Corynebacterium pseudotuberculosis infection in adult ewes by inoculation in the external ear

Caseous lymphadenitis, caused by Corynebacterium pseudotuberculosis, was studied in ewes to devise a reliable method of induced infection that would help to understand mechanisms by which abscesses develop in lymph nodes or internal organs, especially the lungs. Fifteen adult ewes, from a flock where the occurrence of caseous lymphadenitis was unknown, were inoculated SC in the auricle with various doses of a virulent strain of C pseudotuberculosis. Ewes inoculated with at least 10(8) viable corynebacteria develop a strong local reaction that was apparent 4 hours after challenge exposure and lasted 7 to 14 days. Of these ewes, all developed at least 1 abscess and 3 developed multiple abscesses in lymph nodes draining the inoculated auricle and in the lungs. All infected ewes developed a strong antibody response against C pseudotuberculosis exotoxin, which persisted for 3 months. Of 9 ewes infected with 1.2 X 10(8) corynebacteria, 3 developed lesions because of field strains of C pseudotuberculosis and had an increased capacity to limit the dissemination of challenge-inoculum bacteria by developing only one abscess in the draining lymph node (parotid or prescapular lymph node). Seemingly, inoculation in the external ear was a suitable method for obtaining a reproducible infection and indicated that a primary infection might protect against reinfection by limiting dissemination of C pseudotuberculosis.     

Corynebacterium pseudotuberculosis infection in goats. VIII. The effect of vaccination against experimental infection

The effect of an inactivated vaccine against C. pseudotuberculosis infection was tested on castrated male kids from a herd free from caseous lymphadenitis. The animals were divided into 3 groups with 8 animals in each. Group 1 was immunized with crude filtrated C. pseudotuberculosis toxoid and whole killed organisms, while Group 2 in addition was given levamisole. The kids were vaccinated twice at an interval of 4 weeks. Group 3 consisted of unvaccinated animals. All groups were challenged subcutaneously with live bacteria 4 weeks after the last vaccination. Unvaccinated animals showed the most severe course of illness after challenge. Development of abscesses in the regional lymph nodes (Inn. subiliaci) was significantly more common in unvaccinated than in vaccinated kids at necropsy 2 months after challenge. There was, however, no such difference between the vaccinated groups, and there was no difference between any of the groups as regards abscess formation at the inoculation site. In each of the 2 vaccinated groups, there was a titre rise following vaccination in the hemolysis inhibition test, whereas no such rise was seen in the bacterial agglutination test. The titre values in both tests increased significantly after challenge in all the groups, the increase being most rapid in the vaccinated animals. The present investigation indicates that development of caseous lesions in lymph nodes in goats, following subcutaneous inoculation with C. pseudotuberculosis, can be reduced by an inactivated vaccine containing whole organisms and crude toxin.     

The effect of subclinical experimental Cowdria ruminantium infection in ewes on the growth and milk consumption of pre-weaning lambs.

An alternative control option for heartwater (Cowdria ruminantium infection) is the establishment and maintenance of endemic stability which would lessen the existing dependence on acaricides. In an endemically stable state, animals become infected by vaccination or natural challenge at an early age, following which the immunity so created is boosted by continuing tick challenge. In this study, growth rates, health and hematological parameters were monitored at regular intervals for lambs born to two matched groups of ewes until weaning at 4 mo of age. One group of ewes was infected multiple times with Cowdria ruminantium; the other group remained uninfected. The overall mean leucocyte count of lambs born to infected ewes was significantly lower than that of lambs born to uninfected ewes (P=0.04). However, there were few other significant differences in the other hematological data between the two groups. The mean birth weight of single lambs born to uninfected ewes (4.6 kg) was significantly higher than the mean birth weight of single lambs born to infected ewes (4.4 kg) (P=0.02). Trends in milk consumption and growth rates were similar for the two groups, with few significant differences detected. Likewise, there were no significant differences in the incidences of health problems or pre-weaning mortalities between the two groups of lambs. The results of this study indicate that there is no detectable effect on productivity of pre-weaning lambs when their dams are carriers of C. ruminantium--a situation likely to occur in an endemically stable state. Hence, maintenance of endemic stability would be a suitable control option for heartwater.     

Corynebacterium pseudotuberculosis infection in goats. IX. The effect of vaccination against natural infection

A vaccination trial was carried out in 10 infected herds. The trial included 247 female kids, the number of animals in each herd varying from 15 to 38. About half of the animals in each herd were vaccinated twice at 3 to 4 week intervals, the first vaccination being carried out before the age of 4 months. A combination of a crude filtrate of C. pseudotuberculosis toxoid with whole organisms, was used. Overall, the prevalence of animals with superficial swellings was higher in the unvaccinated than in the vaccinated group during the first 1-2 years following immunization. However, in some herds superficial swellings were as common in vaccinated as in unvaccinated animals. An antibody response following vaccination was demonstrated in the hemolysis inhibition test, but not in the bacterial agglutination test. Superficial swellings were more common in vaccinated animals which were negative than in animals which were positive in the hemolysis inhibition test at 1 1/2 months after vaccination. The vaccine used in the present study, was not sufficiently efficacious to be recommended as the only protective measure against caseous lymphadenitis in Norwegian goat herds.     

Serological and bacteriological observations on experimental infection with Salmonella hadar in chickens

Over the past 3 years the frequency of Salmonella hadar infections has increased in Belgium in both poultry and humans. Therefore, the course of infection with S. hadar in poultry was investigated. One day-old and 4 week-old specific pathogen-free chickens were orally infected with one of two S. hadar strains, SH1 or SH2. Mortality was 6% (SH1) and 17% (SH2) in birds infected at 1 day-old. Chickens infected at 1 day-old with SH2 showed a mild diarrhoea. The S. hadar faecal excretion in birds infected at 1 day-old remained high throughout the experiment until 12 weeks post-inoculation (pi). Faecal excretion was lower in older birds. Antibodies to S. hadar were observed from 2 weeks pi (SH2, infected at 1 day-old) or 4 weeks pi (SH1, both groups; SH2, chickens infected at 4 weeks of age). The percentage of chickens with antibodies was higher after infection at 1 day-old than after infection at 4 weeks of age. In a second experiment 1 day-old chicks were infected with SH1 and autopsied at regular intervals until 42 days pi. SH1 was isolated from the caeca from 3 h pi onwards and from the liver and spleen from 18 h until 14 days pi. Serous typhlitis and omphalitis were the main lesions. The number of macrophages in the lamina propria of the caecal tonsils was slightly increased from 18 h until 2 weeks pi. In the liver, inflammation was observed in the portal triads and in the sinusoids. This study indicates that infections with S. hadar lead to intense colonisation of the gut and extensive faecal shedding. It may also cause invasive infections in 1 day-old chickens.     

The effect of experimental infection with Rhodococcus equi on immunodeficient mice

To investigate the pathogenesis of respiratory lesions caused by the facultative intracellular pathogen, Rhodococcus equi, pulmonary clearance was compared in four groups of genetically defined mice, chosen for their specific deficits in immune and inflammatory responses. Complement-deficient A/J, immunodeficient nu/nu (nude), scid/scid.bg/bg (SCID/beige), C57BI/6J.bg/bg (beige) and normal Swiss mice (SW) received approximately 10(7) R. equi intranasally on day 0. Bacterial clearance was assessed in lung, liver and spleen on days 1, 4, 7 and 14. Pulmonary clearance was not significantly different between SW and A/J mice. Beige mice cleared R. equi more rapidly and completely than A/J and SW, indicating that deficits in phagocytic and NK cell function associated with the bg/bg gene did not compromise clearance. Pulmonary clearance in immunodeficient SCID/beige mice paralleled that of the SW and A/J mice initially but bacterial proliferation produced significant differences from SW mice at day 14. Nude mice were unable to clear R. equi from day 1, resulting in the death of two nude mice at day 11. Both SCID/beige and nude mice developed severe pyogranulomatous bronchopneumonia, whereas A/J and SW mice developed transient pulmonary lesions. Beige mice developed minimal lung lesions. Significant systemic bacterial proliferation occurred only in nude and SCID/beige mice. We conclude that deficiencies in complement components, phagocytic and NK cells do not impair the pulmonary clearance of R. equi but that a competent cellular immune system is required to prevent pneumonia and death. The difference in early phase pulmonary clearance in nude and SCID/beige mice indicates two phases are important for clearance. An acapsular mutant of R. equi was completely cleared from the lungs of SCID/beige mice suggesting an important role for the capsule in virulence for mice.     

Histopathology of the early phase during experimental Corynebacterium pseudotuberculosis infection in lambs.

Histological responses during experimental Corynebacterium pseudotuberculosis infection in lambs were investigated in parotid lymph nodes for ten days following inoculation. Lambs were infected by the subcutaneous route into the right eyelid with a virulent strain of C. pseudotuberculosis. Multiple microscopic acute abscesses, predominantly infiltrated with polymorphonuclear (PMN) leucocytes, were seen in the right parotid lymph node on the 1st day post-inoculation (PI). This massive PMN infiltration coincided with a peripheral blood granulocytosis. On day 3 PI, an influx of histiocytes was observed, while the microabscesses became confluent. From day 3 to day 10 PI, these lesions became enlarged and transformed into typical pyogranulomas with a central necrosis and a peripheral mantle of mononuclear cells composed of macrophages, epithelioid cells and lymphocytes; these histological changes were associated with a bacterial dissemination limited to the superficial lymph nodes. A lymphoid hyperplasia with prominent germinal centers was observed in the draining lymph nodes from day 3 PI. These results illustrate the dual role of granulomatous lesions in chronic bacterial infections: although they limit bacterial dissemination, the granulomas do not impair the persistence of infectious organisms in the host, leading to focal tissue damage.     

The effect of experimental infectious mastitis on leukocyte subpopulations and cytokine production in non-lactating ewes.

The interactions between leukocytes and cytokines during the acute response to intramammary infections in the dry mammary gland of sheep were studied. Dry ewes were experimentally infected in one udder half with either Staphylococcus aureus or Escherichia coli, or infused with saline as control. Udder secretion samples, blood samples and udder tissue samples were collected before and 4, 8 and 24 h after infections/infusions. Total and differential leukocyte counts were calculated in both blood and mammary secretions, and flow cytometry was used to detect the presence of CD4+, CD8+, WC1+, IL-2R+, CD18+ or L-selectin + lymphocytes, CD18+ or L-selectin + neutrophils, and CD14+ leukocytes. Moreover, the concentrations of interleukin-1 beta (IL-1 beta), interleukin-8 (IL-8) and granulocyte-macrophage colony stimulating factor (GM-CSF) in udder secretions were measured using ELISA, and RT-PCR was used to detect the presence of corresponding cytokine mRNA in udder tissue biopsies. The results suggest an association between the concentrations of IL-1 beta, IL-8 and the intensity of neutrophil infiltration of the infected gland. Immunologically relevant changes in proportions of lymphocyte subpopulations might also occur in the acute phase of the inflammatory reaction of the udder. Greater cellular and cytokine responses to E. coli infection may have contributed to the milder clinical picture and more rapid resolution of infection than that seen for S. aureus. Enhancing the production of pro-inflammatory cytokines may improve defence against bacterial mastitis.     

Enterocolitis in cattle associated with Yersinia pseudotuberculosis infection

A syndrome in cattle of diarrhoea and death associated with enteric Yersinia pseudotuberculosis infection is described. Outbreaks occurred during winter and early spring in adult cattle grazing pastures waterlogged by recent flooding or persistent heavy rain. Antibiotic therapy was effective early in the course of the syndrome. At necropsy there was severe acute enterocolitis, and bacteria consistent with Y. pseudotuberculosis were observed in the lesions. This organism could usually be isolated from the intestines of affected animals but was recovered less often from other organs. Representative isolates were identified as Y. pseudotuberculosis serotype III. The association of this syndrome with waterlogged pastures and low temperatures suggests that these conditions favour transmission of Y. pseudotuberculosis infection in cattle. The role of Y. pseudotuberculosis as primary pathogen requires confirmation.     

The effects of maedi-visna virus infection on productivity in ewes

A total of five data sets obtained from four separate sheep flocks were used to evaluate the effects of subclinical maedi-visna virus infection (as determined by serological procedures) on productivity in ewes. In general, infection with maedi-visna virus had a detrimental effect on productivity. Infection resulted in a reduction in conception rates, with the odds of an infected ewe conceiving being only approximately 0.67 times those of a non-infected ewe. For ewes which did conceive, there was no detrimental effect of infection on the number of lambs born. The birth weights of lambs born to middle-aged (3–4 years) infected ewes were 3–6% lower than the weights of lambs born to non-infected ewes of the same age. There was very little apparent effect of infection in younger (1–2 years) and older (5 + years) ewes: some factors which may have biased these results are discussed. Lamb weights at 25–50 days of age appeared to be reduced by maedi-visna infection in the ewe but the results were not statistically significant.     

The effect of eprinomectin treatment at calving on reproduction parameters in adult dairy cows in Canada

A clinical trial was carried out in two provinces of Canada to investigate the effect of treatment with eprinomectin at calving on production parameters in adult dairy cattle in 1999-2000. One of the objectives of this study was to evaluate the impact of treatment on reproductive performance as measured by: calving-to-conception interval, calving-to-first service interval and number of services per conception. The ability of an indirect ELISA using a crude adult Ostertagia ostertagi antigen to predict response to treatment also was evaluated. All lactating cows in 20 dairy herds were allocated randomly to receive either eprinomectin pour-on or placebo at calving. Information on reproductive parameters was obtained from computerised cow records. Survival models were used to evaluate the effect of treatment on the two intervals and a Poisson model was used to evaluate the number of services to conception. A total of 549 cows were included in these analyses. A marginally significant treatment effect on calving-to-conception interval was observed (hazard ratio=1.24, P=0.06) but not on calving-to-first service interval. A significant reduction in the number of breedings to conception for treated animals also was observed with a longer effect in cows with short interval to first service. Milk samples from a subset of 109 late-lactation cows were tested for antibodies against O. ostertagi. The ELISA optical-density ratio (ODR) values obtained between 120 days before calving and drying off were categorised as high ODR (>or=0.5) and low ODR (<0.5). Among untreated animals, the hazard of conception was lower (hazard ratio=0.38, 95% CI=[0.19,0.75]) for high-ODR cows compared to low ODR cows suggesting that higher parasite burdens had an adverse effect on reproductive performance. Treated high-ODR cows had a hazard of conception equivalent to the hazard for all cows in the low-ODR group (indicating that treatment prevented the negative effect associated with these higher parasite burdens).     

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.     

Enteritis associated with Yersinia pseudotuberculosis infection in a buffalo

SUMMARY: A week after transport a 4 month old buffalo calf developed diarrhoea. Its condition gradually deteriorated and it died. Necropsy revealed acute haemorrhagic enteritis and enlarged mesenteric lymph nodes. Haemorrhages and numerous microabscesses were detected in the lamina propria of the small intestine associated with colonies of Gram negative bacteria. Yersinia pseudotuberculosis was isolated from the small intestine and from mesenteric lymph node. Enteritis caused by Y pseudotuberculosis does not appear to have been reported previously in buffalo in Australia.