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Bihar Agricultural University, Sabour

Bihar Agricultural University, Sabour established on 5th August, 2010 is a basic and strategic institution supporting more than 500 researchers and educationist towards imparting education at graduate and post graduate level, conducting basic, strategic, applied and adaptive research activities, ensuring effective transfer of technologies and capacity building of farmers and extension personnel. The university has 6 colleges (5 Agriculture and 1 Horticulture) and 12 research stations spread in 3 agro-ecological zones of Bihar. The University also has 21 KVKS established in 20 of the 25 districts falling under the jurisdiction of the University. The degree programmes of the university and its colleges have been accredited by ICAR in 2015-16. The university is also an ISO 9000:2008 certified organisation with International standard operating protocols for maintaining highest standards in teaching, research, extension and training.VisionThe Bihar Agricultural University was established with the objective of improving quality of life of people of state especially famers constituting more than two third of the population. Having set ultimate goal of benefitting society at large, the university intends to achieve it by imparting word-class need based agricultural education, research, extension and public service.

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20171
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Subject: Veterinary Public Health ×
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    ThesisItemOpen Access
    Isolation, characterization and evaluation of Staphylococcus aureus bacteriophage(s) as surface sanitizer
    (Department of Veterinary Public Health & Epidemiology, BAU, Sabour, 2017-09) Archana; Kaushik, P.
    S. aureus is a commensal bacterium capable of colonizing the nose and skin, and can cause life-threatening diseases in different tissues including bones, joints, blood, lungs, heart, and brain. The contaminated objects and surfaces play an important role in S. aureus transmission and once it is established on a surface, can survive for weeks to month. Use of antibiotics has become a routine practice in human as well as in veterinary medicine resulting in development of antibiotic resistance among bacterium. In recent years, there is an increasing consensus about improvement in the disinfection and sanitization of environmental surfaces in healthcare and food processing facilities. In this regard a novel approach, to reduce the need of harsh chemical sanitizers, is the use of lytic bacteriophage(s) as biocontrol or biosanitizing agents. The present study was performed to isolate and characterize wide host range lytic bacteriophages against S. aureus and to evaluate bacteriophage(s) as a surface sanitizing biological agent. A total of 109 S. aureus isolates ( comprising of methicillin sensitive and methicillin resistant S. aureus) were used in the study which were confirmed by biochemical and molecular test. The isolates resulted in the amplification of desired amplicon size of 228bp of target gene (16s rRNA). A total of 32 samples from different sources were processed for isolation of S. aureus and their bacteriophages, but only one lytic phage of S. aureus could be isolated from dog pus sample against the host which was isolated from the same sample. The phage were named as Staphylococcus phage vB_Staph M- BVC1. Host range analysis and lysis profiles of Staphylococcus phage vB_Staph M- BVC1 revealed the high specificity of the phage to the MRSA with a lytic activity of 38.80%. The lysis profile study showed that the phage was not able to lyse any of the Methicillin Sensitive S. aureus isolates of livestock and animal handler’sisindicatesoriginthehigh.specificityTh of phage Staphylococcus phage vB_Staph M-BVC1 to MRSA which can act as a possible agent for the control of MRSA. The Electron micrograph studies demonstrated the novel phage as a member of tailed phage group of order Caudovirales and family Myoviridae. The thermal stability of phage revealed non-significant reduction in phage titre with 100% lytic activity at temperatures when incubated at 4-45oC, however the complete inactivation was observed at 65oC. The SDS-PAGE analysis of structural proteins of Staphylococcus phage vB_Staph M- BVC1 identified 13 major protein bands of 27- 180 kDa as well as 35-51 kDa minor structural proteins. On this basis, it can be suggested that Staphylococcus phage vB_Staph M- BVC1 resembles the phage of the Myoviridae family, which support the finding of transmission electron microscopy. The agarose gel electrophoresis of the phage nucleic acid revealed the presence of genome size of more than 21 kb. The RAPD-PCR of phage with OPL5 primer yielded different patterns of banding consisting of 2 bands between 200 to 2kb whereas RAPD-PCR analysis with RAPD5 primer of phage yielded 10 bands ranging from 200 to 2.0kb. The RAPD-PCR technique appears to be a practical and efficient tool for routine use in high-resolution viral diversity studies by providing assemblage comparisons through fingerprinting, probing, or sequence information. To study the efficacy of phage as surface sanitizer glass cover-slip was used to provide a model system for simulating hard surface contamination in hospital settings. In the study, analysis of the effects of Staphylococcus phage vB_Staph M- BVC1 on glass matrix at room temperature revealed 1 log reduction of contamination at MOI 0.1. The treatment at MOI 1.0, the S. aureus count was reduced by log10 1.1 while 1.5 log reduction was observed at MOI 10. At higher MOI of 100 and 1000, no count or complete reduction of S. aureus was observed for the entire period of study. The findings of the present study suggest that phage treatment can be used as an effective tool to decontaminate MRSA from hard surfaces and the possibility can be explored for its therapeutic use to treat MRSA cases which needs further study.