How the bovine viral diarrhea virus outwits the immune system

The interaction of bovine viral diarrhea virus (BVD virus) with its host has several unique features, most notably the capacity to infect its host either transiently or persistently. The transient infection stimulates an antiviral immune reaction similar to that seen in other transient viral infections. In contrast, being associated with immunotolerance specific for the infecting BVD viral strain, the persistent infection differs fundamentally from other persistent infections like those caused by lentiviruses. Whereas the latter are characterized by complex viral evasion of the host's adaptive immune response by mechanisms such as antigenic drift and interference with presentation of T cell epitopes, BVD virus avoids the immune response altogether by inducing both humoral and cellular immune tolerance. This is made possible by invasion of the fetus at an early stage of development. In addition to adaptive immunity, BVD virus also manipulates key elements of the host's innate immune response. The non-cytopathic biotype of BVD virus, which is capable of persistently infecting its host, fails to induce type I interferon. In addition, persistently infected cells are resistant to the induction of apoptosis by double-stranded RNA and do not produce interferon when treated with this pathogen-associated molecular pattern (PAMP) that signals viral infection. Moreover, when treated with interferon, cells persistently infected with non-cytopathic BVD virus do not clear the virus. Surprisingly, however, despite this lack of effect on persistent infection, interferon readily induces an antiviral state in these cells, as shown by the protection against infection by unrelated viruses. Overall, BVD virus manipulates the host's interferon defense in a manner that optimises its chances of maintaining the persistent infection as well as decreasing the risks that heterologous viral infections may carry for the host. Thus, since not all potential host cells are infected in animals persistently infected with BVD virus, heterologous viruses replicating in cells uninfected with BVD virus will still trigger production of interferon. Interferon produced by such cells will curtail the replication of heterologous viruses only, be that in cells already infected with BVD virus, or in cells in which the heterologous virus may replicate alone. From an evolutionary viewpoint, this strategy clearly enhances the chances of transmission of BVD virus to new hosts, as it attenuates the negative effects that a global immunosuppression would have on the survival of persistently infected animals.     

Relationships between infection with caprine arthritis encephalitis virus, intramammary bacterial infection and somatic cell counts in dairy goats.

Somatic cell counts, the bacteriological condition of the milk and antibodies against caprine arthritis encephalitis virus (CAEV) were measured monthly throughout lactation in 121 lactating goats of the Murcia-Granada breed in four commercial dairy goat herds. The prevalence of bacterial intramammary infection was 5.6 per cent and the prevalence of CAEV infection was 20.6 per cent. An analysis of variance revealed a significant effect of herd, intramammary infection and the interaction between intramammary infection and CAEV on the somatic cell count. In udder halves free of intramammary infection, the somatic cell counts were significantly lower in seronegative goats than in seropositive goats (P<0.05), but the difference was not significant in udder halves persistently infected by bacteria. There was a significant increase in somatic cell counts due to bacterial intramammary infection (P<0.01) in the seronegative goats, but this effect was not present in the seropositive animals.     

Heifer breeding farms as a source for spreading the BVD virus?

It is well known that, in Switzerland, communal grazing of livestock on alpine pastures plays an important role in the spread of BVD virus. Analogously, we might expect that the communal raising on farms specialising in raising heifers of animals born on different farms would also favour the spread of BVDV. This study investigated whether a persistently infected (PI) breeding heifer kept on this type of farm over a period of 26 months would put the other animals at risk of being infected.The PI-animal was in contact with 75 heifers (here defined as contact animals) on this farm. Thirty-two of the contact animals that were probably pregnant (animals at risk of giving birth to a PI-calf) were moved to 8 different breeding farms (here defined as farms at risk). On these 8 farms, 246 calves were found to be at risk of being infected with BVDV. We examined 78 calves and investigated whether the move of the pregnant animals from their original farm had permitted the virus to spread to these 8 other farms.The contact animals had a seroprevalence of 92% and the animals at risk a seroprevalence of 100%. Only one PI-animal was found on the farms at risk.This BVD infection, however, occurred independently of the PI-breeding animal. Seropositive calves were found only on 2 farms. This study did not provide any proof for a spread of BVDV with the PI-breeding animal as a source; likewise, no persistent infection was proven to exist on the farms at risk. This result is likely to be representative for the endemic situation of BVD in Switzerland. Thus, PI-animals present on heifer raising farms infect calves well before servicing. Hence, no new PI-animals are generated, and the infection becomes self-limiting. When we reconstructed the animal movements between the farms and determined the animals to be examined with the aid of the Swiss national animal traffic database (TVD) we found the data of 37% of the heifers to be incomplete and failed to successfully establish the whereabouts of 3 animals.     

Spread of Bovine Virus Diarrhoea virus in a herd of heifer calves

Summary

A calf persistently infected and immunotolerant to Bovine Virus Diarrhoea virus (BVD virus) was, on purpose, introduced to a herd of heifer calves over 4 months of age that had been reared as recipients for embryo transplantation.

All calves were brought in contact with the persistently infected animal. In total, 240 calves were involved in this experiment, 22 of which were serologically negative when introduced. These serologically negative animals developed antibodies against BVD virus within 5 months after introduction. At short distances from the persistently infected BVD virus shedder, negative calves seroconverted within 2 months, but at greater distances the moment of seroconversion was unpredictable.

The calves that had undergone a natural infection with BVD virus received embryos after transportation to an allied farm. In total, 14 calves were born after embryo transplantation, all of which were free of BVD virus, in spite of the presence of BVD‐virus on the latter farm.     

Spread of bovine virus diarrhoea virus in a herd of heifer calves.

A calf persistently infected and immunotolerant to Bovine Virus Diarrhoea virus (BVD virus) was, on purpose, introduced to a herd of heifer calves over 4 months of age that had been reared as recipients for embryo transplantation. All calves were brought in contact with the persistently infected animal. In total, 240 calves were involved in this experiment, 22 of which were serologically negative when introduced. These serologically negative animals developed antibodies against BVD virus within 5 months after introduction. At short distances from the persistently infected BVD virus shedder, negative calves seroconverted within 2 months, but at greater distances the moment of seroconversion was unpredictable. The calves that had undergone a natural infection with BVD virus received embryos after transportation to an allied farm. In total, 14 calves were born after embryo transplantation, all of which were free of BVD virus, in spite of the presence of BVD-virus on the latter farm.     

Establishing a pilot bovine viral diarrhoea virus eradication scheme in Somerset

Beginning in April 2006, 41 farms were recruited onto a pilot Bovine viral diarrhoea virus (BVDV) eradication programme across the south of England with the majority of study herds concentrated in Somerset. Each herd was assessed and where relevant cleared of persistently infected (PI) animals. Seven farms dropped out before whole herd screening could be performed. Of the remaining 34 farms, 20 (59 per cent) were classified as infected although two of these were initially misclassified as BVDV-free. Over the course of three years, 61 PIs were identified across 16 of the 20 infected farms. 72 per cent of PIs indentified on the first herd test were below two years of age. PI prevalence ranged from 0.2 to 3.1 per cent of infected herds and was highest in herds that did not vaccinate. By the end of 2009, 24/34 (71 per cent) of study farms were BVDV-free while 10 (29 per cent) remained infected.     

Reproduction of mucosal disease with cytopathogenic bovine viral diarrhoea virus selected in vitro

Isolates of non-cytopathogenic bovine viral diarrhoea (BVD) virus from 18 persistently infected calves from one herd were compared by using monoclonal antibodies directed against the major viral glycoprotein gp53. All the isolates displayed an almost identical reaction pattern. Based on this antigenic analysis three cytopathogenic BVD and three non-cytopathogenic BVD viruses closely related to the non-cytopathogenic BVD herd isolate were selected. Six of the persistently infected calves were inoculated with a pool of the three closely related cytopathogenic BVD viruses and two with a pool of the three non-cytopathogenic BVD viruses. In addition three animals were infected with one closely related cytopathogenic BVD strain (Indiana) and two animals with the antigenetically different cytopathogenic BVD viral strain A1138/69. Regardless of the inoculation route all the animals superinfected with closely related cytopathogenic BVD viruses developed the characteristic lesions of mucosal disease within 14 days of infection. Animals which were inoculated with non-cytopathogenic BVD viruses which closely resembled the herd isolate, or with cytopathogenic BVD viruses which did not resemble the herd isolate did not develop any signs of disease. However, the latter group produced antibodies to the superinfecting virus.     

Bovine virus diarrhoea (BVD) control programme in an area in the Rome province (Italy)

A BVD control programme based on the identification and removal of persistently infected (PI) animals is being undertaken in an area in the Rome province, where BVD outbreaks had been previously detected. It involves 174 mainly dairy herds, from which blood samples of all bovines older than 1 year are obtained through the national brucellosis and leukosis eradication programme. Samples sufficient to detect the presence of seropositive animals at a prevalence of 5% or more are initially screened for antibodies against BVD virus (BVDV) using an immunoenzymatic assay. Upon identification of seroreagents additional blood samples are tested from the 6-12-month age category not included in the initial samples. Animals are considered immunotolerant if BVDV is demonstrated twice at a minimum 30-day interval. When no seropositive animals are detected during the first serological screening the herd is declared BVD-free if a second testing, preferably carried on the same animals previously tested, confirms the seronegative status of the herd. At present 147 farms have been tested, of which 63 (42.9%) are negative with respect to antibodies against BVDV. Of the 84 remaining herds in which one or more seropositives are detected, 13 are classified as recently infected. In eight of these recently infected herds, 22 PI animals have been identified.     

Disease screening profiles and colostrum management practices on 16 Irish suckler beef farms

Background

Calf output is a key element in determining the profitability of a suckler beef enterprise. Infectious agents such as Bovine Virus Diarrhoea (BVD) virus, colostrum management and parasitic challenge can all affect calf output. Prior to the national BVD eradication programme, there was little published information on either the prevalence or effect of BVD in Irish beef herds. There is little published information on colostrum management practices in Irish commercial beef herds and there have also been few studies published on the prevalence of liver fluke or rumen fluke infection in Irish beef herds. Sixteen farms participating in the Teagasc/Farmers Journal BETTER farm beef programme were used in this study. Fourteen herds were screened for the presence of BVD virus in 2010 using RT-PCR. In 13 herds, blood samples were collected from calves (2–14 days of age) in November 2011 - April 2012 to determine their passive immune status using the zinc sulphate turbidity (ZST) test, while in 12 herds, blood and faecal samples were taken in order to determine the level of exposure to gastrointestinal and hepatic helminths.

Results

The overall prevalence of BVD virus-positive cattle was 0.98% (range 0 - 3% per herd, range 0.6 - 3.0% per positive herd). Eighteen of the 82 calves (22%) sampled had ZST values less than 20 units (herd mean range 17.0 – 38.5 units) indicating a failure of passive transfer. The overall animal-level (herd-level) prevalence of liver fluke and rumen fluke infection in these herds was 40.5% (100%) and 20.8% (75%), respectively.

Conclusions

The potential costs associated with the presence of animals persistently infected with BVD virus through the increased use of antibiotics; the rate of failure of passive transfer of colostral immunoglobulins and the high prevalence of liver fluke infection in these herds highlight that some Irish suckler beef farms may not be realizing their economic potential due to a range of herd health issues. The use of farm-specific herd health plans should be further encouraged on Irish suckler beef farms.     

Viral and viral protein specificity of antibodies induced in cows persistently infected with noncytopathic bovine viral diarrhea virus after vaccination with cytopathic bovine viral diarrhea virus

Neutralizing and nonneutralizing antibodies to bovine viral diarrhea (BVD) virus were detected in 3 cows persistently infected with noncytopathic BVD virus after vaccination with modified-live cytopathic BVD virus. Neutralizing antibodies detected in serum samples from each persistently infected cow at 3 weeks after vaccination were highly specific for certain isolates of cytopathic BVD virus and reacted only with a viral protein with a molecular weight of 53,000. Neutralizing antibodies to 1 of 3 isolates of noncytopathic BVD virus were detected in a serum sample obtained at 12 weeks after vaccination from 1 of 3 persistently infected cows. Nonneutralizing antibodies were detected in all cows at 7 to 12 weeks after vaccination. The nonneutralizing antibodies were less specific for isolates of BVD virus and reacted with viral proteins with molecular weights of 115,000, 80,000, 53,000, and 47,000.     

Mixing of selenium and anthelmintic drench

Extract

Bovine viral diarrhoea (BVD) virus has a worldwide distribution and investigations in various parts of the world have shown that 60%–80% of cattle have neutralising antibodies to the virus⁽¹⁾⁽²⁾. Bovine viral diarrhoea virus infection is also common in New Zealand dairy herds⁽³⁾, and its epidemiology on dairy farms is well understood. It had been considered that the traditional beef cattle population was essentially free from this infection and there was a concern that the rapidly expanding dairy-beef industry may introduce infection into an essentially naive beef cattle population. However, a recent study has shown that BVD virus infection is widespread in beef herds throughout New Zealand⁽⁴⁾. To explore the issue further, we have examined the prevalence of BVD virus antibody- positive animals in selected dairy-beef operations and traditional cow-calf herds, and how BVD-virus infection, if present, is maintained within these cattle populations.     

Cytopathic bovine viral diarrhea viruses (BVDV): emerging pestiviruses doomed to extinction

Bovine viral diarrhea virus (BVDV), a Flaviviridae pestivirus, is arguably one of the most widespread cattle pathogens worldwide. Each of its two genotypes has two biotypes, non-cytopathic (ncp) and cytopathic (cp). Only the ncp biotype of BVDV may establish persistent infection in the fetus when infecting a dam early in gestation, a time point which predates maturity of the adaptive immune system. Such fetuses may develop and be born healthy but remain infected for life. Due to this early initiation of fetal infection and to the expression of interferon antagonistic proteins, persistently infected (PI) animals remain immunotolerant to the infecting viral strain. Although only accounting for some 1% of all animals in regions where BVDV is endemic, PI animals ensure the viral persistence in the host population. These animals may, however, develop the fatal mucosal disease, which is characterized by widespread lesions in the gastrointestinal tract. Cp BVD virus, in addition to the persisting ncp biotype, can be isolated from such animals. The cp viruses are characterized by unrestrained genome replication, and their emergence from the persisting ncp ones is due to mutations that are unique in each virus analyzed. They include recombinations with host cell mRNA, gene translocations and duplications, and point mutations. Cytopathic BVD viruses fail to establish chains of infection and are unable to cause persistent infection. Hence, these viruses illustrate a case of “viral emergence to extinction” – irrelevant for BVDV evolution, but fatal for the PI host.     

Prevalence of ruminant pestivirus infections in Namibia.

Following several clinical cases of suspected bovine virus diarrhoea (BVD) on three Namibian cattle farms, a serological survey was conducted on bovine, ovine, caprine and wild ruminant sera originating from different regions of the country. Neutralizing antibodies to BVD virus (BVDV) were detected in 58% of 1,014 cattle sera, 14% of 618 sheep sera and 4.6% of 1,118 goat sera. Sera from seven of ten wildlife species were positive with kudu, eland and giraffe having prevalence rates greater than 40%. BVDV was isolated from six clinically affected bovines and three healthy heifers persistently infected with BVDV. The survey demonstrated that pestivirus infections are widespread in Namibia in both domestic and wild ruminants.     

Lymphocyte blastogenesis and neutrophil function in cattle persistently infected with bovine viral diarrhea virus

Neutrophil function and mononuclear cell proliferative responses to mitogens were determined in healthy cattle and in cattle persistently infected with bovine viral diarrhea (BVD) virus. Uptake of [3H]thymidine by resting and mitogen-stimulated peripheral blood mononuclear cells was significantly lower in cattle persistently infected with BVD virus than in healthy cattle. Neutrophils from cattle persistently infected with BVD virus had significantly impaired capability to ingest Staphylococcus aureus, but were normal in respect to random migration under agarose, cytochrome C reduction, iodination, and antibody-dependent cell-mediated cytotoxicity. Impairment of neutrophil function in cattle persistently infected with BVD virus differs from impairment of neutrophil function reported in healthy cattle mounting an immune response to recent BVD virus infection.     

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.     

Frequency of persistent bovine viral diarrhea virus infection in selected cattle herds

Sera and blood buffy coat samples were obtained from 3,157 cattle in 66 selected herds. Antibodies to bovine viral diarrhea (BVD) virus were detected in 89% of the serum samples by immunoprecipitation or virus-neutralization tests. Cytopathic or noncytopathic BVD viruses were isolated from blood buffy coat samples from 60 cattle in 6 herds. A second blood buffy coat sample was obtained from 54 of the 60 cattle 2 months after the initial sampling, and BVD virus was isolated again from each cow. The 54 cattle were considered persistently infected with BVD virus. The frequency of persistent infection was 1.7%.     

A survey for BVDV antibodies in cattle farms in Slovakia and genetic typing of BVDV isolates from imported animals

A serological survey for bovine viral diarrhoea virus (BVDV) antibodies on a collection of 1295 serum samples obtained from 6-12 months old cattle originating from 45 farms in Slovakia was carried out. On 13 farms more than 90% of the examined animals were seropositive, on 14 farms 71-90% seroprevalence was observed, on 13 farms only 50-70% animals were found to be positive for BVDV antibodies, while the remaining 5 farms showed fewer than 50% seropositive animals. The average incidence of BVDV antibodies (around 70%) was similar as determined 30 years ago. Of 84 serum samples from seronegative animals originating from 14 farms in which 70-98% seropositivity was observed, six were positive in Ag-BVDV ELISA indicating persistently infected (PI) cattle. On a farm to which animals were imported from abroad, a BVD outbreak was observed. Of 110 animals tested, four were positive in Ag-ELISA indicating the presence of PI cattle on this farm. Genetic typing of two isolates from imported animals performed by RT-PCR (324/326 primers from 5'-UTR), sequencing of PCR products and computer-assisted phylogenetic analysis revealed that they belong to BVDV-1 h group.     

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.