Prevalence of antibodies to bluetongue virus and Anaplasma marginale in Montana yearling cattle entering Alberta feedlots: Fall 2001

A serologic survey was conducted in yearling cattle imported into Alberta feedlots from Montana during October 2001 to estimate the prevalence of antibodies to bluetongue virus (BTV) and Anaplasma marginale in Montana yearling cattle. The apparent prevalence of antibodies to BTV when the competitive enzyme-linked immunosorbent assay (cELISA) was used was 0.37% (21/5608). Test positive cELISA samples were also all positive when tested by virus neutralization (VN) and they reacted to 1 or more BTV serotypes, including 2, 10, 11, 13, and 17. The apparent prevalence of antibodies to A. marginale when a recombinant cELISA (rcELISA) was used with a positive cutoff at 30% inhibition was 1.93% (108/5608). When the rcELISA positive cutoff was at 42% inhibition, the apparent prevalence was 0.73% (41/5608). After the reported sensitivity and specificity of the test had been accounted for, the A. marginale antibody results were consistent with a population that was either free of exposure or had a very low prevalence for A. marginale.     

Prevalence of antibodies to serotypes of Bluetongue virus and Anaplasma marginale in Montana feeder cattle: 2002-2003

In a serologic survey of Montana-source weaned calves and yearling cattle, the apparent prevalence of antibodies to Bluetongue virus was 0.68% and 1.26% in 2002 and 2003, respectively, and to Anaplasma marginale at a positive cutoff at 30% inhibition it was 1.82% and 1.35% in 2002 and 2003, and at a positive cutoff at 42% inhibition it was 0.76% and 0.55% in 2002 and 2003, respectively, suggesting that the risk of importing infected animals was very low.     

Comparison of the complement fixation test and competitive ELISA for serodiagnosis of Anaplasma marginale infection in experimentally infected steers

OBJECTIVE: To compare sensitivity of a complement fixation (CF) test and competitive ELISA (cELISA) for detection of Anaplasma marginale in experimentally infected steers. ANIMALS: 40 crossbred (Angus-Simmental) steers. PROCEDURES: Steers were inoculated with 2.6 x 10(9) A marginale-infected erythrocytes (day 0). Blood samples were collected on days 9, 13, 20, 28, 34, 41, 61, 96, 126, and 156 days after inoculation. The percentage of parasitized erythrocytes (PPE) was determined by microscopic examination of stained blood films, and sera were evaluated with the CF test and cELISA by use of USDA-approved methods. Sensitivity and agreement (kappa statistic) between the 2 methods were determined. Persistent infections were confirmed by inoculation of blood obtained from infected steers into susceptible, splenectomized calves. RESULTS: 9 days after inoculation, sensitivity of the cELISA was 47.5%, whereas the CF test failed to identify seropositive steers. After day 13, sensitivity of the cELISA and CF test was 100% and 20%, respectively. During peak parasitemia (day 20), sensitivity of the cELISA and CF test was 100%. Thereafter, sensitivity of the CF test fluctuated between 7.5% and 37.5%, whereas sensitivity of the cELISA remained at 100%. Overall sensitivity of the cELISA and CF test was 94.8% and 26.5%, respectively (kappa statistic, 0.039). CONCLUSIONS AND CLINICAL RELEVANCE: The cELISA had superior sensitivity for serologic detection of A marginale.The CF test and cELISA each had a high percentage of false-negative results during the prepatent period. These findings are relevant for export certification and anaplasmosis prevention or eradication programs.     

Molecular and serological prevalence of Anaplasma marginale in cattle of North Central Morocco

A cross sectional study was conducted to investigate the epidemiological distribution of Anaplasma marginale in North Central Morocco. Blood samples from five provinces of Morocco were collected from apparently healthy cattle (n=668) and simultaneously analyzed by a nested polymerase chain reaction (nPCR) assay and competitive enzyme-linked immunosorbent assay (cELISA). The overall prevalence of A. marginale was 21.9% by nPCR and 16.5% by cELISA. The Kappa coefficient between nPCR and cELISA indicated a modest level of agreement (0.54). The prevalence of A. marginale varied significantly according to the province and the month of sampling. However age, gender and breed did not have a significant effect on the prevalence of this pathogen. The highest prevalence of A. marginale was found in the Gharb, a sub-humid area while the lowest was reported in the Saiss, a semi-arid area. These results indicate that an A. marginale infection are widespread in the country and suggests that either or both techniques are excellent tools for epidemiological studies and control programs.     

Epidemiologic aspects of bovine anaplasmosis in semiarid range conditions of south central Idaho

The prevalance of Anaplasma marginale-infected cows, as determined by use of the modified rapid card agglutination (MRCA) test, was measured during a 4-year period (1980-1983). The prevalence of A marginale-infected cows, defined as positive reactors on the MRCA test, remained constant (31%-37%). The apparent incidence of A marginale transmission to susceptible cows was approximately 7% from 1980 to 1981, 8% from 1981 to 1982, and no transmission from 1982 to 1983. The occasional MRCA-positive cow became negative on the MRCA test, and 1 cow was determined to be free of A marginale infection by subinoculation of 100 ml of the cow's blood into a susceptible, splenectomized calf. Dermacentor andersoni, a known vector of A marginale, was often found on the cattle and in their environment. However, A marginale was not transmitted to susceptible, splenectomized calves, using collected ticks. Of 56 calves born to MRCA-positive cows, 82% were MRCA-positive within the first 3 months of life. These calves converted to MRCA-negative status and were determined to be free of A marginale infection by subinoculation of their blood into susceptible, splenectomized calves, indicating the passive transfer of colostral antibodies.     

Serological Survey of Babesia bovis and Anaplasma marginale in cattle in Tete Province,Mozambique

A serological survey of bovine babesiosis and anaplasmosis in communal cattle was conducted in the northwestern province of Tete, Mozambique. Blood was collected from cattle ranging from 4 to 15 months old from randomly selected farms from six districts. Thirty-nine per cent of all 478 calves tested in Tete Province were seropositive to the ELISA for Babesia bovis antibodies and 63% of all calves were seropositive in the card agglutination test for Anaplasma marginale. Seroprevalence of B. bovis ranged from 22.8% in Tete City District to 48.1% in Angonia District. For A. marginale, it ranged from 34.4% in Angonia District to 87.3% in Moatize District. The dominant factor affecting seroprevalence for both haemoparasites was district and there was a trend for higher intensity of tick control to be associated with a higher seroprevalence of B. bovis and a lower seroprevalence of A. marginale. The obvious differences were the low prevalence of B. bovis in Tete City Council District and the low prevalence of A. marginale in Angonia District. The levels of exposure to B. bovis seen in our study are well below any that could be considered to be consistent with endemic stability, yet they are sufficiently high to ensure that clinical disease would be a risk. The seroprevalence of A. marginale, however, suggests that endemic stability with respect to this disease could exist in districts other than Angonia. There was no strong and consistent relationship between the intensity of control and the likelihood of seropositivity to either of the diseases.     

Natural transplacental infection of dairy calves with bovine immunodeficiency virus and estimation of effect on neonatal health

Many experimental infection studies with bovine immunodeficiency virus (BIV) have been conducted, but neither virus transmission under natural conditions nor longitudinal clinical effects of naturally occurring infections in non-experimental populations are well explored. We tested the hypotheses that BIV is transmitted across the placenta during gestation and that intragestionally infected calves are at increased risk of neonatal disease. A cohort of 59 dairy cows on one farm were enrolled at parturition and the BIV serostatus of the cows and their pre-colostral calves determined with an indirect fluorescent-antibody assay. Moreover, the enrolled calves were monitored thrice weekly for specific clinical signs through the duration of the 30 day neonatal period and the occurrence of clinical signs analyzed for association with calf pre-colostral BIV serostatus and dam BIV serostatus. Confounding due to calf passive immunity and season of birth were also explored. Forty percent of seropositive cows (14/35) gave birth to seropositive calves but no seropositive calves (0/19) were born to seronegative dams (estimated relative risk 16, 95% exact confidence interval 2.6–5.8×10²⁹). Calf pre-colostral BIV serostatus was not associated with the occurrence or frequency of clinical signs — but dam BIV serostatus was associated with the odds of occurrence of calf hyperthermia and with the frequency of occurrence of calf hyperthermia and hyperventilatory events. This study is inconclusive about the effects of prenatal BIV infection on neonatal health — but it does provide evidence for the natural occurrence of transplacental BIV infection.     

Seroprevalence of anaplasmosis among cattle in Switzerland in 1998 and 2003: no evidence of an emerging disease

Anaplasma marginale infection in Europe has been limited to the Mediterranean and eastern countries, to Austria and to very sporadic cases in Switzerland. There are no reports of its occurrence in the countries north of Switzerland. A severe outbreak of anaplasmosis in August 2002 in a cattle farm in the canton Grisons, Switzerland, north of the Alps, with more than 300 cattle that had to be culled, came unexpected and gave reason to hypothesize presence of an increased yet undetected prevalence of A. marginale in Switzerland. Randomly selected bovine serum samples collected in 1998 and 2003 were tested using a competitive inhibitory ELISA (cELISA) to test the hypothesis. Our validation of the diagnostic sensitivity and specificity of this test, done in the outbreak herd, yielded 99.2 and 83.3%, respectively, probably underestimating the true specificity. The true seroprevalence of anaplasmosis in Swiss cattle determined by cELISA was likely to be zero with upper 95% confidence limits of 2.49% in the canton Grisons and 1.17% in the rest of Switzerland, respectively, in 1998. For 2003, these estimates were even lower. There was no significant difference in apparent prevalences between 1998 and 2003. In search of a possible reservoir, three chamoises out of 46 free ranging wild ruminants from the Swiss National Park, Grisons, tested positive in the cELISA. This reaction is in accordance with A. marginale or a cross reacting agent such as Anaplasma ovis. From our results we conclude that the hypothesis of an increased prevalence of anaplasmosis in cattle in Switzerland must be rejected.     

Prevalence of Neospora caninum and persistent infection with bovine viral diarrhea virus in dairy-breed steers in a feedlot

OBJECTIVE: To determine the prevalence and effect of Neospora caninum infection and persistent infection (PI) with bovine viral diarrhea virus (BVDV) on weight gain, morbidity, and mortality rate in dairy-breed steer calves located on a feedlot in California. DESIGN: Prospective cohort observational study. ANIMALS: 900 dairy-breed steer calves in 2 pens. PROCEDURES: The 3- to 4-month-old calves were evaluated for serum antibodies against N caninum and infection with BVDV at entry to the feedlot. Five months later, sera were again analyzed for anti-N caninum antibodies; calves that were determined to have BVDV infection initially were retested to evaluate PI status. Average daily gain, morbidity, and deaths were recorded for all calves. RESULTS: Among 900 calves, prevalence of N caninum infection was 16.7% (95% confidence interval, 14.3% to 19.3%); prevalence of BVDV-associated PI was 0.2% (95% confidence interval, 0.03% to 0.9%). Morbidity rate and time to first illness were not significantly different between calves that were seropositive or seronegative for N caninum. At the second sample collection, weight and average daily gain of calves that were seropositive for N caninum was less than that of seronegative steers in 1 pen, whereas these measures did not differ between groups in the other pen. Statistical power was insufficient to evaluate the effect of BVDV PI on any outcome measurement. CONCLUSIONS AND CLINICAL RELEVANCE: Although N caninum serostatus had no significant effect on morbidity rate, some seropositive calves had reduced growth, compared with seronegative calves, 5 months after entry to the feedlot.     

Detection of antibodies against Anaplasma marginale in milk using a recombinant MSP5 indirect ELISA

An indirect enzyme linked immunosorbent assay (iELISA) for diagnosis of anaplasmosis using undiluted individual milk samples from dairy cows was developed. The recombinant 19 kDa major surface protein 5 (rMSP5) of Anaplasma marginale was used as antigen. A monoclonal antibody against bovine IgG1 conjugated with peroxidase and the chromogen 3,5,3',5'-tetramethylbenzidine were used in the test. Strong and weak, positive and negative milk samples were set up as reference controls. Results were expressed as percentage of positivity (PP) contrasting with the strongest positive control. The test was evaluated in two groups (G1 and G2) of lactating dairy cows from herds located in A. marginale non-endemic areas of Argentina. The infection status of both groups, G1 (n=128) sampled after anaplasmosis outbreak, and G2 (n=216) free of anaplasmosis was established by polymerase chain reaction (PCR). Serum samples of cows from G1 and G2 were analyzed by card agglutination test (CAT) and competitive ELISA (cELISA), while the novel iELISA was evaluated in their corresponding milk samples. At a cutoff of 42 PP, the ELISA has 98% sensitivity and 95% specificity. A significant difference (P<0.0001) was found between the mean PP value of negative samples from G1 (17.4+/-14.9), and G2 (8.6+/-7.1). The agreement and kappa (kappa) value between iELISA and PCR was 96%, kappa=0.919; between iELISA and CAT was 97%, kappa=0.880; and between iELISA and cELISA was 97%, kappa=0.899. These results strongly support the usefulness of iELISA to detect A. marginale antibodies in milk. Additional studies are necessary to define the ability of the milk iELISA to detect not only acutely infected, but also carrier cattle.     

Validation of an Anaplasma marginale cELISA for use in the diagnosis of A. ovis infections in domestic sheep and Anaplasma spp. in wild ungulates

A commercially available (cELISA) kit for diagnosing Anaplasma marginale infection in cattle was validated for diagnosing A ovis infection in sheep using the bovine serum controls as supplied by the manufacturer (BcELISA) and sheep serum controls from pathogen-free sheep (OcELISA). True positives were identified using two previously established assays, a nested PCR (nPCR) test and an indirect immunofluorescent assay (IFA). The BcELISA was also applied to sera from various species of wild ruminants, comparing the results with the IFA. Receiver operating characteristic (ROC) analysis indicated that the predicted threshold inhibition for the BcELISA was 19.2. The sensitivity for the BcELISA was 98.2% and the specificity was 96.3%. The predicted threshold inhibition decreased to 14.3 for the OcELISA; the sensitivity was 96.5% and the specificity was 98.1%. There was >/=90% concordance between IFA and nPCR, as well as between the BcELISA at 19% inhibition cutoff and either IFA or PCR. Concordance between the cELISA and IFA using sera from elk, mule deer, bighorn sheep, pronghorn antelope, and black-tailed deer ranged from 64% to 100%. This commercially available cELISA test kit can be used very effectively to test domestic sheep for infection with A. ovis using the kit-supplied controls (i.e. the BcELISA) and a 19% inhibition cutoff; the kit may also be useful for detecting intra-erythrocytic Anaplasma infections in wild ruminants.     

Serologic and molecular characterization of Anaplasma species infection in farm animals and ticks from Sicily

Although Anaplasma marginale was known to be endemic in Italy, the diversity of Anaplasma spp. from this area have not been characterized. In this study, the prevalence of Anaplasma spp. antibodies in randomly selected farm animals collected on the island of Sicily was determined by use of a MSP5 cELISA for Anaplasma spp. and an immunofluorescence test specific for Anaplasma phagocytophilum. Genetic variation among strains of Anaplasma spp. from animals and ticks was characterized using the A. marginale msp1alpha and the Anaplasma spp. msp4 genes. Eight species of ticks were collected and tested by PCR. Seropositivity for Anaplasma spp. and A. phagocytophilum was detected in bovine and ovine samples. All the donkeys were seropositive for A. phagocytophilum but not for Anaplasma spp. Four A. marginale genotypes were identified by msp4 sequences from bovine and tick samples. Two new genotypes of Anaplasma ovis were characterized in sheep. The sequences of A. phagocytophilum from three donkeys proved to be identical to the sequence of the MRK equine isolate from California. Six A. marginale genotypes were found in cattle and one tick using the A. marginale msp1alpha sequences. All genotypes had four repeated sequences in the N-terminal portion of the MSP1a, except for one that had five repeats. The Italian strains of A. marginale contained three repeat sequences that were not reported previously. Definition of the diversity of Anaplasma spp. in Sicily reported, herein is fundamental to development of control strategies for A. marginale, A. ovis and A. phagocytophilum in Sicily.     

Determination of the seroprevalence of Neospora caninum in feedlot steers in Alberta

A study was conducted in Alberta to determine the seroprevalence of Neospora caninum in feedlot calves purchased from various auction markets throughout western Canada. Four feedlots (1 feedlot from each of the Airdrie and High River areas and 2 feedlots from the Strathmore area) were selected for sampling. At each feedlot, a random 10% sample of feedlot steer and bull calves entering the feedlot from September 2001 to December 2001 were enrolled in the study until there were a maximum of 500 animals enrolled per feedlot. Blood samples were collected from 1976 male animals at the time of entry to the 4 study feedlots. The animals represented 375 groups purchased from 70 sale points throughout British Columbia, Alberta, Saskatchewan, and Manitoba. Of the 1976 animals tested, 128 animals tested positive for antibodies to N. caninum. The prevalence and adjusted 95% confidence limits for N. caninum in beef calves on entrance to the feedlot in western Canada were 6.5% (95% CI, 5.1 to 8.2). There were no significant (P > or = 0.05) associations between the risk of treatment, the risk of designation as "chronic," and the risk of death and antibodies to N. caninum either before or after adjusting for feedlot, entry weight, entry date, and clustering of disease within lots at each feedlot. In addition, there was no significant (P > or = 0.05) association between serological status and feedlot entry weight or average daily gain. Note that there was no information available on feed conversion because the calves were mixed within existing commercial feedlot pens and the actual feed intake of each animal could not be determined. Adjustment for the concentration of antibodies to bovine viral diarrhea virus on arrival did not change any of the examined associations between N. caninum status and calf health or performance. The results of this study demonstrated that the prevalence of N. caninum in feedlot calves in western Canada was less than the prevalence reported in the United States. Additional studies are required to determine whether the inferior rate of gain and feed efficiency observed in the southern United States with animals testing positive for antibodies to N. caninum also occurs under the management conditions used in western Canada.     

A field trial, of preshipment vaccination, with intranasal infectious bovine rhinotracheitis-parainfluenza-3 vaccines.

A total of 849 calves, 278 controls, 335 vaccinated intranasally with IBR-PI3/TS and 236 vaccinated intranasally with IBR-PI3/PTC were studied in a field trial of preimmunization. All calves were vaccinated in Saskatchewan at least three weeks prior to shipment to feedlots. Four hundred and twenty six calves were not sold within eight weeks of vaccination; however, seven of these died within four weeks of vaccination. Treatment rates varied from 1.0% to 5.2%. There was no significant effect of vaccination on treatment rates. Similarly, there was no significant effect of vaccination in the 74 calves sold to feedlots in Saskatchewan. Three hundred and forty nine calves were sold to feedlots in Ontario. Two of these died from fibrinous pneumonia. Treatment rates varied from 1.7% to 33.3% in different feedlots, but there was no significant effect of vaccination on treatment rates. Therefore, preimmunization is unlikely to significantly reduce the overall treatment rate in calves entering feedlots.     

Effects of subclinical bovine leukemia virus infection on some production parameters in a dairy farm in southern Turkey

Some production parameters of seropositive cows (age, first calving age, 305 day mature equivalent last milk yield production, lifetime mature equivalent milk yield production, lifetime total milk production, lifetime total milking period, lifetime monthly milk production, lifetime daily milk production, lifetime total days of milking, number of inseminations per pregnancy (for last pregnancy), number of calves and calving interval (for last pregnancy)) were analysed in the current study. The study population was clinically healthy Holstein cows from a commercial dairy herd in southern Turkey. Of 109 animals, 65 cows were seropositive by ELISA and the prevalence of bovine leukemia virus (BLV) infection was 59.6%. The prevalence of seropositive cows in 2nd (62.8%), 3rd (64.7%), 4th (61.5%), and 5th (66.6 %) lactations was slightly higher than that of cows in 1st (52.6%) lactations. No statistical differences were observed between BLV seronegative and seropositive cows for production and reproduction parameters analysed in this study (P > 0.05).     

Associations between the prevalence of Mollicutes and Mycoplasma bovis and health and performance in stocker calves

A longitudinal cross-sectional time-series study was carried out to determine the prevalence of nasal mycoplasma carriage, serostatus and seroconversion, and to evaluate the associations between these parameters and health and performance in weaned beef calves during a 42-day feeding period. Nasal swabs and serum were collected on days 0 (arrival), 10, 42 and at the first incidence of bovine respiratory disease complex. The samples were evaluated for Mollicutes (by culture), Mycoplasma bovis (by PCR) and serum antibody to M. bovis. On day 0, 90.4 per cent of the calves were Mollicutes nasal culture-positive. The seroprevalence of M. bovis was 26.6 per cent on day 0 and 98.2 per cent by day 42 (P<0.05). Seroconversion to M. bovis between days 0 and 42 was significantly associated (P=0.04) with lower weight gain. Weight gain was greater in calves that were PCR-negative for M. bovis on day 10 (P=0.01). The percentage of calves seropositive to M. bovis increased throughout the study, indicating exposure and an immunological response to the organism. Although associations with health outcomes were not identified, seroconversion to M. bovis was associated with a decreased rate of weight gain during the study period.     

Factors associated with Neospora caninum serostatus in cattle of 20 specialised Costa Rican dairy herds

Twenty-five specialised Costa Rican dairy farms (located in the Poás area) were used to determine neosporosis seroprevalence and the association of seropositivity with environmental and management factors. The farms involved were selected intentionally and all of them use VAMPP 5.1 (Veterinary Automated Management and Production Control Programme) as management-information system. Holstein–Friesian, Jersey and crosses between them were the most-frequent breeds in these herds. The number of females per farm varied from 41 to 296. Our cross-sectional study had two phases. In the first phase, we determined the presence or absence of seropositivity at herd level. For the second phase, all females in 20 seropositive farms were bled. Serum samples were tested for antibodies to Neospora caninum using an indirect enzyme-linked immunosorbent assay (ELISA). A questionnaire with factors mentioned in the literature was administered to the farmers. Logistic regression (LR with herd as random effect) was used to assess the relationships of the serostatus at the individual level with characteristics of the cows and environmental factors. In the first phase all herds had >20% seropositive females; therefore, all herds were eligible for the second phase. In the second phase, the overall prevalence was 39.7% (1191/3002), and within-herd prevalences were between 25.0 and 70.5%. Age 3–6 years, parity ≤2 of the dam of the cow, Jersey breed and lack of purposive sampling to diagnose abortive infectious disease were associated with positive serostatus; other management and environmental factors did not show significant associations. The lack of association between management and environmental factors with serostatus might be because all farms were exposed to a considerable number of potential factors. That all herds of this study were seropositive for neosporosis and the within-herd prevalence was considerable raises questions about how far the infection is spread in other dairy areas of Costa Rica.     

Radioimmunoassay for Anaplasma marginale antibodies in cattle

A radioimmunoassay is described for use in the detection of Anaplasma marginale antibodies in cattle sera. Optimal sensitivity and specificity were obtained by using 2 antigens, an A marginale antigen and a RBC antigen (obtained before infection was established) from the same calf. In addition, sera were preabsorbed with RBC from healthy cattle and with sonicated Babesia bovis. Of 86 sera obtained from cattle with A marginale infection (as determined by blood smear examination or by results of subinoculation of blood from such infected cattle into splenectomized calves), 85 had positive results by use of this test. Of 100 sera obtained from cattle raised in an anaplasmosis-free area, 98 yielded negative results, and sera obtained from 35 cattle (97 sera) infected with B bigemina and from 18 cattle infected with Theileria orientalis yielded negative results. By use of this test, 99 of 100 sera obtained from cattle with B bovis infection were negative for A marginale. Anaplasma marginale antibodies were detected in 18 cattle that had been pastured in a Boophilus microplus-free area for 2 years after natural infection. After 3 years, 16 of these cattle were still seropositive for A marginale. Sixteen cattle pastured in a Bo microplus-infested area had detectable antibody against A marginale 27 months after initial infection with A marginale. Sensitivity and specificity of the test were assessed as 98.8% for each.     

Anti-Neospora caninum antibodies in milk in relation to production losses in dairy cattle

A comprehensive field study was carried out with the following objectives: (a) to assess the usefulness of individual and bulk tank milk analysis for determining Neospora caninum serostatus in individual cows and herds, and (b) to study the associations between N. caninum infection status (based on milk testing), and several productive and reproductive parameters in the animals. Antibodies were detected with a commercially available ELISA test (Bio K 192/5). Analysis of paired serum and milk samples from 1134 lactating cows on 38 farms revealed that 97.6% of the ELISA results were coincident, irrespective of whether serum or milk samples were used. Moreover, multiple linear regression analysis revealed that 86.0% of the variations in ELISA values in milk were due to variations in the serum. The measurement of antibodies in bulk tank milk was a good estimator of the herd level status of N. caninum infection, and enabled detection of infection in 94.7% herds with ≥10.0% seropositive cows and/or in all herds with >4% highly seropositive cows. The odds ratio for abortion in seropositive animals was 9.1 times higher than in seronegative animals. The infection serostatus was also a significant risk factor, as the odds ratio for abortion was even higher (12.0 times) in cows categorized as highly seropositive. ELISA values for the bulk milk from 387 randomly selected herds were negatively associated with average milk production. Moreover, milk production losses mainly occurred on farms categorized as highly positive (i.e. herds with ≥20.0% seropositive cows).