Immunisation against ovine caseous lymphadenitis: correlation between Corynebacterium pseudotuberculosis toxoid content and protective efficacy in combined clostridial-corynebacterial vaccines

Groups of sheep were dosed with vaccines containing Corynebacterium pseudotuberculosis toxoid combined in varying amounts with 5 clostridial antigens. Resistance of the sheep to infection with C pseudotuberculosis was tested at 1, 6 and 12 months after vaccination by infection with pus from ovine lymph glands actively infected with C pseudotuberculosis. The outcome was assessed 3 months after challenge by slaughter and inspection of the sheep for caseous lymphadenitis lesions. Protection was demonstrated by a significant reduction in the proportion of immunised sheep exhibiting lesions compared with control sheep, and by fewer abscesses in affected immunised sheep than in affected control sheep. A positive correlation was found between amount of C pseudotuberculosis toxoid administered and degree of protection obtained. Chromatographically-purified toxoid induced essentially the same protection, suggesting that anti-toxic immunity is the major factor in protection.     

Immunisation against ovine caseous lymphadenitis: comparison of Corynebacterium pseudotuberculosis vaccines with and without bacterial cells

Sheep were immunised with Corynebacterium pseudotuberculosis vaccines prepared from cell-free toxoid or from toxoid with formalin-killed cells of C pseudotuberculosis added. Resistance of sheep to infection was tested 6 months after immunisation by inoculation with caseous lymphadenitis pus. The outcome was assessed 3 months later by slaughter and inspection of the sheep for lesions of caseous lymphadenitis. immunised sheep were adequately protected against infection as shown by a significant reduction in the number of sheep exhibiting lesions compared with control sheep, and by fewer abscesses in affected vaccinated sheep than in affected control sheep. The protective potency of the vaccines was not improved by the inclusion of cells of C pseudotuberculosis.     

Efficacy of Corynebacterium pseudotuberculosis bacterin for the immunologic protection of sheep against development of caseous lymphadenitis

The efficacy of a Corynebacterium pseudotuberculosis bacterin to protect sheep immunologically against development of caseous lymphadenitis was evaluated in controlled challenge-exposure experiments. Sixty-three mixed-breed, white-faced lambs were used. The lambs were 10 to 12 weeks old and were randomly assigned to 3 groups (21 lambs/group). Group 1 was vaccinated once, using 2 ml of a C pseudotuberculosis bacterin (given subcutaneously) in the right axillary region at the beginning of the study. Group 2 was vaccinated twice; the 1st vaccination was given at the same time that lambs in group 1 were vaccinated and the 2nd vaccination was given 4 weeks later. Group 3 (nonvaccinated controls) was given physiologic saline solution (2 ml, subcutaneously). Each lamb was challenge exposed (ie, given 2 ml of live Corynebacterium pseudotuberculosis inoculum [6 X 10(6) colony-forming units/ml], subcutaneously at 4 different sites) during the 20th week of the study. All lambs were killed and necropsied during week 33. The mean number of abscesses per lamb was 7 for group 1, 4 for group 2, and 32 for group 3. Significant differences in the size of the abscesses were not found between the groups. Results of the study indicated that the vaccine provided immunologic protection of lambs against challenge exposure to Corynebacterium pseudotuberculosis.     

Differential antibody responses to Corynebacterium pseudotuberculosis in sheep with naturally acquired caseous lymphadenitis

Enzyme-linked immunosorbent assays (ELISA) with Corynebacterium pseudotuberculosis cell wall and bacteria-free supernatant with exotoxin preparations as antigens, and hemolysis inhibition tests were used to detect antibodies in the sera of adult range sheep with naturally acquired caseous lymphadenitis (CL). The extent and severity of lesions were quantitated on the basis of a lesion score, derived from an examination of the carcass (peripheral lymphoid tissue) and viscera (including internal lymphoid tissue) at the time of slaughter. The overall prevalence of C pseudotuberculosis-positive CL lesions in 104 sheep was 31.7%. The cell wall ELISA detected antibodies in 96.9% (32/33) of sheep with C pseudotuberculosis-positive CL lesions. The exotoxin ELISA detected antibodies in 84.8% (28/33) of positive sheep in the same group. Both ELISA resulted in a high number of apparent false-positives, with 64.7% and 49.2%, respectively, positive optical density (OD) values in sheep with no gross CL lesions and no apparent C pseudotuberculosis infection. There was no significant relationship between the extent of lesion development (lesion score) and OD values in both cell wall (r = 0.472) and exotoxin (r = 0.464) ELISA. Similarly, there was no significant relationship between the titer of antitoxin antibodies, as measured by the hemolysis inhibition test, and the extent of disease. These investigations indicate that those ELISA that use crude C pseudotuberculosis antigens are of questionable utility in the field, where C pseudotuberculosis infection is endemic in many sheep populations. Furthermore, these studies suggest that antibodies that are reactive with components of C pseudotuberculosis and that develop in response to infection may have little impact on the recovery of the host.     

Prevalence of caseous lymphadenitis and usage of caseous lymphadenitis vaccines in sheep flocks

OBJECTIVE: To estimate the prevalence of caseous lymphadenitis (CLA), determine the current usage of vaccines against CLA and to measure the effectiveness of these vaccines on sheep farms. DESIGN AND POPULATION: A survey was undertaken on 223 sheep flocks in New South Wales, Victoria and Western Australia. METHOD: The prevalence of CLA was measured by conventional inspection techniques at abattoirs in lines of sheep that could be traced back to a farm. Managers of the flocks were sent a questionnaire about their vaccine practices, management practices and knowledge of CLA. RESULTS: The average prevalence of CLA in adult sheep in these flocks was 26% and varied from 20% in Western Australia to 29% in New South Wales. About 43% of sheep producers used CLA vaccines; only 12% used them as recommended. Awareness of CLA was highest in Western Australia. More producers would use CLA vaccine if they knew the prevalence of CLA in their flock and producers obtained most information about CLA from vaccine resellers. CONCLUSIONS: Only 10 to 15% of producers are currently achieving effective CLA control through the use of recommended CLA vaccination programs. In Western Australian flocks more than 25% of effectively vaccinated ewes will be sent to abattoirs in the 2 to 3 years after this study. However, large decreases in the prevalence of CLA can be achieved by about 70% of producers by either making adjustments to their vaccination programs or buying a vaccine with a CLA component. Two or three key facts on effective CLA vaccination could be made available at the point of sale of vaccines and from abattoirs that reported the prevalence of CLA to farmers.     

Use of a toxoid vaccine to protect goats against intradermal challenge exposure to Corynebacterium pseudotuberculosis

Two groups of male, 9-week-old goats (5 goats/group) were vaccinated subcutaneously with formalized exotoxin of Corynebacterium pseudotuberculosis, with Freund's incomplete adjuvant. Each goat was given 2 vaccinations, 2 weeks apart. At each vaccination, each group 1 goat was given 0.5 ml of toxoid, and each group 2 goat was given 1 ml of toxoid. Twenty days after the 2nd vaccination, vaccinated goats and 5 nonvaccinated 12-week-old goats (controls) were inoculated intradermally (challenge exposed) with live C pseudotuberculosis, monitored for 13 weeks, and euthanatized. At necropsy, 5 of the 10 vaccinated goats did not have C pseudotuberculosis lesions, 3 had abscesses limited to the inoculation site and draining lymph node, and 2 had disseminated bacterial lesions. Of the 5 nonvaccinated controls, 4 had disseminated abscesses and 1 had a single abscess in an internal node. Serologically, 9 of the 10 vaccinated goats developed positive (greater than or equal to 1:8) antibody titers against the exotoxin within 1 week after inoculation; the 10th goat seroconverted 2 weeks after inoculation, whereas control goats required 3 weeks to develop a positive antibody response. Therefore, early during an infection with C pseudotuberculosis, antibodies against the exotoxin may protect a goat against spread of the organism. All goats were injected intradermally before challenge exposure, 10 days after challenge exposure, and at 4, 8, and 12 weeks after challenge exposure with a skin-test reagent composed of fragmented bacterial cells.(ABSTRACT TRUNCATED AT 250 WORDS)     

Antigen specificity of antibody responses to Corynebacterium pseudotuberculosis in naturally infected sheep with caseous lymphadenitis.

Constituents of Corynebacterium pseudotuberculosis were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Analysis of sonicated whole bacterial cells and ether-extracted cells revealed more than 35 bands in silver-stained gels. SDS-PAGE analysis of concentrated culture filtrates with exotoxin activity demonstrated more than 15 bands. Sera from sheep with C. pseudotuberculosis-induced disease of variable severity were used to probe immunoblots of electrophoresed ether-extracted cells and culture filtrates. Twenty or more corynebacterial molecules, ranging in molecular weight from 20 to 112 kDa, in ether-extracted cells were recognized by antibodies in the sera of naturally exposed sheep with positive ELISA titers. These sera also recognized up to six molecules, ranging from, 20 to 68.1 kDa, on immunoblots of ammonium sulfate-concentrated culture filtrate. There was no apparent relationship between the stage of disease and the response to specific corynebacterial antigens in these animals.     

Corynebacterium pseudotuberculosis infection in goats. II. The prevalence of caseous lymphadenitis in 36 goat herds in Northern Norway

The precalen-ce of caseous lymphadenitis was surveyed in 36 goat herds in Northern Norway. In each herd, information concerning the occurrence of the disease was obtained from the farmer. Adult animals (1 year of age or older) in 35 herds were examined for superficial swellings, and serum samples were collected from most animals in the herds. The sera were examined for antibodies to Corynebacterium pseudotuber-culosis using the bacterial agglutination test (BAT) and the hemolysis inhibition test (HIT).Gaseous lymphadenitis was diagnosed with certainty in 19 herds. Information from the farmers indicated that the disease indeed oc-curred in these herds, and that the majority had been infected with the disease for many years. The herds had apparently become infected through contact with animals from infected herds. Clinical examina-tions were carried out in 18 of these herds and superficial swellings were found in 26 % of the examined animals. The prevalence of ani-mals with lesions varied from 11 to 40 % among the herds. Of the animals in these herds, 81 % were positive in BAT and 84 % in HIT. The prevalence of positive animals varied from 26 to 99 % in BAT and 28 to 99 % in HIT. The prevalence of seropositive animals was lowest in a herd in which animals were kept separately in stalls.Caseous lymphadenitis could not be diagnosed in 16 herds. In-formation from the farmers indicated that the disease indeed seemed to be absent in 14 of these herds. These 14 herds had no history of contact with animals from herds considered to be infected. However, in the remaining 2 herds, the farmers were somewhat uncertain about the occurrence of the disease. One of these 2 herds had a history of contact with infected herds through participation in a goat “breeding circle”. Only a few of the animals were, however, seropositive and all these had low antibody titres.In 1 newly established herd, a single animal showed a high posi-tive titre in BAT only. All the other animals were negative in both tests. This particular herd consisted of animals obtained both from herds with caseous lymphadenitis and from herds in which the disease was not considered to occur.     

Postal survey of ovine caseous lymphadenitis in the United Kingdom between 1990 and 1999

Members of the Sheep Veterinary Society and three terminal sire breed societies (A, B and C) were surveyed to investigate the prevalence and characteristics of ovine caseous lymphadenitis in the UK. In total, 264 veterinary surgeons and 510 farmers responded. Eighteen per cent of the vets had seen at least one case of the disease, and 45 per cent of the farmers had seen abscesses in their sheep. Relatively few farmers had had the cause of the abscesses investigated, but 24 of the 32 flocks that had submitted samples to a laboratory had had the disease confirmed. The incidence of the disease and abscesses increased from 1990 to 1999. The disease was more likely to be investigated in certain breeds and when male sheep had abscesses. Risk factors for the disease included the presence of sheep of breed B in the flock, the presence of half-bred sheep, showing sheep, and the use of shearing contractors. Dipping sheep appeared to be protective.     

Corynebacterium pseudotuberculosis infection and lymphadenitis (taloa or mala) in the camel

Summary Pure cultures ofCorynebacterium pseudotuberculosis were obtained from 11 cases of lymphadenitis (known locally as taloa or mala) in camels. Camel isolates produced typical taloa in camels experimentally inoculated subcutaneously at the base of the external ear with 10¹⁰ colony forming units. A sheep strain ofC. pseudotuberculosis inoculated into camels produced a local abscess at the site of inoculation but did not produce taloa. Re-infection of camels recovered from experimental inoculation did not produce taloa suggesting the possibility of the development of a vaccine against lymphadenitis in camels.

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Infeccion PorCorynebacterium Pseudotuberculosis Y Linfadenitis (Taloa O Mala) En El Camello

Resumen Se obtuvieron cultivos puros deCorynebacterium pseudotuberculosis a partir de 11 casos de linfadenitis (conocida localmente como taloa o mala) en camellos. La inoculación subcutánea en camellos en la base de la oreja de 10¹⁰ unidades formadoras de colonias procedentes de un cultivo obtenido de camellos infectados causó taola. Sin embargo, una cepa ovina deC. pseudotuberculosis causó un abceso en el lugar de la inoculación pero no produjo taola en camellos. La reinfección de los camellos que se habían recuperado de la inoculación experimental no causó taola, lo que sugiere la posibilidad de desarrollar una vacuna frente a la linfadenitis caseosa del camello.

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Infection ACorynebacterium Pseudotuberculosis Et Lymphadenite (Taloa Ou Mala) Chez Le Chameau

Résumé On a obtenu des cultures pures deCorynebacterium pseudotuberculosis à partir de 11 cas de lymphadénite (dénommée localement taloa ou mala) chez des chameaux. L'inoculation expérimentale des isolements camelins faite en infection sous-cutanée à la base de l'oreille externe avec 10¹⁰ unités formant des colonies a provoqué un taloa typique. Une souche ovine deC. pseudotuberculosis inoculée à des chameaux n'a conduit qu'à la formation d'un abcès local au point d'inoculation mais pas au taloa. La réinfection des chameaux guéris d'une inoculation expérimentale ne provoque pas de taloa, ce qui permet d'augurer la possibilité de développement d'un vaccin contre la lymphadénite cameline.

     

Evaluation of vaccines against ovine dermatophilosis

Intradermal vaccination of live crude filaments (vaccine A) was compared with a vaccine (vaccine B) consisting of a 45 kD zoospore protein and mucoid material coating filaments in its ability to protect sheep from experimental Dermatophilus congolensis infection. Fourteen and 21 days after challenge, vaccine A sheep had fewer lesions (P less than 0.001) than the vaccine B sheep. The lesions on the vaccine A sheep were also less severe 14 and 21 days after challenge (P less than 0.05, P less than 0.01 respectively). In a second study, vaccine A was assessed for its ability to protect against natural challenge. Ten weeks after contact with sheep with active and generalised dermatophilosis no difference was found between the number of lesions present on the vaccine A and unvaccinated sheep and no differences were found in the number of sheep in each group with active lesions.     

A field evaluation of the efficacy of two vaccines against ovine pneumonic pasteurellosis

Two vaccines against pneumonic pasteurellosis were evaluated for efficacy in lambs transported by sea from New Zealand to Saudi Arabia. One vaccine contained whole cell antigens of Pasteurella haemolytica A2 grown under iron restricted conditions. The other contained Pasteurella haemolytica A1 cell surface and leucotoxin antigens. There was no clear evidence of either vaccine leading to a lower pneumonia death or lesion rate than for the control group.     

Molecular genotyping of multinational ovine and caprine Corynebacterium pseudotuberculosis isolates using pulsed-field gel electrophoresis

Caseous lymphadenitis (CLA) is a chronic, suppurative disease, with a worldwide distribution, caused by Corynebacterium pseudotuberculosis. The clinical manifestation of CLA is known to vary between different countries, and has been postulated to be due to differences in the strains present in these countries. Forty-two sheep and goat isolates of C. pseudotuberculosis from Australia, Canada, Eire, The Netherlands and Northern Ireland were characterized by pulsed-field gel electrophoresis (PFGE), biotyping, antimicrobial susceptibility, and production of phospholipase D. The PFGE-determined genotypes of this multicentric collection were then compared with representative ovine and caprine isolates from a previously published panel of PFGE profiles of United Kingdom isolates. Digestion with SfiI generated 16-18 bands in the 48.5 and 290 kb range, and differentiated four distinct pulsotypes amongst the 36 ovine and 6 caprine strains which displayed remarkable homogeneity. Based on these results, it would appear that the genome of C. pseudotuberculosis is highly conserved, irrespective of the country of strain origin.     

Antigen specificity and activity of ovine antibodies induced by immunization with Corynebacterium pseudotuberculosis culture filtrate

Sheep were immunized three times with a vaccine composed of filtrate from a 36 h culture of Corynebacterium pseudotuberculosis and a block polymer adjuvant. Immunization resulted in the development of exotoxin-neutralizing antibody. This corresponded to the recognition of a 31.6 kDa protein on sequential immunoblots of ammonium sulfate-precipitated filtrate. In addition sera from vaccinated sheep recognized at least eight bacterial cellular antigens on immunoblots of ether-extracted C. pseudotuberculosis, including bands of 12, 25.1, 31.6, 36.3, 39.8, 63.1, 70, 75 or 79.4 kDa. Sera from these sheep altered the colony growth characteristics of C. pseudotuberculosis in vitro. These results indicate that immunization with soluble C. pseudotuberculosis in vitro. These results indicate that immunization with soluble C. pseudotuberculosis antigen preparations that have been used in toxoid vaccines induces antibody responses to numerous cellular antigens in addition to exotoxin and suggest that serologically mediated antibacterial effects could be an important component in the protection from disease that has been reported following immunization with C. pseudotuberculosis toxoids.