Neospora caninum serostatus and milk production of Holstein cattle

OBJECTIVE: To determine whether Neospora caninum serostatus was associated with milk production among Holstein cattle in Ontario. DESIGN: Case-control study and cross-sectional observational study. ANIMALS: 3,702 Holstein cows in 83 herds (case-control study) and 3,162 Holstein cows in 57 herds. PROCEDURE: Herds in the case-control study were grouped on the basis of N. caninum abortion status. Herds in the observational study were considered representative of Ontario dairy herds. The N. caninum serostatus of individual cows was determined with a kinetic ELISA. Milk production was modeled to compare seropositive with seronegative animals while controlling for parity, days since parturition, and herd clustering. RESULTS: In the case-control study, 305-day milk production of seropositive cows was significantly less than milk production of seronegative cows in herds with abortions attributable to N. caninum infection and in herds with abortions attributable to pathogens other than N. caninum, but not in herds without abortion problems. In the observational study, 305-day milk production for seropositive cows was not significantly different from milk production of seronegative cows. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the association between N. caninum serostatus and milk production in Ontario Holstein dairy cattle may depend on abortion status of the herd. In herds with abortion problems, regardless of cause, N. caninum-seropositive cattle produced less milk, whereas in herds without abortion problems, N. caninum-seropositive cattle produced the same amount of milk as seronegative cattle.     

Serological evidence of Neospora caninum in dual-purpose cattle herds in Venezuela

Bovine neosporosis is a parasitic disease produced by Neospora caninum that induces abortion in cows, and consequently has a negative impact on the herd's reproductive efficiency. The main objective of this research was to determine the serological evidence of N. caninum in cattle herds from Venezuela using an indirect antibody capture ELISA test. Four hundred and fifty-nine (459) serum samples from crossbred adult cows were collected to be tested for Neospora antibodies. The sampled cows came from 15 large farms located in eight important cattle states that have predominant dual-purpose production systems (cattle from these farms are used for both milk and meat production). Fifty-two cows (11.3%) were seropositive to N. Caninum. Thirteen (86.7%) of 15 studied herds had cows seropositive to N. caninum. The average within-herd seroprevalence was 11.5% (range 3.8-36.7%). Cows that aborted in some of these farms had 2.71 (P: 0.009) greater odds to be seropositive when compared to cows that did not abort. Each one of the eight states represented in our study had seropositive animals. These results are the first evidence of exposure to N. caninum in Venezuelan cattle herds, indicating the possible circulation of this pathogen in the country. Further epidemiological studies should be granted to determine the spread of the disease in the Venezuelan cattle industry and its associated risk factors.     

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.     

Assessment of antibody avidity in aborting cattle by a somatic Neospora caninum tachyzoite antigen IgG avidity ELISA

A Neospora caninum IgG avidity ELISA was carried out on the basis of a somatic N. caninum tachyzoite antigen. The test was validated using experimentally infected calves, where a clear maturation of the IgG avidity over time could be demonstrated. At a maximum of 82 days after infection (d.p.i.), all animals showed antibody avidities ranging above 35%, and respective sera were thus defined as highly avid.Sera of 103 naturally infected seropositive cows with abortion (N. caninum association was provided by a N. caninum PCR-positivity of the fetus in 40 cases) and 139 seropositive animals without abortion history were concurrently examined. Significantly lower avidities were observed in aborting cows when compared to animals without abortion problems (P<0.01). While the avidity of sera collected before abortion remained practically constant until abortion, a significant increase of avidity could be observed in samples collected weeks to months after abortion (P<0.01).The avidities of non-aborting animals from farms with or without abortion problems did not differ significantly with time and were mainly located in the high avidity area. These data indicate that low avidities are not necessarily linked to recent N. caninum infection but can also be an indicator for increased abortion risk in cattle.     

Neospora caninum infection does not affect the fertility of dairy cows in herds with high incidence of Neospora-associated abortions

This present study, was designed to establish whether Neospora caninum infection affects the fertility of high-producing dairy cows, and was based on yearly serological screening for neosporosis and on the confirmation of N. caninum infection in aborted foetuses. Assessment was made of 7518 artificial inseminations (AI) (applied to parous cows) performed in three herds. Of these inseminations, 2540 (33.8%) resulted in pregnancy; 34% of which corresponded to seronegative cows (2226 of 6556 AI performed in seronegative cows) and 32.6% to seropositive animals (314 of 962). Abortion occurred in 97 (30.1%) of the 314 pregnancies recorded in dams seropositive for N. caninum, while only 93 (4.2%) of the 2226 pregnancies in seronegative animals ended in abortion. Logistic regression analysis indicated no significant effects of N. caninum antibody titre, days in milk and milk production at insemination on fertility. Factors found to affect fertility were herd, season of insemination, lactation and insemination number, semen-providing bull and AI technician. Our results indicate that N. caninum infection does not affect the fertility of high-producing dairy cows.     

Evaluation of an iscom ELISA used for detection of antibodies to Neospora caninum in bulk milk

The intracellular parasite Neospora caninum is increasingly recognized as an important cause of abortion and stillbirth in cattle. Presence of specific antibodies indicates infection, and the immunostimulating complex (iscom) enzyme-linked immunoassay (ELISA) has previously been evaluated for use on individual milk and sera. In the present study, this test is investigated for use on bulk milk. In this study, 124 herds were used to analyse the relationship between within-herd prevalences based on individual sera and bulk milk optical densities. The individual test results were translated into a herd-level result, which enabled comparison of the bulk milk test result to the aggregate of individual serum results. The relative contribution of milk from cows with different milk yield and antibody status to the tank, i.e. its composition, was expected to influence the outcome of the bulk milk test. Therefore, sensitivity and specificity were calculated at different cut-off levels, not only using a standard cross-tabulation technique, but also a logistic regression model. By using the latter method, the sensitivity and specificity could be estimated adjusting for milk yield covariates. Specificity was estimated to be high ( approximately 98%) at the 0.20 cut-off, which can be used as a decision threshold to rule in infection. With more equal emphasis on sensitivity and specificity, a lower cut-off should be used. Although infection cannot be completely ruled out, herds with test results below 0.05 are highly likely to be non-infected. The within-herd prevalence of false negative herds is probably less than 10-15% at this level. From what is known about test performance at the individual level and the prevalence of infection, the estimate of the specificity of the bulk milk test should be quite accurate while the sensitivity is likely to be underestimated. We confirmed that the performance of the bulk milk test depends on the milk tank composition. In particular the milk yield of cows with high antibody levels affects the probability of a positive outcome of the bulk milk test.     

Efficacy of a Neospora caninum killed tachyzoite vaccine in preventing abortion and vertical transmission in dairy cattle

A clinical trial was undertaken to assess the efficacy of Bovilis(?) Neoguard, a killed Neospora caninum tachyzoite vaccine on 5 commercial dairy farms in New Zealand with a history of Neospora-associated abortion. Cattle were enrolled in the trial at 30-60 days of gestation and randomly allocated to treatment or control groups. Treatment consisted of 5 mL doses of Bovilis Neoguard administered subcutaneously at enrolment then 4 weeks later. Isotonic saline was administered to the control group. Of 2246 cattle enrolled in the trial, 10.7% of cows and 12.6% of heifers were seropositive to N. caninum. Sampling of a randomly selected proportion of enrolled animals 6 weeks after the second treatment showed that 188/232 (81.0%) vaccinated with Bovilis(?) Neoguard had seroconverted, while 11/130 (8.5%) cows and 10/36 (27.8%) heifers in the control group had seroconverted. Forty-eight vaccinated and 63 control animals aborted. On one farm 12.5% of control animals and 6.1% of vaccinated animals aborted (vaccine efficacy 0.61; p=0.03). On another farm with a high level of abortion 8.4% of control animals and 8.7% of vaccinates aborted. On the remaining 3 farms fewer abortions occurred than expected. A modified Poisson regression approach was used to calculate relative risks for abortion and vertical transmission. Overall vaccine efficacy was 0.25 (p=0.12). Heifer replacement calves from the animals enrolled in the trial were sampled for antibodies to N. caninum at 6-9 months of age. Fourteen of 17 calves from vaccinated, seropositive cows were seropositive as were 13/23 calves from seropositive cows in the control group. The interaction between dam serostatus and treatment group was significant (p=0.05) with vaccination increasing the risk of vertical transmission. It was concluded that vaccination after conception prevented 61% abortions in one of five herds and that vaccination may have increased the risk of early embryonic death.     

Seroprevalence of Neospora caninum in aborting dairy cattle in the Czech Republic

A serological survey for antibodies against Neospora caninum in aborting cattle was carried out in the Czech Republic. Serum samples from 463 aborting dairy cows originated from 137 farms from different parts of the Czech Republic were tested for presence of N. caninum antibodies by use of an enzyme-linked immunosorbent assay (ELISA) and an indirect fluorescent antibody test (IFAT). Antibodies (> or = 1:640) to N. caninum were found in 18 (3.9%) of 463 aborting cows. Farm prevalence in aborting cows was 12.4% (17/137). The antibody titres of cows were 1:200 (9 cows), 1:640 (7 cows), 1:1280 (3 cows), 1:2560 (3 cows), 1:5120 (3 cows), 1:10,240 (2 cows) and 1:20,480 (0 cow). A case-control study was conducted to estimate the association of N. caninum infection and abortion. For this 407 serum samples were collected from cows on five dairy farms with repeated occurrence of endemic and sporadic abortion of unidentified etiology. These samples were obtained from aborting cattle (n=44) and normally calving cattle (control group; n=363) and tested for N. caninum antibodies by an immunofluorescent antibody test (IFAT). Overall, 3.19% (13/407) of cows sampled had positive N. caninum fluorescence with a cut-off titre of 1:200. The prevalence of N. caninum was significantly higher (P<0.05) in the aborting group (13.64%; 95% confidence interval (CI): 5.2, 27.4) than in the control group (1.93%; 95% CI: 0.8, 3.9). A strong association between seropositivity and abortion was found, with seropositive cows being eight times more likely to abort than seronegative cows (odds ratio=8; 95% CI: 2.6, 25.1). This first report on the serological prevalence of N. caninum in cows in the Czech Republic verified a strong association between N. caninum infection and abortions in five dairy farms. Thus, the neosporosis should be considered in differential diagnosis of bovine abortion.     

Validation of a bulk tank milk antibody ELISA to detect dairy herds likely infected with bovine viral diarrhoea virus in New Zealand

AIMS: To assess the sensitivity and specificity of a bulk tank milk (BTM) antibody enzyme-linked immunosorbent assay (ELISA) to detect likely infection of a dairy herd with bovine vi- ral diarrhoea virus (BVDV). The ELISA was subsequently used to estimate the prevalence of likely infected herds in parts of the North Island of New Zealand.

METHODS: BTM samples from 724 randomly selected dairy herds in the Waikato, Bay of Plenty and Northland regions of New Zealand were tested for BVDV antibodies. From this group, 20 herds were again randomly selected from each of the quartiles of the ELISA percentage inhibition (%INH) result. From each participant herd, serum from 15 randomly selected calves aged 6–18 months and 15 cows was collected and tested using an indirect blocking ELISA for BVDV antibodies.

RESULTS: Among serum results from calves from 50 herds available for analysis, 34 (68%) herds were classified as likely non-infected (0-3 seropositive among 15 calves) and 16 (32%) as likely infected (5–15 seropositive among 15 calves). Receiver- operator characteristic (ROC) analysis identified an optimal cut-off for BTM of 80%INH associated with 81% sensitivity and 91% specificity for likely herd infection. The prevalence of BVDV antibodies in cows within herds and %INH for BVDV in bulk milk were positively correlated (p<0.01). The association between bulk milk %INH and the prevalence of BVDV antibodies in calves was stronger than the same association in cows. Based on the threshold of 80%INH, the 95% confidence interval (CI) for prevalence of likely infection in the 724 herds in the Waikato, Bay of Plenty and Northland regions of New Zealand was 12–17%. Vaccination against BVDV was not significantly associated with the likely infection status of the herd based on prevalence of BVDV antibodies among calves.

CONCLUSION: An ELISA test result for BVDV antibodies in BTM ≥80%INH can be used as a threshold to indicate the presence of likely infection with BVDV in dairy herds in New Zealand, with 81% sensitivity and 91% specificity.     

Validation of a bulk tank milk antibody ELISA to detect dairy herds likely infected with bovine viral diarrhoea virus in New Zealand

AIMS: To assess the sensitivity and specificity of a bulk tank milk (BTM) antibody enzyme-linked immunosorbent assay (ELISA) to detect likely infection of a dairy herd with bovine viral diarrhoea virus (BVDV). The ELISA was subsequently used to estimate the prevalence of likely infected herds in parts of the North Island of New Zealand. METHODS: BTM samples from 724 randomly selected dairy herds in the Waikato, Bay of Plenty and Northland regions of New Zealand were tested for BVDV antibodies. From this group, 20 herds were again randomly selected from each of the quartiles of the ELISA percentage inhibition (%INH) result. From each participant herd, serum from 15 randomly selected calves aged 6-18 months and 15 cows was collected and tested using an indirect blocking ELISA for BVDV antibodies. RESULTS: Among serum results from calves from 50 herds available for analysis, 34 (68%) herds were classified as likely non-infected (0-3 seropositive among 15 calves) and 16 (32%) as likely infected (5-15 seropositive among 15 calves). Receiver-operator characteristic (ROC) analysis identified an optimal cut-off for BTM of 80%INH associated with 81% sensitivity and 91% specificity for likely herd infection. The prevalence of BVDV antibodies in cows within herds and %INH for BVDV in bulk milk were positively correlated (p<0.01). The association between bulk milk %INH and the prevalence of BVDV antibodies in calves was stronger than the same association in cows. Based on the threshold of 80%INH, the 95% confidence interval (CI) for prevalence of likely infection in the 724 herds in the Waikato, Bay of Plenty and Northland regions of New Zealand was 12-17%. Vaccination against BVDV was not significantly associated with the likely infection status of the herd based on prevalence of BVDV antibodies among calves. CONCLUSION: An ELISA test result for BVDV antibodies in BTM >/=80%INH can be used as a threshold to indicate the presence of likely infection with BVDV in dairy herds in New Zealand, with 81% sensitivity and 91% specificity.     

Fasciola hepatica - monitoring the milky way? The use of tank milk for liver fluke monitoring in dairy herds as base for treatment strategies

In this study 595 lactating cows originating from 31 carinthian farms were investigated in accordance of liver fluke infection using individual and tank milk as well as individual blood and faecal samples. Two commercial ELISAs were used to test the milk and blood serum, and the results were compared with coproscopy and a commercial copro-antigen ELISA. Receiver operating characteristics (ROC) and two-graph operating characteristics (TG ROC) of tank milk results were conducted based on the individual milk to determine the minimum reliable in-herd antibody prevalence for the predominant condition in the investigation area. In 17.8% of the examined individuals located in 64.5% of the farms eggs were detected by coproscopy. The copro-antigen ELISA delivered 13.4% positive individuals from 54.8% of the farms. The milk ELISAs showed 42.7% (Euroclone) and 44.2% (Pourquier) positive cows on 90.3% of the farms. The blood samples were positive in 43% (Euroclone) and 45.2% (Pourquier) of the individuals from 90.3% to 96.8% of the herds, respectively. Based on the milk and the blood an average in-herd prevalence of 30-45% can be assumed. The serum and milk samples delivered correlating results with kappa values between 0.94 and 0.97, whereas the coproscopy and copro-antigen ELISA did not correlate well with the ELISA results. The two different ELISA tests highly correlated on individual and on herd level. Both showed a reliable minimum in-herd prevalence of ～20%, meaning that one fifth of the individuals in a herd have to be positive to obtain a positive bulk tank milk result. In the investigated area a higher in-herd prevalence is expected, therefore the tank milk is useful as a monitoring tool and can be used as a basis for intervention strategies.     

Neospora caninum and bovine virus diarrhoea virus infections in Swedish dairy cows in relation to abortion

The protozoan parasite Neospora caninum and bovine virus diarrhoea virus (BVDV) are recognized as important causes of bovine abortion and congenital disease worldwide. In this study, serological investigations were performed to estimate the prevalence of N. caninum infection in Swedish dairy cattle, to assess to what extent it may affect abortion rates, and to determine possible effects of coinfection with BVDV. The overall N. caninum seroprevalence in Swedish dairy cows was estimated at 2% (16/780), and the BVDV seroprevalence was 32% (249/780). Among aborting cows from herds with abortion problems, 7% (26/378) had antibodies to N. caninum and 42% (153/378) to BVDV. Seventeen of the N. caninum positive animals also had antibodies to BVDV. There was a statistically significant (P = 0.013) association between presence of antibodies to N. caninum and BVDV. In a case-control study comprising sera from cows in herds without recognized abortion problems, 6% (5/89) and 1% (1/89) of sera from aborting and non-aborting cows, respectively, had antibodies to N. caninum. Two of the N. caninum seropositive aborting cows also had antibodies to BVDV. These results confirm that N. caninum infection is associated with bovine abortion in Sweden and also indicate that there might be concurrent effects of N. caninum and BVDV. It is concluded that Swedish dairy cows have a low prevalence of N. caninum infection which is favourable in relation to possible future control programmes.     

Neospora caninum serostatus and culling of Holstein cattle

OBJECTIVE: To determine whether time until culling or risk of culling was associated with Neospora caninum serostatus among Holstein cattle in dairy herds in Ontario. DESIGN: Retrospective cohort study. ANIMALS: 3,412 cows in 56 herds. PROCEDURE: Blood samples were collected, and serum was tested for antibodies against N. caninum. Information on cows that were culled was collected during the 1- to 2-year period that producers were unaware of serostatus of individual cows in their herds. RESULTS: Herd prevalence of N. caninum-seropositive cows ranged from 0 to 68.3% (median, 7.0%). During the time of the study, 184 of 359 (51.3%) N. caninum-seropositive cows were culled, compared with 1,388 of 3,053 (45.5%) seronegative cows. Mean time from blood sample collection to culling for seronegative cows (289 days; 95% confidence interval, 280 to 299 days) was not significantly different from mean time for seropositive cows (296 days; 95% confidence interval, 269 to 323 days). Survival analysis indicated that N. caninum serostatus was not associated with time to culling or risk of culling. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that N. caninum serostatus of Holstein cows in Ontario was not significantly associated with either time to culling or risk of culling. Thus, N. caninum serostatus alone should not be used to determine whether cows should be culled.     

Epidemiology and impact of Neospora caninum infection in three Queensland tropical dairy herds

This study investigated the epidemiology of Neospora caninum in three tropical dairy herds in North Queensland, Australia. All animals in the herds were bled, and the sera were tested by ELISA for N. caninum antibodies. Herd records were examined, and the number of calves carried to term and the number of abortions which occurred over the lifetime of each animal were recorded to determine the abortion rate for each animal. Pedigrees were constructed for two of the herds to investigate whether vertical transmission was occurring. The seroprevalence of N. caninum ranged from 23% to 34%. The abortion rate in seropositive animals was significantly (p < 0.001) higher than in seronegative animals in all three herds (12-20.1% cf. 3.6-7%). Overall, the probability of a calf being seropositive was 3.5 times higher when the dam was also seropositive than when the dam was seronegative. Subsequent selective breeding employed by one herd reduced the N. caninum seroprevalence from 23% to 5% over a 9-year period. This study shows that N. caninum infection is prevalent in North Queensland dairy cattle, and both post-natal infection and vertical transmission are common.     

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.     

Vertical and horizontal transmission of Neospora caninum in dairy herds in Québec

Neospora caninum is an important cause of abortion in dairy cattle. The objective of this observational study was to estimate the rate of vertical transmission of N. caninum in dairy herds in Québec and to investigate horizontal transmission in the same herds. The genealogy of cows from 23 dairy herds were examined. Prevalence of seropositive animals in herds studied varied from 4.3% to 61.8% (average, 21.9%). The overall rate of vertical transmission was estimated to be 44.4%, varying from 0% to 85.7%. Seven cases of horizontal transmission were identified in 6 of the 23 herds studied. Estimated vertical transmission rate varied from herd to herd, but appeared to be higher in herds with a high prevalence of seropositive animals. Although horizontal transmission was identified in 6 herds, it does not appear to be the major route of infection for N. caninum.     

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.     

A longitudinal study of seroprevalence and seroconversion of Neospora caninum infection in dairy cattle in northeast Thailand

A long-term study was carried out in 11 dairy herds in the Khon Kaen province of northeast Thailand between August 2001 and November 2004. The objective was to investigate seroprevalence dynamics of Neospora caninum infection in the herds and to demonstrate patterns of seroconversion in individual cattle. Each herd was visited once a year, in total four times, and sera from cattle > 3 months of age and farm dogs as well as a sample from the bulk milk were collected. All samples were analysed for presence of specific antibodies by an N. caninum iscom ELISA. The overall percentage of antibody-positive cattle was constant and varied only between 10 and 13% over the 4 years, but the variation in within-herd seroprevalence between herds was substantial. Two herds had > or = 20% seropositive animals at all samplings and consistently high bulk milk OD, whereas two herds had no seropositive animal at the last two samplings and low bulk milk OD. Five herds had a decreasing trend of within-herd seroprevalence, whereas the remaining six herds had a higher portion of test-positive individuals at the end of the study. A total of 424 individuals were sampled more than once; 344 (81%) and 32 (8%) were consistently antibody-negative and antibody-positive, respectively. The proportions of animals that changed from being seronegative to seropositive and from being seropositive to seronegative between the years were 3.9-4.6% and 19-39%, respectively. Apparent vertical and horizontal transmission rates were 58% (95% CI; 44-71%) and 5% (95% CI; 3-7%), respectively. In conclusion, the overall percentage of N. caninum antibody-positive cattle was constant over the years, but the within-herd seroprevalence varied substantially between the herds. Seroconversions were likely to occur in individual cattle although most animals had consistent serological status throughout the study.     

Associations between Neospora caninum specific antibodies in serum and milk in two dairy herds in Scotland

The study evaluated the use of the Mastazyme ELISA for quantification of Neospora caninum (N. caninum) specific IgG in bovine milk and examined the relationship between serum and milk antibodies in two dairy herds. The serum and milk antibodies both had bimodal distributions in each herd. This was mainly due to between cow variation: in both herds, approximately two thirds of cows were either clearly and consistently seropositive or seronegative for N. caninum with one third consistently near the threshold. Milk and serum N. caninum IgG were strongly related. This relationship was modelled using a linear mixed model including a polynomial term for serum, the effect of herd, and between and within cow variance components. The latter gave a significantly better fit to the data than a model that allowed for a different relationship for the positive and negative (according to the serum test) groups of observations. The sensitivity and specificity (based on serum percentage positivity (pp)) of the milk antibody was determined for different milk pp thresholds. In spite of the differences between the relationship of milk to serum seen for the two herds, for those estimates with sufficient precision, sensitivity and specificity greater than 0.73 for both herds were obtained using single thresholds of 14 and 15.5 for milk pp in both herds based on, as our gold standard, serum antibody pp thresholds of 22.5 and 25, respectively. If milk antibody is to be used for detecting persistently infected cows, the higher threshold of 15.5 may be suitable while for epidemiological screening 14 would be preferable. Further validation in a greater number of herds is required, but our results suggest that this test may prove to be a useful adjunct to serum N. caninum IgG assays in the monitoring of N. caninum infection as part of herd health programmes and epidemiological studies.