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  • Author or Editor: Luis E. Samartino x
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Summary

Chorioallantoic membrane (cam) explants were used to determine the in vitro growth and cytotoxic potential of 3 strains of Brucella abortus. Bovine cam explants were inoculated with 2 × 107 colony-forming units of the pathogenic strain 2308, attenuated strain 19, or the rough strain RB51 of B abortus. After inoculation, the explants were harvested and examined at 2 or 4 hours, 12 or 14 hours, and 24 or 26 hours of incubation. Bacterial growth associated with each explant was determined by counting colony-forming units. The degree of cellular damage in each explant associated either with bacterial growth or bacterial toxins was evaluated by morphometric analysis after trypan blue staining.

Significant differences were not detected in the numbers of bacteria of any strain of B abortus in the cam explants at comparable time intervals. The rate of growth of the bacteria in cam explants was higher between 2 and 12 hours after inoculation than between 12 and 24 hours after inoculation. Cytotoxic effects associated with strain 2308 were significantly (P < 0.05) greater than that caused by other strains. Cytotoxic effects associated with strain 19 and rough strain RB51 were similar, and both were significantly (P < 0.05) greater than the phosphate buffer solution control. Chorioallantoic membrane explants inoculated with a filtrate of heat-killed strain 2308 induced minimal cellular damage, compared with that caused by the viable bacteria. These results indicated that the number of B abortus in trophoblasts was independent of the degree of cellular damage.

Free access
in American Journal of Veterinary Research

Abstract

Objective—To develop a novel oral vaccine delivery system for swine, using the rough vaccine strain of Brucella abortus.

Animals—56 crossbred pigs from a brucellosis-free facility.

Procedure—In 3 separate experiments, pigs were orally vaccinated with doses of 1 × 109 to > 1 × 1011 CFU of strain RB51 vaccine. The vaccine was placed directly on the normal corn ration, placed inside a whole pecan, or mixed with cracked pecans and corn.

Results—Oral vaccination of pigs with vaccine strain RB51 resulted in a humoral immune response to strain RB51 and short-term colonization of the regional lymph nodes.

Conclusions and Clinical Relevance—A viscous liquid such as Karo corn syrup in association with pecans that scarify the oral mucosa are necessary when placing the live vaccine directly onto corn or other food rations. Doses of > 1 × 1011 CFU of RB51 organisms/pig in this mixture ensures 100% colonization of regional lymph nodes via the oral route. This method may allow an efficient and economical means to vaccinate feral swine for brucellosis. (Am J Vet Res 2001;62:1328–1331)

Full access
in American Journal of Veterinary Research

Abstract

Objective

To determine shedding and colonization profiles in mature sexually intact bulls and pregnant heifers after vaccination with a standard calfhood dose of Brucella abortus strain RB51 (SRB51).

Animals

6 sexually mature 3-year-old Jersey bulls and 7 mixed-breed heifers in midgestation.

Procedure

Bulls and pregnant heifers were vaccinated IM with the standard calfhood dose of 3 × 1010 colony-forming units of SRB51. After vaccination, selected body fluids were monitored weekly for vaccine organism shedding. Pathogenesis was monitored in bulls by weekly breeding soundness examination and, in heifers, by delivery status of the calf. Vaccine organism colonization was assessed by obtaining select tissues at necropsy for bacterial culture. Serologic analysis was performed by use of numerous tests, including complement fixation, an SRB51-based ELISA, and immunoblot analysis.

Results

After vaccination, none of the vaccinated bulls or heifers shed SRB51 in their secretions. Results of breeding soundness examination for bulls were normal as was delivery status of the pregnant heifers (6 live births, 1 dystocia). At necropsy, SRB51 was not recovered from any of the selected tissues obtained from bulls, heifers, or calves; however, serologic analysis did detect SRB51-specific antibodies in all cattle.

Conclusions and Clinical Relevance

Vaccination with the standard calfhood dose of SRB51 administered IM was not associated with shedding or colonization in sexually mature bulls or pregnant heifers. Also, under conditions of this study with small numbers of animals, IM vaccination with SRB51 does not appear to cause any reproductive problems when administered to sexually mature cattle. (Am J Vet Res 1999;60:722–725)

Free access
in American Journal of Veterinary Research