Bulk milk-estimated seroprevalence of Fasciola hepatica in dairy herds and collecting of risk factor data in East Frisia, northern Germany

The liver fluke Fasciola (F) hepatica is one of the most important trematodes in cattle farming worldwide. Fasciolosis in dairy cows is leading to production losses due to decreased milk yield, liver condemnation and impaired reproduction. The treatment of dairy cows is unsatisfactory, because available drugs are either effective against adult flukes only or have long withdrawal times or in some countries may not be used at all. In the present study the prevalence of F. hepatica in dairy farms located in East Frisia, which is part of the federal state Lower Saxony, was investigated. East Frisia is considered a high risk area for Fasciola hepatica infections, because of its coastal location, high precipitation and moist pastures. About 750 bulk milk samples were collected in January and November 2006 and analysed for F. hepatica antibodies using the Pourquier ELISA. In addition, questionnaires, which were answered by 260 of the participating farmers, were evaluated to analyse management-related factors associated with fasciolosis. In January and November, 52.1% and 53.6% of the bulk milk samples, respectively, showed positive results. Thereby, 88.1% of the examined farms showed an unchanged infection status, whereas 6.2% of the farms became seropositive during the grazing season and 5.8% of the dairy herds turned seronegative. Statistical analysis revealed a significant negative association between average annual milk production and the frequency of infections with F. hepatica.     

Seroprevalence of Dictyocaulus viviparus in first grazing season calves in Sweden

A serological survey was carried out to determine the prevalence and geographical distribution of Dictyocaulus viviparus in calves after their first grazing season in Sweden. A total of 754 animals from 76 randomly selected herds in seven geographical regions were examined between September 24 and December 19, 2001. To get an indication about the geographical distribution of the infection 41 herds with beef-suckler calves were investigated. On each farm, blood was collected from 8 to 10 animals after an average of 26 +/- 24 days post-housing to determine specific IgG1 levels against a possible lungworm sperm antigen that is highly specific against patent infections of D. viviparus. We also investigated the seroprevalence of lungworm infection in relation to cattle management. In one region additional samples were analysed from 35 herds either with: (a) beef-suckling calves that were dewormed at housing, (b) untreated organically raised dairy calves, and finally from conventionally raised dairy calves either, (c) with or, (d) without a prophylactic anthelmintic treatment programme against gastrointestinal parasites on pasture. A questionnaire was used to obtain information about herd size and management, including measures to control nematode parasites on the farm. A total of 86 (11.8%) out of 754 animals had antibodies against D. viviparus, and at least one infected individual was detected in 30 (39.5%) of the 76 herds examined. Lungworm infected animals were found throughout the country and there was no significant differences between regions, although in southern and southwestern Sweden 70.0% of the herds were infected. Furthermore, there were no major differences in the seroprevalence in relation to management. Between 40.0 and 44.4% of the herds were infected irrespective of management, with the exception of calves from organic herds where no seropositive samples were found (0%). This result is in contrast to previous findings of lungworms in Sweden, and indicates that the parasite status on organic farms is diverse.     

The value of a bulk-tank milk ELISA and individual serological and faecal examination for diagnosing (sub)clinical Dictyocaulus viviparus infection in dairy cows

To test the value of a recently developed bulk-tank milk (BTM) ELISA for diagnosing (sub)clinical Dictyocaulus viviparus infection in lactating dairy herds under field conditions, bulk milk samples were collected from farms with or without clinical symptoms suspected to be caused by lungworm infection. Results of the BTM ELISA were compared against individual examinations for lungworm larvae in faeces and lungworm antibodies in serum from up to 20 heifers (parity 1) and up to 20 cows (parity ≥ 2) on the same farms. This also allowed, for the first time, to examine the value of individual faecal and serological examinations in the diagnosis of (sub)clinical lungworm infections. In total, 33 farms participated. Of these, 16 reported clinical symptoms possibly related to lungworm infection (defined as a suspected positive clinical status or CS(+)) and 17 reported having no such symptoms (CS(-)). In total, 503 heifers and 649 cows were sampled. Of all faeces samples positive for lungworm larvae, 94 were from heifers (18.9% of all heifers) and 75 from cows (11.7% of all cows) (P<0.001). Of all sera positive for lungworm antibodies, 130 were from heifers (26.1% of all heifers) and 113 from cows (17.5% of all cows) (P<0.001). Of the CS(-) farms 41% had at least one heifer or cow shedding larvae and 71% had at least one seropositive heifer or cow. Of the CS(+) farms this was 81% and 94%, respectively. There were only 4 farms, all CS(-), where none of the animals were found shedding larvae and all animals tested seronegative. This implies that on 76% of the CS(-) farms lungworm infection circulated unnoticed. On all CS(+) farms the suspicion that lungworm caused the respiratory symptoms was confirmed by the individual faecal and serological examinations, whereas the BTM ELISA confirmed presence of lungworm on half of the CS(+) farms. The latter in particular occurred on farms with the more severe outbreaks. Overall, of 32 available BTM samples 10 tested positive (8 of 15 CS(+) and 2 of 17 CS(-) farms). For diagnosing suspected lungworm disease it was concluded that testing a BTM sample might suffice in case of moderate to severe outbreaks. However, in case of a mild outbreak with just a few animals coughing, examining individual animals has to be preferred over testing a BTM sample. The likelihood to detect lungworm infection is higher if heifers are sampled compared to cows. Sensitivity of the BTM ELISA was 35.7% if the presence of at least one seropositive and/or one larvae shedding animal in the herd was used to define lungworm positive farms. On average, at least 30% of the herd had to be seropositive before the BTM ELISA was found positive for lungworm antibodies. Results indicate that the BTM ELISA in its current form does not appear to be suitable for surveys on the prevalence of lungworm presence on farms. However, this BTM ELISA might be used in large-scale surveys to detect, for instance, annual changes in percentage positive farms, as long as it is recognized that positivity is more closely related to incidence of lungworm disease than to prevalence of lungworm infection.     

Validation of a Dictyocaulus viviparus MSP-ELISA and cut-off adjustment in a one-year longitudinal field study in dairy cattle herds

A one-year field study analysing lungworm seropositivity by use of the MSP-ELISA was performed (1) to investigate the antibody dynamics in individual milk samples following field (re-)infections of dairy cows with the bovine lungworm Dictyocaulus viviparus, (2) to investigate the correlation between individual and bulk tank milk (BTM) antibody titres and (3) to review the current individual as well as BTM cut-off value, which was extrapolated from dilution experiments (Fiedor et al., 2009). Over a one-year period individual and BTM samples were collected monthly on 15 dairy farms. Following a critical review of previous cut-off values, individual and BTM samples were subjected to different cut-off thresholds. Following Receiver-Operating-Characteristics (ROC) analysis, individual milk samples were assessed with the cut-off value 0.573, previously shown to be associated with each 100% sensitivity and specificity. In addition, the present study enabled BTM cut-off adjustment based on field data. To ensure reliable detection of herds with an in-herd prevalence of ≥20% the BTM cut-off was lowered from 0.493 to 0.410, corresponding to 100% sensitivity and 97.32% specificity. Regression analysis showed that the percentage of seropositive animals related to the corresponding BTM ODR correlated moderately (r=0.581, P<0.001), whereas a strong correlation (r=0.764, P<0.001) was found between mean individual and BTM ODR per herd and sampling month. Seasonal antibody pattern became obvious in a single-peaked antibody curve in late summer/early autumn for individual milk whilst BTM showed a two-peaked distribution with an additional spring peak besides the late summer/early autumn peak. This leads to the conclusion that the BTM-ELISA could be a useful tool to detect and control pasture contamination in the spring, following sexual maturation of hypobiotic lungworm larvae harboured by clinically asymptomatic carrier animals. In addition to the knowledge gained on antibody patterns in dairy herds and the relationship of individual and BTM, the present study enabled sensitivity and specificity calculations for the obsolete BTM cut-off value 0.493 to be performed.     

Dictyocaulus viviparus seroprevalence and epidemiology in Costa Rican dairy cattle

A cross-sectional serological survey of Dictyocaulus viviparus was carried out to determine the prevalence of lungworm infections in 28 dairy cattle farms distributed in five selected areas from Costa Rica. The influence of area, farm, host (breed, age and lactation number) and ecological factors (altitude and life zones) on the presence of lungworm infection was analyzed. A sub-sample of 924 sera collected between September 1998 and July 1999 was processed by ELISA (Ceditest). A total of 162 (17.5%) animals from 26 (93.0%) farms showed antibodies against D. viviparus. The overall seroprevalence detected among areas was Poás 25.0%, Cartago 24.3%, Tilarán 22.0%, Alfaro Ruiz 12.0% and San Carlos 12.1%. Using analysis of variance no significant influence of area and host factors on D. viviparus infections was determined, whereas the variable farm within area was highly significant (p<0.001). However, altitude and life zones showed significant association to seropositive animals, when a Chi-square test was applied. In altitudes of 1000-2000 m (p<0.001) and life zones of Lower Montane moist forest and Montane moist forest (p<0.001) D. viviparus infections in bovines were significantly higher. The results obtained in this study indicate a high D. viviparus seroprevalence in the analyzed farms and that the factors farm, altitude and life zones were significantly related to lungworm infections.     

Screening of commercial milk samples using ELISA for immuno-epidemiological evidence of infection by the cattle grub (Diptera: Oestridae)

Pasteurised and ultra high temperature (UHT)-treated milk were tested over the period November 1997-February 1998 in order to evaluate the applicability of an hypodermosis ELISA test on commercial milk samples. Milk samples from six dairy companies were analysed and the development of anti-Hypoderma antibodies recorded for the period from September 1998 to June 1999. For one dairy (no. 3) bulk milk samples were also obtained from the tankers, transporting milk away from the farms, and analysed; the results were compared to the parasitological status of the cows whose milk had been collected on the farms. Out of 32 pasteurised samples tested, 23 were positive, exceeding the cut-off (22%), while 9 samples were negative. UHT milk was always negative. The antibody levels in milk samples from four of the six companies were highest during January and decreased gradually during February to May 1999. The bulk milk samples also had high antibody levels and 47.2% of milk producing cows had lesions of hypodermosis. It was concluded that testing commercial milk for antibodies is an efficient way of detecting the presence of hypodermosis in cattle, especially in those countries for which no data on this disease are available.     

Prevalence of Coxiella burnetii antibodies in Danish dairy herds

During recent years in Denmark higher rates of antibodies to Coxiella burnetii have been detected in animals and humans than previously reported. A study based on bulk tank milk samples from 100 randomly selected dairy herds was performed to estimate the prevalence and geographical distribution of antibody positive dairy herds. Using the CHEKIT Q-Fever Antibody ELISA Test Kit (IDEXX), the study demonstrated a prevalence of 59% antibody positive herds, 11% antibody intermediate herds and 30% antibody negative herds based on the instructions provided by the manufacturer. The geographical distribution does not indicate a relationship between the regional density of dairy farms and the prevalence of antibody positive dairy farms. The result supports the hypothesis of an increase in the prevalence of positive dairy herds compared to previous years.     

Use of an enzyme-linked immunosorbent assay in bulk milk to estimate the prevalence of Neospora caninum on dairy farms in Prince Edward Island, Canada

This study evaluated the use of bulk milk as a diagnostic tool for estimation of herd-level Neospora caninum exposure in Atlantic Canada; it was used to estimate the prevalence of dairy farms with a within-herd N. caninum-seroprevalence > or = 15% in Prince Edward Island (PEI). The variation over time of N. caninum antibodies in bulk milk is also reported. Skimmed bulk milk and individual serum samples were analyzed for N. caninum antibodies by using an enzyme-linked immunosorbent assay (ELISA). Bulk milk samples were collected in May 2004 (n = 235), May 2005 (n = 189), and June 2005 (n = 235). The prevalence of dairy farms with a within-herd seroprevalence > or = 15% on PEI was 6.4% in May 2004. In May and June 2005, respectively, 10.1% and 10.2% of farms had a > or = 15% within-herd seroprevalence. In 11 farms that were considered positive based on bulk milk samples, blood samples were collected from all adult cows in September 2005, in conjunction with a 4th bulk milk sample on the same day. The correlation coefficient between serology and bulk milk ELISA was 0.87. The results of this study demonstrate that the prevalence of N. caninum in dairy farms can be estimated by using a bulk milk ELISA.     

Isolation of Mycoplasma bovis from bulk milk

The prevalence of mycoplasma in Vermont dairy farms was determined by two surveys conducted in March 1983 and January 1984. Bulk tank milk samples representing 74% and 62% of the herds respectively were cultured. Mycoplasma bovis was detected in 3 of the 2,346 bulk samples collected in the initial survey. The infection rate was 1.3 herds per thousand (95% confidence interval: 0.6-2.0 herds per thousand). No positive cultures were obtained in the second survey.     

Failure to eradicate the lungworm Dictyocaulus viviparus on dairy farms by a single mass-treatment before turnout

On two dairy farms it was attempted to eradicate lungworm, Dictyocaulus viviparus, by means of a single mass-treatment of all cattle that had been grazed the previous year(s), before turnout in the spring. Both farms experienced two years of lungworm outbreaks in the adult dairy herd prior to this study. Following confirmation that both herds contained lungworm carriers, all animals older than approximately 6 months were treated with eprinomectin in March 2007. One week after treatment none of the animals were shedding lungworm larvae. Subsequently, animals were pastured according to normal farm routine. From August to November all first-calving heifers were coprologically and serologically monitored for lungworm infection. During 2007 both farms remained lungworm-negative and did not report any clinical sign indicative for a lungworm infection. The following year, on one of the farms replacements grazing on cow pastures, started showing signs of parasitic bronchitis which was serologically confirmed. The other herd remained free of parasitic bronchitis until at least the fourth year after the mass treatment, although some coughing was noticed in 2008 among first-lactation heifers. It was concluded that a single mass-treatment before the grazing season may be useful to break a series of annual lungworm outbreaks. However, it is not a secure method to prevent parasitic bronchitis for more than one year.     

Effect of naturally occurring nematode infections on growth performance of first-season grazing calves

Liveweight of calves on 89 dairy farms was measured at the end of the grazing season and related per herd to the level of exposure to nematode infection during the grazing season. There were significant between-herd variations in antibody titres against Ostertagia spp., Cooperia spp. and Dictyocaulus viviparus as well as in pepsinogen values. All but six herds (93.1%) had gastrointestinal nematode infections, as measured by faecal egg counts in September. Faecal samples of 17 herds (19.3%) contained lungworm larvae in September. Liveweight of calves per herd deviated from -68.1 kg to +84.1 kg from the age-adjusted population mean after their first grazing season. Growth performance up to the time of liveweight measurements was significantly correlated negatively with several serological and parasitological parameters. Data could be fitted by means of both linear and segmented curvilinear regression. Antibody titre against Cooperia spp. and gastrointestinal nematode egg output measured in September accounted for 3.1% (P less than 0.10) and 6.7% (P less than 0.05), respectively, of the variation in growth performance among herds. Certain infection parameters, when combined, accounted for 9.2% of this variation; these were antibody titre against Cooperia spp. and larval counts for both gastrointestinal nematodes and lungworm. Adding certain management factors to these infection parameters resulted in a model explaining 27.6% of the observed variation in growth performance among herds. These factors were supplementary feeding, lungworm vaccination, anthelmintic treatment at housing, date of housing and herd age.     

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.     

Evaluation of enzyme-linked immunosorbent assay using milk samples as a potential screening test of bovine tuberculosis of dairy cows in Korea

An enzyme-linked immunosorbent assay (ELISA) using bulk tank milk samples was evaluated as a screening test for bovine tuberculosis (TB), a contagious chronic disease of cattle. An ELISA with MPB70, a major antigen of Mycobacterium bovis was performed using paired sets of milk and sera samples from 33 tuberculin-positive and 43 tuberculin-negative cattle. Anti-MPB70 antibodies were detected in milk samples and there was a significant correlation between seroreactivities of milk and sera samples (R² = 0.83). Using the tuberculin skin test as the reference test, the sensitivities of ELISA using milk and sera samples were 87.8% and 81.8%, respectively, and the specificities were 97.7% and 100%, respectively.In the screening test using bulk tank milk samples from 931 dairy herds in Whasung, Gyeonggi-do, Korea, the positive rate for anti-MPB70 antibody was 4.5% (42/931) and the tuberculin-positive rate was 2.8% (26/931). Individual milk samples (n = 253) were collected from randomly selected 8 problematic and 3 negative herds (positive and negative in the screening test by MPB70 ELISA using bulk tank milk samples, respectively) and tested by MPB70 milk ELISA. In the problematic herds, positive rates were 10.5% (20/190) for anti-MPB70 antibodies in milk ELISA and 2.1% (4/190) in the tuberculin skin test. More than one dairy cows were positive by milk ELISA among the problematic herds, and all tuberculin-positive dairy cows were positive in the milk ELISA. Further, no positive cows were detected in negative herds both by milk ELISA and tuberculin skin test. These results suggest that an ELISA, using bulk tank milk samples, might be a potential efficient screening test for bovine TB of dairy cows.     

Preventive vaccination of lactating and pregnant heifers against lungworm: safety and protection in three dairy herds

A study of the safety of a vaccine against lungworm was carried out with pregnant and lactating heifers from three dairy herds with a previous history of lungworm outbreaks in adult cows. Half of the heifers were vaccinated while the other half were not. A slight temporary cough following the vaccination was only observed in one herd. No adverse effects on pregnancy or milk production were seen. All heifers were serologically and coprologically examined before the first, before and after the second immunization, 3 months after introduction to pasture and at the end of the grazing season. Serological and faecal examination of the dairy cows before introduction into pasture confirmed the presence of at least one Dictyocaulus viviparus carrier in each herd. Lungworm infection occurred in all herds during the grazing season, most prominently in the herd with the highest number of heifers. In this herd, mild coughing associated with the lungworm infection was noticed, especially in the non vaccinated heifers. No other signs or symptoms were observed. It is concluded that a vaccine against D. viviparus can be used safely in heifers, before they are introduced into the adult herd, and that this vaccine can be used as a preventive measure against lungworm outbreaks in adult cattle.     

An outbreak of dictyocaulosis in lactating cows on a dairy farm

CASE DESCRIPTION: The owner of a herd of 74 Holstein-Friesian cattle reported decreased milk production, weight loss, and coughing among lactating cows. Owner-initiated antimicrobial treatment was unsuccessful; 1 lactating cow died, and 50% of the lactating cows had clinical signs of respiratory distress, such as tachypnea and coughing. CLINICAL FINDINGS: On the basis of history, physical examination findings, and fecal examination results, affected animals were determined to have Dictyocaulus viviparus (lungworm) infestation. The disease history suggested that the herd contained cows with subclinical patent lungworm infestations; after introduction of susceptible heifers, the pastures had become heavily infested with D viviparus and clinical problems subsequently developed in both newly introduced and resident cows. TREATMENT AND OUTCOME: Affected and unaffected heifers and adult cows were treated with a pour-on formulation of eprinomectin (0.5 mg/kg [0.23 mg/lb]). One animal died, but 2 weeks after treatment, clinical signs among affected cattle were markedly improved. Ten weeks after treatment, milk production improved from 23 kg/cow/d (51 lb/cow/d) to 28 kg/cow/d (62 lb/cow/d). CLINICAL RELEVANCE: The outbreak provides additional evidence that dictyocaulosis is becoming more common among adult dairy cattle, rather than almost exclusively affecting young stock. This may be attributable to anthelmintic use and management practices on dairy farms. Combined with anecdotal reports of an increase in the incidence of dictyocaulosis among adult cattle in North America, D viviparus infestation should be included as a differential diagnosis for decreased milk production, weight loss, and coughing among adult dairy cattle.     

Indications of a relationship between buying-in policy and infectious diseases on dairy farms in Wales

During the period February to May 2008, bulk milk samples were collected from 57 dairy farms throughout Wales in the framework of a voluntary somatic cell count project. Bulk milk samples were tested for antibodies to bovine viral diarrhoea virus (BVDV), bovine herpesvirus type 1 (BHV-1) and Leptospira Hardjo, and samples were also tested for the presence of BVDV antigen by PCR. A questionnaire was used to determine whether the herd was open or closed, what the vaccination status was, and to obtain general farm information such as the herd size and average milk yield. Vaccination against BVD, infectious bovine rhinotracheitis and leptospirosis was practised on 37, 12 and 35 per cent of the farms, respectively. The presence of bulk milk antibodies on farms that did not use vaccination was 75 per cent for BVDV, 54 per cent for BHV-2 and 76 per cent for L Hardjo. Open herds had 10 times the odds (95 per cent confidence interval [CI] 1.7 to 59.4)of having bulk milk antibodies for BVDV and 16.7 times the odds (95 per cent CI 2.0 to 49.7) of having bulk milk antibodies to BHV-1 compared with closed herds. A farm with bulk milk antibodies to one disease had significantly higher odds of having bulk milk antibodies to a second disease (P<0.05).     

Determining the Prevalence and Seasonality of Fasciola hepatica in Pasture-based Dairy herds in Ireland using a Bulk Tank Milk ELISA

Background

Fasciola hepatica is a helminth parasite of global importance in livestock, with major economic impact. However information on F. hepatica infections in Irish pasture-based dairy herds is limited. Therefore this study was conducted in order to determine the prevalence, seasonality and management factors associated with F. hepatica. A total of 319 Irish dairy herds were selected for this study. Bulk tank milk (BTM) samples were collected from 290 dairy farms on a quarter year basis, while from a further 29 dairy farms BTM samples were collected on a monthly basis to provide a more detailed pattern of F. hepatica exposure in Irish herds. BTM samples were analysed using a commercially available F. hepatica antibody detection ELISA. Furthermore, within-herd prevalence of F. hepatica was assessed in a subset of these 29 herds (n = 17); both individual serum samples and bulk tank milk samples were collected.

Results

A within-herd prevalence of ≤ 50 % was found for herds with negative bulk tank milk samples. The mean prevalence of the 290 study herds was 75.4 % (Range 52 %–75.1 %), with the highest prevalence being observed in November (75.1 %). The seasonal pattern of F. hepatica shows elevated antibodies as the grazing season progressed, reaching a peak in January. A significant association was found between F. hepatica and age at first calving.

Conclusion

This study demonstrates that F. hepatica is present in a large proportion of Irish dairy herds and provides a basis on which control practices, particularly in adult dairy cows, can be reviewed.     

The suitability of ELISA for the detection of antibodies against Mycobacterium avium ssp. paratuberculosis in bulk milk samples from Rhineland-Palatinate

Paratuberculosis or Johne's Disease, caused by Mycobacterium avium subspecies paratuberculosis (MAP), is a notifiable disease in Germany which produces enormous economical losses in dairy farms. At present,there is no confirmed data about the actual number of infected livestock herds in Germany. A countrywide monitoring program to evaluate the prevalence in dairy herds would only be economically feasible on the basis of bulk milk testing. In this study, we evaluated two ELISA test kits (SVANOVIR Ptb-ELISA, IDEXX-M.pt. Milk test kit) for the detection of antibodies against MAP in bulk milk. First, the Paratuberculosis-status of the herd derived from the history of the farm was used as a gold standard. Paratuberculosis-negative farms were tested negative with each test, but paratuberculosis-positive or Paratuberculosis-serologically-positive farms were detected only in one case (Svanovir) or three cases (IDEXX), respectively. Even if inconclusive results are counted as positive, 82.9 % (Svanovir) or 80 % (IDEXX) of the paratuberculosis-positive or serologically paratuberculosis positive farms were not detected. Nevertheless, a re-validation of both ELISAs by means of ROC and TG-ROC analyses was attempted by searching for ideal cut-offs, optimised for bulk milk. If a high specificity was selected, no acceptable sensitivity could be reached.The best results were obtained using a sensitivity of 32.3 % at a specificity of 100 % (Svanovir). With a small change of the cut-off value, the sensitivity increased to still 57 %, but this reduced the specificity to 67 %. Similar results were obtained with the IDEXX-ELISA. We then evaluated the Svanovir-ELISA for the detection of bulk milk samples on the basis of the current paratuberculosis prevalence within 69 dairy herds from Rhineland-Palatinate using individual milk samples.When the bulk milk samples were tested in two different laboratories using the same ELISA, considerable differences in the results became evident. Nearly all samples were tested with a higher relative test result in one laboratory, which often led to differences in the classification of the prevalence levels.The estimated within-herd seroprevalences ranged between 0 % and 37 %.There was little agreement between the historical paratuberculosis herd status and the within-herd prevalence in milk serum, as reflected in a kappa-index of 0.146.To determine the sensitivity and specificity of the bulk milk ELISA by ROC and TG-ROC analysis, 116 bulk milk samples were used that had been obtained from the 69 dairy herds participating in the study. The optimal ratio of sensitivity (81 %) and specificity (77 %) relative to a "gold standard" was obtained when the cut-off was set at the 10 % level. These values for sensitivity and specificity were better than those obtained in an evaluation of the same ELISA in which the historical Paratuberculosis herd-status was used as a "gold standard." The results of this study question the suitability of the available ELISAs for bulk milk testing.Taking into account that the Svanovir-ELISA for individual milk samples has a sensitivity of 60 96% relative to the blood serum variant of the test, and that the latter has also a limited sensitivity due to the pathogenesis of paratuberculosis, the available test systems examined in this Study do not seem to be suitable for herd diagnosis by using bulk milk samples.     

Between-herd prevalence of Mycoplasma bovis in bulk milk in Flanders, Belgium

Mycoplasma bovis (M. bovis) is a highly infectious pathogen of cattle causing pneumonia, polyarthritis, otitis, and less frequently, subcutaneous abscesses, abortions and meningitis. Ineffective drugs treatments, culling of infected cows and loss of milk production can lead to significant economic loss on dairy farms. The early detection of cows excreting M. bovis bacteria to prevent mastitis outbreaks is warranted. Reports suggest that the risk of M. bovis mastitis is higher in larger dairy herds. The objective of this study is to estimate the herd-level prevalence of M. bovis in Flanders, Belgium by culturing bulk tank milk samples taken from dairy farms. Three bulk tank milk samples per dairy herd were taken over four weeks, with collection intervals of two weeks. Culturing was done after pre-incubation using modified Hayflicks media to increase the chances of recovery of bacteria. For the identification of M. bovis, tDNA intergenic spacer PCR was used. In three herds (1.5%) of the 200 herds sampled, M. bovis was isolated from one of the three consecutive bulk tank milk samples. We conclude that in Flanders in 2009 at least 1.5% of the dairy herds had one or more cows excreting M. bovis in the milk. The frequent monitoring of bulk tank milk to detect the presence of M. bovis, especially in expanding herds on farms that often purchase replacement animals, should be encouraged in order to detect the presence of M. bovis and to monitor the success of control procedures following an outbreak of mycoplasmal mastitis in the herd.     

Dynamics of infections with gastrointestinal parasites and Dictyocaulus viviparus in dairy and beef cattle from Costa Rica

A longitudinal survey was carried out to determine and describe the prevalence and intensity of gastrointestinal parasite infections and Dictyocaulus viviparus in a dairy and a beef cattle farm of two different ecological zones in Costa Rica. The influence of anthelmintic treatment, age and meteorological factors (rainfall, minimum and maximum temperatures) on gastrointestinal nematodes and D. viviparus counts was determined. Calves were subjected to monthly sampling of feces and blood between April 2002 and March 2003. Coprological techniques were used to detect gastrointestinal helminthes, protozoan and D. viviparus. Blood samples were analyzed for antibodies to D. viviparus by ELISA. The most prevalent gastrointestinal parasites detected on both farms (dairy cattle, A; beef cattle, B) were Eimeria spp. (94.7%, 93.7%), Strongylidae (75.0%, 81.4%), Buxtonella sulcata (38.0%, 21.6%) and Strongyloides papillosus (29.8%, 31.7%), whereas Moniezia benedeni (4.8%, 9.1%), Trichuris spp. (7.3%, 13.2%), Toxocara vitulorum (0.0%, 1.8%) and Entamoeba bovis (2.5%, 1.1%) were less prevalent. Mean fecal egg counts (FEC) showed highest values of Strongylidae in April, May and July (>335.3 eggs/g feces) on farm A, and April, May and August (>304.3 eggs/g feces) on farm B. S. papillosus presented low FEC throughout the year on farm A, on farm B the highest values were obtained in April (303.0 eggs/g feces). Trichuris spp. presented maximum FEC values in May (328.6 eggs/g feces) on farm A and in June (157.5 eggs/g feces) on farm B. Treatment and age had significant influence on infection intensity of Strongylidae (farms A and B), S. papillosus (farms A and B) and Trichuris spp. (farm A). Rainfall had significant effect on S. papillosus (farms A and B) and Trichuris spp. (farm B). Maximum temperature showed significant effect on S. papillosus (farm A) and Trichuris spp. (farms A and B). Minimum temperature had significant influence on Strongylidae (farm A), S. papillosus (farms A and B) and Trichuris spp. (farm B). Haemonchus spp. (57%, 66%) and Cooperia spp. (30.0%, 30.7%) were the most prevalent genera identified by coproculture on both farms, in contrast, Trichostrongylus spp. and Oesophagostomum spp. were less frequent. Patent lungworm infections were low on both farms (10.8%, 1.8%). On farm A, high prevalence of antibodies against D. viviparus was determined only at the beginning of the study, in contrast, on farm B the seroprevalence fluctuated throughout the year. Treatment, age and maximum temperature had significant effect on D. viviparus counts on farm A, but not on farm B.