Serological evidence for exposure to bovine viral diarrhoea virus in sheep co-grazed with beef cattle in New Zealand

ABSTRACT

Aims: To determine whether sheep that co-grazed with cattle that were suspected to be positive for bovine viral diarrhoea (BVD) virus had serological evidence of exposure to the virus.

Methods: Eighteen commercial farms that routinely co-grazed cattle and sheep in the same paddocks were recruited through purposive sampling. The recruiting veterinarians identified nine farms with cattle herds that were known or highly suspected to be positive for BVD and nine farms that were considered to be free of BVD. Blood samples were taken from 15 ewes aged 1 year on each farm and samples were submitted to a commercial diagnostic laboratory to test for antibodies against pestiviruses using an ELISA. All samples that were positive were then tested using a virus neutralisation test (VNT)for antibodies against BVD virus.

Results: Of the 270 blood samples, 17 were positive for pestivirus antibodies by ELISA and these originated from two farms that were known or suspected to have BVD virus-positive cattle. None of the samples from the nine flocks co-grazed with cattle herds that were known or suspected to be BVD virus-negative were positive for pestivirus antibodies. Within the two positive farms, 2/15 samples from the first farm and 15/15 samples from the second farm were antibody-positive. When the 17 positive blood samples were submitted for VNT, all 15 samples from the second farm tested positive for BVD virus antibodies with the highest titre being 1:512.

Conclusions and clinical relevance: In this small sample of New Zealand sheep and beef farms with suspected BVD infection in cattle, there was evidence of pestivirus exposure in co-grazed sheep. Although we were unable to confirm the origin of the exposure in these sheep, these findings highlight that farmers who are trying to eradicate BVD from their cattle should be mindful that the infection may also be circulating in sheep, and both populations should be considered a possible risk to each other for generating transient and persistent infections. Further work is needed to estimate the true prevalence of New Zealand sheep flocks that are affected by BVD and the associated economic impacts.     

Outbreak of foetal infection with bovine pestivirus in a central Queensland beef herd

OBJECTIVE: To describe a significant outbreak of foetal infection and subsequent losses due to bovine pestivirus on a 5200 ha beef breeding and fattening property in central Queensland. DESCRIPTION OF THE HERD: The affected herd consisted of 656 cows, including 269 recently purchased cows, and 221 heifers that were joined in December/January 1995/96. There were approximately 2500 cattle on the property. INVESTIGATION: Following the purchase of 269 cows in October 1995, which were mingled with the existing cow herd, losses were experienced due to foetal infection with bovine pestivirus. These losses were recorded between 1996 and 1999 as: reduced pregnancy rates, losses between pregnancy testing (midpregnancy) and branding (calves averaged 3 months-of-age), losses due to pneumonia and ill-thrift between branding and approximately 12 months-of-age, and losses due to ill-thrift and the chronic wasting form of mucosal disease thereafter. All surviving calves were tested for bovine pestivirus in 1997 at an average of 10 months. Fifty-three calves were identified as persistently infected with bovine pestivirus. A further 110 calf losses could reasonably be attributed to bovine pestivirus infection. Persistently infected cattle were always unthrifty compared to their virus negative counterparts. Only one persistently infected calf was identified, on the basis of severe ill thrift, in the 1997 birth cohort and none in 1998. CONCLUSIONS: This outbreak of foetal infection with bovine pestivirus resulted in significant production losses. These losses were recorded over the three years subsequent to the outbreak. Significant numbers of persistently infected calves were not evident among calves born in the two years after this outbreak.     

Severe malformations in calves associated with bovine viral diarrhoea (BVD) virus infection in a dairy cattle herd

During late may 2004, Some dairy cows at Al-Kharj area of central Saudi Arabia, gave birth to severely malformed calves which died, few hours to few days following birth. Samples were collected from the affected calves and their dams of virological and serological investigations. Bovine viral diarrhoea virus was detected by capture enzyme linked immuno-sorbent assay (ELISA) in the brains of affected calves. Serum antibodies were detected in the dams. The present study indicated that in spite of vaccination against BVD in the country, still severe affections of the disease are encountered. Further insight epidemiological studies to elucidate the BVD situation in Saudi Arabia is urgently needed.     

Serological status of cattle to bovine viral diarrhoea virus and bovine herpesvirus 1 at entry to and exit from Australian feedlot backgrounding facilities

A total of 6195 cattle were enrolled in this observational study. Serum antibody concentrations to bovine herpesvirus 1 (BHV1) and bovine viral diarrhoea virus (BVDV) were measured at entry to and exit from backgrounding facilities to assess their statuses on arrival and the extent of seroconversion to these viruses during backgrounding. The backgrounding facilities were contiguous with five feedlots in: Queensland (two sites), New South Wales, South Australia and Western Australia. Cattle were held in the backgrounding facilities for a minimum of 29 days and a median of 34 days. On backgrounding facility entry, 32.7% of the study population was seronegative to BVDV, but 85.7% was seronegative to BHV1. After commingling in the backgrounding facilities, of the cattle that were seronegative on backgrounding facility entry, 33.9% and 30.3% showed a serological increase to BVDV and BHV1, respectively. At backgrounding facility exit, when cattle were placed in their feedlots, 19.6% and 59.1% were seronegative to BVDV and BHV1, respectively, and 0.26% were persistently infected with BVDV. There was a strong association between seroincrease to BVDV and seroincrease to BHV1 (P = 0.005) at animal level in cohorts known to contain an animal persistently infected with BVDV.     

Bovine viral diarrhoea virus in beef cattle herds of Yucatan, Mexico: seroprevalence and risk factors

A survey of bovine viral diarrhoea virus (BVDV) infection was carried out from June 2001 to July 2002 in a non-vaccinated beef cattle population from the livestock region of Yucatan, Mexico, to assess seroprevalence and identify risk factors related to seroprevalence. The aim was also to estimate the intra-herd correlation (r_e) and design effect (D) of BVDV seropositivity. Cattle were selected by a two-stage cluster sampling. Blood samples were collected from 560 animals originating from 40 herds. Sera were tested for antibodies against BVDV using an indirect ELISA test. The sensitivity and specificity of the test was 97.9 and 99.7%, respectively. Risk factors regarding the herd and each animal sampled were recorded through a personal interview at the time of blood sampling. Twenty-four of the 40 herds had at least one seropositive animal. The animal true seroprevalence was estimated as 14%. The marginal logistic regression model used to describe the data found a significant (p < 0.05) association of herd size–cow-origin interaction. The interaction was due to a higher risk of seropositivity in the category of herds with ≤100 animals and purchased cows (OR = 1) as compared to herds with ≤100 animals and cows born in the farm (OR = 0.23). Seropositivity between cows purchased and cows born in the farm was similar for herd sizes of 101–196 and >196 animals. The r_e and D values were 0.17 ± 0.05 and 3.16 ± 0.57, respectively.     

Antibody responses to inactivated vaccines and natural infection in cattle using bovine viral diarrhoea virus ELISA kits: Assessment of potential to differentiate infected and vaccinated animals

Bovine viral diarrhoea virus (BVDV) is one of the most common and economically important viral infections of cattle. As vaccination is common in most European countries, differentiation between infected and vaccinated animals is one of the key challenges facing BVDV eradication campaigns. This study was designed to compare the ability of commercial ELISA kits to differentiate antibodies generated following vaccination with four different commercial inactivated BVDV vaccines from antibodies generated following challenge with virulent BVDV. Although none of the tested vaccine–ELISA combinations was able to differentiate an infected from a vaccinated animal (DIVA) at the individual animal level, p80 blocking ELISAs, in combination with inactivated BVDV vaccines, may have some value under certain circumstances at herd level. In most cases, antibody responses to BVDV vaccines cannot be clearly distinguished from responses seen in the early phase of natural infection. No commercial BVD vaccine showed true marker qualities for DIVA using p80 blocking ELISAs.     

Prevalence of Neospora antibodies in beef cattle in New Zealand

dAim:To estimate the prevalence of Neospora infection in a sample of New Zealand beef cattle.

dMethods: The prevalence of Neospora caninum infection in New Zealand beef cattle was estimated by collecting blood at slaughter from 499 beef cattle from 40 different farms at 2 slaughter plants in the North Island and 1 in the lower South Island. Sera were tested using an ELISA against Neospora tachyzoite antigen.

dResults: The prevalence of seropositive cattle was 2.5% (n=120), 3.6% (n=166) and 2.3% (n=213) at the plants surveyed, the overall prevalence being 2.8%. The serologically positive cattle came from 9 farms, 3 of which had more than 1 positive animal. The highest prevalence recorded amongst animals from 1 farm was 4/13 (31%), in a group of young steers.

dConclusion: Neosporosis appears to be present at a lower level in the New Zealand beef cattle population than in the New Zealand dairy cattle population. Nevertheless, from the high seroprevalence evident amongst young cattle on 1 farm, we suggest that Neospora may be a cause of infertility in beef cattle in this country.     

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.     

Elimination of bovine viral diarrhoea virus in New Zealand: a review of research progress and future directions

The major impacts of bovine viral diarrhoea (BVD) on cattle health and production have prompted many countries to embark on national elimination programmes. These programmes typically involve identifying and removing persistently infected (PI) cattle in infected herds and implementing biosecurity measures, such as pre- or post-movement testing. In order to design a systematic national control programme to eliminate BVD in New Zealand, which achieves the greatest benefits to the industries at the lowest cost to individual farmers, an accurate understanding is necessary of the epidemiology, economics and social motivation for BVD control in New Zealand. In this article we briefly review the pathogenesis of BVD, transmission and diagnosis of BVD virus infection, and effectiveness of vaccination. We summarise the current state of knowledge of the prevalence, risk factors for transmission, and financial impacts of BVD in New Zealand. We describe control programmes in Europe and then discuss the challenges that must be addressed to design a cost-effective national control programme to eliminate BVD in New Zealand.     

Seroprevalence to bovine virus diarrhoea virus and other viruses of the bovine respiratory complex in Venezuela (Apure State)

Six hundred and fifteen serum samples obtained from cows in five districts of Apure State, Venezuela, were tested by ELISA for antibodies to bovine virus diarrhoea virus (BVDV). The same samples were also ELISA-tested for antibodies to bovine herpesvirus type 1 (BHV-1) and bovine respiratory syncytial virus (BRSV). Additionally, the haemagglutination-inhibition (HI) test was used for detecting antibodies to parainfluenza virus type 3 (PIV-3). Overall, seroprevalence to BVDV was 36±7% (SE); seroprevalence varied by district (19–42%). BHV-1 seroprevalence was 67±4%; variation by district was similar to that of BVDV. However, the first 80 serum samples tested by BHV-1 ELISA all had a strong background reaction with the control antigen. Therefore, these sera were adsorbed to a homogenate of non-infected bovine kidney cell line (MDBK) and re-tested by ELISA. The non-specific reactivity was significantly reduced (p < 0.001 by Wilcoxon's signed-rank test). Compared to the virus-neutralisation (VN) test, the adsorbed BHV-1 ELISA showed 94% agreement and gave a κ value of 0.84, indicating that the adsorption did not interfere with test accuracy. Seroprevalence against BRSV was 85±3%, and showed differences across districts. Most of the cows (94±2%) were seropositive to PIV-3, and there were no significant differences among districts.     

Differences in experimental virulence of bovine viral diarrhoea viral strains isolated from haemorrhagic syndromes

In the late 1980s, a new hypervirulent and epidemic form of bovine viral diarrhoea virus (BVDV) infection appeared in North America. A similar but sporadic syndrome was later reported in Europe. To compare the pathogenic characters of the North American and European hypervirulent strains, we inoculated BVDV na?ve calves with BVDV strains isolated from haemorrhagic syndromes originating in Belgium, France and the USA. The experimental procedure comprised daily clinical examination and measurement of blood and virological parameters. The American BVD890/256 strain induced severe thrombocytopaenia, profuse diarrhoea and pneumonia in all calves, indicating that hypervirulent BVDV could be the primary infectious agent of pneumonia. Interestingly, a strong correlation was observed between the intense viraemia and a decreased platelet count. None of the European strains tested induced significant pathological signs, although isolated from cases presenting haemorrhagic syndrome.     

The use of a mass-action model to validate the output from a stochastic simulation model of bovine viral diarrhoea virus spread in a closed dairy herd

The spread of bovine virus diarrhoea virus (BVDV) in a closed dairy herd maintained under typical management conditions is studied using two approaches. In the first instance a stochastic computer model is used to simulate the month-to-month changes in the infection status of each animal. These results are contrasted with the results of a mass-action model which uses three differential equations. A comparison of the two approaches indicates that the results are in broad agreement. The stochastic approach has the benefit of providing an estimate of the probability of the infection becoming extinct and the herd becoming BVDV-free for different herd sizes.     

Evaluation of a blocking ELISA for the detection of bovine viral diarrhoea virus (BVDV) antibodies in serum and milk

The performance characteristics of a blocking ELISA test applied to serum and individual milk for the detection of antibodies to bovine viral diarrhoea virus (BVDV) were assessed using 1189 matched milk/serum samples collected from cows of 42 dairy herds located in Brittany (west of France). This test was based on a monoclonal antibody directed against non-structural protein NS2-3 of pestiviruses. All tests were performed blind. For each type of sample, negative/positive cut-off values were determined using receiver operating characteristic (ROC) analysis. Sensitivity and specificity were estimated using the virus neutralisation test as a reference. For sera, the ROC analysis provided a negative/positive inhibition percentage cut-off value of 50% giving a sensitivity and a specificity of 96.9 and 97.8%. For individual milk samples, the cut-off was fixed at 30%, leading to a sensitivity and a specificity of 96.9 and 97.3%. Using this test, a good overall agreement was found between results obtained on matched milk/serum samples (Kappavalue=0.95). The present results indicate that this blocking ELISA test is reliable enough for use in a mass screening and control scheme on BVDV.     

Genetic and antigenic characterization of bovine viral diarrhoea virus type 2 isolated from cattle in India

Previous studies have shown that bovine viral diarrhoea virus type 1 (BVDV-1) subtype b is predominantly circulating in Indian cattle. During testing for exotic pestiviruses between 2007 and 2010, BVDV-2 was identified by real time RT-PCR in two of 1446 cattle blood samples originating from thirteen states of India. The genetic analysis of the isolated virus in 5′ UTR, N^pro, entire structural genes (C, E^rns, E1 and E2), nonstructural genes NS2-3 besides 3′ UTR demonstrated that the nucleotide and amino acid sequences showed highest similarity with BVDV-2. The entire 5′ and 3′ UTR consisted of 387 and 204 nucleotides, respectively, and an eight nucleotide repeat motif was found twice within the variable part of 3′ UTR that may be considered as a characteristic of BVDV-2. The phylogenetic analysis revealed that the cattle isolate and earlier reported goat BVDV-2 isolate fall into separate clades within BVDV-2a subtype. Antigenic typing with monoclonal antibodies verified the cattle isolate also as BVDV-2. In addition, cross-neutralization tests using antisera raised against Indian BVDV strains circulating in ruminants (cattle, sheep, goat and yak) displayed significant antigenic differences only between BVDV-1 and BVDV-2 strains. This is the first identification of BVDV-2 in Indian cattle that may have important implications for immunization strategies and molecular epidemiology of BVD.     

A scoring system for risk assessment of the introduction and spread of bovine viral diarrhoea virus in dairy herds in Northern Italy

To support a voluntary disease control program, this study aimed to develop an integrated scoring system for the risk assessment of bovine viral diarrhoea virus (BVDV) infection in dairy herds in Northern Italy. Sixty-two dairy herds were classified according to their BVDV serological status. Farmers were interviewed using a questionnaire on potential BVDV risk factors. Scores were used to define risk levels for factors related to (1) BVDV introduction (through livestock trade, attendance of animals at shows/exhibitions and grazing common pasture), (2) within-herd spread of BVDV and (3) the results of initial serological testing. The calculated odds ratios were significant for all categories, except for livestock trade. The application of the screening test, the questionnaire and the related risk assessment showed this to be a practical approach to predicting BVDV herd status.     

Prevalence of serum antibodies to bovine herpesvirus-1 and bovine viral diarrhea virus in beef cattle in Uruguay

Our objective was to determine the prevalence of serum antibodies to bovine herpesvirus-1 (BHV-1) and bovine viral diarrhea (BVD) virus in beef cattle in Uruguay. A random sample of 230 herds selected with probability proportional to population size based on the number of cattle was chosen from a list frame of all registered livestock farms as of June 1999. Sera from up to 10 heifers, cows and bulls (up to 30 sera total per herd) were collected on selected farms between March 2000 and March 2001 and evaluated by means of enzyme-linked immunosorbent assays (ELISAs). Overall, 6358 serum samples were evaluated. We also collected data on previous diagnosis of BHV-1 or BVD infections and on the use of vaccines against these agents.

The estimated prevalence of exposure to BHV-1 and BVD at the herd level for the Uruguayan beef population was 99% and 100%, respectively. Approximately 37% of beef cattle in Uruguay have been exposed to BHV-1 and 69% to BVD virus. Only 3% of beef herds in Uruguay regularly (typically, annually) use vaccines against either of these agents.     

Challenges for bovine viral diarrhoea virus antibody detection in bulk milk by antibody enzyme-linked immunosorbent assays due to changes in milk production levels

Background

Bovine viral diarrhoea (BVD) is considered eradicated from Denmark. Currently, very few (if any) Danish cattle herds could be infected with BVD virus (BVDV). The Danish antibody blocking enzyme-linked immunosorbent assay (ELISA) has been successfully used during the Danish BVD eradication program, initiated in 1994. During the last decade, the cattle herd size has increased while the prevalence of BVDV has decreased. In this study, we investigated how these changes could affect the performance of the Danish blocking ELISA and of the SVANOVIR^®BVDV-Ab indirect ELISA. The latter has successfully been used to eradicate BVD in Sweden.Data (2003–2010) on changes in median herd size and milk production levels, occurrence of viremic animals and bulk milk surveillance were analysed. Additionally, the Danish blocking ELISA and the SVANOVIR ELISA were compared analyzing milk and serum samples. The prevalence of antibody positive milking cows that could be detected by each test was estimated, by diluting positive individual milk samples and making artificial milk pools.

Results

During the study period, the median herd size increased from 74 (2003) to 127 cows (2010), while the prevalence of BVDV infected herds decreased from 0.51 to 0.02 %. The daily milk yield contribution of a single seropositive cow to the entire daily bulk milk was reduced from 1.61 % in 2003 to 0.95 % in 2010 due to the increased herd size. It was observed that antibody levels in bulk milk decreased at national level. Moreover, we found that when testing bulk milk, the SVANOVIR^®BVDV-Ab can detect a lower prevalence of seropositive lactating cows, compared to the Danish blocking ELISA (0.78 % vs. 50 %). Values in the SVANOVIR^®BVDV-Ab better relate to low concentrations of antibody positive milk (R² = 94-98 %), than values in the blocking ELISA (R² = 23–75 %). For sera, the two ELISAs performed equally well.

Conclusions

The SVANOVIR ELISA is recommended for analysis of bulk milk samples in the current Danish situation, since infected dairy herds e.g. due to import of infected cattle can be detected shortly after BVDV introduction, when only few lactating cows have seroconverted. In sera, the two ELISAs can be used interchangeably.     

Outbreak of acute bovine viral diarrhea in Brazilian beef cattle: clinicopathological findings and molecular characterization of a wild-type BVDV strain subtype 1b

When first described in 1946, bovine viral diarrhea (BVD) was characterized as an acute transmissible disease associated with severe leucopenia, high fever, depression, diarrhea, gastrointestinal erosions, and hemorrhages. Recently the severe acute form has been related only to some hypervirulent BVDV-2 strains. This article reports the detection of BVDV-1b associated with an acute and fatal outbreak of BVD in a Brazilian beef cattle herd. Depression, anorexia, watery diarrhea, sialorrhea, and weakness were observed in six steers. One of these animals was evaluated for laboratorial, clinical, and pathological alterations. Laboratory findings were non-specific; clinically, the animal was weak, with dehydration and erosive oral lesions. Pathological alterations were predominant at the tongue, esophagus, and rumen. A RT-PCR assay using primers to partially amplify the 5′ untranslated region (5′UTR) of the BVDV genome was performed and identified BVDV in all clinical samples analyzed. Phylogenetic analysis of BVDV derived from lymph node revealed that this strain was clustered within the BVDV subtype 1b. This differentiating was only possible to be performed by molecular characterization since both clinical presentation and pathologic findings were similar to BVDV-2 infection.