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Detergent solubilised bovine viral diarrhea virus elicits a similar immune response as the inactivated virus.

The immune response to two detergent solubilised preparations of Bovine Viral Diarrhea Virus (BVDV) proteins were compared with the response to a chemically inactivated BVDV preparation. The antibody titres were measured with the standard immunofluorescence assay and with a newly developed enzyme linked immunosorbent assay (ELISA). The range of viral protein specificity of the antibodies was compared using SDS-polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blot analysis. The results clearly demonstrate that the detergent solubilised viral proteins are equally as effective immunogens as the killed BVDV vaccine both in terms of antibody titers and specificity.

Animals↗

A study of some pathogenetic aspects of bovine viral diarrhea virus infection.

The cytopathic (CP) TVM-2 strain of bovine viral diarrhea virus (BVDV) induced in calves a severe disease, characterized by the clinical picture which is usually reported for the acute primary infection observed under natural conditions. In contrast, the calves inoculated with a different biotype of BVDV, the non-cytopathic (NCP) New York-1 strain, remained clinically normal with the only evidence of virus replication in these calves being the recovery of the virus from their pharyngeal swabbings and blood and also the detection of specific neutralizing antibody in their serums. When calves were immunosuppressed with dexamethasone (DMS), they underwent an overt systemic disease of such a severity that in most of the cases it ended with the death of the animals. This result was obtained with either the CP and the NCP strain of BVDV. Finally, the mixed infection that was obtained in the calves with the CP and the NCP BVDV did not result in any particular unexpected pathological situation. It was speculated that the immunosuppressive activity of BVDV could be a property peculiar to certain isolates of the virus.

Animals↗

Effects in calves of mixed infections with bovine viral diarrhea virus and several other bovine viruses.

The objective of this study was to verify whether a mixed infection in calves with bovine viral diarrhea virus (BVDV) and other bovine viruses, such as bovid herpesvirus-4 (BHV-4), parainfluenza-3 (PI-3) and infectious bovine rhinotracheitis (IBR) virus, would influence the pathogenesis of the BVDV infection sufficiently to result in the typical form of mucosal disease being produced. Accordingly, two experiments were undertaken. In one experiment calves were first infected with BVDV and subsequently with BHV-4 and IBR virus, respectively. The second experiment consisted in a simultaneous infection of calves with BVDV and PI-3 virus or BVDV and IBR virus. From the first experiment it seems that BVDV infection can be reactivated in calves by BHV-4 and IBR virus. Evidence of this is that BVDV, at least the cytopathic (CP) strain, was recovered from calves following superinfection. Moreover, following such superinfection the calves showed signs which could most likely be ascribed to the pathogenetic activity of BVDV. Superinfection, especially by IBR virus, created a more severe clinical response in calves that were initially infected with CP BVDV, than in those previously given the non-cytopathic (NCP) biotype of the virus. Simultaneous infection with PI-3 virus did not seem to modify to any significant extent the pathogenesis of the experimentally induced BVDV infection whereas a severe clinical response was observed in calves when simultaneous infection was made with BVDV and IBR virus.

Animals↗

Bovine viral diarrhea virus-infected MDBK monolayer as antigen in enzyme-linked immunosorbent assay (ELISA) for the measurement of antibodies in bovine sera.

A specific ELISA for the detection of IgG antibodies against bovine viral diarrhea virus (BVDV) has been developed and was compared with the seroneutralization (SN) titers for a total of 60 bovine serum samples. A BVDV-infected Madin-Darby Bovine Kidney (MDBK) cell monolayer served as test antigen. The following results were obtained: a coefficient of correlation (r) of 0.63 (P less than 0.01) between BVDV-ELISA and SN titer in BVDV-seroconverted animals and 0.71 for all sera, including that from BVDV vaccinated animals; a sensitivity of 92%, a specificity of 91%, a concordance of 92%; and a demonstration of a parallel increase in BVDV-ELISA and SN titers in animals with seroconversion. The BVDV-ELISA permits a more complete evaluation of the humoral immune response to BVDV.

Animals↗

Competitive and blocking enzyme-linked immunoassay for detection of fetal bovine serum antibodies to bovine viral diarrhea virus.

A competitive blocking enzyme-linked immunoassay (CELIA) was developed to detect bovine viral diarrhea virus (BVDV) antibodies in undiluted fetal bovine serum (FBS). The CELIA was based on competition of serum BVDV antibodies with biotin-labelled anti-BVDV immunoglobulins (Ig) for a limited quantity of solid-phase BVDV antigen. Antigen preparation was simple, FBS could be tested undiluted, and detergent-containing washes were unnecessary. A series of dilutions of postnatal bovine BVDV antiserum prepared in FBS and a set of 147 undiluted abbatoir FBS samples were tested by both CELIA and serum neutralization tests (SNT). CELIA results on both sets of specimens correlated positively with SNT titers (r = 0.99 and r = 0.85). Relative to the SNT, CELIA sensitivity was 100%; specificity was 76%. CELIA detected a level of BVDV antibody below the 1:2-titer threshold detectable with the SNT. Advantages, limitations, and theoretical differences between the CELIA and SNT are discussed. A similar comparison of CELIA with non-competitive enzyme-linked immunoassay approaches to BVDV serodiagnosis is made. It is concluded that the CELIA is valuable in selecting only BVDV-seronegative FBS for use in virologic cell culture media.

Animals↗

Bovine interferon: its biology and application in veterinary medicine.

Investigations of the production and potential use of bovine interferons against viral infections have occurred since the first descriptions of interferons in other systems. The recent advent of recombinant DNA-technology has facilitated such studies and furthered our knowledge about the bovine interferon system in general. This review gives an overview of the biology, antiviral and immunomodulatory activities of bovine interferons. Areas in which the interferons are now applied or have potential application in viral diseases in cattle are described. Finally, the value of studies of the bovine interferon system with respect to comparative interferon research is discussed.

Animals↗

Inactivation of the unconventional agents of scrapie, bovine spongiform encephalopathy and Creutzfeldt-Jakob disease.

Scrapie, bovine spongiform encephalopathy (BSE) and Creutzfeldt-Jakob disease (CJD) are the best known of the transmissible degenerative encephalopathies (TDE) that affect animals and man. Among the unusual properties of the unconventional causal agents is their relative resistance to standard decontamination procedures, and this has resulted in accidental transmission. Scrapie in sheep is the most common of these diseases and, through laboratory studies, is the best understood. As the model for the group, scrapie agent has been used in experiments to devise general standards for decontamination of the agents of the TDE.

Animals↗

Efficacy of an inactivated oil-adjuvanted rotavirus vaccine in the control of calf diarrhoea in beef herds in Argentina.

We have assessed the potency of an inactivated oil-adjuvanted rotavirus vaccine in beef herds in Argentina. Two different vaccine trials were conducted. In a small-scale experimental trial, involving 21 pregnant cows (13 vaccinated and eight unvaccinated controls), a significant increase in neutralizing antibody titres against different serotypes of bovine rotaviruses was found in both the colostrum and serum of vaccinated cows compared with that of unvaccinated controls. Seven days after birth, half of the calves born to vaccinated dams or to control cows were challenged with live virulent virus whereas the other half of both groups were left in contact with the infected calves in order to mimic a natural field challenge. Although no statistically significant differences in the rate of protection were observed among the different groups of animals, a larger number of vaccinated calves were protected in comparison with their controls, particularly where animals in contact with infected calves were concerned. Secondly, a large-scale field trial was carried out in 17 beef herds involving a total of 4066 vaccinated pregnant cows. In 11 farms morbidity and mortality in calves from vaccinated cows were compared with historical data from the previous 3 years at the same locations. In the other six herds, control groups were used to compare data of the same year: 1540 cows were vaccinated and 2700 were left as controls. Taking into account the previous and current incidence of diarrhoea, morbidity and mortality were significantly reduced in 16 of the 17 beef herds tested. Vaccine effectiveness was also evident in farms where other enteropathogens such as cryptosporidium and coronaviruses were present, together with rotavirus.

Adjuvants, Immunologic↗

Toxicity of Diplodia maydis in farm and laboratory animals.

The acute toxicity of maize culture material of eight strains of Diplodia maydis in ducklings, as well as the ability of five of these strains to induce typical diplodiosis (a neuromuscular disease) in cattle and sheep was shown. Typical diplodiosis was induced in 17 sheep and 11 cattle. Two of the five toxic strains were isolated from maize involved in diploidiosis outbreaks, the others from commercial maize. Strains inducing diplodiosis could be isolated from commercial maize from the USA, Argentina and South Africa. There was no correlation between the toxicity of D. maydis strains in ducklings and their ability to induce diplodiosis in cattle and sheep. Some isolates were acutely toxic to ducklings and rats but were unable to induce diplodiosis in either cattle or sheep. Others, equally toxic to ducklings and rats, induced diplodiosis in cattle and sheep at low dose levels. Two doses, each of 5 g/kg, of maize culture material of isolates from the USA, Argentina and South Africa induced diplodiosis in sheep. Culture material incubated for less than 8 wk could not induce diplodiosis in cattle. Acute toxicity in ducklings and rats also increased with longer incubation periods. Cultures of non-sporulating and profusely sporulating strains were equally toxic to ducklings. Heat treatment of culture material for 48 days at 45 degrees C failed to reduce toxicity in ducklings.

Animal Feed↗

Attempted reactivation of latent bovine herpesvirus 1 infection in calves by infection with ruminant pestiviruses.

Three calves, latently infected with bovine herpesvirus 1 (BHV 1), were each inoculated intranasally with 9 strains of ruminant pestivirus (BVDV). All three calves developed a biphasic pyrexia and a lymphopenia followed by a neutrophilia. They did not shed BHV 1 in their nasal secretions in the 14 days following BVDV inoculation, and their BHV 1 antibody levels remained static, as did those of 2 control calves not given BVDV. All five calves were subsequently shown to be latently infected with BHV 1 by the production of recrudescent infections following the administration of dexamethasone. BHV 1 was recovered from nasal secretions and there was a marked rise in BHV 1 antibody titres in the second week after dexamethasone administration.

Animals↗

Congenital infections with nonarbo togaviruses.

The present review deals with the similarities and differences of selected aspects of prenatal pestivirus infections of domestic animals and congenital rubella of man. Hog cholera virus, bovine virus diarrhoea virus and border disease virus are antigenically closely related, but unrelated to rubella virus. The nonarbo togaviruses are capable of producing congenital infections resulting in a wide spectrum of abnormalities. The infected foetus can die in utero, in the neonatal period, or it may be born with teratogenic defects. In addition, apparently healthy progeny can be delivered that develop a late onset disease, months, or years after birth, or remain clinically normal for life. The ultimate outcome of a congenital infection is mainly determined by the stage of foetal development, at which infection occurs. Foetuses exposed to rubella virus raise an antibody response to the virus, whereas domestic animals frequently fail to respond immunologically to a congenital pestivirus infection. In congenital rubella the virus usually disappears from the host's body 1-2 years after birth. However, congenital pestivirus infections may be characterized by a lifelong and widespread persistence of virus in clinically healthy animals. Such animals are of significance in the epizootiology of bovine virus diarrhoea, border disease or hog cholera.

Abortion, Veterinary↗

Comparison by the neutralisation assay of pairs of non-cytopathogenic and cytopathogenic strains of bovine virus diarrhoea virus isolated from cases of mucosal disease.

Neutralising antibody to non-cytopathogenic and cytopathogenic strains of bovine virus diarrhoea virus (BVDV) was assayed in a microtitre test in which cultures of calf testis cells were stained by the immunoperoxidase method to detect viral replication. Fourteen BVDV strains were compared in cross neutralisation tests with antisera prepared in gnotobiotic calves. Ten of the strains comprised five pairs of non-cytopathogenic and cytopathogenic BVDV. Each pair was isolated from an animal with mucosal disease. All five animals were from five separate outbreaks of the disease. Each pair of strains from the same outbreak was found to be antigenically indistinguishable. In contrast, when the coefficient of antigenic similarity was calculated 11 of 45 comparisons between the pairs and 46 of 91 comparisons between all 14 viruses gave R values that distinguished strains. The observations suggest that an antigenic spectrum within a single related group exists for BVDV strains, rather than distinct serotypes. The findings are also consistent with the suggestion that cytopathogenic strains from natural outbreaks of mucosal disease arise by mutation from non-cytopathogenic virus.

Animals↗

A rapid, quantitative assay for titration of bovine virus diarrhoea-mucosal disease virus.

An end point dilution microtitration assay is described that can be used for the titration of both cytopathic and non-cytopathic isolates of bovine virus diarrhoea-mucosal disease virus. Indirect immunofluorescence is used to detect infected MDBK cells in the wells of Terasaki plates. The virus titre is derived from the number of uninfected wells, using the Poisson distribution. The assay is simple, fast and economical. Titres of cytopathic virus determined by the microtitration assay and standard plaque assay are equivalent.

Animals↗

Protection against respiratory infection with bovine virus diarrhoea virus by passively acquired antibody.

Susceptibility to infection with bovine virus diarrhoea virus (BVDV) was compared for calves with varying amounts of specific antibody in their sera passively acquired from the ingestion of colostrum. Challenge consisted of intranasal exposure to a strain of BVDV isolated from an outbreak of respiratory disease. Resistance to infection, as judged by nasopharyngeal shedding of virus, was directly related to the titre of neutralizing antibodies in sera. Besides protecting against infection of the upper respiratory tract, passive antibody, which was mainly IgG1, also protected against viraemia and, to a lesser extent, leukopenia. In the presence of colostral antibody, neutralizing and IgG1 antibody responses were apparently inhibited, but a specific IgG2 response occurred.

Animals↗

In vitro amplification of the 16S rRNA genes from Mycoplasma bovis and Mycoplasma agalactiae by PCR.

Mycoplasma bovis and Mycoplasma agalactiae are two very closely related species which cause mastitis in cows and goats, respectively. M. bovis can also cause arthritis and respiratory disease in cattle. It has recently been shown that the 16S rRNA sequences differ only in 8 nucleotide positions between the two species [J.G. Mattsson, B. Guss and K.-E. Johansson (1994) FEMS Microbiol. Lett., 115: 325-328]. These nucleotide differences are distributed over the molecule in such a way that it is difficult to design specific identification systems, based on PCR only, for M. bovis and M. agalactiae. Two different PCR systems based on 16S rRNA sequence data have, however, been designed for these two species. The forward primers were identical in the two systems and complementary to a segment of the evolutionarily variable region V2. The reverse primers were complementary to the variable region V6, in which there are two nucleotide differences between M. bovis and M. agalactiae. The size of the PCR products, generated with these primers, was 360 bp. Cross-amplification was obtained with the two species in the heterologous PCR systems, but with approximately a 100-fold lower efficiency. Cross-amplification was not obtained with any other bovine or caprine mycoplasma except for Mycoplasma sp. strain A1343 of the caprine group 7. The detection limit of the PCR system for M. bovis with a reference culture was 4 x 10(2) CFU/ml and of the PCR system for M. agalactiae 2 x 10(2) CFU/ml. The M. bovis-PCR system was used to analyze nasal samples of calves from a herd where an outbreak of pneumonia had occured and it proved possible to detect M. bovis in these samples.

Animals↗

Natural killer cells act as early responders in an experimental infection with Neospora caninum in calves.

The intracellular protozoan parasite Neospora caninum is a cause of abortion and congenital disease in cattle worldwide. We have previously shown that natural killer (NK) cells produce IFN-gamma in response to N. caninum tachyzoites in vitro. This study aimed to investigate the role of NK cells and other cellular immune responses in an experimental N. caninum infection model in calves. Phenotyping of peripheral blood lymphocytes showed a drop in the percentage of NK cells at days 4-6 after i.v. inoculation, followed by an increase in the percentage of both NK cells and CD8+ T cells which peaked at days 11-15. A whole blood flow cytometric assay showed that CD4+ T cells were the major IFN-gamma producing cells, but in the early stages of the infection both NK cells and CD8+ T cells contributed to IFN-gamma production. We also compared the ability of two different N. caninum antigen preparations--sonicated soluble antigens and intact heat-inactivated parasites--to induce proliferation and IFN-gamma production in various cell types. Heat-inactivated tachyzoites induced a 3.7 times greater increase in the number of IFN-gamma producing NK cells compared with sonicated soluble antigens. This indicated the presence of some NK cell-stimulating antigens in the intact tachyzoite that were absent from the sonicated soluble antigens. The heat-inactivated whole tachyzoites also inhibited gammadelta T cell proliferation while the soluble antigens from N. caninum did not. We believe this is the first time NK cells have been demonstrated to be early responders in N. caninum infection in calves.

Animals↗