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.     

The detection of Brucella ovis infection in rams

Abstract

Extract

Each year in New Zealand, many rams are palpated for lesions of the epididymes and testes. Although regarded initially as a method of detecting infertile rams (Crawford and James, 1950 Biberstein, E. L. and McGowan, B. 1958. Cornell Vet., 48: 31–31.  [Google Scholar]), the technique is now regarded primarily as a method of detecting infection with the organism for which Buddie (1956 Buddle, M. B. 1956. J.Hyg., 54: 351–351.  [Google Scholar]) proposed the name Brucella ovis.     

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.     

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.     

Production of immunity in rams against Brucella ovis infection

Extract

During earlier investigations in New Zealand into the bacterial and mycotic flora of hedgehogs (Erinaceus europaeus), strains of Salmonella typhimurium were obtained from the gut of a number of animals. It was therefore decided to examine these animals more thoroughly for the presence of salmonellae. This paper embodies the results of a small survey subsequently carried out in the Hamilton suburban area during February, 1964.     

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.     

Trials with an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of subclinical genital infections in rams caused by Histophilus ovis and Actinobacillus seminis

The performance of an enzyme-linked immunosorbent assay (ELISA) was evaluated in the serological diagnosis of subclinical genital infection in 6 naturally infected ram flocks and 2 experimentally infected ram hoggets. The test employs lipopolysaccharide (LPS) antigen prepared by autoclaving from Actinobacillus seminis and Histophilus ovis. A total of 193 sheep (118 unmated virgin rams and 75 mature breeding rams) were examined clinically, serologically (by ELISA) and bacteriologically (semen bacteriology) at the same time. Serum samples from all animals were also tested by an ELISA employing LPS antigen prepared from Brucella ovis in the same way. Shedding of A. seminis and H. ovis did not show close correlation with serological positivity (Table 1), as only 9 (15.0%) out of the 60 A. seminis shedders were ELISA seropositive at the same time. As regards H. ovis only 10 (19.2%) out of the 52 H. ovis shedders were ELISA seropositive at the same time. The results indicate that, when used alone, the ELISA employing LPS antigen is unsuitable for diagnosing subclinical genital infection caused by H. ovis and A. seminis in rams. The authors discussed shortly the employing fields of this ELISA test in the diagnostic work.     

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.     

Lameness in rams following vaccination against Brucella ovis infection

Extract

Since the introduction in 1957 of a vaccination procedure to control Brucella ovis infection in sheep, many thousands of rams have been vaccinated annually throughout New Zealand almost without complications.     

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.     

The serological response of rams to four methods of vaccination against Brucella ovis infection

A comparative study was made of the serological response of rams to an inactivated Brucella ovis saline in-oil vaccine administered either once or twice by either the subcutaneous or the intraperitoneal route. The serological response of rams to two spaced doses of vaccine was more consistent and more persistent than when a single dose of vaccine was used. The rise in titre was more rapid and the final titre of greater magnitude when the subcutaneous route of administration was used in comparison with the intraperitoneal route. On a serological basis, the most satisfactory vaccination technique investigated consisted of two subcutaneous injections of vaccine administered ten weeks apart. The least satisfactory technique was a single intraperitoneal injection.     

The complement fixation test for the diagnosis of Brucella ovis infection in rams

Complement fixation tests using three B. ovis antigen preparations in warm fixation tests (WCFT) and cold fixation (CCFT) tests were done on 541 ram sera. Semen samples from the same rams were examined culturally to identify B. ovis excretors. The CCFT, using an antigen prepared by heat extraction of B.ovis cells, had a sensitivity of 97% in 124 rams which were shedding B.ovis. The specificity was 99% in 144 rams from non-infected flocks. Seventy-seven per cent of 156 rams which reacted to this test were shedding B. ovis in their semen. Tests with other antigens were inferior in sensitivity and/or specificity. The WCFT gave lower titres than CCFT. Vaccination caused large numbers of false positive reactions in 4 flocks.     

Lack of evidence of Brucella ovis infection in rams in Quebec

A study was conducted to estimate the seroprevalence of Brucella ovis infection in rams in the Estrie and Bas-Saint-Laurent regions (Quebec). Rams sera (n = 258) were serologically evaluated from 224 rams in 30 commercial flocks and from 34 rams at 2 slaughterhouses by using an enzyme linked immunosorbent assay. Epididymides and testes were examined by palpation on farms and microscopically for culled rams. No ram was seropositive to Brucella ovis or had lesions suggestive of brucellosis from the farm or slaughterhouse surveys.     

Pathology of experimental Histophilus ovis infection in sheep. II. Pregnant ewes

Intravenous inoculation of Histophilus ovis to pregnant ewes at mid or late gestation resulted in acute bacterial endometritis, placentitis and foetal death. There were no lesions in the foetuses but H. ovis was recovered from the uteri of the ewes and the placentas. Oral or intravaginal administration had no effect.     

Temporal ELISA response of rams to Brucella ovis following experimental infection or vaccination

Sera from rams vaccinated with antigens extracted chaotropically from Brucella ovis by potassium thiocyanate treatment were used to optimise a whole-cell, enzyme-linked immunosorbent assay (CELISA) and to monitor the temporal serological response of rams which had been challenged with infected semen by the intranasal or intrapreputial route. Three patterns of CELISA response were detected. Thirteen of 15 rams intranasally challenged did not respond serologically (pattern 1 or nil response). Only one of 15 rams in the intranasal group exhibited a rise and fall response with CELISA (pattern 2), while another showed a rise and surge response (pattern 3). The numbers of rams in the intrapreputial group which displayed a pattern 1 or 2 or 3 response were four, nine and two, respectively. No ram with a pattern 2 response excreted B ovis in the semen or showed any other evidence of infection, whereas rams with a pattern 3 response excreted B ovis in the semen and developed palpable lesions. Intrapreputially challenged rams that were CELISA-positive consistently mounted an antibody response against B ovis about two to four weeks earlier than intranasally challenged rams.     

Brucella ovis vaccination of rams

Abstract

Extract

An infectious epididymitis of rams caused by Brucella ovis (Buddie, 1956 Buddle, M. B. 1956. J. Hygiene, 54: 351–351.  [Google Scholar]) infection, first described in Australia (Simmons and Boyes, 1953 Simmons, C. G. and Hall, W. J. K. 1953. Aust.vet.J., 29: 33–33. [Crossref] , [Google Scholar]) and New Zealand (Buddie and Boyes, 1953 Buddle, M. B. and Boyes, B. W. 1953. Aust.vet. J., 29: 145–145. [Crossref] , [Google Scholar]) was recognized subsequently in Czechoslovakia (Gdovin et al, 1955 Gdovin, T., Hrudka, F., Chladecky, E. and Koppel, Z. 1955. Shorn, ces. Akad. zemedelsk Ved., 28: 617–617.  [Google Scholar]), the United States (McGowan and Shultz, 1956 McGowan, B. and Shultz, G. 1956. Cornel Vet., 46: 277–277.  [Google Scholar]), South Africa (Van Rensburg et al, 1958 Van Rensburg, S. W. J., Van Heerden, K. M., Le Roux, D. J. and Snyders, A. J. 1958. J.S.Afr. vet. med. Ass., 29: 223–223.  [Google Scholar]), Rumania (Tudoriu, et al, 1958 Tudoriu, C. D., Andrei, M., Draghici, D. and Moldoveanu, P. 1958. Anu. Inst. Pat.Igien. anim. Bucaresti, 8: 5–5.  [Google Scholar]), and South America (Dr Justo Zomara B, 1961, pers. comm.). As the infection can affect ram fertility and, further, can be responsible for abortion in ewes and perinatal mortality in lambs, attention has been directed to the development, evaluation, and application of control measures in a number of important sheep-raising countries.     

The response of vaccinated rams to experimental challenge using Brucella ovis

One hundred and thirty eight rams were allocated to four experimental groups. An inactivated Br. ovis vaccine was administered either once by the intraperitoneal route (1 i/p), twice by the intraperitoneal route (2 i/p), or twice by the subcutaneous route (2 s/c), and the last group was left unvaccinated. They were then challenged by the intravenous inoculation of between 123 and 1.23 × 10⁸ live Br. ovis bacteria. The number of rams that succumbed to infection within the four groups was 4/35 (11%) for the 2 s/c, 7/33 (21%) for the 2 i/p, 9/35 (26%) for the 1 i/p and 18/35 (51%) for the unvaccinated rams. Vaccination reduced the number of rams that succumbed to experimental challenge and although there were differences between the vaccinated groups, these were not statistically significant.

Following challenge, unvaccinated rams were the first to excrete Br. ovis in their semen; three weeks following inoculation. Next, those vaccinated by either one or two doses by the intraperitoneal technique began to excrete the organism (five weeks) and then finally those rams vaccinated twice by the subcutaneous route (seven weeks).