In vitro response of lymphocytes to Micropolyspora faeni extract in cattle.

The in vitro stimulation of small lymphocytes to blast formation, measured by the rate of 3H-thymidine incorporation, was used to study the occurrence of cells sensitive to antigens of Micropolyspora faeni in cattle. M faeni extract induced a significant stimulation index in lymphocytes from the peripheral blood cells of cattle from an endemic area in autumn but rarely in spring. Blood lymphocytes from animals from a non-endemic area tested during the winter period rarely showed a positive reaction or only a relatively weak one. On the other hand, lymph node cells, particularly from bronchial lymph nodes, showed positive results in all investigated animals and even in those from non-endemic areas. In three-months-old calves, positive results were obtained mainly with cells from bronchial lymph nodes. It seems therfore that sensitisation to M faeni antigen is a widespread phenomenon but additional circumstances seem to be required for the clinical manifestation of farmer's lung disease in cattle. The most important factor is probably strong and repeated exposure to the M faeni organism. Whether or not existing reactive lymphocytes against M faeni antigen are directly involved in the pathogenesis of farmer's lung disease in cattle by producing a delayed type reaction remains to be clarified.     

Humoral response of cattle to aerosolized Micropolyspora faeni

Calves (7) were exposed to antigens of Micropolyspora faeni by the aerosol route for 9 weeks. The humoral immune response of calves to M faeni antigens was studied; immunoglobulins (Ig) E, G1, G2, A, and M were measured weekly in serum and nasal secretions by enzyme-linked immunosorbent assay (ELISA). Intradermal injection of antigen was performed during the 6th and 9th weeks; responses were evaluated at 30 minutes, 6 to 8 hours, 24, and 48 hours after injection. Total IgE levels in serum and nasal secretions, evaluated weekly, did not show any elevation. Micropolyspora faeni-specific IgE, IgA, IgG1, and IgG2, but not IgM, were produced by calves exposed to the antigen by the aerosol route; individual variability in magnitude of the response was marked. Thirty-minute skin tests were positive for cytotropic antibody in 2 of 3 aerosol-exposed calves by the 9th week, but delayed-type reactivity was not present. The ELISA test results were compared with those from sera of saline solution aerosol-exposed calves and from a parenterally immunized calf. Comparison of isotype-specific ELISA results obtained from M faeni aerosol-exposed calves with ELISA results from calves exposed to aerosolized ovalbumin according to a similar procedure indicated inherent problems in evaluating immune responses to environmental antigens. Aerosolized M faeni elicited a substantial antibody response. In particular, it is noteworthy that antigen-specific IgE responses were detected.     

Results of experimental immunization of calves with different Pasteurella antigens]

Calves immunised with different Pasteurella antigens (inactivated whole cells, sodium chloride extract) where challenged two weeks after the second immunization with the homologous strain. The intracutaneous application of whole cells of P. haemolytica A1 and P. multocida A was effective. The incidence of pneumonia was reduced and the pneumonic lesions were less severe. The sodium chloride extract was not effective.     

Aerosolized Micropolyspora faeni antigen as a cause of pulmonary dysfunction in ponies with recurrent airway obstruction (heaves)

Ponies with recurrent airway obstruction (principal ponies) and their controls were given aerosolized Micropolyspora faeni antigen via endotracheal tube during a period when the principal ponies were in disease remission. In both groups of ponies, we performed bronchoalveolar lavage (BAL) and measured pulmonary function at base line, and 5 hours after aerosol administration of 30 ml of 0.9% NaCl solution or 30 ml of 1% w/v particulate M faeni antigen in 0.9% NaCl solution. In both groups of ponies, aerosolized M faeni antigen increased WBC count, neutrophil numbers, and albumin concentration in BAL fluid, but macrophage numbers decreased. In the principal ponies, BAL mast cell numbers were decreased 5 hours after administration of M faeni antigen. The M faeni antigen had no effect on the mechanical properties of the lungs or on gas exchange in the control ponies, but did increase respiratory frequency minute ventilation and pulmonary resistance, and decreased arterial oxygen tension in the principal ponies. Changes in pulmonary function were apparent only in the principal ponies, which suggests that neutrophils, per se, do not cause pulmonary dysfunction and that M faeni may be one of the etiologic agents involved in chronic obstructive pulmonary disease.     

Bovine salmonellosis: experimental production and characterization of the disease in calves, using oral challenge with Salmonella typhimurium.

A highly virulent strain of Salmonella tyhimurium was given orally to produce disease experimentally in 21 normal colostrum-fed calves 3 to 9 weeks old. The challenge inoculum varied from 10(4) to 10(11) organisms. The disease was characterized by fever, depressed attitude, and decreased appetite. Many calves given larger challenge dose levels also had diarrheic feces containing mucus, fibrin, and blood. Fecal cultures were positive for salmonella. Septicemia occurred in some calves (9 of 15 calves cultured were positive). Eleven calves died and 10 calves survived challenge exposure. Survival was inversely related to the size of the challenge inoculum and directly related (although to a lesser degree) to age of the calf. White blood cell total and differential counts were variable. Both neutropenia and neutrophilia were observed. Plasma proteins decreased markedly in calves with diarrhea, probably indicating fecal protein loss. Fibrinogen increased during the acute stages of diarrhea.     

Infectious bovine keratoconjunctivitis: experimental induction of infection in calves with mycoplasmas and Moraxella bovis.

Eyes of 14 calves were exposed by conjunctival instillation to cultures of either Mycoplasma conjunctivae (6 calves) or Acholeplasma laidlawii (8 calves). Calves were observed for clinical signs of infectious bovine keratoconjunctivitis (IBK), and eyes were examined for the test organisms by bacteriologic cultural technique for 60 days. Acholeplasma laidlawii became established in the eyes of 5 of 8 calves; M conjunctivae became established in the eyes of 4 of 6 calves. On day 28, eyes of 9 of the 14 calves were exposed by conjunctival instillation to Moraxella bovis, and all developed IBK. Five calves exposed to Moraxenjunctivae or A laidlawii, but not to Mor bovis, did not develop IBK. Four calves not exposed to M conjunctivae or A laidlawii, but exposed to Mor bovis, developed IBK. Mycoplasmas do not have a major role in IBK, but might produce ancillary effects similar to those of infectious bovine rhinotracheitis virus, wind, ultraviolet radiation, dust, and other irritants.     

Bovine pneumonic pasteurellosis: experimental induction in vaccinated and nonvaccinated calves.

A study was undertaken to investigate the effects of the immune response induced by combined aerosol and parenteral vaccination on the lung lesions induced in calves by Pasteurella haemolytica AI. Twenty-four calves, twelve of which had been vaccinated with killed P. haemolytica by aerosol and subcutaneous injection in Freund's complete and incomplete adjuvant were challenged by intratracheal inoculation of live P. haemolytica. Serological response to vaccination was not marked but was best measured by the whole cell agglutination test or by indirect bacterial agglutination rather than by the passive haemagglutination test. Titres of vaccinates were positively correlated with the degree of pneumonic change following challenge while in nonvaccinated controls, titres were negatively correlated with lung lesions. These findings suggest the occurrence of an immunologically mediated hypersensitivity pneumonitis in the lungs of vaccinates and point to the potential efficacy of live bacterial aerosols for stimulation of protective immunity in pneumonic pasteurellosis.     

Colonic anastomosis in calves: an experimental study

Colonic anastomosis was conducted in 12 calves divided equally into four groups. Four suture patterns, viz. single inversion (continuous Connell), double inversion (two rows of continuous Cushing), single eversion (continuous everted mattress), double eversion (single eversion reinforced with simple continuous) were used. In each animal, anastomosis of the same technique was performed at four places. Each technique was evaluated in terms of clinical and gross observations, lumen stenosis and bursting pressure. Observations were made on the 7th, 14th and 28th day after anastomosis. None of the anastomotic techniques showed any untoward incident during the 28 days of postoperative study. Stricture formation at the site of anastomosis was minimal with the single inversion technique whereas it was maximum with the single eversion technique. Pressure for disruption of intestinal segments was higher after inversion than eversion techniques. It was concluded that the one-layer inversion anastomosis technique proved to be the best among the four techniques because (i) of its reduced incidence of adhesions; (ii) it maintains adequate lumen diameter; (iii) it withstands maximum pressure. The double layer inversion technique was conceded to be the second best choice for colonic anastomosis followed by the double eversion and single eversion techniques.     

Sudden cardiac death in calves with experimental heavy infection of Strongyloides papillosus.

For obtaining the preliminary data on the pathogenesis of sudden death in calves naturally heavily infected with Strongyloides papillosus, we monitored 8 Holstein calves experimentally infected with the larvae on electrocardiographic and pneumographic changes. Six calves died suddenly on days 11 to 17 after infection. Sinus tachycardia had been recorded continuously since 1 to 6 days before death. Heart rates increased gradually until death. Since 1 or 2 days before death, various patterns of tachyarrhythmia and bradyarrhythmia had been observed among patterns of sinus tachycardia. Arrhythmias included serious ventricular premature beat, paroxysmal ventricular tachycardia, complete atrioventricular block and so on. The terminal pattern observed suddenly in all of the cases was ventricular arrhythmias consisting of serial ventricular tachycardia, flutter and fibrillation, which were followed by respiratory arrest. Abnormal pneumograms were not obtained before the terminal ventricular fibrillation. Two of 8 calves recovered from the infection, only one of which showed sinus arrest and the second degree of atrioventricular block transiently. We concluded that calves heavily infected with the larvae died due to sudden cardiac arrest.     

Aerosol vaccination of calves with pasteurella haemolytica against experimental respiratory disease.

Three experiments were conducted on calves in which the efficacy of vaccination with live Pasteurella haemolytica in aerosol was tested by challenge with sequential aerosol exposure to bovine herpesvirus 1 and P. haemolytica. Neither single nor multiple aerosol vaccinations protected against the experimental disease. Macroscopically recognizable rhinitis, tonsillitis, tracheitis and pneumonia occurred in both controls and vaccinates. In one experiment as many as three aerosol vaccinations with live P. haemolytica for up to 20 minutes failed to elicit clinical signs in exposed calves. Pasteurella haemolytica was isolated less frequently from tissues of vaccinated calves than from those of nonvaccinated calves. Pasteurella haemolytica was isolated from deep nasal swabs of 4/14 vaccinated calves five and six days after viral exposure. It was concluded that although bovine herpesvirus 1 vaccination has been shown previously to prevent the experimental disease produced by bovine herpesvirus 1-P. haemolytica, live P. haemolytica vaccination by aerosol will not provide the same protection.     

Physiological effects of experimental verminous bronchitis in Friesian calves.

Pulmonary function values were measured in five Friesian calves of five months of age during the patent phase of an experimental moderate lungworm infection and were compared with the pulmonary function values recorded in four control animals. All the nine calves were free of any previous challenge with Dictyocaulus viviparus and were submitted to the same standardized conditions of body conformation, housing, feeding and procedures for pulmonary function testing. A significant increase of respiratory rate, minute ventilation, total pulmonary resistance and power of breathing and a significant decrease of tidal volume, dynamic lung compliance and PaO2 were observed in the infested animals. The absolute intrapleural pressure values were also significantly more negative. The conclusions of the statistical analysis were almost identical when predicted instead of measured pulmonary function values were used in the control group. The clinical, functional and pathological findings in the infested animals were all consistent with the picture of a lower airway obstructive disease.     

Prophylactic use of hyperimmune serum in experimental colisepticaemia in calves.

Evidence is presented to demonstrate the efficacy of an intravenously administered polyvalent hyperimmune serum prepared from bovine serum, in the prevention of experimental colisepticaemia produced by oral challenge with E coli O78: K80(B) in colostrum deprived calves. It is suggested that the protection afforded by this hyperimmune serum is due to high levels of specific antibody demonstrable in the serum of calves post treatment and that protection is given in this instance by antibodies contained in the IgG class of immunoglobulins.     

Hypersensitivity to dietary components in young farm animals: immunisation of calves for IgE antibody responses

An immunisation schedule is described that reproducibly induces long latent period skin sensitising antibody (immunoglobulin) in calves by a combined oral/parenteral route. This response is only directed towards the protein component of the antigen and is boosted upon challenge injection.