Evaluation of highly sensitive indigenous milk ELISA kit with fecal culture, milk culture and fecal-PCR for the diagnosis of bovine Johne's disease (BJD) in India

Country lacks indigenous diagnostic kits against Johne's disease in animals. Indigenous ELISA and IS 900 PCR kits, originally developed for goats and sheep, have been adapted for screening of lactating cows. Multiple diagnostic tests were used to screen 26 lactating dairy cows against Johne's disease. Milk ELISA was evaluated with fecal culture, milk culture and fecal PCR. Of the 26 samples from lactating cows, 84.6, 96.1, 88.4 and 23.0% were positive in fecal culture, milk culture, m-ELISA and m-PCR, respectively. Comparatively milk sediment and milk fat culture detected 84.6 and 76.9% cows positive, respectively. Comparatively fecal culture and milk culture detected 84.6 and 96.1% cows positive, respectively. M-ELISA detected 11.5, 0.0, 11.5, 61.0 and 15.3%, cows as negative, suspected, low positive, positive and strong positive, respectively. There was good correlation between milk and fecal culture with m-ELISA. Three negative cows in m-ELISA were also detected in milk and fecal culture. Of the 26 decontaminated fecal samples, 23.0% cows were positive using specific IS 900 f-PCR. Comparative evaluation of m-ELISA with fecal and milk culture showed agreement in 80.7 and 84.6% cows, respectively. Sensitivity of m-ELISA with respect to fecal and culture was 90.9 and 95.6%, respectively. Comparative evaluation of four tests (milk culture, fecal culture, m-ELISA and f-PCR) showed that only 15.3% cows were detected in all the four tests. In three tests (fecal and milk culture and m-ELISA), 57.6% cows were detected positive. None of the cow was exclusively detected in f-PCR. Of the four diagnostic tests used milk culture was most sensitive (96.15%), followed by fecal culture (86.6%), m-ELISA (76.9%) and IS 900 PCR (23.0%) for the diagnosis of bovine Johne's disease (BJD). Milk ELISA detected only one cow extra, which was negative in milk culture. In view of the simplicity, rapidity and efficacy present milk ELISA kit employing soluble protoplasmic antigen from native Map 'Bison type' genotype of goat origin can be reliable for screening of bovine population against Johne's disease in India.     

Estimated within-herd prevalence (WHP) of Mycobacterium avium subsp. paratuberculosis in a sample of Minnesota dairy herds using bacterial culture of pooled fecal samples

The objective of this study was to describe the estimated within-herd prevalence (WHP) of Mycobacterium avium subsp. paratuberculosis (Map) in a sample of infected dairy herds in Minnesota (N = 66) using test results from bacterial culture of pooled fecal samples. Fecal samples were collected from up to 100 cows in each herd and were tested using bacterial culture in pools of 5 cows based on age order. The mean herd size was 222 (44 to 1500) milking cows; the cows were predominantly Holstein. Using a frequentist approach, the within-herd mean individual fecal prevalence was 10% [95% confidence interval (CI) = 4% to 16%] assuming 70% test sensitivity and 99.5% test specificity. Using Bayesian methods, the estimated true within-herd individual cow prevalence was 14% (95% CI = 7% to 27%). Within-herd prevalence was higher in larger dairy herds than in herds with fewer cows. As Map is the causative agent of Johne's disease (JD), the results of this study could contribute to the success of a nationwide control program for this disease.     

Effect of paratuberculosis on culling, milk production, and milk quality in dairy herds

OBJECTIVE: To determine the effect of paratuberculosis on culling, milk production, and milk quality in infected dairy herds. DESIGN: Cross-sectional study. ANIMALS: 689 lactating dairy cows in 9 herds. PROCEDURE: Milk, blood, and fecal samples were obtained from all cows. Fecal samples were evaluated via mycobacterial culture. Serum samples were tested with a commercially available ELISA for antibodies against Mycobacterium avium subsp paratuberculosis, and preserved milk samples were tested with an ELISA for antibodies against M paratuberculosis. Mixed effect and proportional hazards models were used to determine the effect of paratuberculosis on 305-day milk, fat, and protein production; somatic cell count linear score; and the risk of culling. RESULTS: Cows with positive results of bacteriologic culture of feces and milk ELISA produced less milk, fat, and protein, compared with herdmates with negative results. No difference in 305-day milk or fat production was detected in cows with positive results of serum ELISA, compared with seronegative cows. The 3 survival analyses revealed that cows with positive results of each test were at higher risk of being culled than cows with negative results. Paratuberculosis status, as determined by use of all 3 diagnostic tests, was not associated with milk somatic cell count linear score. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that for the 9 herds in this study, paratuberculosis significantly decreased milk production and cow longevity.     

The association between Mycobacterium avium subsp. paratuberculosis fecal shedding or clinical Johne's disease and lactation performance on two Minnesota, USA dairy farms

Lactation performance of cows infected with Mycobacterium avium subsp. paratuberculosis (Map) was previously studied using only serum ELISA as a diagnostic method. This study evaluated on two dairy farms in Minnesota, USA the lactation performance (measures of health, production, reproduction, and survival) of cows shedding Map in feces before calving and of cows culled with clinical signs consistent with Johne's disease (JD) during the subsequent lactation. Fecal samples were collected from 1052 cows within 21 day before calving and tested for Map with bacterial culture. Producers’ observed signs of clinical disease (milk fever, retained placenta, metritis, ketosis, displaced abomasum, lameness, mastitis, pneumonia, and JD) and production and reproduction data were recorded for each cow. The association between fecal shedding or clinical JD and lactation performance was evaluated. Logistic regression was used to evaluate the association with any clinical and subclinical diseases as the outcome. General linear model was used to evaluate the association with milk production, and survival analysis techniques were used to evaluate the association with days in the study before culling and days from calving to conception. In 84 cows (8% of 1052 cows) fecal samples were positive for Map (46% light, 26% moderate, and 28% heavy shedders). In multivariable analysis, light, moderate, and heavy fecal shedding cows produced on average 537, 1403, and 1534 kg, respectively, less milk per lactation and 1.4, 5.2, and 7.5 kg, respectively, less milk per day than fecal negative cows. Fecal culture positive cows were less likely to be bred and conceive. In the multivariable analysis the 56 cows culled with presumed JD produced approximately 1500 kg/lactation or 5 kg/day less than all other cows. The negative economic impact implied by decreased lactation performance in cows shedding Map or with clinical JD may motivate producers to implement programs to control Map infection and subsequent JD.     

Use of bulk milk for detection of Neospora caninum infection in dairy herds in Thailand

The relationship between the level of Neospora caninum antibodies in bulk milk and the seroprevalence in lactating cows was investigated. Bulk milk was also used to estimate the prevalence of N. caninum infection in dairy herds in the northeast and north Thailand. Bulk milk and individual serum from all lactating cows in 11 herds as well as 220 bulk milk samples from nine milk collection centres were analysed for presence of N. caninum antibodies using an iscom ELISA. In the 11 herds the bulk milk absorbances ranged between 0.04 and 0.89 and the seroprevalences varied between 0 and 46%. Five herds had milk absorbances below 0.20, among those were the two herds housing only seronegative lactating cows. In the remaining three herds with such low bulk milk absorbances one or two cows (5-14%) were seropositive. Six of the investigated herds had bulk milk absorbances above 0.20. In the two herds with the highest bulk milk absorbances more than 30% of the cows were seropositive. Using an absorbance of 0.20 to discriminate between negative and positive herds, 102 (46%) of 220 bulk milk samples were judged positive. There was no significant difference in mean bulk milk absorbance between the milk collection centres within each region. However, the proportion of herds with bulk milk absorbances > or =0.50 in the north was statistically (P < 0.01) higher than that in the northeast. It was concluded that bulk milk antibody testing can be used to identify N. caninum-infected herds and that N. caninum is a common infection in dairy herds in Thailand.     

Prevalence of Coxiella burnetii infection in Dutch dairy herds based on testing bulk tank milk and individual samples by PCR and ELISA

A study using an ELISA and a real-time PCR assay based on the detection of the repetitive transposon-like gene of Coxiella burnetii revealed that infection with the bacterium was widespread among Dutch dairy herds, with antibodies detected in bulk tank milk (BTM) from 268 of 341 herds (78.6 per cent) and bacterial DNA detected in 193 of 341 herds (56.6 per cent). The BTM samples were taken in November and December 2007. Serological and molecular studies in young and adult cattle selected from 100 herds showed that antibodies were present in the blood of 470 of 2936 (16.0 per cent) lactating cows but only in 19 of 1831 (1.0 per cent) young animals. Bacterial DNA was detected in the milk of 254 of 2925 (8.7 per cent) lactating cows; bacterial DNA was not detected in any of the faecal samples obtained from youngstock. The blood and milk samples were taken from the cattle in the period January to April 2008.     

Characteristics and management practices associated with milk production in dairy herds in Ohio enrolled in official Dairy Herd Improvement Association programs

OBJECTIVE: To determine herd characteristics and management practices associated with milk production in dairy herds enrolled in official Dairy Herd Improvement Association (DHIA) programs in Ohio. SAMPLE POPULATION: 186 dairy farms in Ohio. PROCEDURE: All herds in official DHIA programs in 9 counties were invited to participate. Information regarding herd characteristics and management practices was obtained, using a standardized questionnaire. Bulk-tank milk samples were obtained for bacteriologic culture. Official DHIA test-day records were obtained, and associations were identified, using multivariable ANOVA procedures. RESULTS: Of 479 eligible producers, 186 (39%) participated, and consecutive bulk-tank milk samples were available for culture from 172 (36%). Streptococcus agalactiae and Mycoplasma spp were not recovered from bulk-tank milk samples, but Staphylococcus aureus was recovered from 64 (37%) herds. Mean (+/- SD) number of lactating cows in participating herds was 97+/-66, with 123 (66%) herds milking < 100 cows. The RHA was significantly associated with number of cows in milk, estimated percentage of herd detected in estrus, reported annual percentage of heifer calves born alive that died before 8 weeks old, percentage days in milk, use of bovine somatotropin during the preceding 2 years, and sex of the person completing the questionnaire. CONCLUSIONS AND CLINICAL RELEVANCE: In this study, the strongest indicator of milk production was number of cows in milk. However, merely adding cows to a herd should not be considered to guarantee increased milk production, because other management traits could be confounded with increased number of cows in a herd.     

Evaluation of enzyme-linked immunosorbent assays performed on milk and serum samples for detection of paratuberculosis in lactating dairy cows

OBJECTIVE: To determine whether results obtained for milk and serum samples with ELISAs intended for diagnosis of paratuberculosis in dairy cows were comparable to results obtained by means of mycobacterial culture of fecal samples. DESIGN: Cross-sectional study. ANIMALS: 689 lactating dairy cows in 9 Ontario herds. PROCEDURE: Milk, serum, and fecal samples were obtained from all cows. Fecal samples were submitted for mycobacterial culture. Serum samples were tested with a commercially available ELISA for antibodies against Mycobacterium paratuberculosis, and preserved milk samples were tested with an indirect ELISA for antibodies against M paratuberculosis. RESULTS: Results were positive for 130 of the 689 (18.9%) serum samples, 77 of the 689 (11.1%) milk samples, and 72 of the 689 (10.4%) fecal samples. The level of agreement between results for milk and serum samples was only moderate. Proportions of positive results for serum and fecal samples were significantly different, but proportions of positive results for milk and fecal samples were not significantly different. In addition, results for milk samples had a higher level of agreement with results of mycobacterial culture than did results for serum samples. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the indirect ELISA used on milk samples may be a convenient method of detecting paratuberculosis in dairy herds.     

Persistent fecal Salmonella shedding in five dairy herds

OBJECTIVE: To monitor patterns of Salmonella fecal shedding in naturally infected dairy herds, determine the association between fecal shedding and individual animal production measures, and evaluate potential risk factors for shedding of Salmonella organisms among cattle in dairy herds. DESIGN: Longitudinal study. SAMPLE POPULATION: 5 Ohio dairy herds. PROCEDURE: For 3 herds, fecal samples were collected from all mature cows and unweaned calves 7 times during an 18-month period. For the remaining 2 herds, fecal samples were collected from 50 lactating cows 6 times during a 12-month period. Individual animal production records for 3 herds were used to examine associations between individual fecal Salmonella shedding status and 305-day mature-equivalent milk production, somatic cell count, milk fat content, and milk protein content. Multivariable logistic regression was used to test for associations between fecal shedding status and breed, lactation status, lactation number, and duration of lactation. RESULTS: None of the adult animals had clinical signs of salmonellosis, but prevalence of fecal Salmonella shedding at individual collection times ranged from 0 to 99% for cows and from 0 to 67% for unweaned calves. Mature cows were more likely to be shedding Salmonella organisms than were unweaned calves. Within herds, lactation status and duration of lactation for individual animals were associated with Salmonella shedding status. Salmonella fecal shedding status was not associated with individual cow production measures. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that subclinical fecal Salmonella shedding can persist in dairy herds for up to 18 months with no measurable effects on health or production of individual cows.     

Wildlife (Boselaphus tragocamelus)-small ruminant (goat and sheep) interface in the transmission of 'Bison type' genotype of Mycobacterium avium subspecies paratuberculosis in India

Information on Mycobacterium avium subspecies paratuberculosis (MAP) genotypes infecting different animal species in India is limited. Presence of MAP was investigated in free ranging antelopes (locally known as Nilgai/blue bulls/Boselaphus tragocamelus) using direct microscopy, culture, IS900 PCR and IS1311 PCR-REA. IS900 elements of MAP from Nilgai and previously isolated from goats were sequenced and compared to establish inter-species transmission between free ranging Nilgai and closed farm herds and flocks of goats and sheep sharing common grazing and water resources. Fecal samples were collected from two geographical regions (Mathura and Kanpur Dehat districts) separated by 300km, in North India. Of the 42 fecal samples cultured, MAP colonies were recovered from 23.8% samples (Nilgai). Of the 10 positive fecal samples, two were in 'Super shedder' (>1000cfu/g) category and rest were moderate (<10-100cfu/g) shedders. None of the Nilgai from Kanpur Dehat was positive in culture. The 229bp fragment targeting specific IS900 sequence was amplified from template DNA isolated from all the positive MAP cultures of Nilgai. Using IS1311 PCR-REA, MAP colonies were genotyped as 'Bison type'. Goatherds and a sheep flock located at Central Institute for Research on Goats (CIRG), shared 303.52ha of land (Mathura district of Uttar Pradesh) with Nilgai and were endemic for MAP infection. MAP strains isolated from goats and sheep have been genotyped as 'Bison type'. Nucleotide sequence of the insertion elements (900) from MAP 'Bison type' strain (S5) of goat origin and MAP (B42) from Nilgai showed difference of 2 (1%) base pairs at the 11th and 12th position (Genbank accession number EU130943). Study is first report on sharing (inter-species transmission) of a new 'Bison type' genotype of MAP between free ranging wildlife (Nilgai population) and domestic animals (farm goatherds and sheep flocks) in India.     

Reduction in milk yield associated with Mycobacterium avium subspecies paratuberculosis (Map) infection in dairy cows

To assess the profitability of control schemes for Mycobacterium avium subspecies paratuberculosis (Map)-infection implemented in dairy herds, accurate estimates of its production effects are needed. This study aimed at quantifying the variation in milk yield of dairy cows according to their Map-infection status. The cow-status was determined by combining (i) its testing(s)-result(s) (serum ELISA, faecal culture (FC), PCR, Ziehl staining), (ii) the Map-status of its herd, and (iii) its possible vaccination against Map. A total of 15 490 cows in 569 herds located in western France was considered. The effect on test-day milk yield (TDMY) of the cow-status to Map was assessed separately in parity 1, 2 and 3 or more, using mixed linear models, after adjustment for herd-season (random), days in milk and breed. Average TDMY was significantly lower in cows from herds with at least one Map-infected cow (defined as positive herds). Individual TDMY showed a reduction ranging from 1.58 to 3.30, 2.03 to 2.51, 5.36 to 7.20 kg/day (P < 0.001) depending on parity for unvaccinated cows and testing ELISA-positive, PCR- or FC-positive, and Ziehl-positive, respectively, compared to cows in Map-free herds. The loss in milk yield increased with increased parity in ELISA-positive and Ziehl-positive cows. Cows that were both tested ELISA-positive and vaccinated had a smaller loss in TDMY than those that were unvaccinated. The estimates from this study can be used to further assess the economic impact associated with Map-infection in dairy herds or to help in the culling decisions regarding infected cows.     

Effects of sodium monensin on reproductive performance of dairy cattle. I. Effects on conception rates, calving-to-conception intervals, calving-to-heat and milk production in dairy cows

SUMMARY A total of 1061 lactating dairy cows in six different herds were randomly allocated to treatment and control groups. One herd was lot-fed on total mixed rations; three herds were fed on pasture with significant amounts of supplementary concentrates, and two herds were primarily pasture fed. Treated cows received a slow-release, intraruminal capsule containing 32 g sodium monensin within 7 days of calving. Conception rates at first service, days to first oestrus and calving-to-conception interval did not differ significantly between untreated and monensin-treated cows in the 5 herds, 3 herds and 4 herds, respectively, in which these outcomes were examined. Treatment of lactating cows immediately after calving may not be the optimal method to achieve fertility responses with capsules containing sodium monensin. Monensin treatment significantly increased milk yield in one of the six herds. Milk fat or milk protein production was not significantly affected by treatment.     

Bayesian analysis to validate a commercial ELISA to detect paratuberculosis in dairy herds of southern Chile

In Chile, Mycobacterium avium subsp. paratuberculosis (Map) has been isolated on several occasions and clinical cases have been reported. Nevertheless, diagnostic tests have not yet been validated for this agent in the Chilean setting. The objective of the study was to validate a commercial ELISA to detect Map shedding dairy cows in management conditions, prevalence and stages of infection existing in Southern Chile, utilising different statistical approaches. Blood and faeces were collected from 1333 lactating cows in 27 dairy herds (both large commercial and smallholder dairy farms) between September 2003 and August 2004. Within the herds up to a maximum of 100 dairy cows were selected based on age (>or=3 years old) and, if present, clinical signs of a Map infection. In herds with less than 100 cows, all cows >or=3 years old were sampled. Blood samples were tested using a commercial ELISA kit (IDEXX Laboratories, Inc.). Faecal samples were cultured on Herrold's Egg Yolk Medium (HEYM). Latent class models (i.e. maximum likelihood (ML) methods and Bayesian inference) were used to determine the validity of the ELISA. Map was cultured from 54 (4.1%) cows and 10 (37.0%) herds, which were all large, commercial dairy herds. As a result of empty cells in the cross-tabulations, the ML model provided the same results as the validation with faecal culture as the gold-standard. In the Bayesian model, the Se and Sp of the ELISA were estimated to be 26% (95% CI: 18-35%) and 98.5% (95% CI: 97.4-99.4%), respectively. For faecal culture, the Se was 54% (95% CI: 46-62%) and the Sp was 100% (95% CI: 99.9-100%). Interestingly, the prevalence in the smallholder dairy farms was estimated to be 8% even though there were no faecal culture positive cows detected in those herds. There was no significant correlation between the two tests. The advantage of Bayesian inference is that the Se and Sp of both tests are obtained in one model relative to the (latent) true disease status, the model can handle small datasets and empty cells and the estimates can be corrected for the correlation between tests when the tests are not conditionally independent. Therefore, Bayesian analysis was the preferred method for Map that lacks a gold-standard and usually has low cow-level prevalence.     

The Mycobacterium avium subsp. paratuberculosis ELISA response by parity and stage of lactation

Two cross-sectional studies were carried out to determine the enzyme-linked immunosorbent assay (ELISA) response to Mycobacterium avium subsp. paratuberculosis (Map) by cow characteristics and stage of lactation. One of the studies (referred to as "milk-group") used milk samples from all lactating cows (n=7994) in 108 Danish dairy herds. The other study (referred to as "serum-group") used serum samples collected from all cows (n=5323) in a subset of 72 herds from the 108 herds. These samples were analysed using a similar ELISA for detection of antibodies.The results from the ELISAs were interpreted with two cut-off values as the optimal cut-off value is not known, and as several levels are recommended to be used in practice. The results showed that the probability of being ELISA-positive was two to three times lower for cows in parity 1 relative to cows in other parities using both milk and serum ELISA. At the beginning of the lactation the probability of being positive was highest in the milk ELISA. In the serum ELISA the odds of being positive was highest at the end of lactation. The findings are important in the interpretation of ELISA results at cow level with a subsequent tentative diagnosis and correction for parity and stage of lactation should be considered when providing a diagnosis of paratuberculosis. Some issues related to the pathogenesis are also discussed.     

Evaluation of microbial culture of pooled fecal samples for detection of Mycobacterium avium subsp paratuberculosis in large dairy herds

OBJECTIVE: To evaluate sensitivity of microbial culture of pooled fecal samples for detection of Mycobacterium avium subsp paratuberculosis (MAP) in large dairy herds and assess the use of the method for estimation of MAP prevalence. ANIMALS: 1,740 lactating cows from 29 dairy herds in California. PROCEDURE: Serum from each cow was tested by use of a commercial ELISA kit. Individual fecal samples were cultured and used to create pooled fecal samples (10 randomly selected fecal samples/pool; 6 pooled samples/herd). Sensitivity of MAP detection was compared between Herrold's egg yolk (HEY) agar and a new liquid culture method. Bayesian methods were used to estimate true prevalence of MAP-infected cows and herd sensitivity. RESULTS: Estimated sensitivity for pooled fecal samples among all herds was 0.69 (25 culture-positive pools/36 pools that were MAP positive). Sensitivity increased as the number of culture-positive samples in a pool increased. The HEY agar method detected more infected cows than the liquid culture method but had lower sensitivity for pooled fecal samples. Prevalence of MAP-infected cows was estimated to be 4% (95% probability interval, 2% to 6%) on the basis of culture of pooled fecal samples. Herd-level sensitivity estimate ranged from 90% to 100% and was dependent on prevalence in the population and the sensitivity for culture of pooled fecal samples. CONCLUSIONS AND CLINICAL RELEVANCE: Use of pooled fecal samples from 10 cows was a cost-effective tool for herd screening and may provide a good estimate of the percentage of MAP-infected cows in dairy herds with a low prevalence of MAP.     

Unusual history and initial clinical signs of Mycoplasma bovis mastitis and arthritis in first-lactation cows in a closed commercial dairy herd

CASE DESCRIPTION: 9 first-lactation dairy cows in a closed dairy herd had swelling in the forelimbs and forelimb lameness. Mycoplasmal arthritis and mastitis were diagnosed. CLINICAL FINDINGS: Swelling of the carpal joint, diffuse subcutaneous edema from the carpal to metacarpophalangeal joints, and forelimb lameness were evident in 9 first-lactation cows 7 to 21 days after parturition. Diagnostic testing revealed that 3 of 3 bulk-tank milk samples, 3 milk samples from cows with clinical mastitis, 2 fluid samples obtained from arthritic joints, and samples from the lungs and spleen of a cow that had died yielded positive results for Mycoplasma spp. Nucleic acid sequence analysis performed by use of a PCR assay on the joint fluid and lung tissues confirmed infection with Mycoplasma bovis. TREATMENT AND OUTCOME: Affected cows were treated by IM administration of flunixin meglumine and dexamethasone for 3 days. All cows were nonresponsive to treatment (3 cows died, and the other 6 were culled). Follow-up culture for Mycoplasma spp of milk samples from the bulk tank and from all lactating cows was recommended to screen for chronic subclinical carriers. CLINICAL RELEVANCE: Mycoplasmal infections may cause unusual initial clinical signs or an atypical history. When dairy cattle, including those residing in closed herds, have lameness, swelling of the carpal or metacarpophalangeal joints, edema of the distal portions of the forelimbs, or polyarthritis, infection with Mycoplasma spp should be investigated. Delay in diagnosis of mycoplasmal infections in dairy herds can result in substantial financial loss and the establishment of chronic subclinical carriers.     

Production and related variables in bovine leukaemia virus-infected cows

A newly developed milk dot blot test was used to detect anti-bovine leukaemia virus (BLV) antibody in milk samples from 2079 lactating adult cows from among 61 herds. The milk dot blot test was highly repeatable; the concordance rate, compared with the agar gel immunodiffusion test performed on serum, was 83.5%. All herds contained BLV-positive cows; the prevalence rate was 36%. BLV-positive cows tended to come from larger herds and were older and more often later in lactation. Fourteen production and related variables (herd size, age, days open, days in milk, milk somatic cell count, milk, fat, and protein produced in the current lactation, projected production of milk, fat, and protein, and breed class average deviations for milk, fat, and protein) were compared between BLV-positive and BLV-negative cows. Although somatic cell count, milk produced, and projected production of milk and protein were related significantly to BLV status using simple tests of association, once the variables herd size, age and days in milk were controlled, these differences were removed. Further analyses using logistic (outcome: individual cow BLV status) and least-squares regression (outcome:herd proportion of BLV-positive cows) failed to show an association between any of the measured production or related variables and BLV-positivity. We concluded that the effect of BLV on production and related variables in dairy cows was below the sensitivity of our analytical techniques or was non-existent.Abbreviations ABCA herd average breed class average for milk, fat, and protein production - AVGAGE average age of the herd - ADIM herd average for days in milk - AGID agar gel immunodiffusion - AVGSCC herd average milk somatic cell count - BCA breed class average, a milk, fat and protein production index calculated by comparing a cow's actual 305-day lactation production to the corresponding BCA standard for the same breed, age, and month of calving - BLV bovine leukaemia virus - CALVINT calving interval - COWAGE cow age - DBCA breed class average deviation for milk, fat, and protein production, the difference between an individual cow's BCA and the herd average - DIM days in milk - HS herd size corresponding to the number of lactating cows in a herd - LACT actual amount of milk, fat, and protein produced in a cow's lactation - ODHIC Ontario Dairy Herd Improvement Corporation - PCTPOS percentage of herd that is BLV-positive - PROJ projected 305-day production for milk, fat, and protein by fitting to a standard lactation curve adjusted for days in milk and age at calving - RHBCA rolling herd average for breed class average for milk, fat, and protein production, the average for all cows that completed a lactation (cows must have completed a 305-day lactation) during the previous 12 months - SCC milk somatic cell count     

Evaluation of an O antigen enzyme-linked immunosorbent assay for screening of milk samples for Salmonella dublin infection in dairy herds.

Levels of antibodies to the O antigens (O:1,9,12) of Salmonella dublin were tested in 1355 serum, 1143 cow milk and 160 bulk milk samples from dairy herds using an enzyme-linked immunosorbent assay (ELISA). In order to define the background reaction, milk samples from all lactating cows and serum samples from 9 animals were collected in each of 20 salmonellosis-free herds located on the island of Bornholm, where cattle salmonellosis has not been reported. Similar samples were collected from all stalled animals in 10 herds with recent (< 6 months) outbreaks of salmonellosis located in Jutland, where salmonella infection is enzootic. Using herd history of salmonellosis, herd location and clinical status of the herds as criteria, the optimal cutoff in the milk ELISA was determined as being at least 5% of the samples having optical density > 0.5, resulting in herd sensitivity of 1.0 and herd specificity of 0.95. While none of the sera in the herds from Bornholm was ELISA positive, 2 herds had a few reactors in the milk ELISA. Using the same cutoff, all but 1 bulk milk sample from 150 herds on Bornholm was ELISA-negative, and all 10 salmonellosis-positive herds from Jutland were ELISA-positive. A significant correlation was found between ELISA reactions in milk and in serum of cows (34% and 32% respectively, rs = 0.69, P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)     

Characterization of Minnesota dairy herds participating in a Johne's disease control program and evaluation of the program risk assessment tool

The objectives of this study were to: (1) characterize Minnesota dairy herds participating in a Johne's disease control program (JDCP) based on herd size, milk production, and clinical Johne's disease (JD) history, (2) evaluate if change in farm management practices, expressed in risk assessment (RA) total score, is associated with the change between the first and most recent ELISA test herd seroprevalence or change in clinical JD culling rate, and (3) identify farm factors associated with ELISA seroprevalence. A total of 1234 RA, performed between January 2000 and February 2004, were available for analysis from 714 dairy herds. ELISA test results from herd sampling between 2000 and 2004 were obtained from the Minnesota board of animal health (MBAH) database, and were available for 474 herds. Both the first and the most recent ELISA test results for herds with more than one RA were available for 262 herds. Mean herd size and mean annual milk production per cow was higher in JDCP dairy herds (161 milking cows) than either all Minnesota dairy herds or Minnesota dairy herd improvement association (DHIA) herds. For herds with more than one RA available, the most recent RA total score was significantly lower (mean 11% less) than the first. The change in RA total score (and any RA subtotal scores) between the first and most recent RA was not associated with the change between the first and the most recent ELISA within-herd seroprevalence or the change in JD culling rate between the first and most recent RA. The most recent ELISA test results were positively associated with postweaned heifer score and JD culling rate. The RA score was not found to be an effective tool for the prediction of ELISA seroprevalence.     

Bulk milk urea concentrations and their relationship with cow fertility in grazing dairy herds in southern Chile

Milk urea determination is being used as a broad indicator of protein/energy imbalance in dairy herds. The main purpose of this study was to compare blood and bulk milk urea values in grazing herds, to evaluate their seasonal variation under South Chilean conditions, and to examine their potential relationships with herd fertility. The association between herd blood urea concentration (mean of seven lactating cows) and bulk milk urea concentration (tank containing milk from the previous 24 h) was determined in 21 diary herds. Reference values, seasonal and herd variance, and the frequency of herds with values outside a range of 2.5 to 7.3 mmol/l were determined in bulk milk samples obtained monthly for a period of one year from 82 suppliers at two creameries located in southern Chile. Finally, bulk milk urea was measured every two weeks in samples from 24 herds, and the first service conception rate (FSCR) from 2153 dairy cows was determined. Mean bulk urea concentration was highly correlated with mean herd blood urea concentration (r = 0.95; p < 0.01). Mean urea concentration in the bulk milk samples obtained during one year from 82 herds was 4.9 +/- 1.2 mmol/l, with a range of 1.5 to 11.6 mmol/l. The highest values were found during spring and the lowest values during the summer. There was a high seasonal variation (CV = 13-47%) and between-herd variation (CV = 20-31%). Out of a total of 984 samples, 5.4% had urea values > 7.3 mmol/l and 3.8% had values < 2.5 mmol/l. Of the 82 herds, 27% had values outside the reference interval (2.5-7.3 mmol/l) on two or more occasions. FSCR was lower in herds when the bulk milk urea was > 7.3 mmol/l (50.7%) than in cows, where the urea concentration was < 5.0 mmol/l (73.8%) at the time of insemination. The study concluded that bulk milk urea concentrations provided information similar to herd blood urea concentrations in local grazing dairy herds. There was a high frequency of herds with abnormal values, with large variations between herds and between seasons. Increased milk urea concentrations during spring were associated with lower conception rates.