Brucellosis, in particular infections with Brucella abortus, Brucella melitensis or Brucella suis, remains a significant human health threat in many areas of the world. The persistence of pathogenic Brucella spp. in domestic livestock or free-ranging wildlife remains unresolved, despite decades of regulatory efforts worldwide. Although vaccination is probably the most economic control measure, administration of currently available vaccines alone is not sufficient for elimination of brucellosis in any host species. Complacency in brucellosis control programs usually results in failure, or at best, limited reductions in disease prevalence or incidence of human infections. New brucellosis vaccines with high efficacy and safety are needed that address the diversity in host species and can be more widely applied under field conditions. Development of safer and more efficacious vaccines alone, or combined with enhancements or increased emphasis on other regulatory program components, could have tremendous impac...

Essential role of vaccines in brucellosis control and eradication programs for livestock

Brucella spp. are zoonotic, facultative intracellular pathogens, which cause animal and human disease. Animal disease results in abortion of fetuses; in humans, it manifests flu-like symptoms with an undulant fever, with osteoarthritis as a common complication of infection. Antibiotic regimens for human brucellosis patients may last several months and are not always completely effective. While there are no vaccines for humans, several licensed live Brucella vaccines are available for use in livestock. The performance of these animal vaccines is dependent upon the host species, dose, and route of immunization. Newly engineered live vaccines, lacking well-defined virulence factors, retain low residual virulence, are highly protective, and may someday replace currently used animal vaccines. These also have possible human applications. Moreover, due to their enhanced safety and efficacy in animal models, subunit vaccines for brucellosis show great promise for their application in livestock and humans. This review summarizes the progress of brucellosis vaccine development and presents an overview of candidate vaccines.

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Progress in Brucella vaccine development

The recombinant leukotoxin (rLKT) of the bacterium Pasteurella haemolytica A1 was examined for its ability to protect cattle from experimental challenge with logarithmic-phase P. haemolytica. Six different vaccines were utilized in the experiment: P. haemolytica culture supernatant, P. haemolytica culture supernatant enriched with rLKT, rLKT alone, P. haemolytica culture supernatant enriched with Escherichia coli supernatant not containing LKT, E. coli supernatant alone, and phosphate-buffered saline. rLKT alone showed no protective capacity against development of clinical signs of respiratory disease or against development of postmortem lung lesions after experimental challenge. It was, however, shown to enhance the efficacy of the culture supernatant vaccine and decrease clinical signs and pneumonic lesions. The complexity of protective immunity in this disease is emphasized in this study, and, although LKT is an important virulence factor of the organism, an immune response to LKT alone does not protect animals against disease.

https://iai.asm.org/content/iai/59/2/587.full.pdf

Efficacy of recombinant leukotoxin in protection against pneumonic challenge with live Pasteurella haemolytica A1.

Antigens associated with whole Pasteurella haemolytica biotype A serotype 1, a capsular carbohydrate-protein extract of the organism, and P. haemolytica leukotoxin were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Antigens of the electrophoresed preparations were detected by Western blotting (immunoblotting) with sera from cattle which were either nonvaccinated or vaccinated with live or killed P. haemolytica vaccines and had variable degrees of resistance to experimental pneumonic pasteurellosis. Distinct, easily recognizable antigens of these preparations were identified, and the antibody responses to these antigens were quantified by densitometry. To determine their importance to disease resistance, we then compared antibody responses with experimental lesion scores. Antibody reactivity to surface antigens which were significantly correlated with resistance and present in two or more of the preparations were detected at 86, 66, 51, 49, 34, 31, and 16 kilodaltons (kDa). Of these, antibody responses to antigens at 86, 49, and 31 kDa appeared most important based on their concentration and significance levels. Antibody reactivity to leukotoxin antigens which were significantly correlated with resistance and common with important surface antigens were detected at 86, 66, and 49 kDa. Antibody responses to unique leukotoxin antigens which were significantly correlated with resistance were present at 92 and 58 kDa.

Pasteurella haemolytica antigens associated with resistance to pneumonic pasteurellosis.

▪ Nacido en Cartagena (Murcia), en noviembre de 1954. ▪ Licenciado en Ciencias Químicas por la Universidad de Murcia, Catedrático de Física y Química, Divulgador Científico freelance, Escritor, y Conferenciante. ▪ Con 35 años de experiencia docente, ejerciendo como profesor. ▪ A lo largo de su dilatada carrera docente, ha desempeñado cargos de responsabilidad en la gestión de Centros Escolares, tales como Director (7 años), Jefe de Estudios (2 años), Jefe de Departamento de Física y Química (20 años), miembro del Consejo Escolar de Instituto y miembro del Consejo del Centro de Profesores y Recursos de Cartagena (11 años). ▪ Presentación de comunicaciones y ponencias en 28 Congresos, Jornadas y Seminarios. ▪ Ha sido durante 13 años tutor del segundo ciclo del curso de Aptitud Pedagógica de Educación Secundaria, organizado por el Instituto de Ciencias de la Educación, de la Universidad de Murcia y la Universidad Complutense de Madrid. ▪ También ha sido, durante 1 año, tutor de la fase de prácticas del Máster de Profesor de Educación Secundaria, organizado por el Instituto de Ciencias de la Educación, de la Universidad de Murcia.

Curriculum Vitae for

Effects of challenge dose on the clinical and immune responses of cattle vaccinated with reduced doses of Brucella abortus strain 19.

An enzyme-linked immunosorbent assay (ELISA) was developed for detection of bovine serum antibodies to the cytotoxin (leukotoxin) of Pasteurella haemolytica. A partially purified, cytotoxic, and immunogenic protein obtained from supernatants of logarithmic-phase P. haemolytica was used as the ELISA antigen. Preadsorption of sera with various cytotoxic, somatic, and capsular antigen preparations demonstrated that the assay was specific for anticytotoxin antibodies. ELISA anticytotoxin titers had a strong, significant correlation to cytotoxin-neutralizing-antibody titers. The ELISA, however, was more rapid and allowed for greater numbers of samples to be run than did the neutralization technique. ELISA anticytotoxin titers were high in cattle vaccinated with a live P. haemolytica vaccine, whereas unvaccinated cattle and cattle receiving a P. haemolytica bacterin had low ELISA anticytotoxin titers. A significant positive correlation between ELISA titers and resistance to experimental bovine pneumonic pasteurellosis was present.

Enzyme-linked immunosorbent assay for detection of serum antibodies to Pasteurella haemolytica cytotoxin (leukotoxin) in cattle

Two experiments were done on 57 steers. These cattle were allotted to 8 groups (4 groups/experiment) and vaccinated with 1 to 3 X 10(9) colony-forming units of Brucella abortus strain 19. Cattle in 3 of the 4 groups/experiment were given 6 mg of levamisole/kg, subcutaneously, either at the time of vaccination (day 0), 7 days later, or at both times. Serum antibody titers to B abortus were measured sequentially for 28 days in experiment 1 and for 56 days in experiment 2, using the card test, Rivanol test, complement-fixation test, fluorometric immunoassay, and an enzyme-linked immunosorbent assay. In general, the highest mean antibody titers, as determined by all serologic tests, occurred in steers treated with levamisole at 7 days after vaccination or in those treated at the time of vaccination and 7 days later. By the card test on day 56, there was a significantly (P less than 0.05) greater number of seropositive cattle among those given levamisole 7 days after seropositive cattle among those given strain 19 alone. Simultaneous administration of strain 19 and levamisole did not alter antibody responses to B abortus.

S. M. Hall

Papers

Effects of challenge dose on the clinical and immune responses of cattle vaccinated with reduced doses of Brucella abortus strain 19.

Enzyme-linked immunosorbent assay for detection of serum antibodies to Pasteurella haemolytica cytotoxin (leukotoxin) in cattle

Transient enhancement of the serum antibody response to Brucella abortus strain 19 in cattle treated with levamisole.