Agricultural Research Management Information System

Home
Research Summary
Search
About Us
- ARMIS
- Brochure
Contact Us
Report
User Request
Data Input
Help
- Operation Manual
  - PDF
  - Video
- Program Area & Commodity
We have reached 37600 number of research entries at this moment.

- Logout

Research Detail

Home
Research
Detail

The F-34 Stern Strain Anthrax Vaccine Induced Higher Level of Anti-anthrax Igg Antibody Response and Appeared Protective in Mice Model

Provat Chandra Saha
Department of Livestock Services, Ministry of Fisheries and Livestock, Bangladesh

Tahomina Ruba
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh

Umme Kulsum Rima
Department of Medicine, Surgery and Obstetrics, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh

Mohammad Shahidul Islam
Department of Livestock Services, Ministry of Fisheries and Livestock, Bangladesh

Golam Azam Chowdhury
Department of Livestock Services, Ministry of Fisheries and Livestock, Bangladesh

Mohammed Ahasan Habib
Department of Livestock Services, Ministry of Fisheries and Livestock, Bangladesh

Mohammad Zakir Hossain
Department of Livestock Services, Ministry of Fisheries and Livestock, Bangladesh

Priya Mohan Das
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh

Mohammad Abu Hadi Noor Ali Khan
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh

Abstract/ Executive Summary:

Anthrax is a fatal septicemic disease of warm-blooded animals including human and caused by Bacillus anthracis. Anthrax is common in farm ruminants, causes regular mortality and the disease is commonly prevented by using vaccine. The Sterne strain F-34 vaccine has been using in ruminants for more than five decades in Bangladesh but there is little work describing the efficacy of the vaccine in laboratory or farm animal’s. This study was aimed to evaluate efficacy of F-34 Sterne strain anthrax vaccine in mice model and isolate B. anthracis in culture from field outbreaks to identify level of protection in vaccinated mice. Anthrax vaccine containing F-34 Stern strain was obtained from Livestock Research Institute (LRI), Mohakhali, Dhaka. Group C and D female mice were immunized subcutaneously with 0.1ml of vaccine and Group A and B mice were served as non-immunized control. Polymerase chain reaction (PCR) technique was carried out to detect pX01 (210bp) and pX02 plasmid (1035bp) of B. anthracis from the field isolates, vaccine bacteria and challenged mice. Out of 13 field samples tested, B. anthracis was isolated from 05 cases. Following 6 months of immunization the Group B and Group D mice were challenged intraperitoneally with 2x10⁵ colony forming unit (CFU) of virulent field isolate of B. anthracis (isolate of Shahjadpur Upazila, Sirajganj). The vaccine efficacy was evaluated in terms of anti-anthrax IgG antibodies responses in ELISA (A⁴⁵⁰±SD) and protection to challenge in vivo. The early anti-anthrax IgG antibody response was detected in Group C and D mice following week 2 of immunization (0.501±0.167) and reached its peak in study week 4 (1.237±0.257). A steadily higher level of anti-anthrax IgG antibody response was detected until 180 days of study (1.269±0.217, group average±SD). In vivo challenge to vaccinated mice with the virulent B. anthracis bacteria found to confer solid protection following six months of immunization. Non-immunized mice challenged with field isolate of B. anthracis succumbed to death within 18-24hours of infection, showed characteristics lesions of anthrax including black berry jam spleen, wide spread congestion and hemorrhages and bleeding through natural opening after death. B. anthracis was re-isolated from the visceral organs of experimentally infected mice. This study provide evidence that the Sterne strain F-34 Anthrax vaccine is protective in mice model and generated higher level of anti-anthrax IgG antibody response until day 180 of immunization. It needs to study whether the vaccine is effective in farm animal model with longer duration.

Keywords: B. anthracis, Vaccine efficacy, Immunity, Sterne strain, Experimental mice

Location: Tangail sadar, Madhupurl, Gopalpurl, Ghatail and Dhanbari Upazila of Tangail district; Shahjadpur and Belkuchi Upazila of Sirajganj district; Srimongal Upazila, Moulvibazar; Barisal sadar of Barisal district and Moulvibazar sadar of Moulvibazar district, Bangladesh.

Start Date: 00-05-2013

End Date: 00-04-2015

Program Area: Pest Management

Commodity: Vaccine

Objectives:

To investigate experimental anthrax in mice model and to evaluate the pathogenicity of the isolates and protective efficacy of the vaccine in vivo.

Methodology:

Isolation of B. anthraces in culture and preparation of antigens: Isolation and identification of field isolates of B. anthracis was carried out from a total of 13 suspected animal cases in Bangladesh during the period between May 2013 to April 2015. The samples was collected from two infected areas of Tangail sadar, Tangail, Shahjadpur Upazila, Sirajganj and Srimongal Upazila, Moulvibazar. Samples were collected from an infected area of Madhupur Upazila, Tangail, Gopalpur Upazila, Tangail, Ghatail Upazila, Tangail, Dhanbari Upazila, Tangail, Belkuchi Upazila, Sirajganj, Barisal sadar, Barisal and Moulvibazar sadar, Moulvibazar. Cattle suddenly died and discharging tarry colored blood through the natural opening constituted the study materials. The turbinate bone and discharges through rectal opening was collected in sterile vials and transported to the laboratory, Department of Pathology, Bangladesh Agricultural University, Mymensingh. About 1.0gm of turbinate bone or discharges from the dead cattle was crushed on sterile pestle and mortar with 5ml sterile distilled water under a biosafety cabinet level 2 (BSL-2). The suspension was heated in a thermal block at 90? for 05mins to kill contaminating bacteria and other vegetative pathogens. 100μl suspension in 10ml nutrient broth containing 0.05mg/ml Fungizone (amphotericin B, Thermo Fisher Scientific INC, NY) was incubated at 37°C for 12hrs with shaking (160rpm). About 1ml of bacterial growth in nutrient broth in eppendorf tube was centrifuged at 10000 x g for 5 minutes and sediment was smeared on glass slides, stained with Gram’s iodine to visualize the growth of bacteria. In positive cases the bacterial growth in nutrient broth was, therefore, used to isolate B. anthraces on polymyxin-lysozyme-EDTA-thallous acetate (PLET) agar and sheep blood agar media. To isolate vaccine strain of B. anthracis (Sterne strain F-34), a vaccine vial (100ml) was collected from the Livestock Research Institute (LRI), Mohakhali, Dhaka, Bangladesh on July 2013. About 1.0ml of vaccine formulation was taken into an eppendorf tube and centrifuged at 10000 x g for 5mins. The supernatant was discarded and the pellet was washed twice with sterile distilled water by centrifugation at 10000 x g for 5mins. The supernatant was discarded and 100μl sterile distilled water was added to the tube. Using a bacteriological loop the broth was used to grow on sheep blood agar plate at 37°C for 12 hrs. Once the bacterial growth was seen on culture, the bacterial colony was stained with Gram’s iodine (Luna, 1968) and examined under microscope to observe the morphology of the bacteria. PX01 and PX02 plasmid specific PCR was carried out to identify species of B. anthraces. The field bacterial isolates of B. anthraces was used in antigen preparation and to evaluate protective efficacy of the vaccine against challenge infection in mice model. The name of the isolates designated as BD_Vac_2014 (vaccine bacteria, F-34 Stern strain) and FI SPUR_2014 (virulent field isolate of B. anthracis) with the Genbank accession number of KT995458 and KT893481 respectively. The virulent field isolate used in this study was collected and characterize from Shahjadpur Upazila, Sirajganj district, Bangladesh.

Source of Information: J Bangladesh Agril Univ 18(2): 405–414, 2020

Article Link (Online Journal): https://doi.org/10.5455/JBAU.95978

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Out of 13 field samples tested, B. anthracis bacteria was isolated from five cases. The cultural, morphological and PCR assay (targeting pX01 and pX02 plasmids) confirmed that the species of the bacteria isolated in culture was B. anthracis. In two cases bacillus bacteria were isolated, appeared motile, hemolytic in sheep blood agar, were non B. anthracis and thus discarded. The mice model used in immunoefficacy trial, showed fairly a significantly higher level of anti-anthrax IgG antibody response and the response persisted until the end of study (day 180 of immunization). The protective efficacy of anthrax Sterne strain F-34 vaccine was evaluated in mice model. The vaccinated mice were challenged with 2x105 CFU of virulent field isolate of B. anthracis (isolate from Shahjadpur Upazila, Sirajganj district) following 180 days of immunization withstand challenge. The nonvaccinated mice challenged with 2x105 CFU of the virulent field isolate of B. anthracis were died within 1824hours of infection. The Sterne strain F-34 vaccine appeared protective in mice model following a single injection and provide protection up to six months of immunization. It needs to study whether the vaccine is effective for a year and effective in large ruminants as well.

Knowledge Management: Journal