Immune response to bovine viral diarrhea virus (BVDV) vaccines detecting antibodies to BVDV subtypes 1a, 1b, 2a, and 2c.

Bovine viral diarrhea virus (BVDV) represents a major cattle disease with multiple forms including fetal infections resulting in persistently infected (PI) cattle. The objectives of this study were to investigate the immune response to six vaccines, five modified live viral (MLV) and one killed vaccine containing BVDV immunogens as measured by antibodies to BVDV1a, BVDV1b, BVDV2a, and BVDV2c. The predominant BVDV subgenotype in the U.S. is BVDV1b compared to BVDV1a and BVDV2a. There are MLV and killed BVDV vaccines containing BVDV1a and BVDV2a marketed in the U.S. A prior study evaluated immune response to vaccination with BVDV1a and BVDV2a inducing virus neutralizing antibody titers. BVDV1b titers 128 or higher at time of exposure to BVDV1b PI cattle protected heifers against fetal infection. Calves received two doses and postweaning serums were collected and assayed for BVDV antibodies. Antibody titers were expressed as geometric mean averages. Percentages were expressed as proportions of animals within three antibody levels, including targeted level 128 or greater. There were statistical differences among vaccines in each study, particularly to BVDV1a, BVDV1b, and BVDV2a. MLV vaccines containing Singer strain induced higher levels to BVDV1a and BVDV1b than NADL vaccine in all three studies. Two vaccines, both MLV, Vaccine 1 and Vaccine 6 containing Singer strain induced higher proportion of 128 or higher BVDV1b titers than vaccine with NADL. Antibody levels to BVDV2a and BVDV2c were dependent on BVDV2a vaccine strain. This study indicates strain in BVDV vaccines reflects differences in immune response to different BVDV subgenotypes, particularly BVDV1b and BVDV2c.

Bovine viral diarrhea virus 1b fetal infection with extensive hemorrhage.

Bovine viral diarrhea virus (BVDV) 1b was isolated from tissues of a term bovine fetus with petechial hemorrhages noted throughout the body and placenta at autopsy. Fresh lung, kidney, thymus, and liver tissues were examined by direct fluorescent antibody testing and were positive for BVDV antigen and negative for bovine herpesvirus 1 antigen. An organ pool of fresh tissues was positive for noncytopathic (NCP) BVDV-1 by virus isolation. BVDV-1b was identified by sequencing of the 5'-UTR region of the genome. Fixed brain, placenta, thymus, lymph node, lung, kidney, skeletal muscle, liver, and bone marrow were positive for BVDV antigen by immunohistochemistry. Although BVDV hemorrhage and/or thrombocytopenia has been associated historically with NCP strains of BVDV-2, this case adds to more recent reports of BVDV-1 infections and hemorrhage in cattle. This BVDV-1b isolate should be investigated for its potential to cause hemorrhage in postnatal cattle.

Challenge with Bovine viral diarrhea virus by exposure to persistently infected calves: protection by vaccination and negative results of antigen testing in nonvaccinated acutely infected calves.

Calves persistently infected (PI) with Bovine viral diarrhea virus (BVDV) represent an important source of infection for susceptible cattle. We evaluated vaccine efficacy using calves PI with noncytopathic BVDV2a for the challenge and compared tests to detect BVDV in acutely or transiently infected calves versus PI calves. Vaccination with 2 doses of modified live virus vaccine containing BVDV1a and BVDV2a protected the calves exposed to the PI calves: neither viremia nor nasal shedding occurred. An immunohistochemistry test on formalin-fixed ear notches and an antigen-capture enzyme-linked immunosorbent assay on fresh notches in phosphate-buffered saline did not detect BVDV antigen in any of the acutely or transiently infected calves, whereas both tests had positive results in all the PI calves.

Evaluation of diagnostic tests used for detection of bovine viral diarrhea virus and prevalence of subtypes 1a, 1b, and 2a in persistently infected cattle entering a feedlot.

OBJECTIVE: To evaluate diagnostic tests used for detection of bovine viral diarrhea virus (BVDV) and determine the prevalence of BVDV subtypes 1a, 1b, and 2a in persistently infected (PI) cattle entering a feedlot. DESIGN: Prospective study. ANIMALS: 21,743 calves. PROCEDURES: Samples were obtained from calves initially testing positive via antigen capture ELISA (ACE) performed on fresh skin (ear notch) specimens, and ACE was repeated. Additionally, immunohistochemistry (IHC) was performed on skin specimens fixed in neutral-buffered 10% formalin, and reverse transcriptase PCR (RT-PCR) assay and virus isolation were performed on serum samples. Virus was subtyped via sequencing of the 5' untranslated region of the viral genome. RESULTS: Initial ACE results were positive for BVDV in 88 calves. After subsequent testing, results of ACE, IHC, RT-PCR assay, and viral isolation were positive in 86 of 88 calves; results of all subsequent tests were negative in 2 calves. Those 2 calves had false-positive test results. On the basis of IHC results, 86 of 21,743 calves were PI with BVDV, resulting in a prevalence of 0.4%. Distribution of BVDV subtypes was BVDV1b (77.9%), BVDV1a (11.6%), and BVDV2a (10.5%). CONCLUSIONS AND CLINICAL RELEVANCE: Rapid tests such as ACE permit identification and segregation of PI cattle pending results of further tests, thus reducing their contact with the rest of the feedlot population. Although vaccines with BVDV1a and 2a components are given to cattle entering feedlots, these vaccines may not provide adequate protection against BVDV1b.

Bovine viral diarrhoea virus (BVDV) subgenotypes in diagnostic laboratory accessions: distribution of BVDV1a, 1b, and 2a subgenotypes.

The prevalence of bovine viral diarrhoea virus (BVDV) biotypes and subgenotypes was determined from 131 BVDV positive samples from a diagnostic laboratory. The majority of the isolates were from Oklahoma; however, other states including Kansas, Texas, and Arkansas were represented. These BVDV samples were from submissions of 76 live animals and 55 necropsy samples. There were 131 BVDV samples represented by 117 noncytopathic (NCP), 11 cytopathic (CP) and 3 cases with mixed NCP and CP biotypes. The NCP isolates were more common (P < 0.05) than the CP and NCP/CP combination. The BVDV samples were segregated into three subgenotypes by differential PCR and sequencing of a viral genomic region, 5'-untranslated region (5'-UTR). There were more BVDV1b subgenotypes 60/131 (45.8%) than BVDV1a, 37/131 (28.2%) or BVDV2a, 34/131 (26.0%) (P < 0.05). The organ system involvement included the major categories such as respiratory, digestive, mixed/multiple organs, abortions, and persistent infections (PI). All three BVDV subgenotypes were found in persistently infected (PI) cattle and respiratory diseases, both major requests for BVDV diagnosis. Only one of the 131 viruses was genetically similar to the strains present in U.S. vaccines.

Enteric disease in postweaned beef calves associated with Bovine coronavirus clade 2.

Bovine coronavirus (BoCV; Betacoronavirus 1) infections are associated with varied clinical presentations including neonatal diarrhea, winter dysentery in dairy cattle, and respiratory disease in various ages of cattle. The current report presents information on BoCV infections associated with enteric disease of postweaned beef cattle in Oklahoma. In 3 separate accessions from a single herd, 1 in 2012 and 2 in 2013, calves were observed with bloody diarrhea. One calf in 2012 died and was necropsied, and 2 calves from this herd died in 2013 and were necropsied. A third calf from another herd died and was necropsied. The gross and histologic diagnosis was acute, hemorrhagic colitis in all 4 cattle. Colonic tissues from all 4 animals were positive by fluorescent antibody testing and/or immunohistochemical staining for BoCV antigen. Bovine coronavirus was isolated in human rectal tumor cells from swabs of colon surfaces of all animals. The genomic information from a region of the S envelope region revealed BoCV clade 2. Detection of BoCV clade 2 in beef cattle in Oklahoma is consistent with recovery of BoCV clade 2 from the respiratory tract of postweaned beef calves that had respiratory disease signs or were healthy. Further investigations on the ecology of BoCV in cattle are important, as BoCV may be an emerging disease beyond the initial descriptions. Challenge studies are needed to determine pathogenicity of these strains, and to determine if current BoCV vaccines are efficacious against the BoCV clade 2 strains.

Evaluation of health status of calves and the impact on feedlot performance: assessment of a retained ownership program for postweaning calves.

The objective of this study was to evaluate animal health status at entry to a feedlot against feedlot performance and carcass value. There were 24 herds represented by 417 calves in a retained ownership program. The health status at entry was represented by the levels of serum antibody to infectious bovine rhinotracheitis virus (IBRV), bovine viral diarrhea viruses 1 and 2 (BVDV1a, BVDV2), parainfluenza 3 virus (PI3V), bovine respiratory syncytial virus (BRSV), Mannheimia haemolytica, and Pasteurella multocida, as well as by the presence of virus in nasal swabs and blood leukocytes and the presence of bacteria in nasal swabs. The presence or absence of viruses or bacteria at entry did not predict subsequent illness. However, there were predictors of illness severity (number of treatments) and performance parameters of feedlot performance. Herds with a low morbidity rate had higher levels of BVDV1a antibodies than herds with a high morbidity rate. On both an individual-animal and a herd-average basis, calves with low levels of antibody to BVDV1a and BVDV2 had increased total treatment costs. Also, for individual animals and the herd as a whole, low levels of antibody to P. multocida, BVDV1a, and BVDV2 were related to decreased net value to owner (carcass value minus total feedlot cost). Calves treated twice or more had lower levels of antibody to BVDV1a than those treated once or not at all. Differences in herd morbidity rate and treatment costs were more related to appropriate timing of vaccine (last dose at or near delivery of calf) or lack of a 2nd dose of killed vaccine. This was best illustrated by the levels of antibody to BVDV1a. The results of this study were used to formulate recommendations for the subsequent year.

Bovine viral diarrhea virus (BVDV) 1b: predominant BVDV subtype in calves with respiratory disease.

The prevalence of bovine viral diarrhea virus (BVDV) infections was determined in 2 groups of stocker calves with acute respiratory disease. Both studies used calves assembled after purchase from auction markets by an order buyer and transported to feedyards, where they were held for approximately 30 d. In 1 study, the calves were mixed with fresh ranch calves from a single ranch. During the studies, at day 0 and at weekly intervals, blood was collected for viral antibody testing and virus isolation from peripheral blood leukocytes (PBLs), and nasal swabs were taken for virus isolation. Samples from sick calves were also collected. Serum was tested for antibodies to bovine herpesvirus-1 (BHV-1), BVDV1a, 1b, and 2, parainfluenza 3 virus (PI3V), and bovine respiratory syncytial virus (BRSV). The lungs from the calves that died during the studies were examined histopathologically, and viral and bacterial isolation was performed on lung homogenates. BVDV was isolated from calves in both studies; the predominant biotype was noncytopathic (NCP). Differential polymerase chain reaction (PCR) and nucleic acid sequencing showed the predominant subtype to be BVDV1b in both studies. In 1999, NCP BVDV1b was detected in numerous samples over time from 1 persistently infected calf; the calf did not seroconvert to BVDV1a or BVDV2. In both studies, BVDV was isolated from the serum, PBLs, and nasal swabs of the calves, and in the 1999 study, it was isolated from lung tissue at necropsy. BVDV was demonstrated serologically and by virus isolation to be a contributing factor in respiratory disease. It was isolated more frequently from sick calves than healthy calves, by both pen and total number of calves. BVDV1a and BVDV2 seroconversions were related to sickness in selected pens and total number of calves. In the 1999 study, BVDV-infected calves were treated longer than noninfected calves (5.643 vs 4.639 d; P = 0.0902). There was a limited number of BVDV1a isolates and, with BVDV1b used in the virus neutralization test for antibodies in seroconverting calves' serum, BVDV1b titers were higher than BVDV1a titers. This study indicates that BVDV1 strains are involved in acute respiratory disease of calves with pneumonic Mannheimia haemolytica and Pasteurella multocida disease. The BVDV2 antibodies may be due to cross-reactions, as typing of the BVDV strains revealed BVDV1b or la but not BVDV2. The BVDV1b subtype has considerable implications, as, with 1 exception, all vaccines licensed in the United States contain BVDV1a, a strain with different antigenic properties. BVDV1b potentially could infect BVDV1a-vaccinated calves.

Bovine coronavirus (BCV) infections in transported commingled beef cattle and sole-source ranch calves.

This study investigated bovine coronavirus (BCV) in both beef calves direct from the ranch and commingled, mixed-source calves obtained from an auction market. The level of BCV-neutralizing antibodies found in the calves varied among ranches in 2 different studies in a retained-ownership program (ROP), from the ranch to the feedlot. Calves with low levels of BCV-neutralizing antibodies (16 or less) were more likely to be treated for bovine respiratory disease (BRD) than those with higher titers. In 3 studies of commingled, mixed-source calves, BCV was recovered from calves at entry to the feedlot and the infections were cleared by day 8. The BCV was identified in lung samples [bronchoalveolar lavage (BAL) collection] as well as in nasal swabs. Calves with low levels of BCV-neutralizing antibodies at entry were most likely to be shedding BCV. Bovine coronavirus was isolated from both healthy and sick calves, but not from sick calves after 4 d arrival at the feedlot. Bovine coronavirus (BCV) should be considered along with other bovine respiratory viruses in the diagnosis of etiologies in bovine respiratory disease, especially for animals that become sick shortly after arrival. If approved vaccines are developed, it would be best to carry out vaccination programs before calves are weaned, giving them sufficient time to gain active immunity before commingling with other cattle.

Multiple diagnostic tests to identify cattle with Bovine viral diarrhea virus and duration of positive test results in persistently infected cattle.

Several tests for Bovine viral diarrhea virus (BVDV) were applied to samples collected monthly from December 20, 2005, through November 27, 2006 (day 0 to day 342) from 12 persistently infected (PI) cattle with BVDV subtypes found in US cattle: BVDV-1a, BVDV-1b, and BVDV-2a. The samples included clotted blood for serum, nasal swabs, and fresh and formalin-fixed ear notches. The tests were as follows: titration of infectious virus in serum and nasal swabs; antigen-capture (AC) enzyme-linked immunosorbent assay (ELISA), or ACE, on serum, nasal swabs, and fresh ear notches; gel-based polymerase chain reaction (PCR) testing of serum, nasal swabs, and fresh ear notches; immunohistochemical (IHC) testing of formalin-fixed ear notches; and serologic testing for BVDV antibodies in serum. Of the 12 animals starting the study, 3 died with mucosal disease. The ACE and IHC tests on ear notches had positive results throughout the study, as did the ACE and PCR tests on serum. There was detectable virus in nasal swabs from all the cattle throughout the study except for a few samples that were toxic to cell cultures. The serum had a virus titer > or = log(10) 1.60 in all samples from all the cattle except for 3 collections from 1 animal. Although there were several equivocal results, the PCR test most often had positive results. The BVDV antibodies were due to vaccination or exposure to heterologous strains and did not appear to interfere with any BVDV test. These findings illustrate that PI cattle may be identified by several tests, but differentiation of PI cattle from cattle with acute BVDV infection requires additional testing, especially of blood samples and nasal swabs positive on initial testing. Also, calves PI with BVDV are continual shedders of infectious virus, as shown by the infectivity of nasal swabs over the 11-mo study.

Prevalence of bovine viral diarrhea virus (BVDV) in persistently infected cattle and BVDV subtypes in affected cattle in beef herds in south central United States.

The prevalence of bovine viral diarrhea virus (BVDV) in persistently infected (PI) cattle in beef breeding herds was determined using 30 herds with 4530 calves. The samples were collected by ear notches and tested for BVDV antigens using immunohistochemistry (IHC) and antigen capture enzyme-linked immunosorbent assay (ACE). Animals with initial positives on both IHC and ACE were sampled again using both tests and serums were collected for viral propagation and sequencing of a viral genomic region, 5'-untranslated region (5'-UTR) for viral subtyping. Samples were also collected from the dams of PI calves. There were 25 PI calves from 4530 samples (0.55%) and these PI calves were from 5 of the 30 herds (16.7%). Two herds had multiple PI calves and 3 herds had only 1 PI calf. Only 1 of the 25 dams with a PI calf was also PI (4.0%). The subtype of all the PI isolates was BVDV1b. Histories of the ranches indicated 23 out of 30 had herd additions of untested breeding females. Twenty-four of the 30 herds had adult cowherd vaccinations against BVDV, primarily using killed BVDV vaccines at pregnancy examination.

Bovine viral diarrhoea virus antigenic diversity: impact on disease and vaccination programmes.

Bovine viral diarrhoea virus (BVDV) infections in cattle are associated with a variety or "diverse" clinical forms. These include digestive tract, respiratory, foetal (varied, dependent on foetal age), haemorrhagic and systemic diseases such as mucosal disease, and immunosuppression and inapparent infections. The BVDV isolates themselves are "diverse" with genotype differences based on nucleotide sequences, antigenic variability and biotypes (presence or absence of cytopathology in cell culture). Two predominant genotypes are present in the US, BVDV1 and BVDV2. There are subtypes of BVDV1, namely BVDV1a and BVDV1b. Examination of BVDV isolates from cattle derived from diagnostic laboratory submissions indicates that BVDV1b subtype isolates were as prevalent if not more prevalent than BVDV1a isolates. There was an almost equal distribution of BVDV1b and BVDV1a isolates from cattle with history of respiratory disease, and more isolates, 6 versus 2, of BVDV1b than for BVDV1a in necropsy cases of pneumonia. There were significant antibody titre differences in sera from calves receiving modified live virus vaccines containing BVDV1a, with the BVDV1b antibody titres being significantly lower. A survey of the US licensed and marketed BVDV vaccines indicates that only one vaccine contains BVDV1b with the others containing BVDV1a or undesignated BVDV1.

Maternally derived humoral immunity to bovine viral diarrhea virus (BVDV) 1a, BVDV1b, BVDV2, bovine herpesvirus-1, parainfluenza-3 virus bovine respiratory syncytial virus, Mannheimia haemolytica and Pasteurella multocida in beef calves, antibody decline by half-life studies and effect on response to vaccination.

The passive immunity transferred to calves from their dams was investigated in a beef herd to determine half-life of antibody, estimated time to seronegative status and effect on immunization. One hundred two beef calves in a commercial ranch under standard management conditions were utilized. Samples were collected at branding (day 0). This was the first possible date to collect samples postcalving. This was approximately 2 months postcalving, and days 95 and 116. The calves were divided into two groups: vaccinates (51) and nonvaccinates (51). The calves were vaccinated with a commercial inactivated viral vaccine containing bovine viral diarrhea virus (BVDV)1a, BVDV2, bovine herpesvirus-1 (BHV-1), parainfluenza-3 virus (PI-3V), and bovine respiratory syncytial virus (BRSV) on days 0 and 95. Half of the vaccinated and unvaccinated calves also received one dose of an experimental Mannheimia haemolytica and Pasteurella multocida vaccine at day 95. Serums were tested for neutralizing antibody titers to BVDV1a, BVDV1b, BVDV2, BHV-1, PI-3V, and BRSV. Antibodies were detected by ELISA to M. haemolytica whole cell, M. haemolytica leukotoxin, and P. multocida outer membrane protein (OMP). The mean half-life of viral antibodies in nonvaccinated calves to each virus was: BVDV1a, 23.1 days (d); BVDV1b, 22.8 d; BVDV2, 22.9 d; BHV-1, 21.2 d; PI-3V, 30.3 d; and BRSV, 35.9 d. The mean half-life of viral antibodies was greater for vaccinates than for nonvaccinates for all viruses except BRSV. The calculated mean time to seronegative status for nonvaccinates based on titers at day 0 was: BVDV1a, 192.2 d; BVDV1b, 179.1 d; BVDV2, 157.8 d; BHV-1, 122.9 d; PI-3V, 190.6 d; and BRSV, 186.7 d. There was an active immune response after vaccination with two doses to all the viruses, except BRSV. Mean antibody titers of vaccinates at day 116 were statistically higher than nonvaccinates for all viruses except BRSV. However on an individual calf basis there were few seroconversions (four-fold rise or greater to BVDV1a, BVDV1b, BVDV2, PI-3V, or BRSV; or two-fold rise for BHV-1) in the presence of viral antibodies. The predicted time of seronegative status for a group of calves for vaccination programs may not be appropriate as there may be a range of titers for all calves at day 0. In this study the range for BVDV1a was 16-16,384; BVDV1b, 8-8192; BVDV2, 0-8192; BHV-1, 0-935; PI-3V, 8-2048; and BRSV, 8-4096. Using the half-life of 23 d for BVDV1a, the time thereafter for seronegative status would be 46 and 299 d compared to the calculated date of 192.2 d using the mean of estimated time to seronegative status for all the calves. There was an active humoral response in the vaccinated calves to M. haemolytica and P. multocida. Cowherd humoral immunity based on serum antibodies should be monitored as it may relate to transfer of maternal antibodies to calves. Exceptionally high levels of viral antibodies transferred to calves could interfere with the antibody response to vaccination.

Response of calves persistently infected with noncytopathic bovine viral diarrhea virus (BVDV) subtype 1b after vaccination with heterologous BVDV strains in modified live virus vaccines and Mannheimia haemolytica bacterin-toxoid.

Seronegative persistently infected (PI) calves with bovine viral diarrhea virus (BVDV) subtype 1b were vaccinated with each of four modified live virus (MLV) BVDV vaccines and a Mannheimia haemolytica bacterin-toxoid. Nasal swabs and peripheral blood leukocytes (PBL) were collected for virus isolation and serums were collected after vaccination and tested for BVDV1a, BVDV1b, BVDV2, bovine herpesvirus-1 (BHV-1), bovine parainfluenza-3 virus (PI-3V), and bovine respiratory syncytial virus (BRSV) antibodies. M. haemolytica and Pasteurella multocida antibodies were detected using ELISA procedures. None of the PI calves developed mucosal disease (MD) after MLV vaccination. None of the BVDV PI calves seroconverted to BVDV1b after MLV vaccination. Calves receiving MLV vaccines seroconverted to the respective type/subtype in the vaccine. Calves receiving a MLV vaccine with noncytopathic (NCP) BVDV1 (subtype not designated) did not seroconvert to BVDV1a, BVDV1b, or BVDV2. The PI calves were positive for BVDV subtype 1b, in the PBL and nasal swabs throughout the study. Calves receiving each of three vaccines with known BVDV1a strains had BVDV1a positive samples after vaccination, in some but not all calves, up to Day 28. The PI BVDV1b calves did not respond with increased M. haemolytica antibodies after vaccination compared to BVDV negative calves receiving the same M. haemolytica vaccine.