Serodiagnosis of Histophilus ovis-associated epididymitis in rams

An ELISA for the detection of antibodies to Histophilus ovis was used to evaluate the association of epididymal lesions in rams with serologic response to His ovis. Comparison of ELISA results for His ovis in groups of rams with epididymal lesions with ELISA results of clinically normal rams (control group) revealed a significant difference (P less than 0.01) between the control group and those rams from which His ovis was isolated. A significant difference (P less than 0.01) was noticed between the control group and rams with lesions from which an organism other than His ovis or Brucella ovis was isolated. Additionally, a significant difference (P less than 0.01) was noticed in ELISA results between the control group and affected rams from which no organism was recovered and in which the epididymal lesion was not limited to the head of the epididymis. A difference was not detected in the His ovis ELISA results between control rams and rams with lesions associated with a B ovis infection or rams from which no organism was recovered and in which the epididymal lesion was limited to the head of the epididymis. The serologic findings in our study suggest that His ovis is more important in the development of epididymitis in rams than culture results alone would indicate.     

Multiplex PCR for the detection of Brucella ovis, Actinobacillus seminis and Histophilus somni in ram semen

OBJECTIVE: To develop a multiplex polymerase chain reaction (PCR) assay for the rapid detection of Brucella ovis, Actinobacillus seminis, Histophilus somni in fresh ram semen samples. DESIGN: The multiplex assay was based on the single PCR assays published for the detection of A seminis and B ovis, and the forward primer published for the detection of H somni; an alternative reverse primer for H somni was designed in this study. PROCEDURE: Culture and PCR of 295 fresh semen samples were carried out. RESULTS: The multiplex PCR was far more successful in the detection of H somni (45/295) than culture (23/295). A seminis was also detected in more semen samples by multiplex PCR (29/295) than culture (13/295) and B ovis was detected in three samples using both PCR and culture. No amplifications were detected with DNA from a range of bacterial isolates including species associated with epididymitis in rams. CONCLUSION: This PCR could be used as a complementary test, or alternative to culture of ram semen and other biological samples for the detection B ovis, H somni and A seminis.     

Ovine Mastitis Due to Histophilus ovis

The clinical, pathological and bacteriological findings related to ovine mastitis caused by Histophilus ovis are described. A high proportion of the ewes belonging to a flock were infected, but the source of the contamination could not be determined.     

Immunity against genital infection by Histophilus ovis in rams

Rams have been immunized against an infection of their genitalia by Histophilus ovis. An alum-precipitated antigen and an antigen plus Freund's complete adjuvant proved equally effective. An injection of live H. ovis into the epididymal tissue proved to be a better method of challenging immunity than an injection into the vas deferens. It was shown that cell-mediated immunity, as evidenced by tests for lymphocyte transformation, the presence of a macrophage migration inhibition factor and a delayed hypersensitivity skin reaction did not play a role in the resistance, nor did specific IgG antibodies have any protective influence. It was shown that neutrophils play a cardinal role in the immunity against H. ovis infection in so far as they phagocytize and destroy the organisms and are attracted to them by chemotaxis in immune animals.     

Prevalence and causes of ram epididymitis in Utah

Palpation of the testicles and epididymides was used to determine the prevalence of ram epididymitis (RE) in Utah range flocks over a 5-year period. Of 62 range flocks examined, 58 had rams with palpable lesions. A comparison was made of prevalence by right vs left testicle involvement, but no significant difference was found. Culling all rams with palpable lesions from the flock progressively reduced the annual incidence, but did not eliminate RE from any flock. Semen and specimens of reproductive tract tissue were obtained from representative flocks for bacterial culture. Brucella ovis was found commonly in range flocks, but was not isolated from RE-affected rams in virgin, ram-producing flocks. In the ram-producing flocks, a gram-negative, pleomorphic rod-type organism was found. It was concluded that RE should be considered as 2 disease entities, based on the finding of 2 causative organisms and the management systems under which those organisms are found. Brucella ovis was the major cause in persistently infected range flocks, whereas the gram-negative pleomorphic rod-type organism is the apparent cause of RE in virgin, ram-producing flocks.     

Meningoencephalitis and other conditions associated with Histophilus ovis infection in sheep

Histophilus ovis was isolated from 29 sheep in 20 flocks and 2 artificial insemination (AI) centres in southern New South Wales from 1984 to 1990. The clinical and pathological findings were consistent with previous reports and included polyarthritis (7 flocks), epididymo-orchitis (5), meningoencephalitis (3), pneumonia (3), septicaemia (2), mastitis (1) and metritis (1). Six sheep had meningoencephalitis, a syndrome not previously associated with H ovis infection in sheep, which was similar pathologically to thromboembolic meningoencephalitis in cattle, caused by the related organism, Haemophilus somnus. H ovis was isolated from the semen of 12-month-old rams in a flock that had polyarthritis due to H ovis, in 4-month-old ram lambs and from the uterus of a ewe in a flock that had sporadic cases of H ovis septicaemia.     

The acquisition of immunity to Histophilus ovis by sheep in nature

It was demonstrated that skin wound infection with Histophilus ovis elicits an immune response which can protect a ram against a challenge injection of the same organism into its epididymis.     

Clinical findings and pathological changes in Histophilus ovis infections of sheep

The Gram-negative pleomorphic bacterium Histophilus ovis was incriminated as the cause of natural cases of epididymitis in rams, polyarthritis in lambs, mastitis and abortion in ewes. Epididymitis was sporadic in occurrence, unilateral in distribution and varied from acute to chronic in degree. Polyarthritis affected recently born lambs and resulted in acute lameness and subsequent debility. Mastitis was rare in occurrence, unilateral in distribution, acute and gangrenous in type. Abortion was also rare and manifested by a specific haemorrhagic placentitis. These four conditions were reproduced experimentally. The clinical, pathological and histopathological changes associated with field and experimental cases are described.     

Prevalence of Histophilus ovis and Actinobacillus seminis in the genital tract of sheep

Histophilus ovis and Actinobacillus seminis were isolated from the preputial cavity of 6-month-old rams and the vagina of 6-month-old ewes at a substantially higher rate than that in mature (greater than 2 years old) rams and ewes. These organisms appeared to be a transitory component of the ovine genital flora, the prevalence of which was associated with age regardless of gender. Additional evaluation of the recoverability of H ovis and A seminis from the preputial cavity of rams from birth to 1 year of age indicated that the isolation rate from rams and predominance of the organisms in the preputial cavity differed greatly over this age period. These organisms were not recoverable until ram lambs were 12 weeks of age and were most prevalent at 20 weeks of age, after which recoverability of H ovis and A seminis from the preputial cavity steadily decreased, continuing through the time of the last evaluation at 1 year of age. The time period with which these organisms can be isolated from the preputial cavity is closely correlated with the time period when epididymitis associated with these organisms develops, and may be an important factor in the pathogenesis of epididymitis.     

Pathogenicity of different strains of Histophilus somni in the experimental induction of ovine epididymitis

The purpose of this study was to determine any differences in pathogenicity when sheep are experimentally infected with different Histophilus somni isolates: a) 2336 bovine origin strain; b) an isolate from ram orchitis and epididymitis; c) an isolate from the brain of a sheep with neurological signs; d) an isolate from the vagina of a clinically healthy ewe. A total of 20 rams divided in groups of 5 animals each were inoculated in the epididymis with 1 × 10⁷ CFU/mL of H. somni; a negative control group of 5 rams was used. All groups inoculated with H. somni showed some epididymitis, but the most pathology was caused by the epididymitis isolate, followed by the vaginal isolate. It was demonstrated that there is a difference in experimental infection capacity among isolates from different origins, as epididymitis occurred and the bacteria was recovered only from groups inoculated with isolates originating from epididymitis and vaginal exudate.     

Oestrus ovis infestation of a dog in the UK

In March 2011, a dog on a sheep farm in the Cotswolds, UK, expelled a mature live third-stage larva of the sheep nasal botfly, Oestrus ovis, after a violent and traumatic sneezing episode. The dog had been infected with first-stage larvae deposited by an adult fly the previous autumn; larval development had progressed throughout the winter and spring with few apparent clinical signs and possibly masked by ongoing immunotherapy for an unrelated condition. Identification of the parasite at the Liverpool School of Veterinary Science was made from a submitted puparium, the "chrysalis" stage to which the larva had progressed within days of its expulsion from dog's nose. To the authors' knowledge this is the first report of nasal botfly infestation of a dog in the UK.     

Characterisation of Histophilus ovis and related organisms by restriction endonuclease analysis

The banding profiles generated by Bam H1 restriction endonuclease cleavage of bacterial DNA from clinical and reference isolates of Histophilus ovis, Haemophilus somnus and related bacteria were compared. H. ovis, H. somnus and Haemophilus agni isolates were found to have distinct similarities in banding profiles characterised by 10 common bands between 2.0 and 9.6 kilobases (kb). The close taxonomic relationship of these isolates was reinforced by these findings. The reference isolates examined in this study--Actinobacillus lignieresii, Actinobacillus seminis, H. agni, H. somnus, H. ovis, Haemophilus influenzae, Haemophilus parainfluenzae, Haemophilus parahaemolyticus--could be distinguished from each other on the basis of their characteristic banding profiles. Actinobacillus sp were observed to have more bands between 2 and 23 kb compared with the H. ovis and Haemophilus sp isolates studied. Analysis of isolates from an experimental infection trial illustrated the potential of restriction endonuclease analysis in molecular epidemiological applications. It was possible to demonstrate by this means that the post-challenge isolates had identical banding profiles to the challenge (or infecting) isolate which had a distinctly different banding profile from that of pre-challenge H. ovis isolates. Furthermore, restriction endonuclease analysis of H. ovis isolates obtained from follow-up investigations of a recurrent problem of epididymitis in unmated rams, indicated that the H. ovis isolates implicated in epididymitis, were present as a single strain in a number of sheep over a period of time. This suggested that the mechanism of transmission was by perinatal perputial contamination.