Pestivirus in cattle: experimentally induced persistent infection in calves

Twenty-two heifers were infected intranasally with non-cytopathic bovine viral diarrhoea virus (BVDV) between days 74 and 82 of pregnancy. All animals had developed serum antibodies against BVDV 5 weeks later. No clinical effects were seen in the heifers, and they all delivered a live calf. The newborn calves were generally small, appeared unthrifty as typical 'poor doers', and some developed secondary infections with diarrhoea and signs of respiratory disease. Eighteen of the 22 calves were born without antibodies against BVDV and were persistently infected (PI) with the virus. One was weak at birth and died the following day. Four calves were born with serum antibodies against BVDV and with no detectable virus. Three of these showed signs and/or pathological changes indicating disease in the central nervous system. Otherwise, there were no obvious clinical differences between these calves and the PI calves, nor were there any apparent significant differences in blood parameters between these groups. In general, the calves showed low gamma-globulin values and thrombocytopaenia, but moderately increased fibrinogen values and relatively normal lymphocyte numbers.     

Clenbuterol hydrochloride in calves with a natural bovine respiratory syncytial virus infection

The effect of clenbuterol hydrochloride on the course of disease in calves with a natural bovine respiratory syncytial virus infection was examined. Six calves (three to nine months of age) originating from four herds with respiratory tract disease and serological evidence of a bovine respiratory syncytial virus infection were used in this study. The calves were injected intravenously with clenbuterol hydrochloride. The effect of clenbuterol on the course of disease was measured using the PO2 in blood taken from an indwelling canula inserted in the caudal auricular artery and by clinical signs. Clenbuterol did not improve clinical signs. After clenbuterol administration arterial PO2 values decreased significantly in five out of six patients. Six to eight hours after medication the mean arterial PO2 values were higher than initial values. The moderate positive effect of clenbuterol after six to eight hours may be caused by enhancing ciliary activity and by the secretolytic activity of clenbuterol.     

Characterization and identification of a bovine respiratory syncytial virus isolated from young calves.

Two identical viruses designated 371 and 375 were recovered from nasal secretions of 2 of 7 calves in a beef cow-calf herd in which calves (45 to 105 days of age) had signs of acute respiratory tract disease. The cytopathic, morphologic and physico-chemical characteristics of the isolates were those of bovine respiratory syncytial virus. Although a humoral antibody response to bovine respiratory syncytial virus was not observed, it was concluded that this virus probably had a part in the respiratory tract disease in these calves.     

Immunofluorescent Studies of Bovine Para-Influenza 3 Virus II. Experimentally Infected Calves

Fluorescent antibody (FA) studies of tissues from three colostrum deprived calves inoculated intranasally with the SF-4 strain of bovine para-influenza 3 (PI-3) virus indicated that these calves developed a mild upper respiratory infection but infected cells were not identified in the lower respiratory tract. Three other calves inoculated intranasally and intratracheally with PI-3 virus developed more severe clinical signs of infection and virus was identified, by FA techniques, in the upper and lower respiratory tract of all three calves and in the spleen of one calf. PI-3 virus was detected in smears of nasal epithelium from five of six calves at some time during the observation period.     

Experimental production of bovine respiratory tract disease with bovine viral diarrhea virus

Five 6-month-old calves were inoculated with bovine viral diarrhea (BVD) virus (n = 3) or Pasteurella haemolytica (n = 2) endobronchially with a fiberoptic bronchoscope. Five additional calves were inoculated sequentially with BVD virus followed by P haemolytica at a 5-day interval. Blood samples were collected daily from the calves for bacterial isolation. Clinical signs of respiratory tract disease in calves were recorded daily. If the calves survived, they were killed for necropsy 3 or 4 days after inoculation with P haemolytica (or 8 days after inoculation with BVD virus). The extent and nature of pulmonary lesions in the calves were determined, and the lower portion of the respiratory tract (lungs and trachea) was examined for both these organisms. The 3 calves, inoculated with BVD virus only, developed mild clinical signs mainly manifested as fever, nasal discharge, and occasional cough. Approximately 2% to 7% of the total lung capacity of these calves was pneumonic. Mild clinical signs and localized lesions involving about 15% of the lung volume developed in the 2 calves exposed to P haemolytica only. However, severe fibrinopurulent bronchopneumonia and pleuritis involving 40% to 75% of lung volume developed in the 5 calves inoculated sequentially with BVD virus and P haemolytica. The possible role BVD virus may have in bovine respiratory tract disease is discussed.     

Mucosal and systemic isotype-specific antibody responses to bovine coronavirus structural proteins in naturally infected dairy calves.

Blood, feces, nasal secretions, and tears were collected weekly from 5 randomly selected 1- to 8-week-old calves in a large commercial dairy herd. Clinical signs and bovine coronavirus (BCV) shedding from the respiratory and enteric tracts of calves were monitored through the 8-week period by direct immunofluorescence of nasal epithelial cells, protein A-gold immunoelectron microscopy on feces, and ELISA on nasal secretions and feces. All samples were analyzed for antibody isotypes to BCV structural proteins by immunoblotting. All calves had BCV respiratory tract infections and 4 of 5 calves shed virus in feces. Several calves had multiple or prolonged periods of BCV respiratory tract or enteric tract shedding or both. All calves (except 1) had passive IgG1 antibodies to some BCV proteins (mainly the E2 and E3 proteins) in their serum when they were 1 week old. The presence of these passive serum antibodies (mainly to the E2 and E3 BCV proteins) was associated with decreased or delayed systemic and mucosal antibody responses in calves, in particular IgA responses in nasal secretions and tears to the E2 and E3 BCV proteins, but not to the N protein. Moderate amounts of maternal BCV E2- and E3-specific antibodies in serum did not prevent BCV enteric tract or respiratory tract infections in calves, but may have delayed the development of active antibody responses to these BCV proteins. However, calves with BCV respiratory tract or enteric tract infections had no detectable passive antibodies to any BCV proteins in nasal secretions or feces.     

Association of bovine respiratory disease with clinical status and acute phase proteins in calves

Eighty-four calves with respiratory disease from 18 herds in different parts of Finland were chosen for a study evaluating the capacity of different respiratory pathogens to cause changes in different acute phase protein concentrations, white blood cell (WBC) count and clinical signs. The selected acute phase proteins were fibrinogen, haptoglobin, serum amyloid-A, lipopolysaccharide binding protein and alpha1-acid glycoprotein. From each calf, a paired blood sample was obtained for serological studies of bovine parainfluenza virus-3, bovine respiratory syncytial virus, bovine coronavirus, bovine adenovirus-3 and bovine adenovirus-7. Tracheobronchial lavage was performed to detect bacteria and mycoplasma. Isolation of Pasteurella multocida was associated with increased concentrations of all tested acute phase proteins. For other pathogens, no significant relationships were observed. No association was present between viral or bacterial findings and WBC count.     

A paravaccinia virus isolated from cattle.

Isolation of viruses from calves with acute respiratory tract disease were attempted on bovine embryonic lung cell cultures. An isolate obtained from one calf with oral lesions and respiratory disease, designated 44-M-E482, was characterized as a paravaccinia virus on the basis of biological and physical properties. The calf from which the paravaccinia virus 44-M-E482 was isolated did not possess serum neutralizing antibody in its convalescent sera; neither did experimentally inoculated calves possess serum neutralizing antibody to the isolate. However, a low titer of serum neutralizing antibody was produced in one calf after several intravenous injections of the virus. Inoculation of calves with 44-M-E482 into the oral mucosa, skin, nasal cavity and pharynx did not cause any noticeable illness or lesions. The relation of 44-M-E482 to the viruses which cause bovine papular stomatitis and pseudocowpox is discussed.     

Experimental bovine respiratory tract disease with Haemophilus somnus

Eight calves were inoculated into the bronchus with H. somnus. Thirteen calves were inoculated with bovine respiratory syncytial virus (BRSV) and 8 days later with H. somnus. All calves developed necrotizing, suppurative, lobular bronchopneumonia and pleuritis. Clinical signs of disease and pneumonic lesions were significantly more severe in calves that were sequentially inoculated with BRSV followed by H. somnus. Pneumonic lesions in the inoculated calves were similar to those described for naturally occurring H. somnus-associated respiratory tract disease. Control calves inoculated with BRSV alone or sham-inoculated with medium did not develop clinical signs of respiratory tract disease. The BRSV-inoculated control calves developed minimal pneumonic lesions.     

Seroepizootiologic study of bovine respiratory syncytial virus in a dairy herd

A 16-month seroepizootiologic study of bovine respiratory syncytial virus (BRSV) infection was conducted in a dairy herd. Results indicated that antibodies to BRSV present in serum from newborn calves were derived through the ingestion of colostrum. This passive immunity in calves became undetectable in an average of 99 days (SD = 36.5; range = 30 to 208 days). Two epizootics of respiratory tract disease occurred during the study period, and an association with BRSV was demonstrated in both epizootics. In the 2 epizootics, clinical signs of respiratory tract disease were only mildly to moderately severe, with no mortality or evidence of chronic pneumonia occurring. Seemingly, the passive immunity failed to protect calves from infection and disease caused by BRSV. Additionally, it was observed that if active immunity was induced by infection with BRSV, this immunity protected from the development of clinical disease, but not from reinfection upon subsequent exposures to BRSV.     

Quantitation of bovine macrophage colony-stimulating factor in bovine serum by ELISA

We established an enzyme-linked immunosorbent assay (ELISA) system for the quantitation of bovine macrophage colony-stimulating factor (M-CSF) and used it to measure the serum M-CSF levels in bovine fetuses and calves. The average serum M-CSF level was 2.7+/-1.5 ng/ml in 39 calves under 100 days old, and 1.8+/-0.8 ng/ml in 15 cattle between 101 and 418 days old. Fetal sera samples (n = 6) prepared from cattle between 150 and 280 days of gestational age had a higher average level of M-CSF (8.8+/-1.4 ng/ml). Alteration in serum M-CSF levels in each individual calf was also measured. The serum levels of M-CSF in calves at 0-1 day after birth ranged from 0.52 to 7.3 ng/ml. During the period 113-125 days after birth, serum levels were around 1.4+/-0.39 ng/ml. Although serum M-CSF levels generally decreased as the age of calves advanced, differences among individuals, especially among newborn calves, were observed.     

Epidemiologic factors and isotype-specific antibody responses in serum and mucosal secretions of dairy calves with bovine coronavirus respiratory tract and enteric tract infections.

Blood, feces, and nasal swabs specimens were collected 12 to 24 hours after birth and then 3 times/week (blood only once per week) from one group of 10 calves until they were 10 weeks old and from a second group of 10 calves until they were 10 to 20 weeks old. Colostrum was collected from all calves' dams and tears from 5 randomly selected calves in the first group. All fecal and nasal specimens were assayed for bovine coronavirus (BCV) antigens by ELISA. Nasal epithelial cells were examined for BCV antigens by direct immunofluorescence. Isotype antibody titers to BCV in all samples from 5 calves in group 1 were evaluated by ELISA. Zinc sulfate turbidity (ZST) values were determined on the first serum samples taken from all calves in group 1. To determine whether any correlation existed between ZST values, isotype antibody titers to BCV (12 to 24 hours after birth), number of respiratory sick days, number of enteric sick days, or days to first shedding of virus, a Spearman rank order correlation coefficient was done. Bovine coronavirus respiratory tract and enteric tract infections were common on this farm. Most initial infections developed when calves were 1 to 3 weeks old; however, there were also multiple incidences of shedding of viral antigens or seroconversions at later times during the study. Persistence of infection or reinfection of the upper respiratory tract with BCV was common. Colostral antibody titers to BCV (IgG1) were in all cows at moderate amounts; however, calf serum antibody titers and ZST values (12 to 24 hours after birth) were highly variable.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of bovine fibroblast interferon on rhinovirus infection in calves.

The prophylactic/therapeutic activity of natural bovine fibroblast interferon (BoF-IFN) against bovine rhinovirus infection in calves was assessed. Six calves were each given 8 intranasal inoculations of partially purified BoF-IFN (3.25 x 10(5) U at 8 AM, 11 AM, 5 PM, and 8PM on day 1 and 8 AM, 11 AM, 2 PM, and 5PM on day 2), and 6 calves were given placebo. All calves were challenge exposed with 10(5.1) TCID50 of bovine rhinovirus after the first 2 treatments (6 hours after the first IFN or placebo treatment). Nasal excretion of rhinovirus, IFN concentration in the nasal secretions, and nasal secretion and serum rhinovirus antibodies were measured before and at selected times after calves were inoculated. Interferon-treated calves excreted rhinovirus in their nasal secretions in lesser amounts (mean value, 0.84 log10 TCID50/ml vs 1.58 log10 TCID50/ml on postchallenge exposure days 1 and 2; (P less than 0.05) and for a shorter duration (P less than 0.05) than did placebo-treated calves. No calves developed clinical signs of respiratory tract illness. Rhinovirus antibody titer was not significantly different between IFN- and placebo-treated calves.     

Effect of infection with bovine respiratory syncytial virus on pulmonary clearance of an inhaled antigen in calves

OBJECTIVE: To evaluate the effect of infection with bovine respiratory syncytial virus (BRSV) on clearance of inhaled antigens from the lungs of calves. ANIMALS: Eleven 6- to 8-week-old Holstein bull calves. PROCEDURES: Aerosolized (99m)technetium ((99m)Tc)-labeled diethylene triamine pentacetate (DTPA; 3 calves), commonly used to measure integrity of the pulmonary epithelium, and (99m)Tc-labeled ovalbumin (OA; 8 calves), commonly used as a prototype allergen, were used to evaluate pulmonary clearance before, during, and after experimentally induced infection with BRSV or sham inoculation with BRSV. Uptake in plasma (6 calves) and lung-efferent lymph (1 calf) was examined. RESULTS: Clearance of (99m)Tc-DTPA was significantly increased during BRSV infection; clearance of (99m)Tc-OA was decreased on day 7 after inoculation. Clearance time was correlated with severity of clinical disease, and amounts of (99m)Tc-OA in plasma and lymph were inversely correlated with clearance time. Minimum amounts of (99m)Tc-OA were detected at time points when pulmonary clearance of (99m)Tc-OA was most delayed. CONCLUSIONS AND CLINICAL RELEVANCE: BRSV caused infection of the respiratory tract with peak signs of clinical disease at 7 or 8 days after inoculation. Concurrently, there was a diminished ability to move inhaled protein antigen out of the lungs. Prolonged exposure to inhaled antigens during BRSV infection may enhance antigen presentation with consequent allergic sensitization and development of chronic inflammatory lung disease. IMPACT FOR HUMAN MEDICINE: Infection of humans with respiratory syncytial virus early after birth is associated with subsequent development of allergic asthma. Results for BRSV infection in these calves suggested a supportive mechanism for this scenario.     

A bovine virus diarrhea calfhood vaccination trial in a persistently infected herd: effects on titres, health and growth.

A controlled calfhood vaccination trial to prevent bovine virus diarrhea was conducted in a 100 head cow-calf operation with a three year history of annual calf losses due to enteric bovine virus diarrhea (persistently infected herd). Approximately 50% of the calves were vaccinated at six, 12 and 24 weeks of age. Paired serum samples and growth data were collected on three occasions for comparison between vaccinates and controls. Three vaccinated calves died of enteric bovine virus diarrhea in the first year of the trial and one nonvaccinated calf died in the second year. Two of the three vaccinated calves had developed bovine virus diarrhea virus neutralization antibody titres of 2048 or greater before developing clinical signs. The control and third vaccinated calf failed to seroconvert before dying of enteric bovine virus diarrhea. Approximately 90% of the vaccinated calves seroconverted compared to approximately 40% of the controls. Paired serum samples collected from 75% of the cows in the spring, summer and fall of each year of the trial, showed persistent high bovine virus diarrhea virus neutralization titres in all samples. Calf vaccination before 12 weeks of age had little effect on seroconversion due to high levels of passive antibody to bovine virus diarrhea. Growth data showed that there was no improvement in weight gain or rate of growth in the vaccinated calves.     

Bovine herpesvirus-1 and parainfluenza-3 virus interactions: clinical and immunological response in calves.

Calves infected with bovine herpesvirus-1 (BHV-1) or both BHV-1 and parainfluenza-3 virus (PIV-3) developed clinical signs including fever, cough, and nasal and ocular discharges. Animals infected with both viruses appeared more depressed and showed higher rectal temperature, while calves inoculated with PIV-3 alone had a very mild clinical disease. Both BHV-1 and PIV-3 were recovered from nasal secretions up to six to eight days postinoculation. However, the virus titers were lower in calves with mixed infection. An increase in serum antibodies to both BHV-1 and PIV-3 was detected by serum neutralization and enzyme-linked immunosorbent assay. Antibody responses were delayed and significantly lower in calves given mixed infection than in calves infected with a single virus. Interleukin-2 activity in cultures of lymphocytes from BHV-1 and BHV-1 plus PIV-3 infected calves was higher compared to control calves.     

Priming for local and systemic antibody memory responses to bovine respiratory syncytial virus: effect of amount of virus, virus replication, route of administration and maternal antibodies

We studied the conditions under which calves can be primed for mucosal and serum antibody memory responses against bovine respiratory syncytial virus (BRSV), and the relationship between such responses and protection against the virus. Calves were primed via the respiratory tract with a low or high amount of live virus, with killed virus, or intramuscularly with live virus. Calves were challenged via the respiratory tract. Priming with live virus via the respiratory tract induced primary antibody responses in serum and on the mucosae, which were identical after the low and the high amount of virus. These responses were suppressed by maternal antibodies. Intramuscular priming of seronegative calves induced serum IgG1 and sometimes serum IgM and IgG2 responses, but no responses were detected on the mucosae. Sera of calves primed by the intramuscular or the respiratory route recognized the same viral proteins. No responses were observed after priming with killed virus, or after intramuscular priming of calves with maternal antibodies. After challenge, mucosal and serum antibody memory responses developed in calves that had been primed via the respiratory tract with live virus, whether they had maternal antibodies or not. One colostrum-fed calf showed a mucosal memory response, although serum responses were still suppressed by maternal antibodies. None of the calves thus primed shed virus after challenge. Intramuscular priming also primed for mucosal and serum memory responses after challenge, which however started perhaps slightly later and were not associated with protection against virus shedding. Priming with killed virus, or with live virus intramuscularly in the presence of maternal antibodies proved least effective in inducing memory and protection against virus shedding. Thus, protection against virus shedding was afforded by priming with live virus via the respiratory tract, both in calves with an without maternal antibodies. Protection was associated with a strong and rapid mucosal antibody memory response, but the reverse was not necessarily true. Protection against virus excretion had no relationship to titers of serum neutralizing or serum IgG1 or nasal IgA antibodies at the time of challenge.     

Efficacy of a saponin-adjuvanted inactivated respiratory syncytial virus vaccine in calves

The objective of this study was to determine whether a commercially available, saponin-adjuvanted, inactivated bovine respiratory syncytial virus (BRSV) vaccine would protect calves from experimental infection with virulent BRSV. This was a randomized controlled trial comprising 14, 8- to 9-week-old calves seronegative for BRSV Group 1 calves (n = 8) were not vaccinated and group 2 calves (n = 6) were vaccinated on days 0 and 19 with an inactivated BRSV vaccine. All calves were challenged with virulent BRSV on day 46. Clinical signs, arterial PO2, and immune responses were monitored after challenge. Calves were euthanatized on day 54 (8 d after challenge) and lungs were examined for lesions. Vaccination elicited increases in BRSV-specific immunoglobulin (Ig) G and virus neutralizing antibody titers. Challenge with BRSV resulted in severe respiratory tract disease and extensive pulmonary lesions in control calves, but no signs of clinical disease and minimal or no pulmonary lesions in vaccinated calves. Arterial blood oxygen values on day 53 (7 d after challenge) in control calves were significantly lower than those in vaccinated calves, which remained within normal limits. Control calves shed BRSV for several days after challenge, whereas BRSV was not detected on deep nasal swabs from vaccinated calves. In summary, the results indicated that this inactivated BRSV vaccine provided clinical protection from experimental infection with virulent virus 27 d after vaccination and significantly decreased the prevalence and severity of pulmonary lesions. Efficacy was similar to that reported for other commercial inactivated and modified-live BRSV vaccines.     

Efficacy of an inactivated respiratory syncytial virus vaccine in calves.

OBJECTIVE: To determine whether an inactivated bovine respiratory syncytial virus (BRSV) vaccine would protect calves from infection with virulent BRSV. DESIGN: Randomized controlled trial. ANIMALS: 27 nine-week-old calves seronegative for BRSV exposure. PROCEDURE: Group-1 calves (n = 9) were not vaccinated. Group-2 calves (n = 9) were vaccinated on days 0 and 21 with an inactivated BRSV vaccine containing a minimum immunizing dose of antigen. Group-3 calves (n = 9) were vaccinated on days 0 and 21 with an inactivated BRSV vaccine containing an amount of antigen similar to that in a commercial vaccine. All calves were challenged with virulent BRSV on day 42. Clinical signs and immune responses were monitored for 8 days after challenge. Calves were euthanatized on day 50, and lungs were examined for lesions. RESULTS: Vaccination elicited increases in BRSV-specific IgG and virus neutralizing antibody titers and in production of interferon-gamma. Virus neutralizing antibody titers were consistently less than IgG titers. Challenge with BRSV resulted in severe respiratory tract disease and extensive pulmonary lesions in control calves, whereas vaccinated calves had less severe signs of clinical disease and less extensive pulmonary lesions. The percentage of vaccinated calves that shed virus in nasal secretions was significantly lower than the percentage of control calves that did, and peak viral titer was lower for vaccinated than for control calves. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the inactivated BRSV vaccine provided clinical protection from experimental infection with virulent virus and decreased the severity of pulmonary lesions. Efficacy was similar to that reported for modified-live BRSV vaccines.     

Experimentally induced pneumonic pasteurellosis: dose-response relationships and protection against natural reinfection in calves

Calves were inoculated intranasally with 2 X 10(6.2) tissue culture infective doses of infectious bovine rhinotracheitis virus, followed in 7 days by intratracheal inoculations with 1 of 4 challenge doses of pathogenic Pasteurella haemolytica. Severity and duration of the ensuing clinical signs of respiratory tract disease were correlated with the challenge dose of bacteria. Calves given 1 X 10(6) colony-forming units (CFU) of bacteria did not develop reliable clinical evidence of disease, whereas those given 1 X 10(8) CFU or 1 X 10(10) CFU of bacteria developed clinical signs of pneumonic pasteurellosis within 12 to 24 hours of bacterial challenge. Severity of clinical signs was equal at the 10(8) and 10(10) doses of bacteria, but duration of clinical signs was greater in calves given the 10(10) dose. Calves given 1 X 10(12) CFU of bacteria developed relatively severe respiratory tract disease in excess of what was necessary for positive clinical detection. Positive correlations were found between the bacterial challenge dose and the height and duration of increased rectal temperature, amount and duration of increases in ocular and nasal discharges, and the subjective evaluation of depressed attitude and appetite. Correlations were not found between challenge dose and respiratory rate or character, or between challenge dose and complete blood cell count. Convalescent calves were resistant to naturally occurring pneumonic pasteurellosis, which caused severe disease in nontreated calves. Adverse effects of P haemolytica were not observed after the first 4 to 15 days after bacterial administration; however, the bacteria were isolated from nasal secretions of convalescent calves 89 to 116 days after bacterial inoculation.(ABSTRACT TRUNCATED AT 250 WORDS)