Effect of the bovine viral diarrhoea virus (BVDV) infection on dairy calf rearing

The aim of this study was to compare the cumulative incidence of mortality, clinical diarrhoea and respiratory disease in calves, during their first six months of age, in herds with different bovine viral diarrhoea virus (BVDV) infection status. Calves’ health indicators were tested by comparing proportions in 101 farms with dissimilar infection condition. The results indicate that there was a significant relationship between the BVDV status (actively infected herd or not) and the cumulative incidence of mortality and respiratory disorders.     

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.     

Antibody responses of naive cattle to two inactivated bovine viral diarrhoea virus vaccines,measured by indirect and blocking ELISAS and virus neutralisation

Two groups of naive heifers were given primary courses of two inactivated bovine viral diarrhoea (BVD) virus vaccines licensed for use in the UK. Their humoral responses in serum and milk were assayed by means of an indirect ELISA detecting antibodies to structural viral glycoproteins, a blocking ELISA specific for antibodies to the non-structural protein NS2-3 and the virus neutralisation test (VNT). For each assay, the numbers of serum or milk samples testing positive at each sample point and the mean values were determined. In both vaccine groups, serum antibody responses were detected by the indirect ELISA and the VNT, with both the numbers of seropositive animals and mean values peaking five weeks after the second vaccination. In the 23 heifers vaccinated with Bovilis BVD, the mean NS2-3-specific ELISA values remained low throughout the trial, with no serum or milk samples testing positive. In the 24 heifers vaccinated with Bovidec, the mean NS2-3 responses peaked below the level of positivity five weeks after the second vaccination, before declining again; NS2-3-specific antibodies were detected in one serum sample and one milk sample from two heifers in this group. A pooled milk sample from each vaccine group tested negative by both ELISAS 12 weeks after the second vaccination.     

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.     

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.     

Apoptosis inhibitors delay the cytopathic effect of bovine viral diarrhoea virus (BVDV)

Based on their action in cell culture, two biotypes of bovine viral diarrhoea virus (BVDV) can be distinguished. The noncytopathic (ncp) BVDV isolated from persistently infected animals cause no visible damage to cultured bovine cells. In contrast, cytopathic (cp) BVDV induces severe damage and apoptosis in cell cultures. Cp BVDV can be isolated from cattle suffering from mucosal disease (MD) and is associated with the severe lesions that primarily affect the gastrointestinal tract. To get an insight into the molecular events during BVDV induced cytopathic effect (CPE), the effect of three chemical reagents (3-aminobenzamide, ascorbic acid and N-acetyl-leucyl-leucyl-methional) with completely different mode of actions in infected cells was analysed. All three substances were able to delay the cytopathic effect induced in permissive bovine cells.     

Epitopic characterisation of Turkish bovine viral diarrhoea virus (BVDV) isolates using monoclonal antibodies

In this study, 15 bovine viral diarrhoea viruses (BVDV) isolated from the field in Turkey were characterised for their biotype, cloned and eventually analysed for their epitopic composition in terms of glycoprotein E2. Immunoplaque assay, plaque assay, limiting dilution and streptavidin-biotin-peroxidase techniques were used for biotype characterisation, cloning of cytopathic (cp) and noncytopathic (ncp) biotypes and epitope analysis, respectively. While 14 out of 15 BVDV isolates were distinguished as ncp biotype, 1 isolate was found to be containing both biotypes (cp + ncp). According to the reactivity patterns of isolates with 15 monoclonal antibodies, 4 different antigenic groups could be formed. There were no antigenic differences between the isolates derived from the same animal with various time intervals. On the other hand, biotype clones isolated from the same animal exhibited difference in one epitope. This is the first study describing antigenic characterisation of BVDV field isolates in Turkey.     

Principles for eradication of bovine viral diarrhoea virus (BVDV) infections in cattle populations

Systematic eradication of BVDV without vaccination started in Scandinavia in 1993. In principle, the schemes include; (1) identification of non-infected and infected herds using different combinations of serological herd tests such as bulk milk tests and spot tests (sample of animals in a certain age), (2) monitoring/certification of non-infected herds by repeated sampling, applying one of the above-mentioned methods and (3) virus clearance in infected herds aimed at removing persistently infected (PI) animals in a cost- and time-efficient manner. In the virus clearance protocol described, an initial test is performed on all animals with subsequent follow-up of calves born as well as of dams seronegative in the initial test. It is generally recommended to perform an initial antibody test on all samples. This should be done not only to screen for seronegative animals on which virus isolation should be attempted (i.e. possible PI animals), but more in order to identify non-immune animals in reproductive age, that is, the key animals in herd-level persistence of infection. In Sweden, a common finding has been self-clearance, where the infection ceases without any other intervention than controlled introduction of new animals. Other epidemiological observations concern the course of events following virus introduction. Important risk factors for spreading BVDV are discussed, where livestock trade is perceived as the most central to control. Live vaccines, imported semen and embryos constitute special hazards, since they may act as vehicles for the introduction of new BVDV strains. The importance of making farmers aware of herd biosecurity and their own responsibility for it is stressed, and in order to maintain a favourable situation after a scheme has been concluded, effort must be put into establishing such a persisting attitude in the farming community.     

Bovine herpesvirus type 1 (BHV-1) and bovine viral diarrhoea virus (BVDV) infections in dairy herds: self clearance and the detection of seroconversions against a new atypical pestivirus

The epidemiology of bovine herpesvirus type 1 (BHV-1) and bovine viral diarrhoea virus (BVDV) was studied in a population of small dairy herds that had not been vaccinated. Bulk tank milk samples of 186 herds in Thailand were collected four times between 2002 and 2004. Serum samples from individual animals in 11 herds were also taken on three occasions. The prevalence of BHV-1 in the 186 herds was 61% in 2002, decreasing to 48% in 2004 and for BVDV was 91% in 2002, decreasing to 72% in 2004. A BVDV antigen-positive calf was found in one of the 11 herds, and animals in this herd and three other herds seroconverted to a recently described atypical BVDV strain (HoBi). This study showed a significantly decreasing prevalence for both BHV-1 and BVDV due to a self-clearance process. Further studies are needed to find out how the atypical BVDV strain entered the cattle population.     

Natural changes in the spread of bovine viral diarrhoea virus (BVDV) among Estonian cattle

The results of a survey conducted during 1993-2000 to study the spread of bovine viral diarrhoeal virus (BVDV) among Estonian cattle are presented. The BVDV infection status of a representative random sample of cattle herds housing 20 or more dairy cows was established to estimate the prevalence of herds with active BVDV infection [potentially having persistently infected (PI) cattle--suspect PI herds]. The herds investigated comprised approximately 70% of all Estonian dairy cows. The BVDV infection status was established in 315-350 herds (making the sampling fraction about 20%) during three sampling periods: 1993-95, 1997-98, 1999-2000. BVDV antibodies were detected in herd bulk milk samples and/or sera from young stock by a liquid-phase-blocking enzyme-linked immunosorbent assay developed in the Danish Veterinary Institute for Virus Research. The results of the survey demonstrate the reduction in the prevalence of herds with active BVDV infection in the studied fraction of the Estonian cattle population. During the first sampling period (1993-95) a prevalence of 46% (+/- 5%) for suspect PI herds was observed, during the second sampling period this prevalence was 16% (+/- 3%) and in the third period it was 18% (+/- 3%). As there is no control programme for BVDV in Estonia, the observed changes reflect the natural course of the infection in the study population. A possible cause for these changes is the decreased trade in breeding animals as a result of the economic difficulties present in cattle farming during the study period. The farming practices (most large herds are managed as closed herds) and the low density of cattle farms have obviously facilitated the self-clearance of herds from the BVDV infection, diminishing the new introduction of infection into the herds.     

Survivorship of animals persistently infected with bovine virus diarrhoea virus (BVDV)

The survivorship of animals persistently infected with bovine virus diarrhoea virus was studied in ten Danish dairy herds. The ages of 34 persistently infected animals were compared with the ages of non-persistently infected animals in the herds in a cross-sectional study in which the risk rate of removal of animals in the two groups within 1 year was estimated from an exponential probability function of the age. In a cohort study, the 34 persistently infected animals were followed from the date of the initial blood test and onwards and the risk rates of dying or being slaughtered due to unthriftiness or either of these events were calculated.

The attributable risk of leaving the herds within 1 year was 0.35 among persistently infected animals. The risk rate of dying or being slaughtered due to unthriftiness within 1 year was 0.28 and 0.31, respectively; the risk rate of either of these events was 0.5.     

Fatal congenital anaplasmosis associated with bovine viral diarrhoea virus (BVDV) infection in a crossbred calf

Clinical disease resulting from the vertical transmission of Anaplasma marginale has only been reported on 5 occasions despite studies demonstrating successful in utero transmission. During the reported experimental induction of congenital anaplasmosis in calves, the outcome was variable but mostly led to inapparent or mild infection. There are previous case reports of fatal congenital anaplasmosis following natural infection. The clinical findings in a 2-day-old calf presented to the Onderstepoort Veterinary Academic Hospital with clinical signs of congenital anaplasmosis, which was unresponsive to treatment, are described. Subsequent post mortem diagnostic tests revealed that this calf was co-infected with bovine viral diarrhoea virus (BVDV). It is postulated that immunosuppression resulting from BVDV infection predisposed to severe, fatal anaplasmosis in this calf.     

Epidemiological features and economical importance of bovine virus diarrhoea virus (BVDV) infections

Infections with bovine virus diarrhoea virus (BVDV) are widespread throughout the world. Although the prevalence of infection varies among surveys, the infection tends to be endemic in many populations, reaching a maximum level of 1-2% of the cattle being persistently infected (PI) and 60-85% of the cattle being antibody positive. Persistently infected cattle are the main source for transmission of the virus. However, acutely infected cattle as well as other ruminants, either acutely or persistently infected, may transmit the virus. Transmission is most efficient by direct contact. However, as infections have been observed in closed, non-pasturing herds, other transmission routes seem likely to have some practical importance. Differences in BVDV prevalence among regions or introduction of virus in herds previously free of BVDV are often associated with particular epidemiological determinants such as cattle population density, animal trade and pasturing practices. However, on a few occasions there have been no obvious explanations for infection of individual herds. Estimates of economic losses due to BVDV infection vary depending on the immune status of the population and the pathogenicity of the infecting virus strains. Introduction of the infection into a totally susceptible population invariably causes extensive losses until a state of equilibrium is reached. Infection with highly virulent BVDV strains causing severe clinical signs and death after acute infection gives rise to substantial economical losses. At an estimated annual incidence of acute infections of 34%, the total annual losses were estimated as US$ 20 million per million calvings when modeling the losses due to a low-virulent BVDV strain. At the same incidence of infection, the losses due to a high-virulent BVDV strain were estimated as US$ 57 million per million calvings. Low-virulent BVDV infections caused maximum losses at an incidence of 45%, whereas high-virulent BVDV infections caused maximum losses at an incidence of 65%. Thus, cost-benefit analyses of control programs are highly dependent on the risks of new infections under different circumstances and on the strains of the virus involved.     

Immune responses to non-structural protein 3 (NS3) of bovine viral diarrhoea virus (BVDV) in NS3 DNA vaccinated and naturally infected cattle

Immune responses to non-structural protein 3 (NS3) of bovine viral diarrhoea virus (BVDV) were investigated. cDNA encoding NS3 from type 1a BVDV was used to vaccinate five calves, another five calves remained unvaccinated. Three weeks after final vaccination animals were challenged intranasally with heterologous type 1a BVDV. Anti-NS3 antibodies were detected in only one animal post-vaccination. Partial protection from virus challenge was observed in the vaccinates. Virus was not isolated from nasal mucosa of two vaccinates, and virus clearance from nasal mucosa was faster in the vaccinates compared to the controls. While elevated rectal temperatures were evident in both groups 7 days post-challenge, the mean increase in the controls was twice that observed in the vaccinates. In conclusion, NS3 DNA vaccination induced humoral immunity in one calf, and prevented fever and virus establishment in the nasal mucosa in 2/5 calves, demonstrating the efficacy of NS3 vaccination, which may benefit future development of pestivirus and flavivirus vaccines.     

Bovine viral diarrhoea virus (BVDV) diversity and vaccination. A review

BVDV is associated with a range of economically important clinical diseases including reproductive disorders and acute fatal haemorrhagic disease in cattle industry. Vaccination is still the most important control strategy for controlling BVDV infections in many countries of the world. The existence of great genetic and antigenic diversity of BVDV isolates is very important concern for BVDV vaccine development and protective efficacy of current vaccines. In this review, the protective efficacies of the selected examples of BVDV vaccines with regard to BVDV diversity and the novel marker vaccine development studies are discussed.     

An enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to bovine viral diarrhoea virus (BVDV) in cattle sera

A microtitre ELISA has been established for the quantitation of antibodies to bovine viral diarrhoea virus (BVDV). Single dilutions of sera were assayed and units of antibody were calculated from a standard curve. In order to detect the maximum number of responding animals both IgG1 and IgG2 antibody should be assayed, although detection of IgG1 alone was nearly as effective. The ELISA was as sensitive as the virus neutralization test for detection of antibody; comparison of an ELISA that detected IgG1 plus IgG2 antibody to BVDV with the virus neutralization test gave a correlation coefficient (r) of 0.89 (P less than 0.001 for 95 compared sera). Although similar amounts of IgG1 and IgG2 antibodies were present in sera from both experimentally- and naturally-infected cattle, antibody to BVDV in colostrum and in the sera from young calves was predominantly IgG1. The number of adult cows with antibody was 40 out of 41 while 36 of 44 calves reared in a beef unit were found to have produced antibody by the time they were 31.5 weeks old, an indication of the high prevalence of BVDV in the cattle population.