for detection of paratuberculosis in United States Midwest sheep populations

Our objective was to estimate the sensitivity and specificity of the agar-gel-immunodiffusion test (AGID), the ELISA, and the skin test for the detection of Mycobacterium avium subspecies paratuberculosis (MAP) in sheep using Bayesian methods without a gold standard. Fourteen flocks (2 465 sheep) were used. Five flocks (450 sheep) were considered MAP non-infected and 9 flocks (2 015 sheep) had sheep infected with MAP. Sheep were skin tested and blood was collected for AGID and ELISA testing. Results were analyzed using a Bayesian 3-test in 1-population model fitted in WinBUGS. The model allowed for dependence (correlation) between the two serologic tests, but these two tests were assumed to be conditionally independent of the skin test. The estimated specificity was 99.5% (95% PI of 98.9-99.9%) for the AGID; 99.3% (98.4-99.8%) for the ELISA using an optical density measured cutoff of 0.20; 99.2% (98.1-99.8%) using a cutoff of 0.15; 97.5% (95.8-98.7%) using a cutoff of 0.10; and 98.7% (97.3-99.5%) for the skin test. The estimated sensitivities were 8.3% (6.2-10.7%) for the AGID; 8.0% (6.0-10.4%), 10.6% (8.3-13.1%), and 16.3% (13.5-19.4%) for the ELISA using the cutoffs 0.20, 0.15, and 0.10 respectively; and 73.3% (62.3-85.8%) for the skin test. The skin test was specific in non-infected populations and sensitive in infected populations, although in some cases a positive skin test might represent MAP exposure rather than infection. The AGID and ELISA were specific but lacked sensitivity. The AGID and ELISA consistently identified two different populations of infected sheep with only moderate overlap between positive test results.     

Monte Carlo simulation of flock-level sensitivity of abattoir surveillance for ovine paratuberculosis

We used Monte Carlo simulation to estimate distributions for flock-level sensitivity of abattoir-based surveillance for ovine paratuberculosis as currently practised in New South Wales, Australia. Probability distributions were used as input variables for within-flock prevalence, years-infected and individual animal-level sensitivity and specificity of gross pathology as a screening test for the presence of paratuberculosis. Distributions used as inputs for the size of abattoir-slaughter groups were based on existing abattoir-surveillance data from NSW. Predicted flock-level sensitivity depended on within-flock prevalence and the number of animals examined and was sensitive to estimates of animal-level sensitivity and specificity. The median probability of detection of an infected flock based on the examination of one abattoir line was predicted not to exceed 0.95 unless the within-flock prevalence was ≥7%. If the within-flock prevalence was 2%, the probability distribution of flock-level sensitivity had a median of 0.73, with 80% of values lying between 0.55 and 0.84. Improvement in the flock-level sensitivity could be achieved by submitting more than three gross pathology-positive specimens per line, if available—but the degree of improvement depended on the number of sheep slaughtered (line size) and the within-flock prevalence. At 2% prevalence, a median flock-level sensitivity of 0.95 could be obtained in lines of >390 sheep if six gross pathology-positive specimens were submitted. We concluded that abattoir surveillance based on identification of gross pathology as a screening test is not a sensitive tool for detecting recently infected flocks or flocks which have a moderate or lower prevalence of infected animals. But—with relatively minor modifications of the protocol currently in use—it could become a key component of a surveillance programme which included additional testing strategies for small flocks.     

Cross species transmission of ovine Johne's disease from sheep to cattle: an estimate of prevalence in exposed susceptible cattle

OBJECTIVE: To determine the prevalence of infection of cattle with the sheep strain of Mycobacterium avium subsp paratuberculosis at least two years after exposure at < 6 months old. DESIGN: Prospective survey One thousand seven hundred and seventy-four cattle from 12 properties (Farms A to L) were sampled by ELISA and faecal culture to detect evidence of infection with M a paratuberculosis. All properties had a known history of Johne's disease (JD) in sheep, and sampled cattle were likely to be susceptible to JD at the time they were first exposed, being at an age of 6 months or less. In addition, opportunistic investigations were undertaken of ELISA reactor cattle discovered during testing for the Australian Johne's Disease Market Assurance Program for Cattle (Farms M and N). RESULTS: All animals in the survey gave negative results on serology while one animal from a herd of 349 gave a positive faecal culture result. Follow-up faecal culture, post-mortem and histopathology on the latter animal were negative, suggesting that it was a passive faecal shedder or carrier. Two occurrences of OJD transmission to cattle were detected during the opportunistic investigations. CONCLUSION: These observations confirm existing beliefs about the risk of transmission of OJD to cattle, that the risk of transmission is low. However transmission occurs sporadically. An estimated upper limit of prevalence of S strain M a paratuberculosis infection in susceptible exposed cattle in the OJD high prevalence area of New South Wales is 0.8%, assuming a common prevalence within herds.     

Specificity of absorbed ELISA and agar gel immuno-diffusion tests for paratuberculosis in goats with observations about use of these tests in infected goats

OBJECTIVE: To determine the specificity of serological tests that are currently used in veterinary diagnostic laboratories in Australia for detection of Mycobacterium avium subsp paratuberculosis infection in goats. DESIGN: A laboratory study. PROCEDURE: Four tests were studied, comprising AGID with M. a. paratuberculosis antigen derived from cattle isolates of caprine or bovine origin, the EMAI caprine Johne's disease absorbed ELISA and the CSL PARACHEK Johne's absorbed EIA. The specificities of AGID and ELISA for paratuberculosis (Johne's disease) were estimated after examining a panel of 1000 serum samples collected from goats in Western Australia, a region free of paratuberculosis. In addition a comparison was made of test performance in a small number of paratuberculous goats from New South Wales using sera from two archival collections. RESULTS: The specificity of the AGID tests was 100% while the specificities of the two absorbed ELISA were 99.7 to 99.8% at appropriate positive-negative cut-offs. Based on testing the small sample of sera from infected goats, the absorbed ELISA tests detected about twice as many goats with Johne's disease as the AGID. Each test detected paratuberculous animals regardless of whether infection was caused by cattle or sheep strains of M. a. paratuberculosis. CONCLUSIONS: Both ELISA and AGID tests for paratuberculosis have high specificity and can be used in a market assurance program without risk of generating large proportions of false positive test results. However, the results suggested the ELISA is more sensitive for detection of infected goats and should be used in preference to the AGID. The two formats of ELISA evaluated in this study have similar characteristics and could be used in paratuberculosis control programs for the goat industries, but further data on sensitivity would increase confidence in their application.     

Bayesian estimation of true between-herd and within-herd prevalence of Salmonella in Danish veal calves

Specialised veal producers that purchase and raise calves from several dairy herds are potentially at high risk of delivering Salmonella-infected animals to slaughter. However, the true prevalence of Salmonella infected veal producing herds and the prevalence of infected calves delivered to slaughter from infected herds are unknown in Denmark. Due to uncertainties about test sensitivity and specificity, these prevalences are not straightforward to assess. The objective of this study was to estimate the within-herd- and between-herd prevalence of Salmonella in veal calves delivered for slaughter to abattoirs in Denmark. Furthermore, it was investigated to which extent the estimates differed between a setup using both serological tests and faecal culture, compared to just serological tests, and whether the applied sampling scheme in the national surveillance programme in Denmark was sufficient to establish high posterior estimates of freedom from infection in individual herds. We used Bayesian analysis to avoid bias as a result of fixed test validity estimates. Serological test results from 753 animals and faecal culture from 1233 animals from 68 randomly selected Danish veal producing herds that delivered more than 100 calves to slaughter per year were used to estimate the prevalences and estimates of freedom from Salmonella. Serological test results of 7726 animals from 185 herds were used to compare the difference in prevalence estimates between serology alone vs. faecal culture combined with serology. We estimated that 34-57% of specialised veal producing herds were infected with Salmonella. Within the infected herds, 21-49% of the animals were infected. Few herds obtained high posterior estimates for the probability of freedom from infection given the collected data, with only six of 68 herds obtaining posterior probability of being infected less than 10%. Furthermore, this study indicated that serology is sufficiently sensitive and specific to be used for estimating the prevalence of Salmonella-infected specialised veal producing herds.     

Sensitivity and specificity of two serological tests for the detection of ovine paratuberculosis

OBJECTIVE: To determine the sensitivity and specificity of an absorbed ELISA and an AGID test for the detection of clinical and subclinical paratuberculosis in sheep. DESIGN: By testing a panel of sera from 1257 Australian Merino and crossbred sheep greater than 1 year of age, of which 1137 sheep were not infected with Mycobacterium avium subsp paratuberculosis and 120 sheep had paratuberculosis. PROCEDURE: Sera were collected from 457 sheep in Victoria and 800 sheep in Western Australia. Presence of M a paratuberculosis infection in Victorian sheep was determined by histological examination of intestinal tissues, whereas sheep from Western Australia were presumed to be free of Johne's disease. The ability of an absorbed ELISA to discriminate between infected and uninfected sheep was described by test sensitivity and specificity, the distribution of ELISA OD, and the area under a receiver operating characteristic curve. RESULTS: The absorbed ELISA had a specificity of 98.2 to 99.5% (CI) and a sensitivity of 35 to 54% (CI). In sheep from infected flocks in Victoria, the AGID test had a specificity of 99 to 100% (CI) and a sensitivity of 38 to 56% (CI). The sensitivity of serological tests was higher in sheep with a body condition representative of the lower quintile of their flock of origin. CONCLUSION: The AGID test and absorbed ELISA are useful tests for the detection of ovine paratuberculosis. Although the tests had a similar accuracy, they detected different subpopulations of infected sheep with only moderate overlap. The AGID test had a higher specificity than the absorbed ELISA.     

A comparison of methods to estimate the prevalence of ovine Johne's infection from pooled faecal samples

OBJECTIVE: To compare estimates of ovine Johne's infection prevalence produced by several alternate methods based on pooled faecal culture (PFC) results with prevalence estimates based on individual faecal culture (IFC). PROCEDURE: Seven methods for estimating prevalence of infection based on PFC results were incorporated in a computer program, including methods for imperfect test sensitivity and specificity, for variable pool size and a Bayesian method that incorporates prior knowledge about test performance and prevalence. These methods were then used to analyse PFC data at one observation 30 months post-vaccination in a field trial of a killed vaccine for the control of OJD, undertaken on three farms in New South Wales. RESULTS: Prevalence estimates, for three methods that assume a perfect test, were close to the IFC estimate, whereas for three other methods that assume an imperfect test, the estimated prevalence was generally higher than the IFC estimate. In comparison, the Bayesian approach produced more variable estimates that were substantially higher than the IFC estimate when an inappropriately high prior estimate of prevalence was used. CONCLUSION: Despite the limitations of each method, two methods provided accurate and reasonable estimates of the prevalence assessed by IFC in all instances, and are appropriate for the analysis of data from this vaccine trial. One of these methods also has the advantage of allowing for variable pool size. However, further research is needed to develop a method that will simultaneously account for variation in pool size and in test sensitivity and specificity.     

Use of Brucella canis antigen for detection of ovine serum antibodies against Brucella ovis

Brucella ovis causes a genital disease of sheep manifested by epididymitis in rams and placentitis in ewes producing reduced fertility in the flock. Clinical diagnosis is not sensitive enough and bacteriological testing is not feasible for detection of the disease in large numbers of animals. Indirect methods of serological testing are preferred for routine diagnosis, of which agar gel immunodiffusion (AGID), complement fixation (CF) and ELISA tests are recommended as the most efficient. Since B. ovis shares antigenic components with Brucella canis, it would seem that either strain could be used as antigen with the same results; however, the advantage of the B. canis (M-) strain variant is that it can be used to develop a satisfactory antigen for agglutination tests. We present data on AGID and IELISA tests using B. ovis antigen and rapid screening agglutination test (RSAT), 2-mercapto-ethanol RSAT (2ME-RSAT) and IELISA using B. canis antigen. We tested 225 animals. The cut-off values were adjusted by ROC analysis using 51 negative and 32 positive sera; the IELISA-B. canis cut-off value was 39 (%P) and IELISA-B. ovis, 51 (%P), with 100% sensitivity and specificity. Of the 32 positive sera from the infected flock RSAT detected 32 (100%), 2ME-RSAT 29 (91%) and AGID 31 (97%). Of the 142 sera from suspicious flocks, 46 were negative and 56 positive in all the tests; 16 were positive by RSAT, IELISA-B. canis and IELISA-B. ovis, 20 positive only with RSAT and 2 positive only by both IELISAs. RSAT is a very sensitive screening test that, because of its simplicity and easy interpretation, following a study in larger sample, could replace AGID as a screening test for diagnosis of ovine brucellosis caused by B. ovis. The IELISA-B. canis or IELISA-B. ovis could be used as confirmatory tests, since they show equal specificity and sensitivity.     

Evaluation of three serological tests for diagnosis of Maedi-Visna virus infection using latent class analysis

Maedi-Visna virus (MVV) infection in sheep is present in several European countries, including Norway. The current Norwegian surveillance and control programme for MVV infection uses three serological tests: an agar gel immunodiffusion test (AGID) and two commercially available indirect ELISAs (Institut Pourquier, P-ELISA and HYPHEN BioMed, H-ELISA). From 18 flocks with suspected or confirmed MVV infection, sera from naturally infected sheep were obtained, and sensitivity (Se) and specificity (Sp) of the three tests were estimated in absence of a perfect reference test using latent class models in a Bayesian analysis. The AGID had higher Sp (95% posterior credibility interval (PCI) [98.4; 99.9]) than either ELISA (95% PCI: P-ELISA, [95.1; 99.0]; H-ELISA, [91.4; 96.6]), but much lower Se (95% PCI: AGID, [41.4; 59.8]; P-ELISA, [92.7; 100.0]; H-ELISA, [90.9; 99.4]). Currently the P-ELISA is used for screening and positive samples are subsequently confirmed by a setup using all three tests in a serial reading. The Se and Sp of the serial interpretations with and without the H-ELISA were estimated. The results suggested that the H-ELISA could be dropped as a confirmatory test as the Se of the three test serial reading was reduced significantly without adding a significant improvement of the Sp compared to the serial reading of the P-ELISA and AGID alone. However, the perceived cost of false positives versus false negatives will influence this decision. Estimates of the predictive values for the tests and combinations suggested that the P-ELISA is a good choice of screening, but confirmatory tests are needed to achieve acceptable levels of positive predictive values.     

Longitudinal study of the spread of ovine Johne's disease in a sheep flock in southeastern New South Wales

OBJECTIVE: The aim of this study was to apply whole flock testing over time to determine the prevalence, distribution and spread of infection in a recently infected flock, with a view to planning intervention strategies for disease control. PROCEDURE: Serology, pooled faecal culture (PFC) and histology were used to determine the distribution and persistence of infection in a sheep flock in south east New South Wales between 1997 and 2002. Partial flock testing was done up to June 2000, after which annual whole flock testing, using PFC was performed. RESULTS: Faecal shedding of M a paratuberculosis was not detected in home-bred sheep until 7 years after the introduction of infected sheep in 1993. For at least 7 years there was clustering of infection and shedding within two age groups only. The infected groups appeared to have been exposed to infection (mycobacterial contamination) at an early age (<12 months) and commenced shedding at 5 years of age or older. Groups that were exposed to contamination as adults did not shed detectable amounts of M a paratuberculosis during the study period. CONCLUSION: Clustering of detectable infection in age groups of sheep that were exposed as lambs was a feature on this farm, providing indirect evidence of finite duration of survival of M a paratuberculosis on pasture and the influence of age on the susceptibility of sheep to develop detectable M a paratuberculosis infection. Spread of infection occurred very slowly and was probably related to the long incubation period (exposure to shedding interval) of 5 years observed on this farm. The findings suggest that partial flock culling, selective grazing management and vaccination could lead to a reduction in mycobacterial contamination on farm to a level at which patent infection no longer occurs. Better understanding of disease spread within flocks over time through flock profiling using PFC will help in devising surveillance strategies (including testing protocols for market assurance testing) to detect infected flocks where there has been clustering and slow spread of infection.     

Pooled-sample testing as a herd-screening tool for detection of bovine viral diarrhea virus persistently infected cattle.

The study was conducted to develop methodology for least-cost strategies for using polymerase chain reaction (PCR)/probe testing of pooled blood samples to identify animals in a herd persistently infected with bovine viral diarrhea virus (BVDV). Cost was estimated for 5 protocols using Monte Carlo simulations for herd prevalences of BVDV persistent infection (BVDV-PI) ranging from 0.5% to 3%, assuming a cost for a PCR/probe test of $20. The protocol associated with the least cost per cow involved an initial testing of pools followed by repooling and testing of positive pools. For a herd prevalence of 1%, the least cost per cow was $2.64 (95% prediction interval = $1.72, $3.68), where pool sizes for the initial and repooled testing were 20 and 5 blood samples per pool, respectively. Optimization of the least cost for pooled-sample testing depended on how well a presumed prevalence of BVDV-PI approximated the true prevalence of BVDV infection in the herd. As prevalence increased beyond 3%, the least cost increased, thereby diminishing the competitive benefit of pooled testing. The protocols presented for sample pooling have general application to screening or surveillance using a sensitive diagnostic test to detect very low prevalence diseases or pathogens in flocks or herds.     

Intrauterine and transmammary transmission of Mycobacterium avium subsp paratuberculosis in sheep

OBJECTIVE: To investigate intrauterine infection of foetuses with Mycobacterium avium subsp paratuberculosis and the presence of infection in mammary secretions of sheep. DESIGN: A study of 142 late-pregnant ewes and their foetuses from two heavily infected flocks. PROCEDURE: Infection of ewes was determined at necropsy by histopathology and culture of tissues and mammary secretions. Antemortem tests (clinical assessment, faecal culture and serology) were also applied. Foetuses from 59 infected ewes and 47 apparently uninfected ewes were examined by culture and histopathology. RESULTS: Five of five ewes with clinical ovine Johne's disease had infected foetuses. Only one of 54 subclinically affected ewes, and none of 47 uninfected ewes had an infected foetus. M a paratuberculosis was cultured from mammary secretions or mammary glands of only two of 76 ewes, both of which were clinical cases and had infected foetuses. CONCLUSION: Although intrauterine or transmammary transmission of Mycobacterium avium subsp paratuberculosis may occur frequently in clinically affected sheep, these are less common in subclinically infected ewes. Therefore these modes of transmission are unlikely to compromise existing control programs for ovine Johne's disease on most farms, especially if programs include the immediate culling of clinically affected sheep.     

Seroprevalence of Brucella ovis infection in commercial ram flocks in the Tamworth area

Commercial ram flocks in the Tamworth area of northern New South Wales were surveyed to estimate the proportion of flocks, and rams, infected with Brucella ovis. The flock prevalence (percentage of flocks containing seropositive rams) of 9.1% for Merino flocks was significantly lower than that for British-breed flocks (43.8%, p=0.006) and mixed-breed flocks (46.7%, p=0.017). The mean flock prevalence over all flock types was 32.9%. These estimates were supported by data obtained from diagnostic testing for brucellosis carried out during the previous 6 years. The seroprevalence in rams was 10.8% overall, 2.5% for Merinos, 19% for Border Leicester and 26% for Dorset rams. Within infected flocks, the estimated prevalences were 21%, 65% and 67% for Merinos, Border Leicester and Dorset rams respectively. The seroprevalence in Merino rams was significantly lower than that for both other breeds (p<0.001) for all flocks and in infected flocks. There was no apparent association between age and serological status, or age and the prevalence of epididymitis.     

Prevalence and risk factors for Salmonella spp. and Campylobacter spp. caecal colonization in broiler chicken and turkey flocks slaughtered in Quebec, Canada

We conducted an observational study to estimate prevalence and risk factors for Salmonella spp. and Campylobacter spp. caecal colonization in poultry. Eighty-one broiler chicken and 59 turkey flocks selected among flocks slaughtered in the province of Quebec, Canada, were included in the study. Flock status was evaluated by culturing pooled caecal contents from about 30 birds per flock. Exposure to potential risk factors was evaluated with a questionnaire. Odds ratios were computed using multivariable logistic regression.

The prevalence of Salmonella-positive flocks was 50% (95% CI: 37, 64) for chickens and 54% (95% CI: 39, 70) for turkeys, respectively. Odds of Salmonella colonization were 2.6 times greater for chicken flocks which failed to lock the chicken house permanently. In turkeys, odds of Salmonella colonization were 4.8–7.7 times greater for flocks which failed to be raised by ≤2 producers with no other visitors allowed onto the premises, or origin from a hatchery.

The prevalence of Campylobacter-positive flocks was 35% (95% CI: 22, 49) for chickens and 46% (95% CI: 30, 62) for turkeys. Odds of colonization were 4.1 times higher for chicken flocks raised on farms with professional rodent control and 5.2 times higher for flocks with manure heap >200 m from the poultry house, and also increased with the number of birds raised per year on the farm and with the age at slaughter. For turkeys, odds of Campylobacter flock colonization were 3.2 times greater in flocks having a manure heap at ≤200 m from poultry house and 4.2 times greater in flocks drinking unchlorinated water.     

A comparison of whole virus and recombinant transmembrane ELISA and immunodiffusion for detection of ovine lentivirus antibodies in Italian sheep flocks

Sera from two sheep experimentally infected with ovine lentivirus (OLV) and from 186 sheep selected from flocks with known high or low prevalence of infection or on the basis of virological or histopathological examination were simultaneously tested by whole virus (WV) ELISA, recombinant transmembrane (r-TM) ELISA and AGID assay. Antigens for both the WV ELISA and AGID were prepared from an Italian field isolate; recombinant antigen was derived from the N-terminal region of the transmembrane envelope protein of strain K1514. The WV ELISA detected the highest number of seropositives, followed by the r-TM ELISA and AGID test. The sensitivity and specificity of the r-TM ELISA relative to the WV ELISA were 0.66 and 0.95, respectively. Immunoblot analysis of 14 WV ELISA-positive and r-TM ELISA-negative sera showed that the major core protein was immunodominant on WV antigen. It is concluded that the r-TM ELISA was more sensitive than the AGID test but less sensitive that the WV ELISA, particularly for detecting antibodies in the early stages of infection.Abbreviations AGID agar gel immunodiffusion - bp base pair - ELISA enzyme-linked immunosorbent assay - FBS fetal bovine serum - IB immunoblot - kD kilodalton - OLV ovine lentivirus - MEM minimal essential medium - p.i. post-infection - r recombinant - SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis - TM transmembrane - WV whole virus     

Pooled faecal culture for the detection of Mycobacterium avium subsp paratuberculosis in goats

OBJECTIVE: To evaluate pooled faecal culture for herd diagnosis of caprine Johne's disease and relate these findings to faecal shedding rates of Mycobacterium avium subsp paratuberculosis (Map). DESIGN: Radiometric broth culture was applied to several pooling dilutions, and shedding rates were estimated from a regression equation based on bacterial growth rates and known processing losses during radiometric culture. PROCEDURE: Sixteen faecal samples from goats naturally infected with sheep (n = 3) or cattle (n = 13) strains of Map, were diluted in normal goat faeces from 1 in 5 to 1 in 50. Cultures were confirmed by IS900 polymerase chain reaction and restriction endonuclease analysis, and mycobactin dependency. The numbers of viable Map in the culture inocula were determined by endpoint titration (most probable number) of nine samples and related to a cumulative growth index. RESULTS: A pooling dilution of 1 in 25 with an incubation period of 10 weeks detected 13 of 16 culture positive goats, all shedding > or = 2 x 10(4) Map per gram of faeces. Two samples containing very low numbers of Map (< 2 x 10(3)/g) were only culture positive from undiluted faeces. Thirteen of 16 goats were considered to be shedding low to moderate concentrations of Map (< 2 x 10(5)/g faeces). CONCLUSIONS: These data support a pooling dilution of 1 in 25 for application of pooled faecal culture as a diagnostic tool in caprine Johne's disease control. A test based on this dilution would reduce laboratory costs of whole herd testing in goats by approximately 40% relative to serology and 75 to 90% relative to individual faecal culture.