Measurement of isotype-specific antibody responses to Aujeszky's disease virus in sera and mucosal secretions of pigs.

Enzyme-linked immunosorbent assays (ELISAs) for the detection of porcine IgM, IgA, IgG1 and IgG2 antibodies directed against Aujeszky's disease virus (ADV) are described. ADV-specific IgA and IgM were detected in an antibody capture assay, and ADV-specific IgG1 and IgG2 were detected in an indirect double antibody sandwich assay. A selected set of samples was tested in the four ELISAs and in a 24 h virus neutralization assay. Comparison of the results showed that the ELISAs were isotype-specific, sensitive, and reproducible. Samples with ADV antibody of one isotype showed that ADV-specific IgG1, IgG2 and IgM were able to neutralize the virus in vitro. In vitro neutralization of virus can be enhanced by complement. ADV-specific IgA neutralized virus only weakly. ADV-infected cells activated complement in the absence of antibody. Specific IgG2 and IgM enhanced complement activation. Analysis of the time course of antibody responses after infection or vaccination revealed that the isotype-specific ELISAs are suitable to study the humoral antibody response of pigs to the virus in mucosal secretions. Wild-type virus (strain NIA-3) and an attenuated vaccine strain (Bartha) administered intranasally induced mucosal IgM and IgA responses to the virus. In contrast, a killed vaccine (Nobivac) administered intramuscularly induced only weak mucosal IgM responses. The attenuated vaccine strain primed for a mucosal IgA memory response evoked upon challenge infection with wild-type virus.     

A congenital persistent swine fever infection. II. Immune response to swine fever virus and unrelated antigens

After the disappearance of maternal antibodies, no antibodies were detected in five pigs that were congenitally infected with the Bergen strain of swine fever virus. The pigs lived for 2–11 months. One pig showed low levels of precipitating antibody during the last month of its life. The pigs had a normal immune response to sheep red blood cells and to pig parvovirus. These observations suggest the presence of immunological tolerance to swine fever virus in these pigs.Cell mediated immunity, measured by phytohaemagglutinin lymphocyte stimulation, appeared to be normal.     

Mechanistic target of rapamycin is activated in bovine granulosa cells after LH surge but is not essential for ovulation

The LH surge induces functional and morphological changes in granulosa cells. Mechanistic target of rapamycin (mTOR) is an integrator of signalling pathways in multiple cell types. We hypothesized that mTOR kinase activity integrates and modulates molecular pathways induced by LH in granulosa cells during the preovulatory period. Cows were ovariectomized and granulosa cells collected at 0, 3, 6, 12 and 24 hr after GnRH injection. While RHEB mRNA levels increased at 3 and 6 hr, returning to basal levels by 12 hr after GnRH treatment, RHOA mRNA levels increased at 6 hr and remained high thereafter. Western blot analyses revealed increased S6K phosphorylation at 3 and 6 hr after GnRH injection. Similarly, mRNA levels of ERK1/2, STAR and EGR‐1 were higher 3 hr after GnRH treatment. Rapamycin treatment inhibited mTOR activity and increased AKT activity, but did not alter ERK1/2 phosphorylation and EGR1 protein levels in cultured bovine granulosa cells. Rapamycin also inhibited LH‐induced increase in EREG mRNA abundance in granulosa cells in vitro. However, intrafollicular injection of rapamycin did not suppress ovulation. These findings suggest that mTOR is involved in the control of EREG expression in cattle, which may be triggered by LH surge stimulating RHEB and S6K activity.     

Bovine respiratory syncytial virus reinfections and decreased milk yield in dairy cattle

Summary

The influence of Bovine Respiratory Syncytial Virus (BRSV) reinfections on the daily milk yield was studied by evaluating the milk production of 32 BRSV reinfected cows. For the estimation of milk production losses, four lactation curve models were used, including a gamma function, a second degree polynomial, and both of these models with a lag variable. Bovine respiratory syncytial virus reinfections seemed to have only a small effect on the daily milk production. Comparison of the true production with an estimated production according to the gamma function showed that the production for first lactation cows dropped 0.14 kg on average and for cows in their second or later lactation 0.56 kg on average, during 5 consecutive days in the infection period. For the second‐degree polynomial model these values were respectively 0.42 kg and 0.80 kg. All calculated average production losses were relatively small and not significant (P>0.15). The models without lag variable were more suitable than the models with the lag variable to estimate small production losses caused by BRSV reinfections. The power of this study was sufficient to detect a decrease in production of approximately 1 ‐ 1.5 kg milk per cow per day. It was therefore concluded that BRSV reinfections were not associated with an important loss of milkproduction.     

Reactivation of latent bovine herpesvirus 1 in cattle seronegative to glycoproteins gB and gE

Six heifers were vaccinated intranasally with the live bovine herpesvirus 1 (BHV1) temperature-sensitive (ts) vaccine strain RBL106 within 3 weeks of birth. These calves most likely still had maternal antibodies against BHV1. Thereafter, these heifers were vaccinated several times with an experimental BHV1 glycoprotein-D (gD) subunit vaccine. At the age of 3 years these 6 heifers were seronegative in the BHV1 gB and gE blocking ELISAs, but had neutralizing antibodies against BHV1, probably induced by the vaccinations with the gD subunit vaccine. Five of these 6 heifers excreted BHV1 after treatment with dexamethasone. Restriction enzyme analysis of the genome of the excreted viruses revealed that all 5 isolates had a BHV1.1 genotype and that isolates of 3 heifers were not obviously different from the ts-vaccine strain. The restriction enzyme fragment pattern of the isolate of 1 heifer was clearly different from the pattern of the ts-vaccine strain. It is concluded that cattle can be seronegative against BHV1 gB and gE but can still carry BHV1 in a latent form. This finding strongly suggests that there are completely BHV1 seronegative cattle that are latently infected with BHV1. The impact of this finding on BHV1 eradication programmes is discussed.     

A retrospective study into characteristics associated with the seroprevalence of pseudorabies virus-infected breeding pigs in vaccinated herds in the southern Netherlands

Vaccination programs to eradicate pseudorabies virus (PRV) are being considered in several countries. Knowledge of factors that influence PRV transmission within vaccinated breeding herds may contribute to the success of these programs. A multivariate analysis of variance of the PRV-seroprevalence in sows in 209 herds (average seroprevalence 67.0% per herd) in the southern Netherlands revealed the following risk indicators: (1) presence of finishing pigs; (2) production type (producers of finishing piglets had a higher seroprevalence than producers of breeding stock); (3) vaccination of the sows during nursing (in comparison with vaccinating all sows simultaneously at 5 month intervals, or vaccination during the second half of gestation); (4) pig density in the municipality where the herd was located (seroprevalence increased with higher pig density); (5) herd size less than 100 sows; (6) average within-herd parity (seroprevalence increased with higher withinherd parity); (7) replacement pigs raised on the premises; (8) vaccine strain administered to the sows. Purchase policy (breeding pigs purchased between 10 weeks and 7 months of age, or use of home-bred gilts only) did not significantly contribute to the multivariate model.     

Serological response to Actinobacillus pleuropneumoniae serovar 7 infection in a commercial pig herd

Serological responses to Actinobacillus pleuropneumoniae serovar 7 infection were monitored by enzyme-linked immunosorbent assay in a cohort of 66 pigs between weaning and market. Antibody concentrations were high (63/65 seropositive) at 4 weeks of age but declined to low levels from 8 to 12 weeks. Mean antibody concentrations rose significantly (p less than 0.001) between 12 and 23 weeks. Between 8 and 23 weeks of age, 33 (51.5%) of 64 surviving pigs seroconverted to A pleuropneumoniae serovar 7. Peak antibody concentrations in the seroconverting pigs usually (28/33) occurred at 23 weeks. Seroconversion to A pleuropneumoniae during the grower/finisher phase was not significantly associated (p greater than 0.05) with passive antibody concentrations at 4 weeks of age, lack of vaccination against Mycoplasma hyopneumoniae, or weaning weight. Pleuropneumonic lesions were evident at slaughter in 4 (6.3%) of 64 pigs. A pleuropneumoniae serovar 7 was isolated from 2 of 4 lungs with pleuropneumonia and from another lung with lesions considered typical of enzootic pneumonia.