Infectious Bovine Viral Diarrhea Virus Research Articles

Effective utilization of vaccination to protect against viral reproductive pathogens

Bovine viral diarrhea virus (BVDV) and Bovine alphaherpes­virus-1 (BoHV-1) have long been recognized as major causes of reproductive disease in beef and dairy cattle. Enhancing im­munity through prebreeding vaccination and annual revaccina­tion of the cowherd provide important contributions to limiting reproductive losses associated with these viral infections and are also important control procedures to limit transmission of these viruses among cattle populations. While providing efficacy, vaccination should also be safe. Safety concerns are often associated with the BoHV-1 fractions, while efficacy con­cerns are often associated with BVDV fractions of multivalent vaccines. Vaccination against BVDV and BoHV-1 effectively decrease risk of fetal infection, including abortions, birth of per­sistently infected BVDV carriers, and pregnancy loss. Safety con­cerns associated with viral vaccines, more specifically modified-live viral vaccines, should not overshadow the protective effects associated with sound vaccination principles and programs.

"Fading out" - genomic epidemiology of the last persistently infected BVDV cattle in Germany.

Bovine viral diarrhea virus (BVDV) is one of the most important cattle pathogens worldwide, causing major economic losses and animal welfare issues. Disease eradication programs have been implemented in several countries, including Germany where an obligatory nationwide control program is in force since 2011. As molecular epidemiology has become an essential tool to understand the transmission dynamics and evolution of BVDV, 5' untranslated region (UTR) sequences are generated from viruses present in persistently infected animals since the beginning of the BVDV control program. Here, we report the results of the sequence-based subtyping of BVDV strains found from 2018 through 2022 in calves born in Germany. In 2018, 2019 and 2020, BVDV-1d and-1b were the dominant subtypes and cases were spread throughout the area that was not yet officially declared BVDV-free at that time. In addition, BVDV-1a, -1e, -1f and -1h could rarely be detected. From 2021 onwards, subtype 1d clearly took over the dominance, while the other subtypes could be gradually nearly eliminated from the cattle population. The eradication success not only results in a drastic reduction of cases, but also in a marked reduction of strain diversity. Interestingly, before vaccination has been banned in regions and farms with a disease-free status, two live-vaccine virus strains were repeatedly detected in ear tissue samples of newborn calves (n = 14) whose mothers were immunized during gestation. The field-virus sequences are an important basis for molecular tracing and identification of potential relationships between the last outbreaks in the final phase of the German BVDV eradication program, thereby supporting classic epidemiological investigations. Furthermore, the monitoring of the composition of virus subtypes in the cattle population helps to maintain effective diagnostic methods and control measures and is an early warning system for the introduction of new pestiviruses in the naïve cattle population.

Immune Responses to Influenza D Virus in Calves Previously Infected with Bovine Viral Diarrhea Virus 2.

Bovine viral diarrhea virus (BVDV) induces immunosuppression and thymus depletion in calves. This study explores the impact of prior BVDV-2 exposure on the subsequent immune response to influenza D virus (IDV). Twenty 3-week-old calves were divided into four groups. Calves in G1 and G3 were mock-treated on day 0, while calves in G2 and G4 received BVDV. Calves in G1 (mock) and G2 (BVDV) were necropsied on day 13 post-infection. IDV was inoculated on day 21 in G3 calves (mock + IDV) and G4 (BVDV + IDV) and necropsy was conducted on day 42. Pre-exposed BVDV calves exhibited prolonged and increased IDV shedding in nasal secretions. An approximate 50% reduction in the thymus was observed in acutely infected BVDV calves (G2) compared to controls (G1). On day 42, thymus depletion was observed in two calves in G4, while three had normal weight. BVDV-2-exposed calves had impaired CD8 T cell proliferation after IDV recall stimulation, and the α/β T cell impairment was particularly evident in those with persistent thymic atrophy. Conversely, no difference in antibody levels against IDV was noted. BVDV-induced thymus depletion varied from transient to persistent. Persistent thymus atrophy was correlated with weaker T cell proliferation, suggesting correlation between persistent thymus atrophy and impaired T cell immune response to subsequent infections.

Template Entrance Channel as Possible Allosteric Inhibition and Resistance Site for Quinolines Tricyclic Derivatives in RNA Dependent RNA Polymerase of Bovine Viral Diarrhea Virus

The development of potent non-nucleoside inhibitors (NNIs) could be an alternate strategy to combating infectious bovine viral diarrhea virus (BVDV), other than the traditional vaccination. RNA-dependent RNA polymerase (RdRp) is an essential enzyme for viral replication; therefore, it is one of the primary targets for countermeasures against infectious diseases. The reported NNIs, belonging to the classes of quinolines (2h: imidazo[4,5-g]quinolines and 5m: pyrido[2,3-g] quinoxalines), displayed activity in cell-based and enzyme-based assays. Nevertheless, the RdRp binding site and microscopic mechanistic action are still elusive, and can be explored at a molecular level. Here, we employed a varied computational arsenal, including conventional and accelerated methods, to identify quinoline compounds' most likely binding sites. Our study revealed A392 and I261 as the mutations that can render RdRp resistant against quinoline compounds. In particular, for ligand 2h, mutation of A392E is the most probable mutation. The loop L1 and linker of the fingertip is recognized as a pivotal structural determinant for the stability and escape of quinoline compounds. Overall, this work demonstrates that the quinoline inhibitors bind at the template entrance channel, which is governed by conformational dynamics of interactions with loops and linker residues, and reveals structural and mechanistic insights into inhibition phenomena, for the discovery of improved antivirals.

PSX-B-10 Characterization and Comparison of the Bovine Nasal Microbiome to Exposure of Persistently Infected Bovine Viral Diarrhea Virus

Abstract Bovine viral diarrhea virus (BVDV) is a critical disease negatively impacting many facets of the cattle production system. Cattle infected with the persistent (PI) form of BVDV are the primary reservoirs and source of transmission and may be implicated with an increased risk of bovine respiratory disease (BRD) to exposed calves in feedlots. To date, little to no data exists regarding characterization of the effects PI-BVDV has on the bovine nasal microbiome, an important feature for bovine health. To investigate these effects, Angus-crossbred beef calves were selected and randomly assigned to either continuous exposure (E, n = 3) or non-exposure (NE, n = 3) to an auction market sourced PI-BVDV positive (PI, n = 1) calf from weaning to 112 days post-weaning. Nasal swab samples were collected upon weaning, 56 days post-weaning, and 112 days post-weaning. Next generation sequencing was used to measure the effects of PI-BVDV on the nasal microbiome. Alpha diversity, including Shannon Index and Observed OTUs, and beta diversity, including Bray-Curtis and Jaccard distances, were not different (P > 0.05) between treatments. Next, community structure was assessed at the amplicon sequence variant (ASV) level and LEfSe was used to identify the bacterial biomarkers for each treatment. Results indicated that ASV5-Mycoplasma could be a biomarker for the PI and E treatments due to its high relative abundance in each when compared with the NE treatment. Overall, ASV5-Mycoplasma was identified as a potential biomarker for the prediction and diagnosis of persistently infected bovine viral diarrhea virus among both calves infected with and exposed to the virus and may be an additional biomarker for the increased risk of bovine respiratory disease in exposed feedlot cattle.

Co-infection by Neopora caninum and bovine viral diarrhea virus in cattle from Rio Grande do Sul, Brazil, destined to exportation

ABSTRACT: Reproductive tests in cattle are of great economic importance, given the impact it can have on the production system and may be caused by agents. Neospora caninum and Bovine Viral Diarrhea virus (BVDV) are considered of great importance as reproductive and should be considered responsible for keeping animals persistently infected. The present study included 479 calf serum samples for export in the state of Rio Grande do Sul (RS). All samples were screened for BVDV by an ELISA antigen. BVDV antigen-positive ELISA samples were isolated from BVDV in cell culture. An indirect immunofluorescence (IFT) technique was used to detect anti-N. caninum antibodies. Of the 479 export-treated serum samples, 361 were positive for BVDV antigens by ELISA and/or viral isolation test (361/479-75.36%), and 109 IFT-positive samples for N. caninum (109/479-22.75%). Despite detection of antibodies anti-N. caninum did not differ statistically between naturally infected BVDV and non-BVDV infected animals suggesting that there is no interference of BVDV infection on infection or detection rate of animals with N. caninum, positive animals in viral isolation and high DO in BVDV-Ag ELISA. may present active disease and consequent immunosuppression, contributing to a potential reactivation of N. caninum.

Attenuated porcine-derived type 2 bovine viral diarrhea virus as vector stably expressing viral gene

Infectious bovine viral diarrhea virus (BVDV) cDNA clones have been used for the expression of classical swine fever virus (CSFV) genes for immune prevention and control. However, can it be used for the expression of an allogenetic fragment? To answer this question, a BVDV chimeric virus expressing the spike (S) antigen fragment of porcine epidemic diarrhea virus (PEDV) was constructed. Antigen S499-602 was inserted into pig-derived BVDV-2 infectious cDNA clone pASH28 in tandem by overlapping PCR, located between the seventh and eighth amino acids at the N-terminus of the capsid (C) protein of BVDV. Indirect immunofluorescence assay confirmed that the chimeric virus vASH-dS312 containing double S499-602 sequences was successfully assembled, which could react with the monoclonal antibody (MAb) against BVDV E2 and PEDV S proteins. Further western blot analysis confirmed that the exogenous S499-602 double protein could be stably expressed. Next, the chimeric virus vASH-dS312 was administered to BALB/C mice either orally or by intramuscular injection. The immunized mice were healthy and showed no signs of toxicity. IgG against BVDV and PEDV antibodies could be detected in the mice administered vASH-dS312 by intramuscular injection, which had neutralization activity against BVDV and PEDV. Thus, this study reported a new insertion site in the BVDV infectious cDNA clone that could successfully express an allogenetic antigen.

Determination of Infectious Bovine Viral Diarrhea Virus in Bovine Lung Lavages by a Combination of Virus Propagation in Cell Culture and Quantitative Real-Time PCR

Material of bovine origin is often used in biotechnological applications. Bovine viral diarrhea virus (BVDV) is one of the major viral contaminants, and not only detection and inactivation but also quantification of the viral load in bovine starting material is required by the regulatory agencies. Here, we investigated combined virus propagation in cell culture and quantitative real-time PCR (qRT-PCR) for the applicability to detect and estimate low BVDV titers in bovine lung lavages, the source material for manufacturing pulmonary surfactant. qRT-PCR analyses of the crude lung lavages were performed and qRT-PCR calibration curves based on infective viral doses (TCID50/mL) were generated with a detection limit of 100 TCID50/mL. Lung lavages were inoculated on susceptible MDBK cells and cell culture samples were again analyzed by qRT-PCR. Immunofluorescence staining was performed to prove qRT-PCR results. Interestingly, initial BVDV contaminations in lung lavages were below qRT-PCR detection limit. An amplification step in cell culture enabled BVDV propagation to levels detectable by qRT-PCR. In comparison with the qRT-PCR calibration curve and control experiments with defined inoculation doses, the estimation of minor BVDV contaminations in lung lavages was possible. Both techniques can be successfully combined to estimate the viral load in dilute sample material.

Embryos produced from fertilization with bovine viral diarrhea virus (BVDV)-infected semen and the risk of disease transmission to embryo transfer (ET) recipients and offspring

Bovine diarrhea virus (BVDV) causes a variety of economically important enteric and infertility problems in cattle. For that reason, several countries have eradicated the disease, and some others have schemes in progress to achieve freedom. Although there is a considerable amount of information about the risk of BVDV transmission through contaminated semen used for artificial insemination (AI), there is no evidence to indicate whether the resulting embryos, when used for embryo transfer, can lead to the transmission of BVDV to recipients or offspring. For this experiment, semen from a bull persistently infected with BVDV (105 50% tissue culture infective doses/mL NY strain) was used for insemination (two times at estrus) of BVDV-seronegative, superovulated cows (N = 35). Embryos were collected 7 days after insemination and subsequently were washed according to the International Embryo Transfer Society recommendations or left unwashed. Out of 302 collected oocytes and embryos, 173 (57%) were fertilized and the remaining 129 (43%) had degenerated. Infectious BVDV was detected in 24% (17/71) of unwashed and 10% (8/77) of washed embryos, and in all (N = 11) follicular fluid samples, oviductal epithelial cells, endometrium, and corpora lutea tissues as determined by the virus isolation test. After transfer of 39 washed embryos to 27 BVDV-seronegative recipients, 12 (44%) cows became pregnant and 17 calves free of BVDV and BVDV antibodies, including five sets of twins, were born. After embryo transfer, all pregnant and nonpregnant recipients remained free of BVDV and antibodies. In conclusion, results herein suggest that BVDV can be transmitted by AI resulting in the production of some proportion of contaminated embryos. However, it appears that such embryos, when washed according to International Embryo Transfer Society and the World Organization for Animal Health guidelines do not cause BVDV transmission to recipients or their offspring.

Weaning management of newly received beef calves with or without continuous exposure to a persistently infected bovine viral diarrhea virus pen mate: Effects on rectal temperature and serum proinflammatory cytokine and haptoglobin concentrations1

Exposure to animals persistently infected (PI) with bovine viral diarrhea virus (BVDV) results in immunomodulation in cohorts. It is hypothesized that the extent of modulation differs for low-risk, preconditioned (PC) vs. high-risk, auction market (AM) beef cattle. Our objective was to compare immune responses of PC or AM calves in the presence (PI) or absence (CON) of a PI-BVDV pen mate. Crossbred PC steers (n = 27) from a single ranch origin were weaned, dewormed, vaccinated against respiratory and clostridial pathogens, tested for PI-BVDV, and kept on the ranch for 61 d. Subsequently, PC steers were transported to a receiving unit (RU), weighed, stratified by d -1 BW, and assigned randomly to treatment (PCPI or PCCON) with no additional processing. Simultaneously, crossbred AM calves (n = 27) were assembled from regional auction markets and transported to the RU. The AM calves were weighed, stratified by gender and d -1 BW, processed under the same regimen used for PC steers at their origin ranch, except bull calves were castrated, then assigned randomly to treatment (AMPI or AMCON). Treatment pens were arranged spatially so that PI did not have fence line contact with CON. Blood samples were collected on d 0, 1, 3, 7, and 14 to determine serum concentrations of haptoglobin (Hp), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), IL-4, and IL-6. Rectal temperature (RT) was recorded concurrent with blood sampling. In AM calves, RT and Hp increased (management effect; P < 0.001) sharply on d 1; however, exposure to a PI-BVDV pen mate did not affect either variable (P ≥ 0.79) during the 14-d evaluation period. Serum concentrations of TNF-α tended to increase (P = 0.09) for the PI cohort. A treatment × day interaction (P ≤ 0.05) was observed for IFN-γ on d 7 and 14 and IL-6 on d 14; these indices were greatest for AMPI. Results indicate weaning management and PI-BVDV exposure alter the immune status of newly received beef cattle. These main effects may be additive because proinflammatory cytokine concentrations were greatest for AMPI. Therefore, results further indicate that potential health or growth consequences in cohorts exposed to a PI-BVDV pen mate are impacted by previous management and health history.

Seroprevalence of some bovine viral respiratory diseases among non vaccinated cattle in Saudi Arabia

Aim: Four viral pathogens, bovine viral diarrhea virus (BVDV), and bovine herpes virus type 1 (BHV-1), bovine parainfluenza type 3 virus (PI-3V), bovine respiratory syncytial virus (BRSV) are mainly associated with bovine respiratory diseases that cause major economic losses in the dairy cattle industry. This study aimed to document exposure of cattle in Saudi Arabia to infectious BVDV, BHV-1, PI-3V and BRSV viruses in non vaccinated cattle in order to obtain epidemiological and immunological information. Materials and Methods: In the present study, 460 random serum samples obtained from non vaccinated cattle in five districts (Riyadh, Eastern Province, Jizan, Najran, Asir) of Saudi Arabia between January to March 2011. These samples were tested for presence of antibodies against BVDV, BHV-1, BRSV and PIV-3 by commercial indirect ELISA kits. Results: Our findings displayed that Seropositivity rates were 26 % for BVD, 17.4 % for BHV-1, 69.1 % for PI-3V and 75.6 % for BRSV in the sampled population. In addition, coinfections with more than one virus were considerably common among non-vaccinated dairy cattle. Conclusion: These results indicate that exposure to these agents is common within the study areas. Preventive and control measures against these infectious agents should therefore be adopted.

Weaning management of newly received beef calves with or without continuous exposure to a persistently infected bovine viral diarrhea virus pen mate: Effects on health, performance, bovine viral diarrhea virus titers, and peripheral blood leukocytes1

ABSTRACTExposure to animals persistently infected (PI) with bovine viral diarrhea virus (BVDV) results in immunomodulation of cohorts that may have health and growth consequences; however, effects may differ in low-risk, preconditioned (PC) vs. high-risk, auction market (AM) beef cattle. Our objective was to compare health and performance of PC or AM management systems with (PI) or without (CON) presence of a PI-BVDV pen mate using a 2 × 2 factorial arrangement. Four shipment blocks of crossbred PC steers (n = 236) from 3 ranch-origins were weaned, dewormed, vaccinated, tested for PI-BVDV, and kept on the ranch for ≥42 d. Subsequently, PC steers were transported to a stocker receiving unit (RU), weighed (251 ± 2 kg), blood sampled, stratified by d −1 BW, and assigned randomly to treatment (PCPI or PCCON) with no additional processing. Simultaneously, 4 blocks of crossbred AM calves (n = 292) were assembled from regional auction markets and transported to the RU ± 36 h from PC arrival. The AM calves were weighed (245 ± 1.3 kg), stratified by gender and d −1 BW, processed under the same regimen used for PC steers at their origin ranch except bull calves were castrated, and then assigned randomly to treatment (AMPI or AMCON). Treatment pens (0.45 ha) were arranged spatially such that PI did not have fence-line or water source contact with CON. Calves were fed identically and followed the same antibiotic treatment protocol. Daily BW gain for the entire 42-d receiving trial was greater (P < 0.001) for PC (1.2 kg) compared with AM (0.85 kg). There was an exposure effect (P = 0.002) on ADG from d 28 to 42; CON gained 1.12 kg vs. 0.90 kg BW for PI cohort. Morbidity was markedly greater (P < 0.001) in AM (70%) vs. PC (7%), resulting in (P < 0.001) an antibiotic treatment cost of $20.52 and $2.48/animal, respectively. Treatment with a third antibiotic occurred more often (P = 0.04) for PI cohort, and the percentage of chronically ill cattle was greatest (P = 0.06) for AMPI. Upon arrival, BVDV type 1a, 1b, and 2a titers were greater for PC (treatment × day, P < 0.001), and the percentage seropositive to BVDV type 1a on d 0 was 100% for PC vs. 23% in AM. Platelets increased transiently (P < 0.001) with greater platelets observed in AM (P < 0.001). Results indicate that PC calves gain faster and require fewer antibiotic treatments during the receiving period. Exposure to PI reduced BW gain from d 28 to 42, increased the number of calves treated thrice, and increased chronically ill cattle for AM.

Methods to select suitable fetal bovine serum for use in quality control assays for the detection of adventitious viruses from biological products

Production of biological products, especially vaccines, usually requires materials derived from animals, and there are always risks that animal pathogens derived from these materials could contaminate the final products. Detection of adventitious agents is performed by quality control tests. In these biological assays, animal derived materials are also used and another problem arises, as fetal bovine serum (FBS) is used as an ingredient in tissue culture media. FBS contaminated with bovine viral diarrhea virus (BVDV) or other bovine pathogens, as well as antibodies against these pathogens may lead to false results in quality control assays. In this study, in order to determine the actual status of commercial FBS, we performed quality tests on various FBS samples. As a result, in 28 of 49 FBS samples (57.1%), pestivirus genes were detected by pan-pestivirus reverse transcription-polymerase chain reaction assay. Furthermore, two samples contained infectious BVDV. Neutralizing antibodies against BVDVs were detected in 48 of 49 samples (97.6%) by the virus neutralization test based on the serum-dilution or virus-dilution methods. Antibodies against other bovine pathogens were detected rarely in these samples. From our results, we recommend methods to select FBS that are focused on detection of BVDV and neutralizing antibodies against BVDV.

Characterization and purification of recombinant bovine viral diarrhea virus particles with epitope-tagged envelope proteins

Bovine viral diarrhea virus (BVDV) belongs to the genus Pestivirus within the family Flaviviridae. The lipid membrane of the virions is supposed to contain the three glycosylated envelope proteins E(rns), E1 and E2, but detailed studies of virus assembly are complicated because no efficient purification method for pestiviruses has been described so far. In this study, we generated infectious BVDV with N-terminally FLAG-tagged E(rns) or E2 proteins, respectively. The expression of the epitope-tagged E(rns) and E2 proteins could be shown by immunofluorescence and Western blot experiments. Furthermore, an affinity tag purification protocol for the isolation and concentration of infectious BVDV was established. In the preparation with a titre of 10(8.75) TCID(50) ml(-1), spherical particles with a diameter of 43-58 nm (mean diameter: 48 nm) could be detected by negative staining electron microscopy, and immunogold labelling located both E(rns) and E2 proteins at the virus membrane.

Bovine Viral Diarrhea Virus (BVDV) Infection in Relation to Fertility in Heifers

In this study, blood serum and leukocyte samples were collected from 400 Holstein heifers, all of which appeared to be healthy. Antibodies (Ab) against bovine viral diarrhea virus (BVDV) were detected in 57 serum samples, and BVDV antigen (Ag) was detected in 38 leukocyte samples. There were statistically important differences between the average first insemination ages (FIT) of the BVDV (Ag-/Ab+) heifers (p<0.0001) (pregnant p<0.05, nonpregnant p<0.0001) and BVDV (Ag-/Ab-) heifers. The average conception rates (CR) of BVDV (Ag-/Ab+) heifers and BVDV (Ag-/Ab-) heifers were not significant statistically. There were statistically important differences in average FIT between persistent infected (PI) BVDV (Ag+/Ab-) heifers (p<0.0001; PI pregnant p<0.05, PI nonpregnant p<0.0001) and BVDV (Ag-/Ab-) heifers. No significant differences in average CR between PI BVDV (Ag+/Ab-) heifers and BVDV (Ag-/Ab-) heifers were found. The differences in average FIT between BVDV (Ag+/Ab+; p<0.0001; nonpregnant p<0.0001) and BVDV (Ag-/Ab-) heifers were important statistically. Although there were no BVDV (Ag+/Ab+) pregnant heifers, the differences in average CR between BVDV (Ag+/Ab+) pregnant heifers and BVDV (Ag-/Ab-) heifers were found to be statistically important (p<0.0001). We conclude that fertility is affected in heifers with BVDV (Ag-/Ab+, Ag+/Ab- and Ag+/Ab+).

Experimental infection of early pregnant cows with bovine viral diarrhea virus: Transmission of virus to the reproductive tract and conceptus

The objective of this study was to chronologically investigate the effect of bovine viral diarrhea virus (BVDV) infection on early pregnant cows before placenta formation. Six cows were intravenously inoculated with either BVDV (treated, n = 4) or growth medium (control, n = 2) (day 0) at day 26 of pregnancy. Two treated cows and one control cow were euthanized on day 3 post-infection and the remaining animals were euthanized on day 6. BVDV was isolated from maternal tissues such as lymphoid or reproductive tissues of treated animals on days 3 and 6 post-infection. Additionally, one treated cow autopsied on day 6 post-infection had evidence of infectious BVDV in the allantoic membranes, allantoic fluid and embryos. In three treated cows, a significant decline in progesterone concentration was also observed post-infection while in control cows they remained constant. Therefore, BVDV can infect bovine embryos before placenta formation and may affect progesterone profiles in cows during early pregnancy.

Bovine respiratory disease research (1983–2009)

Bovine respiratory disease (BRD) research has provided significant understanding of the disease over the past 26 years. Modern research tools that have been used include monoclonal antibodies, genomics, polymerase chain reaction, immunohistochemistry (IHC), DNA vaccines and viral vectors coding for immunogens. Emerging/reemerging viruses and new antigenic strains of viruses and bacteria have been identified. Methods of detection and the role for cattle persistently infected bovine viral diarrhea virus (BVDV) were identified; viral subunits, cellular components and bacterial products have been characterized. Product advances have included vaccines for bovine respiratory syncytial virus, Mannheimia haemolytica and Pasteurella multocida; the addition of BVDV2 to the existing vaccines and new antibiotics. The role of Mycoplasma spp., particularly Mycoplasma bovis in BRD, has been more extensively studied. Bovine immunology research has provided more specific information on immune responses, T cell subsets and cytokines. The molecular and genetic basis for viral-bacterial synergy in BRD has been described. Attempts have been made to document how prevention of BRD by proper vaccination and management prior to exposure to infectious agents can minimize disease and serve as economic incentives for certified health programs.

Prevalence of Infectious Agents in Free-ranging White-tailed Deer in Northeastern Mexico

The objectives of this study were to determine the prevalence of antibodies against brucellosis, leptospirosis, infectious bovine rhinotracheitis virus, and bovine viral diarrhea virus (BVDV) in white-tailed deer (Odocoileus virginianus) in northeastern Mexico. Deer (n=521) were captured from helicopter using a netgun on 15 ranches covering 62,114 ha in the states of Coahuila, Nuevo Leon, and Tamaulipas during spring 2004. The prevalence of antibodies against Leptospira, infectious bovine rhinotracheitis, BVDV, and brucellosis were 5.6, 41.1, 63.5, and 0%, respectively, indicating that white-tailed deer and cattle may share disease agents when cohabiting in northeastern Mexico.

Bovine viral diarrhea virus is inactivated when whole milk from persistently infected cows is heated to prepare semen extender

Bovine viral diarrhea virus (BVDV) can be present in cryopreserved bovine semen and be transmitted through artificial insemination. Because BVDV can be shed in milk, the virus might also be introduced as a contaminant of milk-based semen extenders. Thus, the purpose of this study was to evaluate the epidemiologic risk of using heated, BVDV-contaminated milk to prepare semen extender. Milk was obtained from cows free of and persistently infected (PI) with BVDV. Six replicates of milk samples were processed by heating (85–92.2 °C, 10 min). Samples of milk collected before and after heating were assayed for BVDV. Additionally, milk was injected intravenously into eight BVDV seronegative calves to monitor for seroconversion and viral infection. Virus was not detected in any milk samples from negative animals. Virus was consistently isolated from unheated milk samples from PI cows by passage of somatic cells, ultracentrifugation, and animal inoculation. Virus was usually detected in these samples by RT-nPCR (reverse transcription nested polymerase chain reaction). In heated milk samples from PI cows, no infectious BVDV was detected using any technique, but viral RNA was detected using RT-nPCR in four of six replicates. Bovine viral diarrhea virus in milk from PI cows was inactivated by heating. Therefore, properly heated milk used in semen extenders will not result in transmission of infectious BVDV. Although RT-nPCR detected the presence of viral RNA in milk samples after heating, the virus was not infectious as demonstrated by lack of replication despite using multiple sensitive techniques.

Impact of Persistent Bovine Viral Diarrhea Viral Infection on the Duration and Level of Shedding ofEscherichia coliO157 in Calves

The goal of this study was to determine whether calves persistently infected with bovine viral diarrhea virus (BVDV) and inoculated with Escherichia coli O157 will shed the organism at a higher concentration and for a longer duration than calves not infected with BVDV. Nine calves, 6-8 weeks old, persistently infected with noncytopathic BVDV and eight calves not infected with BVDV obtained from separate cow-calf operations were used in this study. BVDV status, positive or negative, of all calves was confirmed by repeated testing throughout the study. Both groups were orally inoculated with 10(9) colony-forming units (CFU) of five nalidixic acid-resistant strains of E. coli O157. All calves were examined daily, and fecal samples were collected three times a week for 55 days for detection and enumeration of the nalidixic acid-resistant E. coli O157. Calves were then necropsied, and samples from the gastrointestinal tract were taken for the detection of the nalidixic acid-resistant E. coli O157. Data analysis indicated no statistical difference in the concentration of E. coli O157 shed or the duration of shedding between the persistently infected BVDV calves and the control calves throughout the length of the study. Our data indicate that persistent infection with noncytopathic BVDV does not play a role in the level or duration of shedding of E. coli O157 in cattle.