Endophytic Bacillus species isolated from mangrove plants, and their antagonestic effects against some pathogenic bacterial strains
Keywords:
Antimicrobials, Mangroves, Bacillus, BiotechnologyAbstract
This study aimed to isolate bacterial endophytes with potential antimicrobial effects from five mangrove plants and to investigate the stability of the most active metabolites under different PH and temperature. Bacterial colonies were isolated from the collected plant parts. Bacterial strains with potential antagonistic interactions were subjected to idnetification by VITEK 2 system or sequencing of the 16S rRNA. MIC values of the crude substances were determined against Bacillus cereus, Streptococcus uberis, Escherichia coli, Klebsiella pneumonia and Salmonella typhiumurium using the the serial dilution technique.The stability of the most active crude extracts was examined under different, PH (2-7) and temperature (60 -90 °C). Twenty one of the isolated strains showed potential antagonistic effects. Of these, 16 identified as Bacillus species. Best antimicrobial effects were recorded for Bacillus tequilensis against, B. cereus and S. typhimurium (MIC value=65 µl/m1), S.uberis (MIC value 78 µl/ml), E.coli and K.pneumonia (MIC value=125 µl/m1). Both Bacillus subtilis and Brevibacillus brevis inhibited growth of S. typhimurium with MIC value of 78 and 60 µl/m1 respectively. The metabolites of B.subtilis and B.tequilensis appeared to be stable under PH2-9. Product of B.subtilis showed stability under high temperature (90 °C) against S. typhimurium. The isolated endophytes possessed wide range of antimicrobial activities against the selected pathogens. Bacillus tequilensis and B.subtilis produced the most active metabolites. This is the first report on isolation of endophytic strains of B.tequilensis. The crude extracts obtained from B.tequilensis and B.subtilis in this study could be further developed as food preservatives.
References
Eldeen IMS.Isolation of 12 Bacterial endophytes from some mangrove plants and determination of, antimicrobial properties of the isolates and the plant extracts. Int. J Phytomed.2014; 6(3):425-432.
Ryan RP, Germaine K, Franks A, Ryan DJ, Dowling DN. Bacterial endophytes: recent developments and applications. FEMS Microb Lett .2008;278: 1-9.
Eldeen IMS, Effendy MAW. Antimicrobial agents from mangrove plants and their endophytes. In: Méndez-Vilas 9ed.Microbial pathogens and Strategies for Combating them: Science, Technology and Education. 2013; P: 872-882. Spain, Badajoz FORMATEX Microbiology Book Series.Formatex Research Centre.
Baruzzi F, Quintieri L, Morea M, Caputo L. Antimicrobial compounds produced by Bacillus spp. and applications in food. In: Méndez-Vilas (ed). Science against microbial pathogens: communicating current research and technological advances. 2011; P: 1102-1111.Spain, Badajoz FORMATEX Microbiology Book Series.Formatex Research Centre.
Griesbach E, Lattauschke E, von Bertragung U. Clavibacter michiganensis subsp. michiganensis in Tomaten-Hydroponikkulturen und Möglichkeiten zur Bekämpfung des Erregers. Nachrbl. Dtsch. Pflanzenschutzd 1999; 43: 69–73.
Zokaeifar H, Luis Balca´zar J, Kamarudin MS, Sijam K, Arshad A, Saad C.R. Selection and identification of non-pathogenic bacteria isolated from fermented pickles with antagonistic properties against two shrimp pathogens. J Antibiot .2012; 65: 289–294.
Nath S, Chowdhury S, Dora K.C. Application of Bacillus sp. as a biopreservative for food preservation. J Engineering Res and Applic .2015; 5 (4): 85-95.
Gray EJ, Lee KD, Souleimanov AM, Di Falco M.R. et al. A novel bacteriocin, thuricin 17, produced by plant growth promoting Rhizobacteria strain Bacillus thuringiensis NEB17: Isolation and classification. J Appl Microbiol 2006; 100: 545-554.
Shin MS, Han SK, Ji AR, Kim KS, Lee WK. Isolation and characterization of bacteriocin-producing bacteria from the gastrointestinal tract of broiler chickens for probiotic use. J Appl Microbiol .2008; 105: 2203-2212.
Kayalvizhi N, Gunasekaran P. Purification and characterization of a novel broad-spectrum bacteriocin from Bacillus licheniformis MKU3. Biotech and Bioproc Engin .2010; 15:365-370.
Abriouel H, Charles Franz MAP, Omar NB, Alvez, A.GDiversity and applications of Bacillus bacteriocins. FEMS Microbiol . 2010; 35: 201-232.
Smith S, Bhat SGThermostable bacteriocin BL8 from Bacillus licheniformis isolated from marine sediment. J Appl Microbiol . 2013; 114(3): 688-694.
Nithya B P. 16S rRNA phylogenetic analysis of actinomycetes isolated from Eastern Ghats and marine mangrove associated with antibacterial and anticancerous activities. African J. of Biotech .2012; 11(60): 12379-12388.
Nguyen T, Quach T, Hoang T, Doan H, Santa R J.Detection of tetradotoxin–producing Providencia rettgeri T892 in Lagocephalus pufferfish. World J. Microbial Biotechnol. 2014; 1-7.
Eloff JN. A sensitive and quick microplate method to determine the minimal inhibitory concentration of plant extracts for bacteria. Planta Med 1998; 64:711-713.
Graf B, Adam T, Zill E, Göbel U. Evaluation of VITEK2 system for rapid identification of yeast and yeast-like organisms. J Clinic Microbiol 2000; 38(5):1782-1785.
Ling TKW, Liu ZK, Cheng AFB Evaluation of the VITEK 2 system for rapid direct identification and susceptibility testing of Gram-negative Bacilli from positive blood cultures. J Clin Microbiol . 2003; 41(10): 4705–4707.
Fontana C, Favaro M, Pelliccioni M, Pistoia ES. Use of the MicroSeq 500 16S rRNA Gene-Based Sequencing for Identification of Bacterial Isolates That Commercial Automated. J Clinic Microbiol .2005; 43: 615–619.
Chakravorty S, Helb D, Burday M, Connell N, Alland DA .Detailed analysis of 16S ribosomal RNA gene segments for the diagnosis of pathogenic bacteria. J Microbiol Meth .2007; 69: 330–339.
Gatson JW, Benz BF, Chandrasekaran C. Bacillus tequilensis sp. nov., isolated from a 2000-year-old Mexican shaft-tomb, is closely related to Bacillus subtilis. Int J Syst Evol Micr .2006; 56: 1475–1484.
Chatterjee S, Lad SJ, Phansalkar MS, Rupp RH, Ganguli BN, Fehlhaber HW, Kogler H.) Mersacidin, a new antibiotic from Bacillus: fermentation, isolation, purification and chemical characterization. J Antibiot .1992; 45: 832–838.
Permpoonpattana P, Hong HA, Khaneja R, Cutting SM. Evaluation of Bacillus subtilis strains as probiotics and their potential as a food ingredient. Benef Microb. 2012;1-10.
Tiwari BK1, Valdramidis VP, O'Donnell CP, Muthukumarappan K, Bourke P, Cullen PJ. Application of Natural Antimicrobials for Food Preservation. J Agric Food Chem 2009; 57:5987–6000.
Olmos J, Paniagua-Michel JP. Bacillus subtilis a potential probiotic Bacterium to formulate functional feeds for aquaculture. Microb & Biochem Technol. 2014; 6 (7): 361-365.
