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Six agglutination and two complement fixation tests were compared with respect to specificity, sensitivity and relative sensitivity for the serodiagnosis of bovine brucellosis. Based on 1051 sera from brucellosis free herds, the specificity of the tests was 98.9% for the buffered plate antigen test (BPAT), 99.2% and 99.3% for the standard tube and plate agglutination tests (STAT and SPAT), respectively, and 99.8% for the 2-mercaptoethanol test (2MET). On this small sample, the rose bengal plate test (RBPT), card test (CARD) and the complement fixation test (CFT) correctly classed all sera as negative. On a sample of 167 culture positive cattle, the sensitivities of the tests were CFT: 79.0%, BPAT: 75.4, RBPT: 74.9%, CARD: 74.3%, SPAT: 73.1%, STAT: 68.9%, and 2MET: 59.9%. All tests combined detected only 82% of these infected cattle. Analysis of the relative sensitivity of the six agglutination tests gave the following ranking: BPAT greater than RBPT greater than CARD greater than SPAT greater than STAT. The 2MET ranked between the BPAT and RBPT or between the RBPT and CARD depending on the analysis used. The use of the BPAT as a screening test is recommended provided that a test of high specificity and sensitivity such as the CFT is used to confirm screening test reactions.

The most common clinical manifestations of brucellosis in livestock are associated with reproduction. This paper reports the result of a cross-sectional study conducted between October, 2007 and April, 2008 in Western Tigray, North Ethiopia, with the objectives of assessing the effect of Brucella infection on reproduction conditions of female breeding bovine and to explore the presence of Brucella seroreactors in vulnerable humans. A total of 1,354 and 246 sera were collected from female cattle and humans, respectively. The sera were screened using Rose Bengal plate test (RBPT), and positive samples were confirmed by complement fixation test (CFT). Reproductive conditions for female cattle and risk to human brucellosis seropositivity were tested by using logistic regression analysis. The result indicated that the overall prevalence in female cattle was 6.1%. The study showed 1.2% prevalence among human risk groups, all of which were herdsmen. Logistic regression identified parity status, calving interval, abortion history, and abortion period were significantly associated with seropositivity. The association was not significant with reproductive status and parity number. Proper hygienic practices and team work between veterinary and health personnel should improve the efforts to combat disease transmission.

We investigated the seroprevalence and risk factors for Brucella seropositivity in cattle in Jordan. The sera from 671 cows were randomly collected from 62 herds. The antibodies against Brucella were detected using a Rose Bengal plate test and indirect ELISA. A structured questionnaire was used to collect information on the cattle herds' health and management. A multiple logistic regression model was constructed to identify the risk factors for Brucella seropositivity. The true prevalence of antibodies against Brucella in individual cows and cattle herds was 6.5% and 23%, respectively. The seroprevalence of brucellosis in cows older than 4 years of age was significantly higher than that in the younger cows. The seroprevalence of brucellosis in cows located in the Mafraq, Zarqa and Ma'an governorates was significantly higher than that of the other studied governorates. The multiple logistic regression model revealed that a larger herd size (odd ratio = 1.3; 95% CI: 1.1, 2.6) and mixed farming (OR = 2.0; 95% CI: 1.7, 3.7) were risk factors for cattle seropositivity to Brucella antigens. On the other hand, the use of disinfectants (OR = 1.9; 95% CI: 1.1, 2.1) and the presence of adequate veterinary services (OR = 1.6; 95% CI: 1.2, 3.2) were identified as protective factors.

A cross-sectional epidemiological study was conducted to determine the seroprevalence and to identify risk factors for bovine brucellosis seropositivity in traditional and smallholder dairy cattle production systems in the Tanga region of North-eastern Tanzania. The study populations comprised 246 indigenous and 409 crossbred cattle, randomly selected from 105 smallholder dairy and 25 traditional managed herds, respectively. Individual animal and herd-level data were collected using a structured questionnaire. Serum samples were screened for Brucella antibodies using the Rose Bengal Plate Test The overall seroprevalence of Brucella antibodies in the smallholder dairy and traditional managed cattle was 4.1% and 7.3% respectively. The corresponding overall herd prevalence was 10.5% and 20% respectively. Using multivariate logistic regression analysis, closeness to stock route, access to surface drinking water and location were identified as the major risk factors for individual herd seroprevalence. Older animals (≥6 years) were associated with increased risk of sero-positivity compared to animals of age category of ≤6 years. The results showed that brucellosis is prevalent and widely distributed locally, underscoring the need for further studies including surveillance and institution of preventive and control measures particularly among female young-stock and the general public who are at high risk of contracting brucellosis.

Background

Brucellosis is known to cause debilitating conditions if not promptly treated. In some rural areas of Tanzania however, practitioners give evidence of seeing brucellosis cases with symptoms of long duration. The purpose of this study was to establish health-seeking behaviour of human brucellosis cases in rural Tanzania and explore the most feasible ways to improve it.

Methods

This was designed as a longitudinal study. Socio-demographic, clinical and laboratory data were collected from patients who reported to selected hospitals in rural northern Tanzania between June 2002 and April 2003. All patients with conditions suspicious of brucellosis on the basis of preliminary clinical examination and history were enrolled into the study as brucellosis suspects. Blood samples were taken and tested for brucellosis using the Rose-Bengal Plate Test (RBPT) and other agglutination tests available at the health facilities and the competitive ELISA (c-ELISA) test at the Veterinary Laboratory Agencies (VLA) in the UK. All suspects who tested positive with the c-ELISA test were regarded as brucellosis cases. A follow-up of 49 cases was made to collect data on health-seeking behaviour of human brucellosis cases.

Results

The majority of cases 87.7% gave a history of going to hospital as the first point of care, 10.2% purchased drugs from a nearby drug shop before going to hospital and 2% went to a local traditional healer first. Brucellosis cases delayed going to hospital with a median delay time of 90 days, and with 20% of the cases presenting to hospitals more than a year after the onset of symptoms. Distance to the hospital, keeping animals and knowledge of brucellosis were significantly associated with patient delay to present to hospital.

Conclusion

More efforts need to be put on improving the accessibility of health facilities to the rural poor people who succumb to most of the diseases including zoonoses. Health education on brucellosis in Tanzania should also stress the importance of early presentation to hospitals for prompt treatment.

Bovine viral diarrhoea, caused by the bovine viral diarrhoea virus (BVDV) in the Pestivirus genus of the Flaviviridae, is one of the most important diseases of cattle world wide causing poor reproductive performance in adult cattle and mucosal disease in calves. In addition it causes immunosuppression and increased susceptibility to other infections, the impact of which is uncertain, particularly in sub-Saharan Africa where animals are exposed to a much wider range and higher intensity of infections compared to Europe. There are no previous estimates of the seroprevalence of BVDV in cattle in Cameroon. This paper describes the serological screening for antibodies to BVDV and antigen of BVDV in a cattle population in the Adamawa Region of Cameroon in 2000. The estimates of herd-level and within herd seroprevalences adjusted for test imperfections were 92% and 30% respectively and 16.5% of herds were classed as having a persistently infected calf (PI) in the herd within the last year based on the “spot” test approach. There was evidence of clustering of herds with PI calves across the north and west of the Region which corresponds with the higher cattle density areas and of self-clearance of infection from herds. A multivariable model was developed for the risk of having a PI calf in the herd; proximity to antelope, owning a goat, mixing with 10 other herds at grazing and the catchment area of the veterinary centre the herd was registered at were all significant risk factors. Very little is known about BVDV in sub-Saharan Africa and these high seroprevalences suggest that there is a large problem which may be having both direct impacts on fertility and neonate mortality and morbidity and also indirect effects through immunosuppression and susceptibility to other infections. Understanding and accounting for BVDV should be an important component of epidemiological studies of other diseases in sub-Saharan Africa.

Studies have been done on public health significance of brucellosis using serology with little or no emphasis to risk factors. Therefore, this study was designed to investigate seroprevalence of brucellosis and assess epidemiological variables associated with human brucellosis. After obtaining verbal consent, 241 peripheral blood samples were collected from occupationally exposed groups with and without pyrexia of unknown origin. A structured questionnaire was prepared to gather risk factors, such as occupation, age, sex, history of consuming raw milk and other unpasteurised dairy products, direct contact with domestic animals, general knowledge about the route of transmission and awareness level. Purposive sampling was used to select the key informants. All serum samples were first screened by Rose Bengal Plate Test (RBPT) and further analysed by Standard Tube Agglutination Test (STAT). The results revealed that 24.5% were positive by RBPT and diagnosis was established in 26.6% using STAT with a titre range between 80 and 1,280 IU/ml. Among occupational groups, prevalence was 17.8% in veterinarians and pharmacists but was not statistically significant. The most common clinical symptoms at presentation were fever, headache, back pain, arthralgia and myalgia. No female reactor was found and the mean age and standard deviation of seropositive patients was 34.69±10.97 years. Risk factors such as residence in rural area, participation in vaccination of animals and eating during working hours were significantly associated (P<0.05) with brucellosis by univariate and multivariate analysis. In conclusion, to deal with occupation-related disease like brucellosis, awareness on risk factors must be part of extension education campaign. Besides, regular surveillance of the disease needs to be integrated into control and prevention programme at a local and national level.

Brucellosis is a disease with worldwide distribution affecting animals and human beings. Brucella abortus is the causative agent of bovine brucellosis. The cross-reactions of currently available diagnostic procedures for B. abortus infection result in false-positive reactions, which make the procedures unreliable. These tests are also unable to differentiate Brucella-infected and -vaccinated animals. The present work is focused on the use of a nonlipopolysaccharide (LPS) diagnostic antigen, a recombinant 10-kDa (r10-kDa) protein of B. abortus, for specific diagnosis of brucellosis. The purified recombinant protein was used as a diagnostic antigen in plate enzyme-linked immunosorbent assay (p-ELISA) format to screen 408 bovine serum samples (70 presumptively negative, 308 random, and 30 vaccinated), and the results were compared with those of the Rose Bengal plate agglutination test (RBPT) and the standard tube agglutination test (STAT). Statistical analysis in presumptive negative samples revealed 100 and 98.41% specificity of p-ELISA with RBPT and STAT, and an agreement of 91.43% with the tests using Cohen's kappa statistics. In random samples, the agreement of p-ELISA was 77.92% and 80.52% with RBPT and STAT, respectively. p-ELISA investigation of vaccinated samples reported no false-positive results, whereas RBPT and STAT reported 30% and 96.6% false-positive results, respectively. The data suggest that p-ELISA with r10-kDa protein may be a useful method for diagnosis of bovine brucellosis. Furthermore, p-ELISA may also be used as a tool for differentiating Brucella-vaccinated and naturally infected animals.

The antibodies active in the Rose Bengal plate test (RBPT) for bovine brucellosis have been studied. The results of fractionation experiments showed that RBPT activity was associated with fractions containing immunoglobulin of the IgG1 class; other immunoglobulin classes were inactive in this respect although active in other tests. These results were confirmed by inhibition tests with specific antisera and by elution of the antibody from agglutinated RBPT antigen.

The major proportion of the serum complement-fixing activity was also present in the IgG1 fraction and it is suggested that the RBPT and CF reactions are probably mediated by the same antibodies.

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Background

Prior to the present study, the seroprevalence of leptospirosis in Irish suckler herds was unknown. In this study, we describe the herd and animal-level prevalence of Leptospira Hardjo infection in the Irish suckler cattle population. For the purposes of the study, the 26 counties of the Republic of Ireland were divided into 6 regions from which a representative number of herds were selected. A herd was considered eligible for sampling if it was not vaccinating against leptospirosis and if it contained ≥ 9 breeding animals of beef breed ≥ 12 months of age. In total, 288 randomly selected herds were eligible for inclusion in the seroprevalence dataset analysis. Serological testing was carried out using a commercially available monoclonal antibody-capture ELISA, (sensitivity 100%; specificity 86.67%).

Results

Herds were categorised as either “Free from Infection” or “Infected” using the epidemiological software tool, FreeCalc 2.0. Using this classification, 237 herds were “Infected” (82.29%). The South West and South East regions had the highest herd prevalence. The regional effect on herd prevalence was largely mirrored by breeding herd size. A true animal-level prevalence of 41.75% was calculated using the epidemiological software tool, TruePrev. There was a statistically significant regional trend, with true prevalence being highest in the South East (P < 0.05). The median Breeding Herd Size (BHS), when categorised into quartiles, had a statistically significant influence on individual animal true seroprevalence (P < 0.001); true seroprevalence increased with increasing BHS.

Conclusions

Leptospirosis is a widespread endemic disease in the Republic of Ireland. It is possible that economic losses due to leptospirosis in unvaccinated Irish suckler herds may be underestimated.

Kyrgyzstan reported 77.5 new human brucellosis cases per 100,000 people in 2007, which is one of the highest incidences worldwide. In Kyrgyzstan, the currently used diagnostic tests in humans and animals are the Rose Bengal Test and the Huddleson test. A national representative cross-sectional study using cluster sampling proportional to size in humans, cattle, sheep, and goats was undertaken to assess the apparent seroprevalence in humans and animals. A total of 4,936 livestock sera and 1,774 human sera were tested in Naryn, Chuy, and Osh Oblasts. The overall apparent seroprevalences of brucellosis were 8.8% in humans (95% CI 4.5–16.5), 2.8% (95% CI 1.6–4.9%) in cattle, 3.3% (95% CI 1.5–6.9%) in sheep, and 2.5% (95% CI 1.4–4.5%) in goats. Naryn Oblast had the highest seroprevalences in humans and sheep. More men than women were seropositive (OR = 1.96; P < 0.001). Human seroprevalence was significantly associated with small ruminant seroprevalence but not with cattle seroprevalence. Annual incidence of human brucellosis exposure, measured by serological tests, was more than ten times higher than the annual incidence of reported clinical brucellosis cases. This indicates an under-reporting of human brucellosis cases, even if only a fraction of seropositive people have clinical symptoms. In conclusion, this study confirms the high seroprevalence of brucellosis in Kyrgyzstan and warrants rapid effective intervention, among others, by mass vaccination of sheep and goats but also of cattle.

Brucellosis, leptospirosis and Q fever are important infections of livestock causing a range of clinical conditions including abortions and reduced fertility. In addition, they are all important zoonotic infections infecting those who work with livestock and those who consume livestock related products such as milk, producing non-specific symptoms including fever, that are often misdiagnosed and that can lead to severe chronic disease. This study used banked sera from the Adamawa Region of Cameroon to investigate the seroprevalences and distributions of seropositive animals and herds. A classical statistical and a multi-level prevalence modelling approach were compared. The unbiased estimates were 20% of herds were seropositive for Brucella spp. compared to 95% for Leptospira spp. and 68% for Q fever. The within-herd seroprevalences were 16%, 35% and 39% respectively. There was statistical evidence of clustering of seropositive brucellosis and Q fever herds. The modelling approach has the major advantage that estimates of seroprevalence can be adjusted for the sensitivity and specificity of the diagnostic test used and the multi-level structure of the sampling. The study found a low seroprevalence of brucellosis in the Adamawa Region compared to a high proportion of leptospirosis and Q fever seropositive herds. This represents a high risk to the human population as well as potentially having a major impact on animal health and productivity in the region.

The serology of 27 heifers found to be positive to culture after inoculation with Brucella abortus strain 544, was studied. Eighteen heifers had previously been vaccinated with strain 19 or strain 45/20 and nine were unvaccinated. Post-infection serum samples were tested for Brucella antibodies by radioimmunoassay (RIA), complement fixation test (CFT), indirect haemolysis test (IHLT) and Rose Bengal plate test (RBPT). All of the unvaccinated heifers showed strong humoral responses to experimental infection in the RIA, CFT, IHLT and RBPT. The CFT and RBPT became positive sooner after infection than the other tests in the unvaccinated heifers. However, in vaccinated heifers the RIA was the most sensitive test early in infection and the results of the RBPT were variable. Three of the vaccinated heifers showed weak and inconsistent humoral responses and, in these animals, the RIA gave fewer false negative reactions than the other tests.

Sera were collected using a systematic random sampling from 348 cattle herds in Ontario, in proportion to the cattle population in different areas. One cow in five from 296 dairy herds and one in three from 52 beef herds were sampled. The sera were analyzed for prevalence of antibodies to Leptospira interrogans serovar grippotyphosa, hardjo, icterohaemorhagiae and pomona using the microscopic agglutination test. Herd seroprevalence (one or more animals with titer greater than or equal to 80) in beef and dairy herds combined was grippotyphosa 2%, hardjo 13.8%, icterohaemorrhagiae 10.1% and pomona 25.8%; 39% of all herds showed evidence of leptospiral infection with one or more serovars; 44.2% of 52 beef herds had serological evidence of infection with serovar hardjo compared to 8.4% of 296 dairy herds (P less than 0.0001). Seroprevalence of other serovars was not significantly different between beef and dairy herds. The proportion of beef animals seropositive for hardjo and for pomona increased with age, particularly for hardjo; 26.5% of beef animals aged nine years or over were seropositive for hardjo. Dairy animals showed a significant rise of hardjo but not pomona titers with age. The seroprevalence of pomona infection was significantly higher in dairy cattle in eastern Ontario than in other regions. Thirty-four (6.1%) of 553 aborted bovine fetuses had leptospires detected by immunofluorescence techniques. Sixty-five percent of these fetuses were from submissions made between November and January. Leptospires were identified as serovar hardjo by specific immunofluorescence. There appeared, however, to be a paradoxical serological response in that eight aborting cows had antibody titers to pomona rather than hardjo.(ABSTRACT TRUNCATED AT 250 WORDS)

Data from cattle herds infected with brucellosis and from control (noninfected) herds were collected and analyzed using case control techniques. It appeared that herds located close to other infected herds and those herds whose owners made frequent purchases of cattle had an increased risk of acquiring brucellosis, particularly those who made purchases from other herds or from cattle dealers. Infected herds had a lower level of vaccination than noninfected herds. However, the percentage vaccinated was highly variable in each group. Vaccination per se did not appear to adversely influence the interpretation of serological test results nor did it appear to protect the individual animal. Once infected, the time required to become free of brucellosis was increased by large herd size and/or loose housing. Closed herds also took longer to become brucellosis free than more open herds. The percentage of animals removed from the herd was increased by active abortion. Those herds with multiple serological reactors (positives and questionables) at the first herd test after the imposition of quarantine had the highest percentage of cattle removed.

A cross-sectional study was conducted to determinate the seroprevalence rate of equine brucellosis in the state of Tamaulipas, Mexico. Serum samples from 420 equines were analyzed with the Rose Bengal test at cell concentrations of 3% (RBT-3%) and 8% (RBT-8%), and positive results were confirmed with the Rivanol test (RT). Risk factors were determined with the prevalence ratio (PR) and the use of variables generated from a questionnaire administered to the animals’ owners. Serum from 1 stallion had positive results with both the RBT-8% and the RT, for a seroprevalence rate of 0.238%. Drinking of water from a pond that was also used by cattle and dogs was the only associated risk factor for this animal (PR = 0.25). However, the results were considered false-positive, because the results for other horses in the same environmental conditions were negative. Although brucellosis is considered endemic in ruminants in the study area, the results obtained suggest that equines are not a reservoir of brucellosis and do not play an important role in the epidemiologic patterns of this disease in northeastern Mexico.

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%.

Background

Human brucellosis has been found to be prevalent in the urban areas of Kampala, the capital city of Uganda. A cross-sectional study was designed to generate precise information on the prevalence of brucellosis in cattle and risk factors for the disease in its urban and peri-urban dairy farming systems.

Results

The adjusted herd prevalence of brucellosis was 6.5% (11/177, 95% CI: 3.6%-10.0%) and the adjusted individual animal prevalence was 5.0% (21/423, 95% CI: 2.7% - 9.3%) based on diagnosis using commercial kits of the competitive enzyme-linked immunosorbent assay (CELISA) for Brucella abortus antibodies. Mean within-herd prevalence was found to be 25.9% (95% CI: 9.7% - 53.1%) and brucellosis prevalence in an infected herd ranged from 9.1% to 50%. A risk factor could not be identified at the animal level but two risk factors were identified at the herd level: large herd size and history of abortion. The mean number of milking cows in a free-grazing herd (5.0) was significantly larger than a herd with a movement restricted (1.7, p < 0.001).

Conclusions

Vaccination should be targeted at commercial large-scale farms with free-grazing farming to control brucellosis in cattle in and around Kampala city.

Background

Neosporosis caused by the protozoan parasite Neospora caninum, is an economically important cause of abortion, stillbirth, low milk yield, reduced weight gain and premature culling in cattle. Consequently, a seroepidemiological study of N. caninum antibodies was conducted in England with 29,782 samples of blood taken from 15,736 cattle from 114 herds visited on three occasions at yearly intervals. Herds were categorised into lower (< 10%) and higher (≥ 10%) median herd seroprevalence. Hierarchical models were run to investigate associations between the sample to positive (S/P) ratio and herd and cattle factors.

Results

Ninety-four percent of herds had at least one seropositive cow; 12.9% of adult cattle had at least one seropositive test. Approximately 90% of herds were seropositive at all visits; 9 herds (8%) changed serological status between visits. The median N. caninum seroprevalence in positive herds was 10% (range 0.4% to 58.8%). There was a positive association between the serostatus of offspring and dams that were ever seropositive. In the hierarchical model of low seroprevalence herds there was no significant association between S/P ratio and cattle age. There was a significantly lower S/P ratio in cattle in herds that were totally restocked after the foot-and-mouth epidemic of 2001 compared with those from continuously stocked herds and cattle purchased into these herds had a higher S/P ratio than homebred cattle. In the model of high seroprevalence herds the S/P ratio increased with cattle age, but was not associated with restocking or cattle origin.

Conclusion

There were no strong temporal changes in herd seroprevalence of N. caninum but 90% of herds had some seropositive cattle over this time period. Vertical transmission from seropositive dams appeared to occur in all herds. In herds with a high seroprevalence the increasing S/P ratio in 2–4 year old cattle is suggestive of exposure to N. caninum: horizontal transmission between adult cattle, infection from a local source or recrudescence and abortions. Between-herd movements of infected cattle enhance the spread of N. caninum, particularly into low seroprevalence herds. Some restocked herds had little exposure to N. caninum, while in others infection had spread in the time since restocking.

A study was performed to evaluate the previously described PCR (C. Romero, C. Gamazo, M. Pardo, and I. López-Goñi, J. Clin. Microbiol. 33:615-617, 1995) for the diagnosis of brucellosis in dairy cattle. Milk samples from 56 Brucella milk culture-positive cattle and from 37 cattle from Brucella-free herds were examined for Brucella DNA by PCR and for specific antibodies by an indirect enzyme-linked immunosorbent assay (ELISA). The specificities of both tests were 100% when testing the milk samples from Brucella-free cattle. The milk samples from 49 infected cattle were positive by PCR (87.5% sensitivity), and 55 were positive by ELISA (98.2% sensitivity). A PCR-positive sample was negative by ELISA, and 7 ELISA-positive samples were PCR negative, yielding an observed proportion of agreement of 0.91 for the two tests. Although the results suggest that ELISA is a better screening test than PCR, the combined sensitivity of the two assays was 100%, and their simultaneous application could be more useful than one test alone for a rapid screening of brucellosis in dairy cattle.

The results are presented of testing untreated producer—retailer herd milk samples for the presence of Brucella abortus during the period 1965-1972 in the North Lancashire region.

There was a steady decline in the incidence of infected herds from 22% in 1965 to 12% in 1971. A sharp fall to 5% in 1972 suggests that the introduction of the Brucellosis Incentives Scheme and the eradication programme has helped to reduce the practice of selling brucella-infected cattle in the open market which was prevalent in the period 1965 to 1970.

This practice of selling brucella-infected cattle may also be a prime factor in the changing pattern of distribution of the biotypes of B. abortus which was observed during the period 1965 to 1970.

A comparison of the two areas in the region show that the incidence of herd infection was always greater in the area containing flying herds than in the area in which self-contained herds predominated.

Bulk milk samples from 220 dairy herds were collected at 9 public milk collection centres in the northeastern and northern Thailand, and a subset of 11 herds was selected for individual testing. The samples were tested for presence of antibodies to BVDV and BHV-1 using an indirect ELISA. The results from the bulk milk testing demonstrated a moderate level of exposure to BVDV and BHV-1 (73% and 67%, respectively). However, the low proportion of herds with high BVDV antibody-levels (13%) and the low within-herd seroprevalence of BVDV and BHV-1 in the 11 herds (24% and 5%, respectively), particularly among the young stock (15% and 0%, respectively), demonstrated a low prevalence of active BVDV infection and a low rate of reactivation of latent BHV-1. The presence of a self-clearance process was also indicated by the results from the individual testing. Moreover, a surprisingly low prevalence of BVDV and BHV-1 antibody-positive herds at one of the milk centres was found. This centre was established 5–10 years before the others. Our impression is that this reflects the self-clearance process, where consecutive replacement of imported infected animals without further spread has resulted in a nearly total elimination of the infections.

Based on our experiences and on these results we are convinced that this process can continue if there is awareness of herd biosecurity. This is especially important in the context of a future intensification of the dairy production.

The purpose of this study was to describe the responses of sera from five groups of cattle to an enzyme-linked immunosorbent assay (ELISA) for paratuberculosis by using serum absorbed with Mycobacterium phlei at a single working dilution. The infection status of the cattle was determined by fecal culture. Cattle with different levels of exposure (high versus low prevalence and test negative) and disease manifestation (clinically suspect infection versus subclinical infection) were examined, as follows: (i) two paratuberculosis-negative herds; (ii) a fecal culture-confirmed, clinically suspect cases of paratuberculosis; (iii) cows from a paratuberculosis-infected herd with a high infection rate, as determined by fecal culture, but with no clinical cases at the time of sampling; (iv) cows from three paratuberculosis-infected herds known to have paratuberculosis diagnosed on the farm (low infection rate determined by fecal culture); and (v) one fecal culture-negative herd with known serologically positive cattle. Results generally showed a decreased ELISA response when absorbed rather than nonabsorbed serum from each animal was used. The results of the fecal culture confirmed clinically suspect cases, which were analyzed in relation to the amount of colonies isolated from the animals on fecal culture (0, +, ++,+++ , ++++, and above). There was a significant increase in the ELISA response for animals with heavy Mycobacterium paratuberculosis shedding ( ++++ or above), when both unabsorbed and absorbed sera were used, compared with the response in animals that were fecal culture negative or that shed M. paratuberculosis at lower levels (less than +) (P less than 0.05). The effects on sensitivity and specificity by using different cutoff points for the five groups of cattle with different levels of exposure is described, since sera were not discretely segregated into distinct groups of positive and negative samples. The specificity of the ELISA in the two fecal culture-negative herds was 100% at an ELISA cutoff of an optical density (OD) of 0.1 and above for absorbed serum. For unabsorbed serum the specificity was 62.9% at a similar cutoff value. Similarly, the specificity of the fecal culture-negative, serologically positive herd increased from 37.5 to 72.2 at an ELISA cutoff value of 0.1 to 0.2 (OD) by using absorbed versus unabsorbed serum from 75.0 to 94.4 at an ELISA cutoff value of 0.2 to 0.3 (OD).

Five diagnostic tests based on enzyme-linked immunosorbent assay (ELISA) technology for bovine paratuberculosis were evaluated by using individual serum or milk samples from 359 dairy cattle in seven paratuberculosis-free herds and 2,094 dairy cattle in seven Mycobacterium paratuberculosis-infected dairy herds. Three independent laboratories using three different culture procedures completed fecal cultures for M. paratuberculosis on these cattle and found 417 cows to be shedding M. paratuberculosis in their feces. An animal that was fecal culture positive for M. paratuberculosis by any of the three laboratories was considered a confirmed case of infection. The specificity of three ELISAs (two on serum and one on milk) was ≥99.8%. The specificity of the remaining two ELISAs, both done on serum, was 94.9 and 84.7%. Four of the five ELISAs evaluated produced similar sensitivity in detecting fecal culture-positive cattle (27.8 to 28.9%). Serum ELISA “D” had the lowest specificity (84.7%) and the highest sensitivity (44.5%), but if the cutoff value defining a positive test was changed from 125 to 250% (of the positive control) the sensitivity and specificity, 31.8 and 97.5%, respectively, were comparable to those of the other four assays. If the case definition for M. paratuberculosis infection was based on the culture results of a single laboratory instead of the combined results of three laboratories, ELISA sensitivity estimates were 45.7 to 50.0%. With the exception of ELISA D, assay agreement was high (kappa 0.66 to 0.85) for categorical assay interpretations (positive or negative), but linear regression of quantitative results showed low correlation coefficients (r2 = 0.40 to 0.68) due to the fact that ELISA results for some cows were high in one assay but low in another assay. Likelihood ratio analysis showed a direct relationship between the magnitude of ELISA result and the odds of a cow shedding M. paratuberculosis in its feces. If used judiciously and interpreted quantitatively, these ELISAs are useful tools in support of paratuberculosis control programs in dairy herds.

Background

Bovine viral diarrhoea (BVD) is an infectious disease of cattle with a worldwide distribution. Herd-level prevalence varies among European Union (EU) member states, and prevalence information facilitates decision-making and monitoring of progress in control and eradication programmes. The primary objective of the present study was to address significant knowledge gaps regarding herd BVD seroprevalence (based on pooled sera) and control on Irish farms, including vaccine usage.

Methods

Preliminary validation of an indirect BVD antibody ELISA test (Svanova, Biotech AB, Uppsala, Sweden) using pooled sera was a novel and important aspect of the present study. Serum pools were constructed from serum samples of known seropositivity and pools were analysed using the same test in laboratory replicates. The output from this indirect ELISA was expressed as a percentage positivity (PP) value. Results were used to guide selection of a proposed cut-off (PCO) PP. This indirect ELISA was applied to randomly constructed within-herd serum pools, in a cross-sectional study of a stratified random sample of 1,171 Irish dairy and beef cow herds in 2009, for which vaccination status was determined by telephone survey. The herd-level prevalence of BVD in Ireland (percentage positive herds) was estimated in non-vaccinating herds, where herds were classified positive when herd pool result exceeded PCO PP. Vaccinated herds were excluded because of the potential impact of vaccination on herd classification status. Comparison of herd-level classification was conducted in a subset of 111 non-vaccinating dairy herds using the same ELISA on bulk milk tank (BMT) samples. Associations between possible risk factors (herd size (quartiles)) and herd-level prevalence were determined using chi-squared analysis.

Results

Receiver Operating Characteristics Analysis of replicate results in the preliminary validation study yielded an optimal cut-off PP (Proposed Cut-off percentage positivity - PCO PP) of 7.58%. This PCO PP gave a relative sensitivity (Se) and specificity (Sp) of 98.57% and 100% respectively, relative to the use of the ELISA on individual sera, and was chosen as the optimal cut-off since it resulted in maximization of the prevalence independent Youden’s Index.

The herd-level BVD prevalence in non-vaccinating herds was 98.7% (95% CI - 98.3-99.5%) in the cross-sectional study with no significant difference between dairy and beef herds (98.3% vs 98.8%, respectively, p = 0.595).

An agreement of 95.4% was found on Kappa analysis of herd serological classification when bulk milk and serum pool results were compared in non-vaccinating herds. 19.2 percent of farmers used BVDV vaccine; 81% of vaccinated herds were dairy. A significant association was found between seroprevalence (quartiles) and herd size (quartiles) (p < 0.01), though no association was found between herd size (quartiles) and herd-level classification based on PCO (p = 0.548).

Conclusions

The results from this study indicate that the true herd-level seroprevalence to Bovine Virus Diarrhoea (BVD) virus in Ireland is approaching 100%. The results of the present study will assist with national policy development, particularly with respect to the national BVD eradication programme which commenced recently.