RESUMEN
Plasmids play a crucial role in the evolution of bacterial genomes by mediating horizontal gene transfer. In this work, we sequenced two plasmids found in a Brazilian Bacillus thuringiensis serovar israelensis strain which showed 100% nucleotide identities with Bacillus thuringiensis serovar kurstaki plasmids.
RESUMEN
During the fermentation process, Bacillus thuringiensis (Bt) phages can result in bacterial death and decreased yield. In this work, we describe the genome of a new phage related to the Siphoviridae viral family from a Brazilian strain of Bt which showed high nucleotide sequence identity to the genomes of phages phi4l1 and BtCS33.
RESUMEN
The aim of the present study was to compare the potency and safety of vaccines against Clostridium botulinum (C. botulinum) type C and D formulated with chitosan as controlled release matrix and vaccines formulated in conventional manner using aluminum hydroxide. Parameters were established for the development of chitosan microspheres, using simple coacervation to standardize the use of this polymer in protein encapsulation for vaccine formulation. To formulate a single shot vaccine inactivated antigens of C. botulinum type C and D were used with original toxin titles equal to 5.2 and 6.2 log LD50/ml, respectively. For each antigen a chitosan based solution of 50 mL was prepared. Control vaccines were formulated by mixing toxoid type C and D with aluminum hydroxide [25% Al(OH)3, pH 6.3]. The toxoid sterility, innocuity and potency of vaccines were evaluated as stipulated by MAPA-BRASIL according to ministerial directive no. 23. Encapsulation efficiency of BSA in chitosan was 32.5-40.37%, while that the encapsulation efficiency to toxoid type C was 41,03% (1.94 mg/mL) and of the toxoid type D was 32.30% (1.82 mg/mL). The single shot vaccine formulated using chitosan for protein encapsulation through simple coacervation showed potency and safety similar to conventional vaccine currently used in Brazilian livestock (10 and 2 IU/mL against C. botulinum type C and D, respectively). The present work suggests that our single shot vaccine would be a good option as a cattle vaccine against these C. botulinum type C and D.
Asunto(s)
Vacunas Bacterianas/administración & dosificación , Botulismo/prevención & control , Quitosano , Animales , Anticuerpos Antibacterianos/inmunología , Vacunas Bacterianas/efectos adversos , Vacunas Bacterianas/química , Vacunas Bacterianas/inmunología , Clostridium botulinum/inmunología , Preparaciones de Acción Retardada , Cobayas , Vehículos Farmacéuticos , Potencia de la VacunaRESUMEN
The aim of the present study was to evaluate and standardize the ToBI test in vitro as a substitute for the serum neutralization test in mice for quality control of clostridial vaccines. The ToBI test in vitro was used to evaluate 40 serum samples of known antibody content, obtained from rabbits immunized against clostridiosis with experimental vaccine. The correlation between epsilon antitoxin titers in rabbit sera, determined by the ToBI test and serum neutralization in mice, ranged from 0.222% to 0.452% in polyvalent vaccines and from 0.154% to 0.387% in monovalent vaccines. Interplate coefficients of variation were not significant, reaching 0.350% in polyvalent vaccines and 0.400% in monovalent vaccines, indicating high homogeneity. In conclusion, the ToBI test in vitro is suitable for assessing the potency of clostridial vaccines and may be used as an alternative method able to replace current in vivo tests.
Asunto(s)
Antitoxinas/sangre , Toxinas Bacterianas/antagonistas & inhibidores , Clostridium perfringens/inmunología , Pruebas Serológicas/métodos , Animales , Toxinas Bacterianas/inmunología , Vacunas Bacterianas/inmunología , Vacunas Bacterianas/normas , Infecciones por Clostridium/diagnóstico , Infecciones por Clostridium/inmunología , Infecciones por Clostridium/prevención & control , Enterotoxemia/diagnóstico , Enterotoxemia/inmunología , Enterotoxemia/prevención & control , Inmunización , Técnicas In Vitro , Ratones , Pruebas de Neutralización/métodos , Control de Calidad , Conejos , Pruebas Serológicas/normasRESUMEN
The aim of the present study was to evaluate and standardize the ToBI test in vitro as a substitute for the serum neutralization test in mice for quality control of clostridial vaccines. The ToBI test in vitro was used to evaluate 40 serum samples of known antibody content, obtained from rabbits immunized against clostridiosis with experimental vaccine. The correlation between epsilon antitoxin titers in rabbit sera, determined by the ToBI test and serum neutralization in mice, ranged from 0.222% to 0.452% in polyvalent vaccines and from 0.154% to 0.387% in monovalent vaccines. Interplate coefficients of variation were not significant, reaching 0.350% in polyvalent vaccines and 0.400% in monovalent vaccines, indicating high homogeneity. In conclusion, the ToBI test in vitro is suitable for assessing the potency of clostridial vaccines and may be used as an alternative method able to replace current in vivo tests.
