Plasmid Mediated Antibiotic and Heavy Metal Resistance in Bacillus Strains Isolated from Soils in Rize, Turkey

Year 2015, Volume: 19 Issue: 2, 133 - 141, 02.09.2015

Elif Sevim Ali Sevim

Abstract

Fifteen Bacillus strains which were isolated from soil samples were examined for resistance to 17 different antibiotics (ampicillin, methicillin, erythromycin, norfloxacin, cephalotine, gentamycin, ciprofloxacin, streptomycin, tobramycin, chloramphenicol, trimethoprim-sulfamethoxazole, tetracycline, vancomycin, oxacilin, neomycin, kanamycin and, novabiocin) and to 10 different heavy metals (copper, lead, cobalt, chrome, iron, mercury, zinc, nickel, manganese and, cadmium) and for the presence of plasmid DNA. A total of eleven strains (67%) were resistant to at least one antibiotic. The most common resistance was observed against methicillin and oxacillin. The most resistance strains were found as Bacillus sp. B3 and Bacillus sp. B11. High heavy metal resistance against copper, chromium, zinc, iron and nickel was detected, but mercury and cobalt resistance was not detected, except for 3 strains (B3, B11, and B12) which showed mercury resistance. It has been determined that seven Bacillus strains have plasmids. The isolated plasmids were transformed into the Bacillus subtilis W168 and it was shown that heavy metal and antibiotic resistance determinants were carried on these plasmids. These results showed that there was a correlation between plasmid content and resistance for both antibiotic and heavy metal resistance

Keywords

Bacillus, Plasmid, Antibiotic resistance, Metal resistance, Transformation

Rize’deki Topraklardan İzole Edilen Bacillus Suşlarında Plazmit Kaynaklı Antibiyotik ve Ağır Metal Direnci

Abstract

Toprak örneklerinden izole edilen 15 adet Bacillus suşu 17 farklı antibiyotiğe (ampisilin, metisilin, eritromisin, norfloksasin, sefalotin, gentamisin, siprofloksasin, streptomisin, tobramisin, kloramfenikol, trimetoprimsulfametaksozol, tetrasiklin, vankomisin, oksasilin, neomisin, kanamisin ve novabiosin) ve 10 farklı ağır metale (bakır, kurşun, kobalt, krom, demir, cıva, çinko, nikel, manganez ve kadmiyum) karşı dirençliliği ve plazmit DNA içeriği bakımından incelenmiştir. Toplam olarak 11 suş (%67) en azından bir antibiyotiğe dirençli bulunmuştur. En yaygın direnç metisilin ve oksasilin’e karşı gözlemlenmiştir. En yaygın dirençli suşlar ise Bacillus sp. B3 ve Bacillus sp. B11 olarak tespit edilmiştir. Bakır, krom, çinko, demir ve nikel’e karşı direnç belirlenmesine rağmen, cıva direnci gösteren 3 suş hariç (B3, B11 ve B12) cıva ve kobalta karşı direnç belirlenmemiştir. Yedi adet Bacillus suşunun plazmit içerdiği tespit edilmiştir. İzole edilen plazmitler Bacillus subtilis W168 suşuna transforme edilmiş, ağır metal ve antibiyotik direnç belirleyicilerinin plazmit üzerinde taşındığı bulunmuştur. Bu sonuçlar hem antibiyotik hem de ağır metal direnci ile plazmit içeriği arasında bir ilişki olduğunu göstermektedir.  

Keywords

Bacillus, Plazmit, Antibiyotik direnci, Metal direnci, Transformasyon

References

• Amsterdam, D., 1996. Susceptibility testing of antimicrobials in liquid media, in Loman, V. (Ed.),
• Antibiotics in laboratory medicine 4th ed., Williams and Wilkins, Baltimore, MD, USA, pp. 52-111. Austin, C.B., Wright, M.S., Stepanauskas, R., McArthur, J.V., 2006. Co-selection of antibiotic and metal resistance.TRENDS in Microbiol. 14, 176-182.
• Bauer, A.W., Kirby, W.M.M., Sherries, J.C., Truck, M., 19 Antibiotic susceptibility testing by a standardized single disc diffusion method. Am. J. Clin. Pathol. 45, 493-496. Belliveau, B.H., Starodub, M.E., Trevors, J.T., 1991.
• Campos, J., Martinez-Pacheco, M., Cervantes, C., 19 Hexavalent-chromium reduction by a chromate-resistant Bacillus sp. strain. Antonie van Leeuwenhoek 68, 203-208. Chaturvedi, M.K., 2011. Studies on chromate removal by chromium-resistant Bacillus sp. isolated from tannery effluent. J. Environ. Protec. 2, 76-82.
• Choudhury, R., Srivastava, S., 2001. Zinc resistance mechanisms in bacteria. Curr. Sci. 81, 768-775.
• Christopher, M., Paul, O., Hamadi, B., 2014.
• Characterization of five chromium-removing bacteria isolated from chromium-contaminated soil. Water Air Soil. Pollut. 225, 1904-1914.
• Hookoom, M., Puchooa, D. 2013. Isolation and identification of heavy metals tolerant bacteria from industrial and agricultural areas in mauritius. Curr.
• Res. Microbiol. Biotechnol.1, 119-123. InceYilmaz, E., 2003. Metal tolerance and biosorption capacity of Bacillus circulansstrain EB1. Res. Microbiol. 154, 409–415.
• Kamala-Kannan, S., Lee, K.J., 2008. Metal tolerance and antibiotic resistance of Bacillus species isolated from suncheon bay sediments, South Korea. Biotechnol.7, 149-152.
• Kunst, F., Msadek, T., Rapoport, G., 1994. Signal transduction network controlling degradative enzyme synthesis and competence in Bacillus subtilis, in Piggot, P.J., Moran, C.P., Youngman, P.
• (Eds.), Regulation of bacterial differentiation, ASM Press, Washington, DC, USA, pp. 1-20. Mahler, I., Levinson, H.S., Wang, Y., Halvorson, H.O., 19 Cadmium- and mercury-resistant Bacillus strains from a salt marsh and from Boston Harbor. App. Environ. Microbiol. 52, 1293- 1298.
• Mathema, V.B., Thakuri, B.C., Sillanpa, M., 2011.
• Bacterial mer operon-mediated detoxiŞcation of mercurial compounds: a short review. Arch. Microbiol. 193, 837–844. Matyar, F., Kaya, A., Dincer, S., 2008.Antibacterial agents and heavy metal resistance in Gram-negative bacteria isolated from seawater, shrimp and sediment in Iskenderun Bay. Sci. Total Environ. 407, 279–285.
• Montuelle, B., Latour, X., Volat, B., Gounot, A.M. 19 Toxicity of heavy metals to bacteria in sediments. Bull. Environ. Contam. Toxicol. 53, 753- 7
• Nies, D.H., 1999. Microbial heavy metal resistance.
• Appl. Microbiol. Biotechnol. 51, 730-750. Nies, D.H., 2004. Metals and their compounds in the environment, in Anke, K., Ihmat, M., Stoeppler, M.
• (Eds.), Part II. The Elements: Essential and Toxic Effects on Microorganisms. Weinheim. Nieto, J.J., Fernandez-Castillo, R., Marquez, M.C., Ventosa, A., Quesada, E., Ruiz-Berraquero, F., 1989.
• Survey of metal tolerance in moderately halophilic eubacteria. Appl. Environ. Microbiol. 55(9): 2385- 23
• Sevgi, E., Coral, G., Gizir, A.M., Sangun, M.K., 2010.
• Investigation of heavy metal resistance in some bacterial strains isolated from industrial soils. Turk. J. Biol. 34, 423-431. Sevim, E., AlpayKaraoglu, S., Sevim, A., Canakcı, S. 20 Antimicrobial activity of Bacillus strains isolated from spring water and a novel bacteriocin: RS J. Pure App. Microbiol. 7, 2757-2765.
• Şensoy Karaoğlu, Ş., Sevim, A., Sevim, E. 2014.
• Production and characterization of bacteriocin-like peptide produced by Bacillus amyloliquefaciens B10. J. Ins. Sci. Technol. Erciyes Uni. 30(5): 338-345. Sinha, S., Chatterjee, R., Singh, D., Dimri, G., Aggarwal, M.L., 2013. Studies on heavy metal tolerance and antibiotic resistance patterns of bacterial population isolated from effluent treated water of Delhi. J. Biomed. Pharma. Res. 2, 69-76.
• Smith, K., Novick, R.P., 1972. Genetic studies on plasmid-linked cadmium resistance in
• Staphylococcus aureus. J. Bacteriol. 112, 761-772. Trajanovska, S., Britz, M.L., Bhave, M., 1997.
• Detection of heavy metal ion resistance genes in gram-positive and gram-negative bacteria isolated from a lead-contaminated site. Biodegradation 8, 113- Velusamy, P.Y., Awad, Y.M., Abd El-Azeem, S.A.M., Ok, Y.S., 2011. Screening of heavy metal resistant bacteria isolated from hydrocarbon contaminated soil in Korea. J. Agricul. Life Environ. Sci. 23, 40-43.
• Voskuil, M.I., Chambliss, G.H., 1993. Rapid isolation and sequencing of purified plasmid DNA from
• Bacillus subtilis. App. Environ. Microbiol. 59, 1138- 1 Abbreviations DNA kg m-3
• MHA Muller Hinton Agar CLSI Clinical and laboratory Standards Institute NCCLS National Committee for Clinical Laboratory Standards µg MIC µl rpm nm °C PAL LB pH mM mL OD UV WT Wild Type