Determination of Antibiotic and Heavy Metal Resistance in Paenibacillus lautus 51ATA

Microbial and chemical contamination is one of the biggest environmental problems since exposure to pathogenic microorganisms, such as food poisoning bacteria and their biological by-products, are often associated with disease and allergic reactions. Paenibacillus lautus is an aerobic or facultative anaerobic opportunistic bacterial pathogen, which is found in a variety of sources. In this study, we isolated and characterized P. lautus from Niğde Province, which is a wastewater disposal area and therefore presents a great source of contamination. The bacterial isolate was grown, colony morphology and Gram-reactions were observed. Bacteria were identified by 16S rDNA sequencing analysis, which was performed using DNA samples isolated from pure cultures and amplified by polymerase chain reaction (PCR). Sequencing results were compared against known bacterial strains using BLAST (Basic Local Alignment Search Tool) across databases. Antibiotic resistance against 20 different antibiotics and heavy metal resistance against different concentrations of zinc, copper, iron, cobalt, chromium and silver containing compounds were tested via disk diffusion method. Inhibition zones were examined and minimum inhibitory concentrations were recorded. Our results indicated that P. lautus was resistant to a certain set of antibiotics, while it was sensitive against others. Similarly, the bacteria were resistant against high concentrations of most metals tested. In summary, this study provided the first data regarding the isolation and characterization of Paenibacillus lautus strain from a local source in Turkey, which will account as preliminary data and guide our future efforts to fight against microbial contaminations.

PDF

___

Abskharon RNN, Hassan SHA, Gad El-Rab SMF, Shoreit AAM. 2008. Heavy metal resistant of E. coli isolated from wastewater sites in Assiut City, Egypt. Bulletin of Environmental Contamination and Toxicology, 81:309-315.

Bhullar K, Waglechner N, Pawlowski A, Koteva K, Banks ED, Johnston MD, Barton HA, Wright GD. 2012. Antibiotic resistance is prevalent in an isolated cave microbiome. PLoS One. 7(4):e34953. doi: 10.1371/journal.pone.0034953.

Bionda N, Pitteloud JP, Cudic P. 2013. Cyclic lipodepsipeptides: a new class of antibacterial agents in the battle against resistant bacteria. Future Med Chem., 5(11):1311-30.

Celandroni F, Salvetti S, Gueye SA, Mazzantini D, Lupetti A, Senesi S, Ghelardi E. 2016. Identification and Pathogenic Potential of Clinical Bacillus and Paenibacillus Isolates. PLoS One. 11(3): e0152831. doi: 10.1371/journal. pone.0152831.

Chronopoulos J, Haidouti C, Chronopoulou A, Massas I. 1997. Variations in plant and soil lead and cadmium content in urban parks in Athens, Greece. Sci Total Environ., 196:91–98.

Colak F, Olgun A, Atar A, Yazicioglu D. 2013. Heavy metal resistances and biosorptive behaviors of Paenibacillus polymyxa: Batch and column studies. Journal of Industrial and Engineering Chemistry, 19, 863–869.

Díaz-Ravina M, Baath E, Frostegard A. 1994. Multiple heavy metal tolerance of soil bacterial communities and its measurement by a thymidine incorporation technique. Appl Environ Microbiol., 60(7):2238-47.

Frank JA, Reich CI, Sharma S, Weisbaum JS, Wilson BA, Olsen GJ. 2008. Critical Evaluation of Two Primers Commonly Used for Amplification of Bacterial 16S rRNA Genes. Appl Environ Microbiol., 74(8):2461-70.

Govarthanan M, Mythili R, Selvankumar T, Kamala-Kannan S, Rajasekar A, Chang Y-C. 2016. Bioremediation of heavy metals using an endophytic bacterium Paenibacillus sp. RM isolated from the roots of Tridax procumbens. Biotech., 6, 242, 1-7.

Guzzo J, Diorio DC, Alexander DC and DuBow MS. 1999. Toward understanding metal stress in environment microbial flora. Proceedings of the 8th International Symposium on Microbial Ecology, Canada.

He Z, Kisla D, Zhang L, Yuan C, Green-Church KB, Yousef AE. 2007. Isolation and identification of a Paenibacillus polymyxa strain that coproduces a novel lantibiotic and polymyxin. Appl Environ Microbiol., 73:168–178.

Hussein H, Ibrahim SF, Kandeel K, Moawad H. 2004. Biosorption of heavy metals from waste water using Pseudomonas sp. Electronic Journal of Biotechnology, 1, 7, 38-46.

Igiri BE, Okoduwa SIR, Idoko GO, Akabuogu EP, Adeyi AO, Ejiogu IK. 2018. Toxicity and Bioremediation of Heavy Metals Contaminated Ecosystem from Tannery Wastewater: A Review. J Toxicol., 2018:2568038.

Khare S, Ahmed N, Pant S, Das R. 2010. Characterizationand Evaluation of heavy metal tolerance of bacterialspecies from soil of waste area near Riyan steel rolling mills,Muzaffarnagar, India. Journal of Applied and Natural Science, 2: 88-9 2.

Lane TW, Morel FM. 2000. A biological function for cadmium in marine diatoms. Proc Natl Acad Sci U S A. 97(9):4627-31.

Lima de Silva AA, de Carvalho MA, de Souza SA, Dias PM, da Silva Filho RG, de Meirelles Saramago CS, de Melo Bento CA, Hofer E. 2012. Heavy metal tolerance (Cr, Ag AND Hg) in bacteria isolated from sewage. Braz J Microbiol., 43(4):1620-31. doi: 10.1590/S1517-838220120004000047.

Loong SK, Ishak SN, Lim FS, Khoo JJ, Tan SN et al., 2018. Paenibacillus lautus, an opportunistic bacterial pathogen, isolated from Ixodes granulatus Supino (Acari: Ixodidae) collected from a Müller’s giant Sunda rat (Sundamys muelleri) Systematic & Applied Acarology 23(4): 597–602. http://doi.org/10.11158/saa.23.4.2

Marais L. 2012. Characterization of bacteria isolated from a platinum mine tailings dam, Environmental Sciences, School of Biological Sciences, North-West University (thesis)

Marzan LW, Hossain M, Mina SA, Akter Y, Chowdhury AMMA. 2017. Isolation and biochemical characterization of heavy-metal resistant bacteria from tannery effluent in Chittagong city, Bangladesh: bioremediation viewpoint. Egypt. J. Aquat. Res., 43 pp. 65-74.

Raza W, Shen Q. 2010. Growth, Fe3+ reductase activity, and siderophore production by Paenibacillus polymyxa SQR-21 under differential iron conditions. Curr Microbiol. 61(5):390-5.

Rehman W, Zeb A, Noor N, Nawaz M. 2008. Heavy metal pollution assessment in various industries of Pakistan. Environ Geol., 55: 353–358.

Singh R, Gautam N, Mishra A, Gupta R. 2011. Heavy metals and living systems: An overview. Indian J Pharmacol. 43(3):246- 53. doi: 10.4103/0253-7613.81505.

Trevors JT, Oddie KM, Belliveau BH. 1985. Metal resistance in bacteria. FEMS Microbiol. Rev.32 (1): 39-54.

Wilkins TD, Holdeman LV, Abramson IJ, Moore WEC. 1972. Standardized single disc method for antibiotic susceptibility testing of anaerobic bacteria. Antimicrob Agents Chemother., 1(6):451-9.

Xie JB, Du Z, Bai L, Tian C, Zhang Y, Xie JY, et al. 2014. Comparative genomic analysis of N2-fixing and non-N2- fixing Paenibacillus spp.: organization, evolution and expression of the nitrogen fixation genes. PLoS Genet.;10:e1004231.

Xie J, Shi H, Du Z, Wang T, Liu X, Chen S. 2016. Comparative genomic and functional analysis reveal conservation of plant growth promoting traits in Paenibacillus polymyxa and its closely related species. Sci Rep. 6:21329. doi: 10.1038/srep21329.