A single-cohort study of Cryptosporidium bovis and Cryptosporidium ryanae in dairy cattle from birth to calving.

In order to investigate the infection dynamics of Cryptosporidium bovis and C. ryanae, a two-year prospective cohort study was performed on a dairy farm known to be free of C. parvum. Sixteen calves were recruited when newborn. Faecal samples were collected weekly until calves were nine weeks old, then monthly until calving or culling. Samples (n = 455) were examined by fluorescence microscopy, and when positive the species were determined by DNA sequencing. In calves up to nine weeks, C. bovis was found in 58.5% of the samples, C. ryanae in 9.2%, and both C. bovis and C. ryanae in 3.1%. The prevalence of shedding calves peaked at 87.5% in week five, which is earlier than many international studies have shown for C. bovis. The cumulative incidence of C. bovis reached 100% when the calves were five weeks old. In four calves, the species detected changed from C. bovis to C. ryanae or the other way around, and two samples were a combination of both species. Several individuals shed oocysts sporadically up to 16 months of age. The highest oocysts per gram faeces count was seen in week three (3.6 × 106 OPG). Diarrhoea was not associated with oocyst shedding.

Infection dynamics of Cryptosporidium bovis and Cryptosporidium ryanae in a Swedish dairy herd.

In order to investigate the infection dynamics of Cryptosporidium bovis and Cryptosporidium ryanae, a Swedish dairy farm known to be free of C. parvum was recruited for a one-year study. Fecal samples were collected every four weeks (a total of 13 occasions) from 20 randomly selected calves younger than 65 days. A total of 238 feces samples were collected directly from the rectum and then examined with immunofluorescence microscopy; those positive for Cryptosporidium spp. oocysts were further processed by molecular species determination. Oocysts were found in 92 samples, and in 72 of these the species was successfully determined: 87.5% C. bovis, 9.7% C. ryanae and 2.8% a mix of both species. No C. parvum was found. The prevalence of shedding calves was highest at ages 4 and 5 weeks; however, the feces with the highest oocyst per gram (OPG) were registered at 2-4 weeks of age. This is earlier than what has been found in most other studies, indicating that the infection dynamics of C. bovis/C. ryanae may be affected by the presence of C. parvum. The OPG for the positive calves ranged from 200 to 1.1 × 106 (geometric mean 6.88 × 103). The youngest calf in which C. bovis was identified was 5 days old, and the youngest calf in which C. ryanae was identified was 15 days old. Calves' housing type and seasonality were not associated with differences in the shedding of oocysts. Furthermore, there was no association between the presence of diarrhea and oocyst shedding.

Infection dynamics of Cryptosporidium bovis and Cryptosporidium ryanae in a Swedish dairy herd.

In order to investigate the infection dynamics of Cryptosporidium bovis and Cryptosporidium ryanae, a Swedish dairy farm known to be free of C. parvum was recruited for a one-year study. Fecal samples were collected every four weeks (a total of 13 occasions) from 20 randomly selected calves younger than 65 days. A total of 238 feces samples were collected directly from the rectum and then examined with immunofluorescence microscopy; those positive for Cryptosporidium spp. oocysts were further processed by molecular species determination. Oocysts were found in 92 samples, and in 72 of these the species was successfully determined: 87.5% C. bovis, 9.7% C. ryanae and 2.8% a mix of both species. No C. parvum was found. The prevalence of shedding calves was highest at ages 4 and 5 weeks; however, the feces with the highest oocyst per gram (OPG) were registered at 2-4 weeks of age. This is earlier than what has been found in most other studies, indicating that the infection dynamics of C. bovis/C. ryanae may be affected by the presence of C. parvum. The OPG for the positive calves ranged from 200 to 1.1 × 106 (geometric mean 6.88 × 103). The youngest calf in which C. bovis was identified was 5 days old, and the youngest calf in which C. ryanae was identified was 15 days old. Calves' housing type and seasonality were not associated with differences in the shedding of oocysts. Furthermore, there was no association between the presence of diarrhea and oocyst shedding.

Disinfection with hydrated lime may help manage cryptosporidiosis in calves.

Diarrhea is common in young calves and is often caused by Cryptosporidium parvum infection. The aim of this study was to investigate if disinfection of calf pens with hydrated lime would reduce contamination of C. parvum oocysts and improve calf health in herds with C. parvum associated diarrhea problems. Four dairy herds with ongoing C. parvum associated calf diarrhea problems each participated in the study over six to seven months. During the study period, all pens/huts for young calves were cleaned according to the usual farm routine before a new calf entered. Hydrated lime was then used to disinfect half of the pens/huts. Diarrhea incidence was recorded by the farmers and by veterinarians, who clinically examined the calves every second month. In total, 402 calves participated in the study. The farmers detected diarrhea in 214 (53%) calves, with similar proportions in calves kept in lime disinfected and control pens. Age at diarrhea onset was significantly higher in lime disinfected pens than in control pens, 9.0 days and 7.6 days, respectively. There was no difference between the groups regarding duration or severity index recorded by the farmers. The body condition score in 6-8 week old calves was significantly higher in calves that had been kept in lime disinfected pens during their first weeks of life, indicating that calves in disinfected pens/huts were less affected by their infections. Faecal samples from 5 to 21 day old calves, were collected on four occasions at each farm (n = 95). Cryptosporidium positive samples were found at all samplings in all four herds. Cryptosporidium spp. was detected in 79 (83%) samples with no difference between lime disinfected and control pens. C. parvum was the dominant species. Two different C. parvum subtypes were found; IIaA16G1R1b in three herds and IIaA16G1R1b_variant in one herd. Only one subtype was found in each herd. Disinfection of calf pens with slaked lime delayed onset of diarrhea and improved the body condition in the calves, but did not affect diarrhea incidence or duration. Although lime disinfection alone will not be sufficient to control Cryptosporidium associated diarrhea in herds with extensive calf diarrhea problems, these results suggest that it can be a valuable complement to other measures.

Detection of Treponema phagedenis-like antibodies in serum and bulk milk from cows with and without digital dermatitis.

Digital dermatitis (DD) in dairy cows is a widespread disease linked to infection with Treponema. The traditional diagnostic method is clinical inspection, which is subjective and laborious. We explored the performance of 4 different immunogenic proteins from Treponema phagedenis in a new antibody ELISA for analysis of serum or milk. Analysis of samples from 390 cows in 25 herds showed that the ELISA could distinguish the majority of cows with DD from healthy cows. By changing the cutoff and applying parallel or serial testing, high sensitivity or specificity could be achieved. The investigation indicated that aggregated test results can be useful in the assessment of a herd's DD status. In addition, analysis of bulk tank milk samples showed good agreement with results from individual cows. The test system could be useful in research on the epidemiology and immunology of DD.

Impact of meteorological and environmental factors on the spatial distribution of Fasciola hepatica in beef cattle herds in Sweden.

BACKGROUND: Fasciola hepatica is a parasite with a significant impact on ruminant livestock production. Previous studies in north-west Europe have described its geographical distribution and determined potential predictors of fasciolosis using geographical information system (GIS) and regression modelling. In Sweden, however, information about the distribution of fasciolosis is limited. This study examined the geographical distribution of F. hepatica and identified high-risk areas for beef cattle in Sweden and sought to characterise potential predictors. Beef cattle serum samples were collected during winter 2006-2007 from 2135 herds which were examined for F. hepatica antibodies by enzyme-linked immunosorbent assay (ELISA). Fasciolosis distribution maps were created using GIS based on postcode location of seropositive herds. Spatial scan analysis (SaTScan) was performed to determine high-risk areas. Using datasets on animal density, temperature, precipitation and Corine land cover data, including soil type and soil mineral concentrations in Sweden, bivariate and multiple logistic regression analyses were carried out in R software to reveal potential predictors of F. hepatica infection. RESULTS: Overall herd seroprevalence of F. hepatica in beef cattle was 9.8 % (95 % CI: 8.6-11.1). An irregular spatial distribution of F. hepatica, with two main clusters, was observed in south-west Sweden. The most northerly occurrence of F. hepatica in the world was documented. The final model explained 15.8 % of the variation in F. hepatica distribution in study herds. Absence of coniferous forest was the variable with the highest predictive value. Precipitation in July-September, Dystric Cambisol, Dystric Regosol, and P and Cu concentrations in soil were other negative predictors. Beef cattle herd density, Dystric Leptosol and Fe concentration were positive predictors. CONCLUSIONS: The spatial distribution of F. hepatica in Swedish beef cattle herds is influenced by multi-factorial effects. Interestingly, absence of coniferous forest, herd density, specific soil type and concentration of some soil minerals are more important predictors than climate factors.

Cryptosporidium parvum IId family: clonal population and dispersal from Western Asia to other geographical regions.

In this study, 111 Cryptosporidium parvum IId isolates from several species of animals in China, Sweden, and Egypt were subtyped by multilocus sequence typing (MLST). One to eleven subtypes were detected at each of the 12 microsatellite, minisatellite, and single nucleotide polymorphism (SNP) loci, forming 25 MLST subtypes. Host-adaptation and significant geographical segregation were both observed in the MLST subtypes. A clonal population structure was seen in C. parvum IId isolates from China and Sweden. Three ancestral lineages and the same RPGR sequence were shared by these isolates examined. Therefore, the present genetic observations including the higher nucleotide diversity of C. parvum IId GP60 sequences in Western Asia, as well as the unique distribution of IId subtypes (almost exclusively found in Asia, Europe, and Egypt) and in combination with the domestication history of cattle, sheep, and goats, indicated that C. parvum IId subtypes were probably dispersed from Western Asia to other geographical regions. More population genetic structure studies involving various C. parvum subtype families using high-resolution tools are needed to better elucidate the origin and dissemination of C. parvum in the world.

Application of network analysis parameters in risk-based surveillance - examples based on cattle trade data and bovine infections in Sweden.

Financial resources may limit the number of samples that can be collected and analysed in disease surveillance programmes. When the aim of surveillance is disease detection and identification of case herds, a risk-based approach can increase the sensitivity of the surveillance system. In this paper, the association between two network analysis measures, i.e. 'in-degree' and 'ingoing infection chain', and signs of infection is investigated. It is shown that based on regression analysis of combined data from a recent cross-sectional study for endemic viral infections and network analysis of animal movements, a positive serological result for bovine coronavirus (BCV) and bovine respiratory syncytial virus (BRSV) is significantly associated with the purchase of animals. For BCV, this association was significant also when accounting for herd size and regional cattle density, but not for BRSV. Examples are given for different approaches to include cattle movement data in risk-based surveillance by selecting herds based on network analysis measures. Results show that compared to completely random sampling these approaches increase the number of detected positives, both for BCV and BRSV in our study population. It is concluded that network measures for the relevant time period based on updated databases of animal movements can provide a simple and straight forward tool for risk-based sampling.

Identification of immunogenic proteins in Treponema phagedenis-like strain V1 from digital dermatitis lesions by phage display.

Digital dermatitis (DD) is a contagious claw disease causing lameness in cattle, affecting both animal welfare and economics. In this study, shotgun phage display was used to identify immunogenic proteins in a strain (V1) of the Treponema phylotype closely related to Treponema phagedenis, indicated as a key agent in the pathogenesis of DD. A genomic phage library was constructed and selected against antibodies from a rabbit immunized with live strain V1 bacteria. A homolog to the immunogenic protein TmpA of Treponema pallidum subsp. pallidum was identified, as well as a putative phage tail tape measure protein (Ttm), and a putative proline-rich repeat lipoprotein (PrrA). The complete amino acid sequences of these proteins were predicted from a genomic sequence of strain V1 generated by 454 Sequencing™. The presence of these genes in ten Treponema spp. field isolates was investigated by PCR. The tmpA and ttm genes were detected in all T. phagedenis-like isolates while prrA was detected in four out of seven. None of the genes were detected in the three Treponema pedis isolates investigated. Recombinant proteins were produced and used in indirect ELISAs. For all three proteins, a majority of serum samples from cattle with DD (n=8) showed higher optical density values than samples from cattle without DD (n=7).

Prevalence of antibodies against Toxoplasma gondii and Neospora caninum in moose (Alces alces) and roe deer (Capreolus capreolus) in Sweden.

Toxoplasma gondii and Neospora caninum are two coccidian parasites with a worldwide distribution. T. gondii is one of the more common parasitic zoonoses in the world and in young children and immunocompromised persons, infection can lead to severe disease and death. N. caninum is an important cause of abortions in cattle. Wildlife have been identified as reservoirs and transmitters for both parasites. The purpose of this study was to investigate the seroprevalences of T. gondii, and N. caninum in moose (Alces alces), and roe deer (Capreolus capreolus) in Sweden. Blood samples were collected from 417 moose during 2000-2005 and from 199 roe deer during 1990-2007. The samples were investigated for presence of antibodies by a T. gondii direct agglutination test and a N. caninum iscom ELISA. Because the iscom ELISA has not been validated for moose or roe deer, sera that gave a positive result were further investigated by immunoblot analysis to verify presence of antibodies. Antibodies to T. gondii were detected in 85 (20%) and 68 (34%) moose and roe deer sera, respectively. In moose the seroprevalence was higher in south and central Sweden than in the north, whereas there was no difference between the regions for roe deer. Adult moose and roe deer had higher odds of being seropositive than young animals but there were no difference in seroprevalence between males and females. One roe deer was positive by immunoblotting and was regarded as N. caninum positive, whereas all moose sera were negative. The results show that T. gondii infection is widely spread in the Swedish moose and roe deer populations. Precautions should therefore be taken when handling internal organs and carcasses of harvested cervids. Proper handling and cooking of game meat also is important to prevent toxoplasmosis in humans.

Cryptosporidium infection in herds with and without calf diarrhoeal problems.

A case-control study was designed to investigate the role of different Cryptosporidium spp. in Swedish dairy herds with and without calf diarrhoeal problems. Faecal samples were collected from preweaned calves, young stock and cows. Cryptosporidium oocysts were detected by sodium chloride flotation and epifluorescence microscopy. Molecular diagnostics were used to identify Cryptosporidium species. Samples containing C. parvum were further analysed to determine subtypes. Calf faecal samples were also analysed for rotavirus, coronavirus and Escherichia coli F5+. Total protein was assessed in 1- to 8-day-old calves. A questionnaire was used to identify differences in management routines. Cryptosporidium infection was diagnosed in all herds, with equal prevalence in case and control herds in all three age groups. Cryptosporidium parvum, Cryptosporidium bovis, Cryptosporidium ryanae and Cryptosporidium andersoni were all identified, as were rotavirus, coronavirus and E. coli F5+. C. ryanae and C. andersoni were only detected in non-diarrhoeal samples, whereas the other pathogens were detected in both diarrhoeal and non-diarrhoeal samples. Diarrhoea was more common in case herd calves. Disinfection of single pens was more common in case herds and several other management routines seemed to differ although results were not significant.

Spatial patterns of bovine corona virus and bovine respiratory syncytial virus in the Swedish beef cattle population.

BACKGROUND: Both bovine coronavirus (BCV) and bovine respiratory syncytial virus (BRSV) infections are currently wide-spread in the Swedish dairy cattle population. Surveys of antibody levels in bulk tank milk have shown very high nationwide prevalences of both BCV and BRSV, with large variations between regions. In the Swedish beef cattle population however, no investigations have yet been performed regarding the prevalence and geographical distribution of BCV and BRSV. A cross-sectional serological survey for BCV and BRSV was carried out in Swedish beef cattle to explore any geographical patterns of these infections. METHODS: Blood samples were collected from 2,763 animals located in 2,137 herds and analyzed for presence of antibodies to BCV and BRSV. Moran's I was calculated to assess spatial autocorrelation, and identification of geographical cluster was performed using spatial scan statistics. RESULTS: Animals detected positive to BCV or BRSV were predominately located in the central-western and some southern parts of Sweden. Moran's I indicated global spatial autocorrelation. BCV and BRSV appeared to be spatially related: two areas in southern Sweden (Skaraborg and Skåne) had a significantly higher prevalence of BCV (72.5 and 65.5% respectively); almost the same two areas were identified as being high-prevalence clusters for BRSV (69.2 and 66.8% respectively). An area in south-east Sweden (Kronoberg-Blekinge) had lower prevalences for both infections than expected (23.8 and 20.7% for BCV and BRSV respectively). Another area in middle-west Sweden (Värmland-Dalarna) had also a lower prevalence for BRSV (7.9%). Areas with beef herd density > 10 per 100 km(2) were found to be at significantly higher risk of being part of high-prevalence clusters. CONCLUSION: These results form a basis for further investigations of between-herds dynamics and risk factors for these infections in order to design effective control strategies.

Molecular characterisation of Cryptosporidium isolates from Swedish dairy cattle in relation to age, diarrhoea and region.

Cryptosporidium positive samples from 176 preweaned calves, young stock and cows of 48 herds were subjected to molecular characterisation of the 18S rRNA gene to determine which species are present in Swedish dairy cattle. In addition, samples characterised as Cryptosporidium parvum were further analysed at the GP60 gene to investigate distribution and zoonotic potential of subtypes. The 18S rRNA gene was successfully sequenced in 110 samples, with Cryptosporidium bovis in 83, C. parvum in 15, Cryptosporidium ryanae in 10, and Cryptosporidium andersoni in two samples. C. bovis was the most common species, being identified in 74% of calf samples, in 77% of young stock samples and in 100% of cow samples. The youngest calves infected with C. bovis were 7 days old, showing that the prepatent period is shorter than the previously stated 10 days. C. parvum was detected in 15 calves from nine farms, and samples were clustered in the southern parts of Sweden. Diarrhoeic calf samples contained C. parvum, C. bovis or C. ryanae. Sequencing of the GP60 gene was successful in 13 of the C. parvum samples. Eight subtypes, including three novel ones, were detected. Four of the subtypes have previously been identified in humans. This indicates that there is a zoonotic potential in C. parvum infected Swedish dairy calves.

Investigation of Chlamydiaceae in semen and cauda epididymidis and seroprevalence of Chlamydophila abortus in breeding bulls.

BACKGROUND: Reproductive disorders associated with chlamydial infection have been reported worldwide in cattle and there are indications of potential venereal transmission. METHODS: Semen samples from 21 dairy bulls and cauda epididymidis tissue samples from 43 beef bulls were analysed for chlamydial agent by real-time polymerase chain reaction (PCR) including an internal amplification control (mimic). Additionally, presence of antibodies against Chlamydophila (Cp.) abortus among the bulls was investigated with the commercial Pourquier ELISA Cp. abortus serum verification kit. RESULTS: No chlamydial agent was detected by PCR in either the semen samples or in the tissue samples. Additionally, no antibodies against Cp. abortus were detected. CONCLUSIONS: The results suggest that Cp. abortus is very rare, or absent in Swedish bulls and thus the risk for venereal transmission of chlamydial infection through their semen is low. However, because Chlamydophila spp. infection rates seem to differ throughout the world, it is essential to clarify the relative importance of transmission of the infection through semen on cattle fertility.

Seroprevalence and spatial distribution of Neospora caninum in a population of beef cattle.

A cross-sectional serological survey for Neospora caninum was carried out in Swedish beef cattle in order to estimate the seroprevalence and investigate any geographical patterns of the infection. Blood samples were collected from 2754 animals in 2130 herds and analysed for presence of antibodies to N. caninum. The study included 18.5% of all Swedish beef herds. Spatial autocorrelation was assessed by calculating Moran's I and cluster detection was performed using LISA- and spatial scan statistics. The seroprevalence of N. caninum was 2.8% (95% CI: 2.18-3.44). Positive samples were found only in the southern half of Sweden. Moran's I was positive (I=0.045, p<0.01) and indicated global spatial autocorrelation. Using the spatial scan test based on a Bernoulli model, a cluster comprising 16 postal code areas and including 19 seropositive animals was identified. The results indicate that there is a cluster of higher risk of N. caninum (relative risk=3.95) in the southern part of Sweden, in the counties of Kronoberg and Skåne. This is the first report on the seroprevalence of N. caninum in beef cattle in Sweden.

Systematic review and meta-analyses of the effects of halofuginone against calf cryptosporidiosis.

Halofuginone seems to reduce diarrhoea and oocyst shedding in calves with cryptosporidiosis, but provides no complete cure. To develop more precise estimates of the effects of halofuginone on calf cryptosporidiosis, meta-analyses were performed, including studies on prophylactic and therapeutic treatment. Meta-analysis increases statistical power because several trials are evaluated together, increasing the effective sample size and possibility of detecting true effects. In total, 20 cohort or clinical studies (in 16 publications) investigating halofuginone treatment in calves were identified. One study was excluded because treated calves and control calves were not investigated in parallel. Four studies (three publications) were excluded because only abstracts were available. Thus, 15 studies from 12 publications, with 10-311 calves were included for data extraction. Of these, five studies from three publications could not be used for meta-analysis because they did not report the data needed. Effects on infection prevalence, diarrhoeal prevalence and mortality were investigated. For prophylactic treatment, halofuginone had an effect on infection and diarrhoeal prevalence on study days 4 and 7, but the control group had significantly lower infection prevalence than the halofuginone treated group on study day 21. Heterogeneity was detected on study days 14 and 21 and publication bias was detected on study days 7 and 14. Mortality was not affected. For therapeutic treatment, a shortage of studies in combination with heterogeneity made interpretations uncertain, and we could not determine if halofuginone treatment benefits calves.

Prevalence and associated management factors of Cryptosporidium shedding in 50 Swedish dairy herds.

Cryptosporidium parvum is a protozoan parasite causing diarrhoea in young calves. This cross-sectional study was performed to estimate the prevalence of Cryptosporidium infected herds in a sample of Swedish dairy herds and to identify potential risk factors associated with shedding of oocysts. Fifty dairy herds, selected by stratified random sampling, were included. The herds were visited once during the indoor seasons of 2005-2006 and 2006-2007. Faecal samples were collected from 10 calves, 10 young stock and 5 cows in each herd. Clinical observations of sampled animals and environmental status were recorded, and farmers were interviewed about management procedures. Faecal samples were cleaned by sodium chloride flotation and detection of oocysts was made by epifluorescence microscopy. Cryptosporidium parvum-like oocysts were found in 96% of the herds. Prevalence was 52% in calves, 29% in young stock and 5.6% in cows. Three two-day-old calves shed oocysts. Cryptosporidium andersoni was found in seven animals from four different herds. Factors associated with prevalence of shedders among sampled animals in a herd were age at weaning, cleaning of single calf pens, placing of young stock, system for moving young stock, and year of sampling. Factors associated with shedding in calves were age, placing of young stock, routines for moving young stock and time calf stays with the cow. The only significant factor in young stock was age. In cows, number of calves in the herd and type of farming (organic vs. conventional) affected shedding.

Spatial analysis of Neospora caninum distribution in dairy cattle from Sweden.

The national herd prevalence and spatial distribution of Neospora caninum infected dairy herds in Sweden were investigated. The study was based on a bulk milk survey comprising samples from 2,978 herds. Test-positive herds were found in all parts of Sweden and the overall prevalence of test-positive herds was 8.3% (95% confidence interval = 7.3-9.3%). The presence of spatial autocorrelation was tested using the Moran's I test. Possible clusters of test-positive herds were identified by applying the local indicator of spatial association (LISA) test statistic and the spatial scan statistic. Analysis based on data aggregated by postal code areas as well as analysis based on exact coordinates identified significant clusters of high prevalence in the middle parts of Sweden and low prevalence in the south. This was not expected considering the results from other European studies of N. caninum in cattle. However, the findings are supported by the distribution of previously known case herds.

Investigation of Chlamydophila spp. in dairy cows with reproductive disorders.

BACKGROUND: Reports worldwide indicate high prevalence of Chlamydophila spp. infection in cattle. To assess the prevalence in Sweden, 525 cows in 70 dairy herds with reproductive disorders was investigated. METHODS: To detect antibodies two commercially available kits were used. Moreover, 107 specimens, including vaginal swabs, organ tissues and milk were analysed by Polymerase Chain Reaction (PCR). RESULTS: Two (0.4%) cows were seropositive in the Pourquier Cp. abortus ELISA. The seroprevalence with the Chekit ELISA was 28% with no difference between cases and controls. Five specimens were positive in real-time PCR and further analysed by nested PCR. Cp. pecorum was confirmed by partial omp1 DNA sequencing of the nested PCR product of vaginal swabs from control cows. CONCLUSION: The results suggest that Cp. abortus infection is absent or rare in Swedish cows whereas Cp. pecorum is probably more spread. They also suggest that Chlamydophila spp. are not related to reproduction disorders in Swedish cattle.

Prevalence of Neospora caninum and bovine viral diarrhoea virus in dairy cows in Southern Vietnam.

The main purpose of this study was to investigate the prevalence of Neospora caninum and bovine viral diarrhoea virus (BVDV) in some dairy herds in Southern Vietnam, and to ascertain whether there were differences in seroprevalences between herds with imported and locally bred cows. Serum samples collected on five state farms and 97 smallholder herds were analysed for the presence of antibodies to N. caninum and BVDV. All BVDV antibody-negative sera were further tested by antigen-ELISA in order to identify persistently infected individuals. The N. caninum prevalence varied between 16% and 53% in the state herds, and was higher in the four herds that had imported cows than in the herd that only had locally bred cows. Nineteen percent of the samples collected on smallholder farms, which all had only locally bred cows, had antibodies to N. caninum. The BVDV seroprevalence varied between 58% and 93% on the state farms. In smallholder herds, the prevalence of BVDV among the sampled cows was 18% and even lower on the state farms. Despite the high seroprevalence for BVDV in the state herds, no persistently BVDV infected cows were found. Given the high prevalence for Neospora and BVDV among herds with imported cows, it seems advisable to test for both infections before cattle are imported into the country.