ABSTRACT
PURPOSE: Foot-and-mouth disease (FMD) and anthrax are important diseases in sheep. Vaccination is a favorable strategy against both infections. Simultaneous administration of vaccines does generally not impede the immune responses of each other, although there are some exceptions, and it may help reduce the labor and costs of vaccination as well as distress on animals. Although oil adjuvant FMD vaccine has been tried with live anthrax vaccine in cattle, there are no reports on the simultaneous use of both vaccines in sheep. MATERIALS AND METHODS: In this study, FMD seronegative sheep were used to investigate the impact of the simultaneous vaccination of FMD and anthrax on FMD antibody titers of sheep. Virus neutralization test and liquid phase blocking enzyme-linked immunosorbent assay were used to determine the antibody response to the FMD vaccine. RESULTS: The results demonstrated that both vaccines can be used simultaneously without any interference with the FMD response. Moreover, the simultaneous administration with anthrax vaccine had a stimulating effect on the early (day 7 post-vaccination) virus neutralization antibody response to the FMD vaccine. CONCLUSION: The simultaneous use of the FMD and anthrax vaccines did not hinder the response to the FMD vaccine in sheep.
Subject(s)
Animals , Cattle , Anthrax Vaccines , Anthrax , Antibody Formation , Enzyme-Linked Immunosorbent Assay , Foot-and-Mouth Disease , Neutralization Tests , Sheep , Vaccination , VaccinesABSTRACT
On December 14, 2017, a faculty member of a university in Hunan Province reported that an anthrax vaccine strain might have recovered virulence during an undergraduate experiment and potential exposure could not be ruled out for the students involved. Upon receiving the case report, the CDC, health bureaus, and local governments at the county, prefectural, and provincial levels promptly organized experts in different fields (including epidemiologists, biosafety experts, and laboratory testing experts) for case investigation, evaluation, and response. As the investigation results showed, no virulence recovery was identified in the involved anthrax vaccine strain; and no contamination of Bacillus anthracis was detected at the involved areas. Thus, the university returned to normal functioning.
Subject(s)
Humans , Anthrax Vaccines , Bacillus anthracis , Virulence , China , Containment of Biohazards , Laboratories , VirulenceABSTRACT
Anthrax is a zoonosis caused by Bacillus anthracis, which seriously affects human health. In recent years, a special phenomenon is found that the metabolic active of a bacterium remains after it is killed. To development of a KBMA (killed but metabolically active) Bacillus anthracis vaccine candidate strain, a plasmid pMAD and a recombinase system Cre-loxP were used to knockout the uvrAB gene of B. anthracis AP422 which lacks both of two plasmids pXO1 and pXO2. The results of PCR and RT-PCR shows that uvrAB genes were deleted from B. anthracis AP422 chromosome successfully. The constructed B. anthracis AP422deltauvrAB was inactivated by photochemical treatment (PCT) including an exposure in a long-wave-length ultraviolet (UVA) light and a treatment of 8-Methoxypsoralen (8-MOP), then the metabolic activity were detected by the method of MTS. The results showed that the killed B. anthracis AP422deltauvrAB maintained a highly metabolic activity for at least 4 hours, showing a state of KBMA. The KBMA strain of B. anthracis AP422deltauvrAB provides the prospective vaccine candidate strain for anthrax.
Subject(s)
Anthrax , Allergy and Immunology , Microbiology , Anthrax Vaccines , Genetics , Allergy and Immunology , Radiation Effects , Bacillus anthracis , Genetics , Allergy and Immunology , Gene Knockout Techniques , Methoxsalen , Pharmacology , Ultraviolet Rays , Vaccines, Inactivated , Genetics , Allergy and ImmunologyABSTRACT
An expression plasmid carrying anthrax protective antigen (PA) gene was constructed, which has an OmpA signal sequence attached to the 5' end of PA gene. The plasmid was transformed into E. coli and induced to express recombinant PA (rPA) . The recombinant protein, about 10% of the total bacterial protein in volume, was secreted to the periplasmic space of the cell. After a purification procedure including ion-exchange, hydrophobic interaction chromatography, and gel filtration, about 15 mg of 95 % pure rPA was obtained from 1-liter culture. The bioactivity of rPA was proved by in vitro cytotoxicity assay. The polyclonal antiserum from rabbits immunized with rPA could inhibit the action of anthrax lethal toxin in vitro, which suggests that antibodies against rPA can provide high passive protection against anthrax. The results reported here may be helpful to develop a safe and efficacious recombinant PA vaccine against anthrax.
Subject(s)
Animals , Mice , Rabbits , Amino Acid Sequence , Anthrax Vaccines , Allergy and Immunology , Antigens, Bacterial , Chemistry , Genetics , Allergy and Immunology , Toxicity , Bacterial Toxins , Chemistry , Genetics , Allergy and Immunology , Toxicity , Base Sequence , Molecular Sequence Data , Plasmids , Recombinant Proteins , Allergy and Immunology , Vaccines, Synthetic , Allergy and ImmunologyABSTRACT
BACKGROUND & OBJECTIVE: Anthrax has been reported from almost every country and India is endemic for this disease. There is considerable under reporting of the disease because of lack of microbiological facilities and diagnostic reagents. In India only conventional methods which have limitations, are being used to diagnose the disease. Hence the aim of this study was to isolate and purify protective antigen (PA) using different protocols and to use this PA for detection of anti-PA antibodies from sera samples. METHODS: Protective antigen was isolated and purified from the Sterne strain of Bacillus anthracis. B. anthracis lacking pXO1 and pXO2 transformed with pYS5 (B. anthracis pYS5) and recombinant Escherichia coli transformed with pQE30 containing PA gene using hydroxyapatite (HA), Q-sepharose fast protein liquid chromatography (FPLC) and nickel-nitrilotriacetic acid (Ni-NTA) chromatographic methods, respectively. A mixture of PA and edema factor (EF) was injected subcutaneously into rabbits to test the biological activity of PA. The immunogenicity of PA was tested by inoculating the protein into rabbits along with adjuvant. Using this PA, 20 bovine sera samples (pre- and post-vaccinated) were tested by Western blotting (WB) for the presence of anti-PA antibodies. RESULTS: The 83 kDa PA protein was obtained from all the bacteria with the yields of 13, 50 and 9.0 mg/l from Sterne B. anthracis, B. anthracis pYS5 and recombinant Esch. coli, respectively. Formation of edematous ulcers at the site of PA+EF injection clearly confirmed the retention of biological activity of the proteins. Of the 10 post-vaccination sera tested, 9 showed clear positive by WB whereas none of the pre-vaccination sera showed the reaction. INTERPRETATION & CONCLUSION: The purified PA preparations obtained in the present study may possibly be utilized for detection of anti-PA antibodies in the sera of anthrax patients for timely diagnosis of the disease and, might also be tested for their efficacy and use as human anthrax vaccine.
Subject(s)
Animals , Anthrax Vaccines/immunology , Antigens, Bacterial , Bacillus anthracis/metabolism , Bacterial Toxins/blood , Biocompatible Materials/pharmacology , Blotting, Western , Cattle , Chromatography, Liquid , Durapatite/pharmacology , Electrophoresis, Polyacrylamide Gel , Escherichia coli/metabolism , Viper Venoms/metabolismABSTRACT
Anthrax is caused by Bacillus anthracis, an encapulated and spore-forming bacillus. The disease is usually contracted through uptake of spores that remain viable in the contaminated soil for many years. Anthrax is primarily a disease of herbivorous animals and is uncommon in humans who may get the infection through contact with contaminated animals or their products. Anthrax spores germinate after entering the body through skin abrasions (cutaneous anthrax) or by inhalation (inhalation anthrax) or ingestion (gastrointestinal anthrax) and multiply to produce two exotoxins which determine the virulence along with capsule. Although most cases occur within 48 hours of exposure, germination of spores may occur upto 60 days later. While inhalation anthrax is almost always fatal, intestinal anthrax results in death in 25% to 60% of cases. Upto 20% of cases having cutaneous anthrax may die. Antibiotics are effective if the disease is recognised early and treated appropriately. Penicillin is the drug of choice when disease occurs in natural setting. Ciprofloxacin is recommended when aerosols of anthrax spores are used as bioweapon, prophylactic antibiotics should not be prescribed until risk of exposure is considered real by experts.
Subject(s)
Animals , Anthrax/diagnosis , Anthrax Vaccines , Anti-Bacterial Agents/therapeutic use , Bacillus anthracis/pathogenicity , Bioterrorism , Cattle , Diagnosis, Differential , Guidelines as Topic , HumansABSTRACT
Anthrax toxin consists of three separate proteins, protective antigen (PA), edema factor (EF), and lethal factor (LF). PA binds to the receptor on mammalian cells and facilitates translocation of EF or LF into its cytosol. PA is the primary component of anthrax vaccines. In this study we purified PA from culture filtrates of Bacillus anthracis. The purification involved sequential chromatography through hydroxylapatite, DEAE-Sepharose CL-4B, followed by Mono-Q. The purified PA was judged to be homogeneous on SDS-PAGE, and consisted of a single polypeptide chain with a relative molecular weight of 85,000.