ABSTRACT
Salmonella choleraesuis C500 strain is an attenuated vaccine preventing piglet from paratyphoid and can also be used as a live vector of other DNA vaccines. Through mucosal immunization, immune response to specific antigens carried by it can be induced. To enhance the immune efficiency of DNA vaccine it carried, promoter Ptrc was inserted into the down stream of the human cytomegalovirus (CMV) immediate early promoter of eukaryotic expression plasmid pEGFP-C1. Then transcription terminator rrnbT1T2 was inserted into down stream of the multiple clone sites of pEGFP-C1, and the dual-promoter expression vector pEGFPPtrcR was constructed. Using 1xTSS method, we transformed the recombinant plasmid into C500, and obtained C500/pEGFPPtrcR. We used SDS-PAGE and Western blotting to detect the expression of report gene EGFP. Strong green fluorescence was observed under fluorescent microscope. The stable passages of this recombinant bacterium were at least 20 generations in vitro. Using liposome we transfected plasmid pEGFPPtrcR into Vero cell. After 24 h, green fluorescent was observed, showing the expression of EGFP in nuclei and endochylema. The construction of dual-promoter expression vector pEGFPPtrcR was successful. The foreign gene was expressed in Salmonella strain C500 and somatocytes, resulting in increased antigen expression. This research provides a foundation for the research of new DNA vaccines which use Salmonella C500 as carrier.
Subject(s)
Animals , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , Metabolism , Plasmids , Genetics , Promoter Regions, Genetic , Genetics , Recombinant Proteins , Genetics , Metabolism , Salmonella Vaccines , Genetics , Allergy and Immunology , Salmonella arizonae , Genetics , Allergy and Immunology , Metabolism , Swine , Vaccines, DNA , GeneticsABSTRACT
In the current work, the fusion gene including somatostatin (SS) and the hepatitis B surface antigen gene was cloned into a balanced lethal system plasmid (pYA3493), and then transformed into asd- attenuated Salmonella choleraesuis C500 strain, the positive transformant without antibiotic resistance gene was confirmed by restriction analysis and DNA sequencing, designated as pYA-SS. The expression and immunogenicity of fusion protein were detected by SDS-PAGE and Western blot analysis. These results show that the recombinant prokaryotic expression plasmid pYA-SS could express the SS fusion protein with good immunogenicity in C500 strain. In above all, this study could provide reliable materials to develop novel, good and safe vaccine in enhancing the growth of animals.
Subject(s)
Animals , Humans , Artificial Gene Fusion , Cloning, Molecular , Hepatitis B Surface Antigens , Genetics , Plasmids , Genetics , Prokaryotic Cells , Metabolism , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Salmonella arizonae , Genetics , Metabolism , Somatostatin , Genetics , Allergy and ImmunologyABSTRACT
Descrição de caso de endoftalmite endógena pelo Arizona sp., microorganismo negativo raramente reconhecido como patógeno humano. Relata-se o caso de um homem com 45 anos, banco, internado devido a um quadro de endocardite bacteriana e sepsis pela Salmonella Arizona sp. O paciente apresentava diminuição da acuidade visual e dor em olho direito. O aspirado vítreo confirmou a etiologia. Vancomicina e Amicacina pelas vias endovenosa e intravítrea foi a opção de manejo adotada. Resposta sist6emica favorável, porém evoluçãocom atrofia do globo ocular em 30 dias. A endoftalmite metastática pelo Arizona sp. é incomum, sem qualquer aspecto clínico específico e de prognóstico muito desfavorável.
Subject(s)
Male , Middle Aged , Humans , Endocarditis, Bacterial/diagnosis , Endophthalmitis/microbiology , Salmonella Infections , Salmonella arizonae/isolation & purificationABSTRACT
Classical Swine Fever Virus (CSFV) E2 protein eukaryotic expression plasmid pVAXE2 was constructed. The plasmid pVAXE2 was transformed into Salmonella choleraesuis C500 (S. C500) attenuated vaccine strain by electroporation to generate Salmonella choleraesuis engineering strain S. C500/pVAXE2. The characterization of S. C500/pVAXE2 in morphology, growth, biochemistry and serology indicated that it retained the same properties as its original strain S. C500 with exception of kanamycin resistance originated from the plasmid pVAXE2. The plasmid stable in the bacteria after 15 passages. Kunming mice and rabbits were vaccinated three times at two weeks interval with S. C500/pVAXE2 in oral and intramuscular routes at the dosage of 1 x 10(8) CFU for mice and 2 x 10(9) CFU for rabbits each time. The specific antibody response against CSFV and Salmonella choleraesuis was detected by ELISA. Two weeks after the third boost the immunized rabbits were challenged with 20 ID50 of hog cholera lapinized virus (HCLV), followed by a virulent strain of Salmonella choleraesuis two week later than HCLV challenge. The results showed that all immunized mice and rabbits produced significant antibodies against CSFV and Salmonella choleraesuis, and the immunized rabbits demonstrated the effective protection against the challenge of HCLV and virulent Salmonella choleraesuis. These results indicated the potential of developing multiplex swine DNA vaccine by using this bacteria as the vector.
Subject(s)
Animals , Mice , Rabbits , Classical Swine Fever , Allergy and Immunology , Virology , Classical Swine Fever Virus , Genetics , Allergy and Immunology , Salmonella arizonae , Genetics , Swine , Vaccines, DNA , Allergy and Immunology , Viral Envelope Proteins , Genetics , Allergy and Immunology , Viral Vaccines , Allergy and ImmunologyABSTRACT
Avaliou-se a qualidade microbiológica de 30 amostras de queijo prato e parmesäo ralado, coletadas em diferentes supermercados da cidade de Caxias do Sul, Rio Grande do Sul(Brasil). Os resultados mostraram índices de contaminaçäo acima do permitido pela legislaçäo em 11 amostras de queijo prato e em 4 de queijo parmesäo. Em uma amostra de queijo prato ralado foi detectada a presença de Salmonella arizonae.
Subject(s)
Cheese , Food Analysis , Food Contamination , Food Microbiology , Salmonella arizonaeABSTRACT
Introducción: la meningitis es una complicación rara de la infección por Salmonella. Caso clínico: lactante masculino de nueve meses de edad, no inmunocomprometido, con antecedentes de convivencia con perros, conejos, aves de corral y lagartijas. Una semana antes de su ingreso al hospital presentó diarrea que se autolimitó en 24 horas. Evoluciona con deterioro neurológico, fontanela anterior abombada, rigidez de nuca y Kerning; se cultivó Salmonella arizonae del líquido cefalorraquídeo (LCR). El manejo inicial fue con ampicilina y cloranfenicol; al conocer la sensibilidad antibiótica se continuó con cloranfenicol y cefotaxima. El LCR se negativiza después de tres días. Una tomografía computarizada mostró atrofia cerebral sin evidencia de absceso o empiema subdural. Egresa a los 31 días con hemiparesia izquierda, tono muscular aumentado y clonus, los cuales desaparecen dos meses después.Conclusiones: este es el segundo caso reportado a nivel mundial, y el primero en México, de meningitis por S. arizonae en niños.
Subject(s)
Humans , Male , Infant , Ampicillin Resistance , Meningitis, Bacterial/etiology , Salmonella arizonae/isolation & purification , Salmonella arizonae/pathogenicity , Cefotaxime/therapeutic use , Chloramphenicol/therapeutic use , Gastroenteritis/complicationsABSTRACT
The septicemia caused by the Arizona group organism is rare and usually observed in adults with underlying diseases. In Korea, Salmonella infection is common, but a report of Arizona infection is unknown. We isolated S. entercia subsp. diarizonae from blood of a 6-month-old infant. The serovar was determined as 28:z10:-, a rare one in America. The isolate was susceptible to ampicillin, chloramphenicol, cotrimoxazole and others. The patient rapidly recovered with ampicillin and gentamicin therapy. Clinical laboratories should consider that the infection exists in Korea and should attempt to isolate and identify Arizona organism in certain patients.