MOLECULAR ANALYSIS OF STAPHYLOCOCCUS AUREUS ISOLATED FROM INFECTED DAIRY GOATS IN RAWALPINDI PAKISTAN
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Author:
MUHAMMAD ASGHAR QAZI
Citable URI :
https://vspace.vu.edu.pk/detail.aspx?id=140
Publisher :
Virtual University of Pakistan
Date Issued:
8/27/2018 12:00:00 AM
Abstract
Staphylococcus aureus is the main cause of mastitis in dairy goats. This organism is recognized as major mastitis causing pathogen in dairy animals worldwide. In humans, it also caused various diseases such as food poisoning and Staphylococcal enterotoxins. This bacterium mostly infected the dairy herds due to poor sanitary conditions and untreated groundwater used to wash utensils during milking practice. The purpose of this study was to determine the prevalence of Staphylococcus aureus among infected dairy goats. Hundred milk samples were collected from infected dairy goats and grown on a blood agar. A total of 58/100 (58%) were positive for S.aureus that was further confirmed on the basis of biochemical tests namely catalase positive and coagulase positive. The positive isolates of S. aureus strains were analyzed using a molecular technique. DNA of positive S. aureus isolates was extracted using Macherey-Nagel kit method. The extracted isolates of S. aureus were run on agarose gel electrophoresis. Ethidium bromide dye was used to visualize DNA bands using UV illuminator. The amplification of genes encoding 16S rRNA of positive S. aureus isolates was performed by Polymerase chain reaction. Primers (Forward primer 5'-AGA GTT TGA TCC TGG CTC AG & Reverse primer 5'-AAG GAG GTG ATC CAG CCG CA) were designed using Primer 3 software. PCR clean up method was used in purification of sequences. These amplified gene products were further sequenced by using Sanger method. The sequenced results of these bacterial species were confirmed by comparing the sequences present in BLAST program at National Centre of Biotechnology Information. MEGA 7 software was applied for the construction of Phylogenetic tree. Some other bacterial species such as Staphylococcus epidermidis, Staphylococcus hominis, Enterobacter cloacae and Acinetobacter baumannii along with Staphylococcus aureus were also identified due to their genetic differences. The evolutionary history was inferred using the Neighbor-Joining method. The evolutionary distances were computed using the p-distance method and in the units of the number of base differences per site. The analysis involved 14 nucleotide sequences. There were a total of 371 positions in the final dataset. Antibiotics such as penicillin, erythromycin or tetracycline are used in the treatment of mastitis in goats. It is concluded that Staphylococcus aureus was more prevalent in the milk samples of infected goats and is an etiologic agent of mastitis in dairy goats. The consumption of raw milk may infect humans and lead to various infections.
URI :
https://vspace.vu.edu.pk/details.aspx?id=140
Citation:
Qazi, M.A(2017), MOLECULAR ANALYSIS OF STAPHYLOCOCCUS AUREUS ISOLATED FROM INFECTED DAIRY GOATS IN RAWALPINDI PAKISTAN.Virtual University of Pakistan.(Lahore, Pakistan).
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Final Version
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