In vitro interferon production by bovine tissues: induction with infectious bovine rhinotracheitis virus.

The interferon-inducing ability of infectious bovine rhinotracheitis (IBR) virus was determined in tissue cultures of bovine origin inoculated with untreated and ultraviolet (UV) irradiated IBR viruses. Interferon was assayed by the plaque-reduction method in bovine fetal kidney (BFK) cell cultures, using vesicular stomatitis virus as challenge virus. Highest interferon concentrations were produced by cultures of bovine fetal (BF) spleen cells and aveolar macrophage cultures derived from adult cattle. Moderate interferon concentrations were produced by peripheral blood leukocyte (PBL) suspension cultures from adult cattle with serum-neutralizing antibodies against IBR virus. Cultures of PBL from 1 cow without detectable serum-neutralizing antibodies against IBR virus did not produce detectable interferon in response to IBR virus. Cultures of PBL from cattle with or without detectable serum-neutralizing antibodies against IBR virus produced interferon when stimulated with phytohemagglutinin (PHA). Low levles of viral inhibitors were detected infrequently in monolayer cultures of BFK and BF nasal mucosa inoculated with UV-irradiated IBR virus and in BF tracheal organ cultures inoculated with untreated IBR virus. Interferon was not detected in fluids collected from IBR virus-exposed monolayer cultures of primary and secondary BF lung, secondary BF tracheal mucosa, secondary BF liver, secondary BF adrenal, and PBL in the 4th and 7th passages. The antiviral inhibitors from BF spleen, bovine alveolar macrophage, and PBL cultures induced with IBR virus, as well as inhibitors from PBL cultures induced with PHA, had the usual properties of interferon.     

Isolation of infectious bovine rhinotracheitis virus from a latently infected feral pig

Infectious bovine rhinotracheitis (IBR) virus was isolated from the trigeminal ganglion of a feral pig after dexamethasone treatment. Three pigs inoculated intranasally with the IBR virus did not respond clinically or serologically. The virus was re-isolated from tonsillar swabs from two animals on Post-Infection Day (PID) 3.     

Effects of infection with parainfluenza-3 virus and infectious bovine rhinotracheitis virus on neutrophil functions in calves

Calves were challenge exposed in separate experiments with parainfluenza-3 (PI-3) virus or infectious bovine rhinotracheitis (IBR) virus. Blood neutrophils were assayed for functional activity every other day for at least 3 weeks by random migration, Staphylococcus aureus ingestion, antibody-dependent cell-mediated cytotoxicity, cytochrome-c reduction, iodination, and native chemiluminescence. Exposure to PI-3 virus resulted in a brief febrile response and no other clinical signs. Alterations in total or differential WBC counts were not detected. Chemiluminescence and iodination activities were reduced from activities before exposure. Exposure to IBR virus resulted in mild clinical signs and a febrile response of several days' duration. Total WBC and mononuclear cell counts were reduced. Random migration was reduced, whereas S aureus ingestion was enhanced. We concluded that infection of calves with IBR virus and PI-3 virus might directly or indirectly result in alterations of neutrophil function. The functional alterations apparently are different for each virus. These virus-induced alterations in neutrophil function might predispose calves to secondary bacterial pneumonia.     

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.     

Nonspecific immunostimulation against viruses

A trypsinized preparation from chromogenic selected strain of Mycobacterium phlei (NSI) stimulated the recipient immune system non-specifically against a variety of viruses viz. Rabies virus (RNA virus). Marek's disease (DNA virus) and Foot and Mouth disease virus (RNA virus) in phylogenetically different hosts like mice, chicks and guinea pigs respectively. Investigation into mechanisms of such nonspecific immunostimulation revealed that there was induction of strong cell mediated immune response (CMIR) against both specific as well as to nonspecific viral antigens as evinced by LMIT, lymphokine (LyIF) assay and lymphocyte transformation test etc. There was induction of appreciable quantity of "Infectivity inhibiting substance(s)" (IIS) which was not the classical antibody (Ig) in the serum of NSI inoculated animals and birds. This substance(s) neutralized FMD, IBR, rabies and Newcastle disease virus in cell culture, mice and in embryonated eggs. Electrophoretic separation of the NSI induced serum revealed an increase in beta fraction of globulin.     

Evaluation of multiple immune parameters after vaccination with modified live or killed bovine viral diarrhea virus vaccines

The development of immunity to vaccine antigen was examined using three prime/boost strategies and the progression of immune activities was evaluated over the course of 8 weeks. Calves were vaccinated and multiple immune parameters were evaluated using several methods to assess humoral or cellular immunity from the same samples in parallel. The three vaccination protocols used were a killed vaccine followed by a killed boost (killed/killed), MLV vaccine and boost (MLV/MLV), or a MLV vaccine and killed boost (MLV/killed). All the vaccines used included modified live IBR/PI3 viruses to make the bystander context as similar as possible. The Singer strain of BVDV was used as the source antigen in the killed vaccine, and the NADL strain of BVDV was used in the MLV vaccine. Controls received a vaccine containing only MLV IBR/PI3. The assessment panel measured SN titers, as well as lymphocyte proliferation, cytokine mRNA expression, intracellular cytokine production, and released IFN-gamma after in vitro stimulation with three strains of BVDV virus. MLV/MLV and MLV/killed groups developed significant SN titers to the type 1 BVDV virus strains, Singer and NADL, and low crossover titers were also seen to the type 2 strain, 890 over the evaluation period. These two groups showed significant proliferation in response to the NADL virus as compared to controls. Multiple immune assessments were conducted simultaneously to attempt to provide a broader, more in depth evaluation of immune response to these BVDV vaccination protocols. We observed that the correlation among most of the assays conducted were weak; the correlation between SN titers and cellular proliferation assays demonstrated a moderate correlation.     

Active and Passive Immunity to Bovine Viral Respiratory Diseases in Beef Calves After Shipment

Fifteen steers were vaccinated after shipment with a modified live virus vaccine containing infectious bovine rhinotracheitis (IBR), bovine virus diarrhea (BVD), and bovine myxovirus parainfluenza-3 (PI3), and 16 unvaccinated steers were kept as controls. Geometric mean titers one month after vaccination were highest to BVD, followed by PI3 and IBR. Weight gains were higher during 30 days after vaccination in the controls. One case of acute respiratory disease developed in one vaccinated calf. Revaccination 79 days after the first dose increased antibody to PI3 and BVD virus but not IBR. In a second trial, no clinical respiratory disease developed after shipment of 13 heifers that received an antibacterial-antiviral antiserum or in the 12 controls. Weight gains 30 days after shipment were identical in both groups.     

Effects of supplemental chromium on antibody responses of newly weaned feedlot calves to immunization with infectious bovine rhinotracheitis and parainfluenza 3 virus.

The objective of this study was to determine the effect of supplemental dietary chromium (Cr) on antibody responses of feedlot calves. Fifty-five newly weaned calves were divided into two groups, 28 that received supplemental Cr and 27 that did not, and were immunized with a commercial vaccine against bovine infectious rhinotracheitis virus (IBR) and bovine parainfluenza virus type 3(PI-3). Sera harvested from blood sampled preimmunization, and at days 14 and 28 postimmunization (PI), were assayed for anti-IBR and anti-PI-3 antibody titers. Individual calves were also scored as seroconverters if day 14 or 28 PI titers were > or = 3 times the value of the preimmunization titer. Thirty-five calves did not seroconvert to either antigen. Of 20 IBR seroconverters, 15 calves were from the Cr-supplemented group while only five calves were controls (p = 0.007). There was no treatment difference in the number of PI-3 seroconverters. Least squares analysis of actual antibody titers revealed that Cr supplementation increased the magnitude of the peak antibody response to the IBR (p = 0.003), but had no effect on anti-PI-3 antibody titers. These data confirmed and extended our previous observations that supplemental Cr can be immunomodulatory in cattle.     

In utero infection of swine fetuses with infectious bovine rhinotracheitis virus (bovine herpesvirus-1)

The pathogenesis of infectious bovine rhinotracheitis (IBR) virus (bovine herpesvirus-1) was studied in porcine fetuses after in utero inoculation. Laparotomies were performed on 8 seronegative pregnant sows at 34 to 86 days of gestation, and all fetuses in 1 uterine horn of each sow were exposed to IBR virus via inoculation into the amniotic sacs. Fetuses in the other horn served as controls. Clinical signs of infection were not observed in the sows, except for 2 sows that aborted at postinoculation days (PID) 11 and 15. Fetuses of the remaining 6 sows were collected at slaughter on PID 15 to 28. Fetuses were examined for gross abnormalities, presence of IBR virus in tissues, and the formation of neutralizing antibodies to IBR virus. Of 33 inoculated fetuses from 6 sows, 10 were mummified, 11 were hemorrhagic and/or edematous, and 12 were alive. Necrotic lesions were observed on the skin and in the liver of dead and live fetuses. Virus was recovered from 29 of 33 inoculated fetuses. Infectious bovine rhinotracheitis virus was isolated from fetal skin, liver, lungs, kidney, spleen, stomach contents, brain, amniotic fluid, and placenta. Virus was isolated from 4 of 11 fetuses recovered from 1 aborting sow. Antibodies to IBR virus were not detected in sera from the sows. However, antibodies were detected in 6 of 15 fetuses inoculated at 63 to 86 days of gestation and collected at slaughter at 86 to 112 days of gestation. The youngest fetus with detectable IBR antibody was estimated to be 74 days of gestation by measuring crown-rump length of the fetus.     

Anamnestic immune response of pigs to pseudorabies virus: latent virus reactivation versus direct oronasal and parenteral exposure to virus.

The humoral antibody response of pseudorabies-immune pigs to reactivation of latent pseudorabies virus (PRV) was compared with the response following direct exposure to virulent PRV. Nine pigs that had been vaccinated for pseudorabies and later exposed to virulent virus to establish latent infection were given dexamethasone to reactivate latent virus (3 pigs), were exposed oronasally and parenterally to virulent virus (3 pigs), or were kept as nontreated controls (3 pigs). Sera collected from all 9 pigs just before and 3 weeks after treatment were tested by virus neutralization and radioimmunoprecipitation. The 3 pigs exposed directly to virulent virus and 2 of the 3 pigs given dexamethasone had a 4-fold or greater increase in neutralizing antibody titer. All 6 of these pigs had an increase in precipitating antibody activity. Precipitation patterns changed both quantitatively and qualitatively, especially for virus-coded proteins of relatively low molecular weight (less than 46 K). There were some differences in the precipitation patterns associated with sera of individual pigs. However, there was no clear indication of any difference between the 2 treatment groups and therefore no evidence that reactivation of latent virus is associated with any unique immunologic response that could be detected by radioimmunoprecipitation and used diagnostically. Clinical signs were limited to the 3 pigs that were exposed oronasally and parenterally to virulent virus even though the dexamethasone-treated pigs shed more virus for much longer than did those exposed directly to virus.     

A live Pasteurella haemolytica vaccine efficacy trial

A live Pasteurella haemolytica serotype 1 vaccine was used in an efficacy trial conducted on 100 lightweight feeder calves purchased from a Florida ranch. Forty-one calves were inoculated with the vaccine intradermally in the neck. Fifty-nine calves served as nonvaccinated controls. Fourteen days later, the calves were shipped to an order buyer in eastern Tennessee, where the calves were mixed with 60 local calves in a community sale barn for 72 hours. After 3 additional days, the calves were shipped to a research feedlot in Bushland, Tex. They remained in the feedlot for 56 days, and the test was concluded 76 days after vaccination. The P haemolytica vaccine had no significant effect on performance, morbidity, or mortality. There was no significant difference between the vaccinated and nonvaccinated calves in the number of times Pasteurella was isolated. The calves became seropositive to bovine viral diarrhea virus, respiratory syncytial virus, and infectious bovine rhinotracheitis (IBR) virus during the 76-day experiment. All calves initially were seropositive to parainfluenza-3 virus. A virulent outbreak of IBR occurred 30 days after the calves arrived at the feedlot. Before the onset of IBR, the isolation of P haemolytica serotype 1 from nasal turbinates was rare (2 of 500 nasal swabs). After the IBR outbreak, P haemolytica serotype 1 was isolated from 40 of 92 calves.     

Detection of bovine rhinotracheitis virus antibody by neutralizing test and ELISA in experimentally infected rabbits.

The antibody response of rabbit to infectious bovine rhinotracheitis (IBR) virus was examined. Two rabbits were inoculated with IBR virus strain Los Angeles into the trachea and intravenously, and intravenously two times, respectively. The patterns of antibody titers in the rabbits measured by neutralization test and ELISA were similar to those in the case of bovine. The antibody was detected after the inoculation, and much more antibody was detected after the second inoculation. The fact suggests that rabbits are a very useful laboratory animal for IBR virus infection studies.     

Swine Interferon II. Induction in Pigs with Viral and Synthetic Inducers

The production of interferon by pigs in response to viral and synthetic inducers was studied. The inducers used included polyriboinosinic-polyribocytidylic acid (Poly I:C), swine influenza virus and pseudorabies virus. Following intravenous inoculation of pigs with the inducers, sera were examined for interferon by the plaque-reduction method in porcine kidney (PK₁₅) cell cultures using vesicular stomatitis virus as the challenge inoculum. It was shown that pigs can produce interferon in response to each of these inducers. The pseudorabies virus used in this investigation was found to be a better interferon inducer than the swine influenza virus.

The interferon produced in pigs was identified as an interferon because it was pH stable, non-dialyzable, sensitive to trypsin, non-sedimentable and possessed broad-spectrum antiviral activity as well as host-species specificity.

     

Studies on the ability of a 98-kilodalton pseudorabies virus diagnostic antigen to detect latent infections induced by low-dose exposure to the virus

The effect of low-dose challenge of immunity with pseudorabies virus (PRV) on subunit-vaccinated pigs was studied in 2 experiments. In the first experiment, we studied the effect of challenge dose on the antibody response to an early excreted 98-kilodalton PRV-glycoprotein that was used as a diagnostic antigen in the ELISA. In the second experiment, we studied the effect of low doses of virus on the establishment of latent infections in subunit-vaccinated pigs. The relationship of virus exposure dose and vaccine dose to the response of pigs to diagnostic antigen was studied in 18 pigs. Two groups of 3 pigs were vaccinated with a total of 200 micrograms of a lectin-derived PRV subunit vaccine over a 5-week period. Two groups of 3 pigs were similarly vaccinated with a total of 100 micrograms. Two groups of 3 pigs served as nonvaccinated controls. One group of pigs from each of the preceding categories was intranasally exposed to 10(6.0) and 10(2.7) plaque-forming units (PFU) of virus. Antibody to diagnostic antigen was detected by the ELISA and radioimmunoprecipitation 3 to 7 days earlier in pigs exposed to 10(6.0) PFU, demonstrating that the size of the virus challenge dose affects the antibody response to diagnostic antigen. The establishment of latent infections by low PRV doses and the ability to detect these infections was studied in 10 subunit-vaccinated pigs. Each pig was intranasally exposed to 10(2.3) PFU of virus (day 0).(ABSTRACT TRUNCATED AT 250 WORDS)     

Attempts to reactivate bovid herpesvirus-2 in experimentally infected calves

A study was carried out to determine whether bovid herpesvirus-2 (BHV-2) is able to induce a recurrent infection in experimentally infected calves. Twelve calves infected with the virus were treated with dexamethasone (DMS) beginning 69 days after the infection, ie, several weeks after the animals had recovered from the disease and were negative for BHV-2. The stress induced by DMS treatment failed to reactivate the clinical condition or to induce shedding of BHV-2. However, treatment with DMS reactivated a latent infectious bovine rhinotracheitis (IBR) virus infection in all calves previously inoculated with BHV-2, and also in 2 noninoculated controls. The reactivation of IBR virus occurred without any clinical evidence of the disease, but the virus was isolated from nasal and pharyngeal swabbings and from the organs. A proliferative ganglionitis of the trigeminal ganglion was also observed. Because of the interference by IBR virus, this study did not resolve the question as to whether BHV-2 can induce a recurrent infection.     

Clinical infectious bovine rhinotracheitis in cattle fed organic iodine and urea.

The feeding of ethylenediamine dihydriodide (EDDI) at the dose levels of 50 and 500 mg/animal/day and urea at the dose level of 45 g/animal/day did not affect duration of clinical signs, body weight gain, magnitude or duration of fever, serum concentration of glutamic oxalacetic transaminase, packed cell volume, and differential white blood cell counts in feeder cattle experimentally infected with infectious bovine rhinotracheitis (IBR) virus. However, coughing and abundance of nasal discharge were significantly greater in calves fed EDDI before and during primary IBR virus infection. Those calves fed 500 mg of EDDI/day coughed more, had greater nasal discharge, and exhibited greater lacrimation than did those given the smaller dose. These 3 clinical signs were considered to reflect both the expectorant action of EDDI and the pathogenic effects of IBR virus. In all calves, including controls, the coughing, nasal discharge, and lacrimation were most prominent during the period of peak infection (7 to 14 days after the calves were given intranasal inoculation) of the IBR virus. Total serum iodine concentration became maximal (mean of 1,400 ng/ml) in 8 calves after they had been fed the larger dose of EDDI for 2 weeks. This value was maximal (about 300 ng/ml) in another 8 calves after 3 weeks' feeding of the smaller dose (50 mg/day). When EDDI exposure was maintained at the dose level of 50 mg/day for 5 weeks longer, mean serum iodine values remained at about 275 ng/ml, and those of control calves averaged 140 ng/ml.     

Persistence of antibodies and anamnestic response in calves vaccinated with inactivated infectious bovine rhinotracheitis virus and parainfluenza-3 virus vaccines

Persistence of antibodies in calves vaccinated with 2 types of inactivated infectious bovine rhinotracheitis (IBR) virus and parainfluenza-3 (PI-3) virus vaccines were determined. Calves seronegative for IBR and PI-3 viruses were inoculated with 2 doses of inactivated IBR virus-PI-3 virus vaccines administered 2 weeks apart. Blood samples were obtained from the calves for serum at 2 weeks, 6 months, and 1 year after vaccination. The serums were tested by serum-neutralization tests. Antibody response to the vaccines persisted on a declining scale for 1 year. The anamnestic responses to the vaccines were determined by inoculating the same calves with a booster dose of vaccine 1 year after the original 2 doses were given. Blood samples were obtained from the calves for serum 2 weeks later. The serums were tested by serum-neutralization tests. The single booster dose of vaccine elicited an anamnestic response to both IBR and PI-3 viruses.     

Ovarian lesions in heifers exposed to infectious bovine rhinotracheitis virus by non-genital routes on the day after breeding

Twelve heifers were exposed to either a Colorado infectious bovine rhinotracheitis (IBR) virus isolate or an Iowa IBR isolate obtained from a bovine respiratory disease outbreak. All inoculations were made on the day after the heifers had been in estrus and bred by an IBR virus-negative bull. Pairs of heifers were inoculated with each virus isolate intravenously, intramuscularly or exposed by aerosol. The heifers were killed 11-15 days after inoculation and their reproductive tracts and ovaries subjected to virological and pathological study. Virus was isolated from the ovaries of all 4 heifers inoculated intravenously and from 3 of the 4 heifers inoculated intramuscularly, but not from the ovaries of heifers exposed by aerosol. Virus isolations and lesions were, with only 1 exception, confined to the ovary containing the corpus luteum. In ovaries from which IBR virus was isolated, lesions in the corpus luteum ranged from focal necrosis and infiltration of mononuclear cells to diffuse hemorrhage and necrosis. Most of these ovaries also had necrotic follicles and a diffuse mononuclear cell accumulation in the stroma. Lesions were not found in ovaries from which IBR virus was not isolated. It was concluded that lesions are readily induced in the ovaries of post-estrus heifers as a result of hematogenous spread of IBR virus and suggest that the differences in lesion development observed with the 3 routes are related to whether or not a viremia occurred.     

A bovine respiratory virus vaccination trial

A respiratory virus vaccination trial was carried out in a commercial calf-rearing unit with a history of virus pneumonia. The effects of vaccination on the incidence of virus respiratory disease and growth rate were assessed. Forty-four bought-in calves were allocated to groups and treated as follows: A, unvaccinated controls; B, intranasal temperature-sensitive infectious bovine rhinotracheitis (IBR) vaccine at three and 10 weeks; C, intranasal temperature-sensitive combined IBR and parainfluenza-3 (PI3) vaccine at three and 10 weeks; D, intranasal temperature-sensitive combined IBR and PI3 vaccine at three and 10 weeks plus live attenuated bovine respiratory syncytial (BRS) virus vaccine intramuscularly at seven, 10 and 16 weeks. Two outbreaks of virus pneumonia occurred, one at three to four months of age associated with BRS virus and the other at four to five months of age with PI3 virus. During these outbreaks the incidence of pneumonia was lower and the number of days of elevated temperature and the number of treatments were significantly less in groups vaccinated against the associated virus. Despite these findings there were no significant differences between the growth rates of the groups either during the outbreaks of virus pneumonia or during the 10 month period to slaughter.     

A Controlled Field Study Using Live Virus Vaccines and an Antiserum in a Preconditioning Program

Thirty-five vaccinates and 29 control beef calves from five farms were studied. Vaccinates in group 1 received a modified live virus vaccine against infectious bovine rhinotracheitis (IBR) and bovine virus diarrhea (BVD) 30 days after shipment; vaccinates in groups 2, 3 and 4 received live virus vaccines agains IBR and bovine parainfluenza 3 (PI3) seven to 17 days before shipment. Half of group 5 were given bovine origin antiserum containing antibodies against IBR, BVD and PI3. Three weeks later, the animals that had received serum were given a live modified vaccine containing IBR, BVD and PI3. In group 1, WBC counts were lower in the vaccinates than in the controls for two weeks after vaccination. WBC counts in groups 3 and 4 were higher in vaccinates than in controls after addition to the feedlot. Seroconversions to BVD virus occured in all groups. Clinical disease apparently due to BVD affected one vaccinated calf in group 2 and eight calves in group 5. Combined weight gains were significantly higher in three groups of calves vaccinated before shipment compared to unvaccinated control animals after addition to the feedlot. Vaccination with IBR and PI3 live virus vaccines should be given at least 17 days before shipment to feedlots containing infected cattle. Antiserum containing antibodies against the three viruses showed no apparent advantage in preventing clinical respiratory disease over control calves not receiving the serum.