Screening and identification of aquatic bacteriocinogenic Bacillus strains inhibiting clinical MRSA and VRE from Pakistan


  • Zaid Ahmed Pirzada University of Karachi


Objective: The purpose of current study is to screen and identify novel bacteriocinogenic strains from marine, fresh and estuarine water bodies which can serve as alternatives to the antibiotics, as the antibiotic resistance is on the rise.

Methodology: Altogether seven fresh, brackish and marine water samples from various regions of Sindh were collected All the aquatic bacterial strains were isolated by standard plate count and overlay methods. The bacteriocinogenic strains were screened by spot agar, stab and overlay, cross streak and agar well-diffusion techniques.

Results: Out of the total 146 isolates, 41 (28%) were bacteriocinogenic, among which 28 (68%) were isolated from freshwater, 12 (29%) from marine and 1 (3%) from brackish water. The most significant bacteriocinogenic strains amongst the isolates were identified as B coagulans, B simplex, B macerans and B circulans. When tested against 17 significant pathogens, these strains could inhibit various clinical drug resistant Gram positive pathogens like Staphylococcus aureus and Methicillin resistant Staphylococcus aureus (MRSA), Streptococcus pyogenes, S agalactiae and Vancomycin resistant Enterococcus (VRE) and C diptheriae. Furthermore, Escherichia coli and Klebsiella spp. were observed to be sensitive against two bacteriocinogenic strains of B coagulans. Furthermore, 59% of the aquatic isolates were sensitive against one of the  bacteriocinogenic strain indicating competitive antagonism phenomenon.

Conclusion: This research study reveals the bacteriocinogenic potential of indigenous aquatic isolates against drug resistant clinical pathogens as well as against fish pathogens.


Christopeit T, Øverbø K, Danielson UH and Nilsen IW (2013) Efficient screening of marine extracts for protease inhibitors by combining FRET based activity assays and surface plasmon resonance spectroscopy based binding assays Mar Drugs. 11(11):4279-93. doi: 10.3390/md11114279

Klaenhammer TR (1988) Bacteriocins of lactic acid bacteria. Biochimie 70:337-349.

Meseguer I, Rodriguez-Valera F (1985) Production and purification of halocin H4. FEMS Microbiol Lett 28:177–182.

Cheung J, Danna KJ, O'Connor EM, Price LB, Shand RF (1997) Isolation, sequence, and expression of the gene encoding halocin H4, a bacteriocin from the halophilic archaeon Haloferax mediterranei R4. J Bacteriol 179:548-551.

Pirzada Z, Ali SA, Khan BM, Rasool SA (2004) Production and physico-chemical characterization of bacteriocins-like inhibitory substances from marine bacterium ZM81. Pak J Biol Sci 7:2026-2030.

Smitha S and Bhat SG (2013) Thermostable Bacteriocin BL8 from Bacillus licheniformis isolated from marine sediment J Appl Microbiol.114(3):688-94. doi: 10.1111/jam.12097

Longeon A, Peduzzi J, Barthélemy M, Corre S, Nicolas JL, Guyot M (2004) Purification and partial identification of novel antimicrobial protein from marine bacterium Pseudoalteromonas species strain X153. Mar Biotechnol 6: 633-641.

Aunpad R, Na-Bangchang K (2007) Pumilicin 4, a novel bacteriocin with anti-MRSA and anti-VRE activity produced by newly isolated bacteria Bacillus pumilus strain WAPB4. Curr Microbiol 55:308-313.

Rosa-Garcia DL, Muñoz-García AA, Barahona-Pérez, LF, Gamboa-Angulo, MM (2007) Antimicrobial properties of moderately halotolerant bacteria from cenotes of the Yucatan peninsula. Lett Appl Microbiol 45:289-294.

Nowroozi J, Mirzaii M, Norouzi M (2004) Study of Lactobacillus as Probiotic Bacteria. Iran J Pub Heal 33:1-7.

Schillinger U, Kaya M, Lucke FK (1991) Behaviour of Listeria monocytogenes in meat and its control by a bacteriocin-producing strain of Lactobacillus sake. J Appl Bacteriol 70: 73-78.

Uzair B, Ahmed N, Ahmad VU, Mohammad FV, Edwards DH (2008) The isolation, purification and biological activity of a novel antibacterial compound produced by Pseudomonas stutzeri. FEMS Microbiol Lett 279:243-250.

Cooper D, James R (1984) Two new E colicins, E8 and E9, produced by a strain of Escherichia coli. J Gen Microbiol 130:209-215.

Irshad S., Mahmood M. and Perveen (2012) F. In-Vitro Anti-Bacterial Activities of Three Medicinal Plants Using Agar Well Diffusion Method. Res J Biol 2(1):1-8.

Uzair B, Ahmed N, Kousar F, Edwards DH (2006) Isolation and characterization of Pseudomonas strain that inhibit growth of indigenous and clinical isolate. Inter J Microbiol 2: DOI:10.5580/12c9

Bost AL (2004) Isolation of bacteriocins from Vibrio Spp. and Pseudomonas spp. attached to aquatic particulate material. (MSc. Thesis. University of North Carolina, Wilmington)

Abriouel H, Franz CM, Ben ON, Gálvez A (2011) Diversity and applications of Bacillus bacteriocins. FEMS Microbiol 35:201-232.

Mannanov RN, Sattarova RK (2001) Antibiotics produced by Bacillus bacteria. Chem Nat Compd 37:117-123.

Hammami I, Rhouma A, Jaouadi B, Rebai A, Nesme X (2009) Optimization and biochemical characterization of a bacteriocin from a newly isolated Bacillus subtilis strain 14B for biocontrol of Agrobacterium spp. strains. Lett Appl Microbiol 48:253-260.

Cherif A, Quazri H, Daffonchio D, Cherif H, Siama, BK, Hassen A, Japua S, Boudabous A (2001) Thurin 7: A novel bacteriocin produced by Bacillus thuringenesis BMG 1.7, a new strain isolated from soil. Lett Appl Microbiol 32:243-247.

Saleem F, Ahmad S, Yaqoob Z, Rasool SA (2009) Comparative Study of Two Bacteriocins Produced by Representative Indigenous Soil Bacteria. Pak J Pharm Sci 22:252-258.

Bizani D, Brandelli A (2002) Characterization of a bacteriocin produced by a newly isolated Bacillus sp. Strain 8 A. J Appl Microbiol 93:512-519.

Cladera OF, Caron GR, Brandelli A (2004) Bacteriocin-like substance production by Bacillus licheniformis strain P40. Lett Appl Microbiol 38:251-256.

Parada JL, Caron CR, Medeiros ABP, Soccol CR (2007) Bacteriocins from Lactic Acid Bacteria: Purification, Properties and use as Biopreservatives. Braz Arch Biol Technol 50:521-542.

Schulz D, Marques RF, Muller IC, Batista CRV (2005) Bacillus amyloliquefaciens as potential biopreservative of food: determination of the profile of protein and aminogram. Aliment Nutricao 16:65-70.



How to Cite

RIZVI, R. Z., A. . WAHAB, and Z. A. Pirzada. “Screening and Identification of Aquatic Bacteriocinogenic Bacillus Strains Inhibiting Clinical MRSA and VRE from Pakistan”. Asian Journal of Pharmaceutical and Clinical Research, vol. 7, no. 5, Nov. 2014, pp. 53-56,



Original Article(s)