Isolation of Haemophilus agni from Six Alberta Ram Lambs with Septicemia

Six ram lambs were submitted to a veterinary diagnostic laboratory for necropsy. Clinical signs included sudden illness or death with or without observed depression, reluctance to move, scours or fever. Gross findings and histopathology revealed evidence of bacterial septicemia. Haemophilus agni was isolated from brain, spleen, lung and lymph node.     

Haemophilus somnus isolated from a urachal abscess in a calf

A urachal abscess was diagnosed in a 2-month-old, crossbred heifer that was presented for a distended abdomen and clinical signs of choking. Cultures of the mucopurulent exudate, obtained from within the mass on necropsy, yielded Haemophilus somnus. This is the first known documented report of H. somnus isolated from a urachal abscess.     

Isolation of pathogenic strains of Haemophilus somnus from the female bovine reproductive tract.

The prevalence of Haemophilus somnus in the genital tract of slaughtered and live cows in southern Ontario was investigated. The vagina and uterus of slaughtered cows were swabbed separately. Live cows were examined and sampled in two field surveys: Centre A and Centre B. In the former, aspirated mucus secretions and in the latter, specimens obtained by guarded swabbing were examined bacteriologically. Haemophilus somnus was isolated from 28 genital tracts of 461 slaughtered (6.1%), and seven of 199 live (3.5%) cows during the centre B survey. The isolates were recovered from both normal and diseased reproductive tracts. Fourteen strains isolated from genital organs were examined for pathogenicity in vivo to test the occurrence of pathogenic isolates. In the initial stage of the in vivo study on pathogenicity, each of the fourteen isolates was examined on one calf using an intracisternal inoculation. Subsequently, one pathogenic and one nonpathogenic strain were inoculated into five calves each to statistically confirm their pathogenic potential. Of 14 genital isolates of H. somnus examined in an intracisternal calf assay, six (43%) caused a fatal peracute neurological disease, while eight were nonpathogenic. A comparative pathological study of pathogenic and nonpathogenic isolates showed that the former caused a severe fatal suppurative meningoencephalitis whereas the latter caused no lesions whatsoever or a mild leukocytic leptomeningitis. The salient data obtained in this study indicate that there are pathogenic strains of H. somnus in the genital tract of apparently normal cows as well as of those with inflammatory disease.     

Haemophilus somnus complex: antigenicity and specificity of fractions of Haemophilus somnus.

Five Haemophilus somnus type 8025 preparations (whole cell, sonicate, crude polysaccharide, purified polysaccharide, and protein) were produced for studies of their antigenicity in rabbits. Bacterial agglutination and passive hemagglutination tests were used to assess the level of antibody produced in rabbits inoculated with the different antigenic preparations. Cross-reactions were seen between the antiserums against the H sumnus 8025 antigens and a variety of related and unrelated bovine pathogens. The strongest cross-reaction occurred between antiserums against H somnus 8025 whole cell and crude polysaccharide antigens and Haemophilus agni and Actinobacillus lignieresii cell suspensions.     

Ovine Haemophilus somnus: experimental intracisternal infection and antigenic comparison with bovine Haemophilus somnus.

Experimental infection was produced by two of four isolates of ovine Haemophilus somnus given by intracisternal inoculation into two to three-month-old lambs. Isolate 2041 (originally obtained from a septicemic lamb in Alberta) caused lethal infection in eight of nine lambs, isolate 67p from the prepuce of a normal lamb produced less acute disease in four of nine lambs, and the other two isolates (93p and 1190) caused no detectable disease. Significant lesions were limited to the brain and spinal cord. Purulent meningitis was characteristic but vasculitis or septicemia were not detected, perhaps due to the route of inoculation. Since a difference in virulence was noted among strains, we analyzed surface proteins thought to be virulence factors of bovine H. somnus. Protein profiles of bovine and ovine H. somnus done by sodium dodecyl sulphate-polyacrylamide gel electrophoresis showed similar patterns for virulent bovine isolates and ovine septicemic isolates. Preputial isolates showed a lower molecular mass major outer membrane protein than septicemic isolates. Antigenic analysis revealed that outer membrane proteins p270, p78, p76, p40, and p39 were detected in both ovine and bovine isolates except for 1190, which was probably not a true H. somnus isolate. Thus the preputial and septicemic isolates of ovine H. somnus were similar to bovine H. somnus in pathogenicity and in surface antigens.     

Development of a defined medium for Haemophilus somnus isolated from cattle

Nutritional factors that influence the growth of Haemophilus somnus were examined, and a defined medium was developed. Optimal growth of H somnus in broth occurred under conditions of maximum aeration. Nutritional components required for or enhanced growth of most H somnus isolates in the defined medium included uracil, D-glucose, isotonic NaCl, Na2HPO4, nicotinamide, flavin mononucleotide, pantothenic acid, pyridoxine, and a variety of salts and amino acids. The defined medium supported optimum growth of 18 of 21 isolates of H somnus from cattle.     

Haemophilus somnus Mastitis in a Dairy Cow

A clinical case of peracute bovine mastitis is described in which the most severely affected quarter yielded a heavy growth of Haemophilus somnus on culture.     

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.     

Haemophilus somnus as a cause of bronchopneumonia in calves

A bacteriological examination of the pulmonary tissues of calves suffering from bronchopneumonia was complemented by a microaerophilic or anaerobic cultivation. Thirty-two strains (23.7%) of a microorganism hitherto not described in Czechoslovakia were isolated during the examination of 135 calves with disorders of the respiratory system. On the basis of the growth, morphological, biochemical and serological properties, the microorganism was identified as Haemophilus somnus. Chocolate agar supplemented with yeast extract, and blood agar were used for the cultivation. Incubation took place in a microaerophilic medium with 10% carbon dioxide or in an anaerobic medium. Serums against the collection strain of Haemophilus somnus were prepared in rabbits for the serological diagnosis. These antiserums reacted with all tested strains during test tube agglutination but did not enter cross reactions with other bacterial species. The coagglutination test, which was verified, did not provide reliable results.     

Microscopic lesions associated with the isolation of Haemophilus somnus from pneumonic bovine lungs

Sixty-one of 68 sets of bovine lungs from which only Haemophilus somnus was isolated had microscopic lesions of purulent bronchiolitis and bronchopneumonia. In 37 of 61 lungs, the bronchiolar exudates were markedly necrotic with accompanying necrosis of the adjacent bronchiolar epithelium. Bronchiolitis obliterans was prominent in 23 of 28 lungs affected with chronic lesions with abscesses present in seven. Alveolar filling with inflammatory cells (neutrophils with fewer macrophages) was limited to peribronchiolar alveoli in 25 of 61 lungs and was multifocal to diffuse in the other 36. Lesions in the remaining lungs (7 of 68) were classified as fibrinous pneumonia with bronchiolitis (2), fibrinous pleuritis (2), suppurative interstitial pneumonia with vasculitis (2), and diffuse congestion (1).     

Aerosol challenge of calves with Haemophilus somnus and Mycoplasma dispar

The aim of the study was to examine the ability of Haemophilus somnus and Mycoplasma dispar to induce pneumonia in healthy calves under conditions closely resembling the supposed natural way of infection, viz. by inhalation of aerosol droplets containing the microorganisms. The infections were investigated by recording clinical data, cytokine expression of peripheral blood cells and pathology. Twelve calves were included in the study: Three animals were exposed to H. somnus only, and two to M. dispar only, whereas five were challenged to M. dispar followed by exposure to H. somnus 11-14 days later. Also, one calf was exposed to M. dispar followed by exposure to a sterile saline solution 11 days later, and one calf was only exposed to a sterile saline solution. Just one animal, only challenged with H. somnus, developed a focal necrotizing pneumonia, from which H. somnus was isolated. Thus, the ability of H. somnus and M. dispar to act as primary pathogens under these conditions were minimal and inconsistent.However, a transient rise in body temperature, a marked granulocytosis and increased levels of interleukin-8 in peripheral blood after inoculation with H. somnus indicated a clear systemic response, probably as a consequence of the natural non-specific local and systemic defence mechanisms acting in healthy calves.     

A Comparison of Various Haemophilus somnus Strains

Sixty-eight Haemophilus somnus strains isolated from the bovine in Canada and the U.S.A. were compared. In media enriched with 5% ovine serum, 5% bovine serum and 10% yeast extract, H. somnus fermented glucose, levulose, maltose, mannitol, mannose, sorbitol, trehalose and xylose, but failed to ferment arabinose, dulcitol, galactose, inositol, lactose, raffinose, rhamnose, salicin and sucrose. The organisms acidified litmus milk, produced cytochrome oxidase, indole and hydrogen sulfide (H₂S) and reduced nitrates to nitrites. The motility, methyl-red, acetylmethyl-carbinol urease catalase, citrate, malonate, lysine, ornithine and arginine tests were negative. Haemophilus somnus was resistant to lincomycin, neomycin and triple sulfa, but susceptible to ampicillin, chloramphenicol, streptomycin, penicillin and tetracycline. No antigenic differences were noted between strains when tested against rabbit antisera of eight strains using agglutination, complement-fixation, immunodiffusion and counterimmunoelectrophoresis tests. Low titre cross-reactions were found in the agglutination tests with some of the anti-H. somnus rabbit sera with Actinobacillus lignieresi and Moraxella bovis. No distinct antigenic similarities to nine other species of pathogenic bacteria of animal origin were found. No difference was observed between H. somnus isolates from Ontario and those from western Canada and the U.S.A.     

Epidemiology of Haemophilus somnus in young rams

The prevalence of Haemophilus somnus in the prepuce of young rams was examined. Of 473 rams entering Record of Performance (ROP) stations at 50 days of age, 43 (9.1%) were positive. Average daily gain was not affected by Haemophilus status, but was influenced by breed of ram. Suffolks were predicted to gain 0.515 kg daily compared to 0.427 kg for a group combining all other breeds. Using logistic regression to identify risk factors for individual H. somnus infection, rams in 1989 were 0.382 times as likely to be infected as rams in 1988, and Suffolks were 0.314 times as likely to be infected as the other breeds group, but these factors were not significant at the flock level. Of 80 eligible flocks of origin, 22 (27.5%) were classified as infected with H. somnus, based on rams submitted to the ROP station. Infected flocks contributed 133 rams, 43 (32.3%) of which were positive. There was no association between H. somnus status and lambing percent of the percent of abortions and stillbirths, but there was a statistically significant association with the percent of ewes which failed to lamb. In the model developed, 6% of the bred ewes in noninfected flocks failed to lamb, compared to a rate of 12% in infected flocks. These results suggest H. somnus may influence ewe fertility earlier, rather than later in gestation. Purchasing replacement animals and having cattle on the farm were risk factors for Haemophilus infection in the flock. Where replacements had been purchased within the previous year, the risk of flock infection rose 8.5 times, and on farms having cattle as well as sheep, the risk rose 13.2 times.(ABSTRACT TRUNCATED AT 250 WORDS)