The effect of vaccination of lambs with live parainfluenza virus type 3 on pneumonia produced by parainfluenza virus type 3 and Pasteurella haemolytica

Groups of 20 lambs were vaccinated with parainfluenza virus type 3 PI₃ by the intranasal (I.N.) or intramuscular route (I.M.).Approximately 6 weeks later, vaccinated and non-vaccinated lambs were challenged sequentially with PL₃, and Pasteurella haemolytica. At slaughter, 10 to 12 days after challenge, 65% of non-vaccinates and 45% of I.M. vaccinates had pneumonic lesions whereas lesions were not observed in any of the I.N. vaccinates.     

Efficacy of an intranasal modified live bovine respiratory syncytial virus and temperature-sensitive parainfluenza type 3 virus vaccine in 3-week-old calves experimentally challenged with PI3V

Two experimental parainfluenza type 3 virus (PI3V) challenge studies were undertaken to evaluate the efficacy of a single intranasal dose of an attenuated live vaccine containing modified live bovine respiratory syncytial virus (BRSV) and temperature-sensitive PI3V in 3-week-old calves. In the first study, vaccine efficacy was evaluated in colostrum deprived calves. Nasal shedding of PI3V was highly significantly reduced in vaccinated calves challenged 10 days or 21 days after vaccination. In the second study, vaccine efficacy was assessed in calves with maternal antibodies against PI3V by challenge 66 days post-vaccination. Vaccination also significantly reduced PI3V excretion after challenge in this study. In both studies, clinical signs after challenge were very mild and were not different between vaccinated and control calves.     

Effects of 4-ipomeanol on bovine parainfluenza type 3 virus-induced pneumonia in calves.

Previous research has demonstrated that 4-ipomeanol toxicosis can enhance the severity of para-influenza virus-induced pneumonia in mice. The objectives of this study were to determine whether calves are susceptible to 4-ipomeanol-induced enhancement of parainfluenza type 3 viral pneumonia and to determine whether 4-ipomeanol alters pulmonary replication of parainfluenza virus. Male Holstein calves were injected with either 4-ipomeanol (3 mg/kg) or vehicle (polyethylene glycol) 3 days prior to intratracheal inoculation with either parainfluenza virus or sham inoculum of culture medium. Calves in the four treatment groups (ipomeanol-parainfluenza, ipomeanol-medium, vehicle-parainfluenza, and vehicle-medium) were necropsied at 5 days after inoculation with parainfluenza virus or medium. The lungs were studied by correlated methods of light and electron microscopy, digitizing morphometry and pulmonary lavage to quantitate the severity of pneumonia. Pulmonary viral titers were determined, and viral antigen was identified in the lung by immunoperoxidase technique. The calves in the ipomeanol-virus treatment group had over a 9-fold higher (P less than 0.05) volume density of virus-induced interstitial pneumonia than did the calves in the other three treatment groups. This 4-ipomeanol-enhanced viral pneumonia was associated with significantly greater (P less than 0.05) numbers of pulmonary macrophages and neutrophils in the lavage fluid and higher (P less than 0.05) pulmonary titers of pulmonary infectious parainfluenza virus. Four-ipomeanol-enhanced viral pneumonia was characterized in part by extensive hyperplasia of type II alveolar epithelial cells and by dense aggregates of macrophages and neutrophils in alveolar spaces and interalveolar septa. The results indicate that 4-ipomeanol exacerbates interstitial pneumonia in calves induced by bovine parainfluenza type 3 virus.     

An inactivated parainfluenza virus type 3 vaccine: the influence of vaccination regime on the response of calves and their subsequent resistance to challenge.

Calves maintained in insolated pens were vaccinated with an inactivated parainfluenza virus type (3) (pi3) vaccine usingparenteral and local route singly and in combination. The calves were subsequently monitored for serum antibody response and challenged intranasally with live virus to assess the protection derived from vaccination. Calves receiving one subcutaneous dose of vaccine in oil adjuvant produced a marked antibody response and were partially protected against challenge. Those receiving two successive subcutaneous doses produced a much greater antiboyd response and were completely protected against challenge. One intranasal dose of aqueous vaccine failed elicit a significant serum antibody response or protection against challenge. However, there was some evidence that intranasal vaccination following a single subcutaneous vaccination produced more effective immunity than one subcutaneous dose alone. Thus a vaccination regime was established which protected calves against experimental challenge and which could thefore be used in the field to assess the role of Pi3 virus in calf respiratory disease.     

Observations on outbreaks of respiratory disease in calves associated with parainfluenza type 3 virus and respiratory syncytial virus infection.

In four outbreaks of indoor calf pneumonia, dyspnoea was a prominent clinical finding. At necropsy it was associated with pneumonia involving the cranial lobes of the lung and severe pulmonary emphysema. Histological examination of lung tissue revealed bronchiolitis and alveolitis with alveolar epithelial cell hyperplasia and multinucleate syncytium formation. Intraalveolar haemorrhage, intra-alveolar oedema and hyaline membrane formation were also noted. In all cases parainfluenza type 3 (PI3) virus was isolated from the lungs. In each of the four outbreaks there was evidence of PI3 virus and respiratory syncitial virus (RSV) infection.     

Studies of the effect of recombinant human-alpha 1 interferon on experimental parainfluenza type 3 virus infections of the respiratory tract of calves

The effect of the administration of recombinant human interferon on the severity of clinical disease and the extent of pneumonic lesions in calves infected experimentally with bovine parainfluenza 3 (PI3) virus was studied in two experiments. In the first, three pairs of calves aged seven to 10 days were used; one of each pair was injected intramuscularly with 10(6) units of interferon/kg bodyweight for three consecutive days, and the other was left untreated. On the day after the first injection of interferon all the calves were challenged with PI3 virus, a different dose being administered to each pair. There was no evidence of any protective effect from the treatment with interferon. The second experiment used eight, six-week-old calves; four were inoculated in the same way with interferon and all the calves were challenged with the same dose of PI3. Again, there was no evidence of a reduction either in the severity of clinical disease or in the extent of lung consolidation in the calves treated with interferon.     

Immunofluorescence in the serological diagnosis of parainfluenza type 3 and respiratory syncytial virus infection in calves

The fluorescent antibody (FA) test is compared with the haemagglutination inhibition (HI) test for parainfluenza virus type 3 (PI-3) and virus neutralisation (VN) test for respiratory syncytial (RS) virus for detection and titration of virus-specific antibodies. In experimentally inoculated calves PI-3 and RS virus FA tests detected seroconversion at the same time as HI and VN tests, however, in serially diluted sera, the FA test was positive to higher dilution. In studies with paired samples from calves from four farms with respiratory problems, the FA test gave similar results to PI-3 HI and RS virus VN tests. Large increases in antibody titre to RS virus detected by FA and VN tests indicated this was the problem on two of the farms. Individual animals showed large rises to PI-3 by FA and HI test on three farms. It is concluded that the FA test provides a rapid and sensitive alternative to the more conventional serological tests for respiratory viruses.     

Serological studies of parainfluenza type 3 virus, bovine adenovirus type 3 and bovine respiratory syncytial virus infection in beef calves

Six serum samples were taken at monthly intervals from birth to weaning from each of 41 newborn calves in the autumn and spring calf crops of a beef cow--calf herd. The serum hemagglutination-inhibition (HI) antibody titres to parainfluenza type 3 virus (PIV-3), virus-neutralization (VN) antibody titres to bovine adenovirus type 3 (BAV-3) and bovine respiratory syncytial virus (BRSV) were determined using microtitration techniques. There was serological evidence of a significantly higher incidence of infection with BAV-3 in the fall calves than in the spring calves. Serological responses to BAV-3 were not detected in calves with VN titres of greater than 1/256. Serological evidence of subclinical infection with PIV-3 occurred mainly in late February or early March during a period of marked environmental temperature fluctuations. Serological evidence of a high incidence of infection with BRSV was obtained for both the fall and spring calf crops. Serum antibody appeared to be unable to prevent infection with BRSV. An association between infection with BRSV and clinical pneumonia was found in 3 out of 9 calves. BAV-3 infection was related to pneumonia in only 1 instance; however, there was simultaneous evidence of BRSV infection in this calf. PIV-3 infection was found to be related to pneumonia in only 1 instance. There was serological evidence of infection with BAV-3 in association with the occurrence of diarrhea in 3 calves.     

Effect of experimentally induced type II bovine viral diarrhea virus infection on platelet function in calves

OBJECTIVE: To evaluate platelet aggregation responses in calves experimentally infected with a thrombocytopenia-inducing type II bovine viral diarrhea virus (BVDV) isolate (BVDV 890). ANIMALS: 9 neonatal male Holstein calves. PROCEDURE: 5 calves were inoculated with BVDV 890, and 4 were used as controls. Platelet aggregation studies and attempts to isolate BVDV from platelets were performed 2 days before, the day of, and every 2 days for 12 days after inoculation. Platelet function was assessed by means of optical aggregometry, using adenosine diphosphate and platelet-activating factor as agonists. Bovine viral diarrhea virus was isolated from purified platelet preparations by use of an immunoperoxidase monolayer assay. RESULTS: Maximum percentage aggregation and slope of the aggregation curve decreased over time in calves infected with BVDV. Bovine viral diarrhea virus was not isolated from platelets from control calves, but it was isolated from infected calves from 4 through 12 days after inoculation. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that platelet function may be depressed in calves infected with type II BVDV. Although the mechanism for altered platelet function was not determined, it likely involved an increase in the percentage of aged platelets in the circulation, a direct virus-platelet interaction, or an indirect virus-platelet interaction. Platelet dysfunction, in addition to thrombocytopenia, may contribute to the hemorrhagic syndrome associated with acute type II BVDV infection in calves.     

Effects of vaccination of calves against induced Haemophilus somnus pneumonia

The efficacy of a killed whole-cell Haemophilus somnus bacterin against induced H somnus pneumonia was examined in 10-week-old male calves. Twenty calves were assigned to 1 of the 3 following groups: group 1, nonvaccinated controls (n = 4); group 2, vaccinated once (n = 8); and group 3, vaccinated twice 14 days apart (n = 8). The serum antibody response to vaccination and challenge exposure was evaluated by the bacterial agglutination test and solid-phase immunoassay (SPIA). Vaccinating calves twice, 14 days apart, significantly (P less than 0.05) reduced the severity of clinical signs of pneumonia and gross lesions. Deaths occurred in 1 of 4 nonvaccinated controls, 1 calf vaccinated once, and none of the calves vaccinated twice, 14 days apart. Postvaccination bacterial agglutination titers measured 14 days after the final vaccination were not significantly different between groups 2 and 3, but SPIA titers were significantly (P less than 0.05) higher in groups 2 and 3, compared with those in group 1. The less severe clinical signs of pneumonia observed in group-3 calves, compared with those in calves in groups 1 and 2, were significantly (P less than 0.01) correlated to higher SPIA titers, indicating the protective value of vaccinating twice.     

Clinical and pathologic studies of experimentally induced Pasteurella haemolytica pneumonia in calves

Pasteurella haemolytica pneumonia of the right caudal lung lobe was experimentally induced in 2-week-old Holstein calves (n = 11) by endobronchial inoculation of 7.9 x 10(10) colony-forming units of 6-hour log-phase bacteria. Calves were studied for 72 hours after inoculation. The challenge procedure consistently induced a lesion in the right caudal lung lobe, which was consistent radiographically with results of pathologic examination and a similar volume of bronchography contrast medium. Clinically, the calves developed a significant increase in rectal temperature within 24 hours after inoculation. Seventy-two hours after inoculation, the total WBC counts, absolute band neutrophil counts, monocyte counts, and blood fibrinogen concentrations were significantly higher than normal and albumin concentration was significantly decreased. Necropsy revealed a circular to oblong lesion that was congested, edematous, and firm and occupied 20 to 40% of the right caudal lung lobe. Histologic examination revealed a severe acute inflammatory reaction characterized by cellular exudate and proteinaceous fluid in the alveoli, interlobular septa, and pleura.     

Aerosol stability of bovine parainfluenza type 3 virus.

Aerosols of bovine parainfluenza type 3 virus were generated with a Devilbiss 40 nebulizer from Eagle's minimum essential medium and nasal secretion from a non-infected calf and stored in a rotating drum at temperatures of 6 degrees C or 32 degrees C and relative humidities of 30% or 90%. The aerosols were sampled at seven minutes, one, two and three hours after the start of generation with an all glass impinger (AGI-30) and titrated for infectivity in cell cultures. Physical decay was determined by a rhodamine tracer technique. Media, temperature or relative humidity had little effect on the survival of parainfluenza type 3 virus during spraying (zero to seven minutes). During aging of aerosols at 32 degrees C and 30% relative humidity, parainfluenza type 3 virus was less stable in Eagle's minimum essential medium than in nasal secretion from a noninfected calf, but at 6 degrees C and 30% relative humidity, the virus was more stable in Eagle's minimum essential medium. At 32 degrees C, the virus was less stable during aging at 90% relative humidity than at 30% relative humidity. The virus was consistently more stable during aging of aerosols at 6 degrees C than at 32 degrees C.     

Characterization of parainfluenza type 3 virus isolated from the lung of a lamb with pneumonia

A virus designated DH-1 was isolated from the lung of a lamb that died during the course of an epizootic of acute respiratory tract disease. The virus contained ribonucleic acid and was sensitive to heat (56 C), lipid solvent, and acid (pH 3.0). Inoculated cell cultures absorbed and culture fluids agglutinated guinea pig erythrocytes. Morphogenic and morphologic characteristics of the isolate were compatible with those of the genus Paramyxovirus of the family Paramyxoviridae. Serologic results indicated that the virus was indistinguishable from prototype parainfluenza virus type 3.     

Prophylactic efficacy of buparvaquone in experimentally induced Theileria annulata infection in calves

The antitheilerial activity of buparvaquone (BW 720C) was evaluated in experimentally induced Theileria annulata infections in cross-bred male calves. T. annulata infections were induced by injecting a suspension of infected ground tick tissue suspension (GUTTS) equivalent to two ticks subcutaneously into each calf. Buparvaquone at a dose of 2.5 mg kg-1 body weight was given as a single injection (intramuscularly) on Day 0 (Group 1), Day 8 (Group 2) and Day 12 (Group 3) post-infection. The animals in Groups 4 and 5 were untreated and challenged controls, respectively. All of the recovered animals from Groups 1-4 were challenged with a lethal dose of T. annulata at 6 weeks post-infection. The immunized animals were resistant to the homologous challenge, which killed three of four control animals (Group 5); the controls showed typical antemortem and post-mortem lesions of theileriosis.     

Experimentally induced parainfluenza type 3 virus infection in young lambs: pathologic response

Pulmonary changes in five 1-week-old, colostrum-deprived lambs transtracheally inoculated with parainfluenza type 3 virus were studied by immunofluorescent, microscopic, and ultrastructural techniques. The lambs were killed at postinoculation days (PID) 3, 5, and 7. Immunofluorescence specific for parainfluenza type 3 virus was first seen in small airways and alveolar epithelium and later in the lumens of airways and alveoli and, to a lesser extent, in the interstitium of the lungs. Grossly, there were multifocal areas of consolidation in all lobes of the lungs. These areas were characterized microscopically by bronchiolitis and interstitial pneumonitis. The bronchiolitis involved the terminal airways and consisted of necrosis and sloughing of epithelial cells followed by hyperplasia of the epithelium. The interstitial lesion comprised extensive infiltration of alveolar septa and alveoli with macrophages and the necrosis of alveolar epithelium. This was followed by hyperplasia of the epithelium. Degenerated bronchiolar and alveolar epithelium contained numerous intracytoplasmic inclusions early in the infection, but such inclusions were not seen in the lambs killed at PID 7. The degenerated changes were also seen with the electron microscope, as were numerous inclusions of viral nucleoprotein and a few viral buds at PID 3 and 5. Viral inclusions and buds were seen in ciliated and nonciliated bronchial epithelial cells and type I and type II alveolar epithelial cells.     

Protection against respiratory disease in calves induced by vaccines containing respiratory syncytial virus, parainfluenza type 3 virus, Mycoplasma bovis and M dispar

A field trial to assess the ability of two vaccines to protect calves against respiratory disease was carried out on a large beef rearing unit in southern England over the two winters of 1983 to 1984 and 1984 to 1985. A quadrivalent vaccine containing the killed antigens of respiratory syncytial virus, parainfluenza virus type 3, Mycoplasma bovis and M dispar or a vaccine containing only the respiratory syncytial virus component were inoculated into 246 and 245 calves, respectively; 245 calves remained as unvaccinated controls. The calves were reared in seven batches and outbreaks of disease occurred in five; significant protection was achieved in the four batches in which disease was associated with respiratory syncytial virus and M bovis infection, together or independently. The death rate from pneumonia was 9 per cent in the control group, 2 per cent in the calves inoculated with the quadrivalent vaccine (P less than 0.001), a protection rate of 77 per cent, and 3 per cent in the calves inoculated with the respiratory syncytial virus vaccine (P less than 0.01), a protection rate of 68 per cent. The proportion of calves receiving treatment for respiratory disease was 38 per cent in the control group, 25 per cent in the calves inoculated with the quadrivalent vaccine (P less than 0.001) and 27 per cent in the calves inoculated with the respiratory syncytial virus vaccine (P less than 0.01). The results show that protection against respiratory disease can be achieved by parenteral vaccination of calves with the appropriate inactivated microorganisms.     

Efficacy of a modified live intranasal bovine respiratory syncytial virus vaccine in 3-week-old calves experimentally challenged with BRSV

Two experimental bovine respiratory syncytial virus (BRSV) challenge studies were undertaken to evaluate the efficacy of a single intranasal dose of a bivalent modified live vaccine containing BRSV in 3-week-old calves. In the first study, vaccine efficacy was evaluated in colostrum deprived (maternal antibody negative) calves 5, 10 and 21 days after vaccination. Nasal shedding of BRSV was significantly reduced in vaccinated calves challenged 10 or 21 days after vaccination. Virus excretion titres were also reduced in vaccinates challenged 5 days after vaccination but reduction in duration of shedding and total amount of virus shed were not statistically significant. Clinical disease after challenge in this study was mild. In the second study, vaccine efficacy was assessed in calves with maternal antibodies against BRSV by challenge 66 days post-vaccination. Vaccination significantly reduced nasal shedding after challenge and the severity of clinical disease was also reduced.