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Updated: Jul 20, 2026

Use of the Soft-agar Overlay Technique to Screen for Bacterially Produced Inhibitory Compounds
Published on: January 14, 2017
Coliphage hsa as a model for antiviral studies/spectrum by some indigenous bacteriocin like inhibitory substances
Humaira Qureshi1, Sadia Saeed, Samia Ahmed
1Laboratory of Molecular Genetics, Department of Microbiology, University of Karachi, Pakistan.
Abstract:
Coliphage HSA was isolated from a raw sewage sample (collected from a local sewage treatment plant). The phage was analyzed by spot and tube lysis followed by plaque assay. Phage titre (plaque forming units i.e. PFU) was found to be 4.2 x 103 PFU/mL. Further purification of the phage was achieved by acid-precipitation method. Genomic identification of the coliphage HSA (done by fluorescent staining using acridine orange) revealed it to be a dsDNA bacterial virus. Staphylococcin188, Enterocins AAR-71, AAR-74, and Erwiniocin NA4 were screened for their antiphage activity by plaque assay. Accordingly, all the bacteriocin preparations possess demonstrable antiphage activity witnessed as a reduction in PFU after treatment. In the case of Staphylococcin 188, the number dropped up to 40 PFU/mL, Enterocin AAR-71 and Erwiniocin NA4 treatment reduced it to a zero PFU level, while Enterocin AAR-74 could reduce PFU to 50 (after addition of a constant volume, 500uL, of each of the crude bacteriocin preparations). Transmission Electron Microscopy studies revealed the phage to have an icosahedral head with a long tail and tail fibers.
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