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Volume 2, Issue 3

Principal component analysis exploring the association between antibiotic resistance and heavy metal tolerance of plasmid-bearing sewage wastewater bacteria of clinical relevance Open Access

Abstract

This paper unravels the occurrence of plasmid-mediated antibiotic resistance in association with tolerance to heavy metals among clinically relevant bacteria isolated from sewage wastewater. The bacteria isolated were identified following conventional phenotypic and/or molecular methods, and were subjected to multiple-antibiotic resistance (MAR) profiling. The isolates were tested against the heavy metals Hg, Cd, Cr and Cu. SDS-PAGE and agarose gel electrophoretic analyses were performed, respectively, for the characterization of heavy metal stress protein and R-plasmid among the isolated bacteria. Principal component analysis was applied in determining bacterial resistance to antibiotics and heavy metals. Both lactose-fermenting () and non-fermenting ( and ) Gram-negative bacterial strains were procured, and showed MAR phenotypes with respect to three or more antibiotics, along with resistance to the heavy metals Hg, Cd, Cr and Cu. The Gram-positive bacteria, , isolated had ‘ampicillin–kanamycin–nalidixic acid’ resistance. The bacterial isolates had MAR indices of 0.3–0.9, indicating their (, , and ) origin from niches with high antibiotic pollution and human faecal contamination. The Gram-negative bacteria isolated contained a single plasmid (≈54 kb) conferring multiple antibiotic resistance, which was linked to heavy metal tolerance; the SDS-PAGE analysis demonstrated the expression of heavy metal stress proteins (≈59 and ≈10 kDa) in wastewater bacteria with a Cd stressor. The study results grant an insight into the co-occurrence of antibiotic resistance and heavy metal tolerance among clinically relevant bacteria in sewage wastewater, prompting an intense health impact over antibiotic usage.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): 16s RNA gene sequence , co-resistance , heavy metal tolerance , multiple-antibiotic resistance phenotypes , plasmid and sewage wastewater bacteria
Funding
This study was supported by the:
  • No funding (Award Not applicable)
    • Principle Award Recipient: Not Applicable
© 2020 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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2020-02-10
2025-05-14
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References

  1. Sommer MOA, Church GM, Dantas G, George MC, Gautam D. The human microbiome harbors a diverse reservoir of antibiotic resistance genes. Virulence 2010; 1:299–303 [View Article][PubMed]
     [Google Scholar]
  2. Baker-Austin C, Wright MS, Stepanauskas R, McArthur JV. Co-selection of antibiotic and metal resistance. Trends Microbiol 2006; 14:176–182 [View Article][PubMed]
     [Google Scholar]
  3. Eze E, Eze U, Eze C, Ugwu K. Association of metal tolerance with multidrug resistance among bacteria isolated from sewage. J Rural Trop Public Health 2006; 8:25–29
     [Google Scholar]
  4. Galvin S, Boyle F, Hickey P, Vellinga A, Morris D et al. Enumeration and characterization of antimicrobial-resistant Escherichia coli bacteria in effluent from municipal, hospital, and secondary treatment facility sources. Appl Environ Microbiol 2010; 76:4772–4779 [View Article][PubMed]
     [Google Scholar]
  5. Le T-H, Ng C, Chen H, Yi XZ, Koh TH et al. Occurrences and characterization of antibiotic-resistant bacteria and genetic determinants of hospital wastewater in a tropical country. Antimicrob Agents Chemother 2016; 60:7449–7456 [View Article][PubMed]
     [Google Scholar]
  6. Das SN, Mandal M, Mandal S. Plasmid mediated antibiotic and heavy metal co-resistance in bacterial isolates from Mahananda river water (Malda, India). Transl Med 2016; 6:185
     [Google Scholar]
  7. Das SN, Mandal M, Mandal S. Detection of mercury and cadmium resistance among multiple antibiotic resistant enteric bacteria from municipal sewage water in Malda, India. Int Res J Pharm 2018; 9:171–176 [View Article]
     [Google Scholar]
  8. Das SN, Mandal M, Mandal S. Heavy metal tolerance in association with plasmid mediated multiple antibiotic resistances among clinical bacterial isolates. Biosci Biotechnol Res Commun 2018; 11:612–618 [View Article]
     [Google Scholar]
  9. Wright GD. The antibiotic resistome: the nexus of chemical and genetic diversity. Nat Rev Microbiol 2007; 5:175–186 [View Article][PubMed]
     [Google Scholar]
  10. Zowawi HM, Harris PNA, Roberts MJ, Tambyah PA, Schembri MA et al. The emerging threat of multidrug-resistant gram-negative bacteria in urology. Nat Rev Urol 2015; 12:570–584 [View Article][PubMed]
     [Google Scholar]
  11. Holt JG. Bergey’s Manual of Systematic Bacteriology Baltimore: Williams and Wilkins; 1984
     [Google Scholar]
  12. Forbes BA, Sahm DF, Bailey WAS. Scott’s Diagnostic of Microbiology, 12th Edition. USA: Mosby (Elsevier); 2007
     [Google Scholar]
  13. Zhang Z, Schwartz S, Wagner L, Miller W, Scott SLW. A greedy algorithm for aligning DNA sequences. J Comput Biol 2000; 7:203–214 [View Article][PubMed]
     [Google Scholar]
  14. Bauer AW, Kirby WM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol 1966; 45:493–496 [View Article][PubMed]
     [Google Scholar]
  15. CLSI Performance standards for antimicrobial susceptibility testing. CLSI supplement M100, 28th Ed. Wayne, Pa: Clinical and Laboratory Standards Institute; 2018
     [Google Scholar]
  16. Nandi S, Mandal S. Bacteriological profiling of commercially available eye cosmetics and their antibiotic susceptibility pattern. Transl Biomed 2016; 7:1–8 [View Article]
     [Google Scholar]
  17. Adefisoye M, Okoh A. Ecological and public health implications of the discharge of multidrug-resistant bacteria and physicochemical contaminants from treated wastewater effluents in the eastern Cape, South Africa. Water 2017; 9:562 [View Article]
     [Google Scholar]
  18. Krumperman PH. Multiple antibiotic resistance indexing of Escherichia coli to identify high-risk sources of fecal contamination of foods. Appl Environ Microbiol 1983; 46:165–170 [View Article][PubMed]
     [Google Scholar]
  19. Tambekar DH, Hirulkar NB, Waghmare AS. Mar indexing to discriminate the source of faecal contamination in drinking water. Nat Environ Poll Tech 2005; 4:525–528
     [Google Scholar]
  20. Kaneene JB, Miller R, Sayah R, Johnson YJ, Gilliland D et al. Considerations when using discriminant function analysis of antimicrobial resistance profiles to identify sources of fecal contamination of surface water in Michigan. Appl Environ Microbiol 2007; 73:2878–2890 [View Article][PubMed]
     [Google Scholar]
  21. Ismail INA, Noor HM, Muhamad HS, Radzi SM, Kader AJA et al. Protein produced by Lactobacillus plantarum ATCC 8014 during stress. World J Sci Technol Res 2013; 1:174–181
     [Google Scholar]
  22. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227:680–685 [View Article][PubMed]
     [Google Scholar]
  23. Kado CI, Liu ST. Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol 1981; 145:1365–1373 [View Article][PubMed]
     [Google Scholar]
  24. Maniatis T, Fritsch EF, Sambrook J. Molecular cloning: a laboratory manual, cold Spring Harbor laboratory. New York 1982
     [Google Scholar]
  25. Anjanappa M, Horbola PC. Elimination (curing) of R plasmids in Salmonella gallinarum and their transconjugants. Ind Vet J 1993; 70:10–13
     [Google Scholar]
  26. Mandal S, Deb Mandal M, Pal NK. Plasmid encoded UV resistance and UV induced ciprofloxacin resistance in Salmonella enterica serovar Typhi. Internat J Integrat Biol 2008; 2:43–48
     [Google Scholar]
  27. Jolliffe IT. Principal Component Analysis New York: Springer; 1986
     [Google Scholar]
  28. Rodríguez-Gómez F, Romero-Gil V, Bautista-Gallego J, Garrido-Fernández A, Arroyo-López FN. Multivariate analysis to discriminate yeast strains with technological applications in table olive processing. World J Microbiol Biotechnol 2012; 28:1761–1770 [View Article][PubMed]
     [Google Scholar]
  29. Jayaseelan T, Damodaran R, Ganesan S, Mani P. Biochemical characterization and 16S rRNA sequencing of different bacteria from textile dye effluents. J. Drug Delivery Ther. 2018; 8:35–40 [View Article]
     [Google Scholar]
  30. Abo-Amer AE, El-Shanshoury AE-RR, Alzahrani OM. Isolation and Molecular Characterization of Heavy Metal-Resistant Alcaligenes faecalis from Sewage Wastewater and Synthesis of Silver Nanoparticles. Geomicrobiol J 2015; 32:836–845 [View Article]
     [Google Scholar]
  31. Alm EW, Zimbler D, Callahan E, Plomaritis E. Patterns and persistence of antibiotic resistance in faecal indicator bacteria from freshwater recreational beaches. J Appl Microbiol 2014; 117:273–285 [View Article][PubMed]
     [Google Scholar]
  32. Harding CM, Hennon SW, Feldman MF. Uncovering the mechanisms of Acinetobacter baumannii virulence. Nat Rev Microbiol 2018; 16:91–102 [View Article][PubMed]
     [Google Scholar]
  33. Kim SE, Park S-H, Park HB, Park K-H, Kim S-H et al. Nosocomial Pseudomonas putida Bacteremia: High Rates of Carbapenem Resistance and Mortality. Chonnam Med J 2012; 48:91–95 [View Article][PubMed]
     [Google Scholar]
  34. Fernández M, Porcel M, de la Torre J, Molina-Henares MA, Daddaoua A et al. Analysis of the pathogenic potential of nosocomial Pseudomonas putida strains. Front Microbiol 2015; 6:871 [View Article][PubMed]
     [Google Scholar]
  35. Fournier P-E, Vallenet D, Barbe V, Audic S, Ogata H et al. Comparative genomics of multidrug resistance in Acinetobacter baumannii . PLoS Genet 2006; 2:e7 [View Article][PubMed]
     [Google Scholar]
  36. Kempf M, Rolain J-M. Emergence of resistance to carbapenems in Acinetobacter baumannii in Europe: clinical impact and therapeutic options. Int J Antimicrob Agents 2012; 39:105–114 [View Article][PubMed]
     [Google Scholar]
  37. Kümmerer K. Resistance in the environment. J Antimicrob Chemother 2004; 54:311–320 [View Article][PubMed]
     [Google Scholar]
  38. Devarajan N, Köhler T, Sivalingam P, van Delden C, Mulaji CK et al. Antibiotic resistant Pseudomonas spp. in the aquatic environment: A prevalence study under tropical and temperate climate conditions. Water Res 2017; 115:256–265 [View Article][PubMed]
     [Google Scholar]
  39. Blair JMA, Webber MA, Baylay AJ, Ogbolu DO, Piddock LJV. Molecular mechanisms of antibiotic resistance. Nat Rev Microbiol 2015; 13:42–51 [View Article][PubMed]
     [Google Scholar]
  40. Adelowo OO, Vollmers J, Mäusezahl I, Kaster A-K, Müller JA. Detection of the carbapenemase gene bla VIM-5 in members of the Pseudomonas putida group isolated from polluted Nigerian wetlands. Sci Rep 2018; 8:15116 [View Article][PubMed]
     [Google Scholar]
  41. Karkman A, Do TT, Walsh F, Virta MPJ. Antibiotic-resistance genes in waste water. Trends Microbiol 2018; 26:220–228 [View Article][PubMed]
     [Google Scholar]
  42. Frigon D, Biswal BK, Mazza A, Masson L, Gehr R. Biological and physicochemical wastewater treatment processes reduce the prevalence of virulent Escherichia coli . Appl Environ Microbiol 2013; 79:835–844 [View Article][PubMed]
     [Google Scholar]
  43. Odonkor ST, Addo KK. Prevalence of Multidrug-Resistant Escherichia coli Isolated from Drinking Water Sources. Int J Microbiol 2018; 2018:17 [View Article][PubMed]
     [Google Scholar]
  44. Harwood VJ, Staley C, Badgley BD, Borges K, Korajkic A. Microbial source tracking markers for detection of fecal contamination in environmental waters: relationships between pathogens and human health outcomes. FEMS Microbiol Rev 2014; 38:1–40 [View Article][PubMed]
     [Google Scholar]
  45. Aminov RI. Horizontal gene exchange in environmental microbiota. Front Microbiol 2011; 2:158 [View Article][PubMed]
     [Google Scholar]
  46. Tacão M, Moura A, Correia A, Henriques I. Co-resistance to different classes of antibiotics among ESBL-producers from aquatic systems. Water Res 2014; 48:100–107 [View Article][PubMed]
     [Google Scholar]
  47. Fraser AS. 2018; Enterococcal infections. Medscape . https://emedicine.medscape.com/article/216993-print on December 13, 2018
  48. Guardabassi L, Dalsgaard A. Occurrence and fate of antibiotic resistant bacteria in sewage. Danish E P Agency 2002; 722:1–59
     [Google Scholar]
  49. Mounaouer B, Nesrine A, Abdennaceur H, Achour N, Hassen A. Identification and characterization of heavy metal-resistant bacteria selected from different polluted sources. Desalination Water Treat 2014; 52:7037–7052 [View Article]
     [Google Scholar]
  50. Kaur UJ, Preet S, Rishi P. Augmented antibiotic resistance associated with cadmium induced alterations in Salmonella enterica serovar Typhi. Sci Rep 2018; 8:12818 [View Article][PubMed]
     [Google Scholar]
  51. Malik A, Aleem A. Incidence of metal and antibiotic resistance in Pseudomonas spp. from the river water, agricultural soil irrigated with wastewater and groundwater. Environ Monit Assess 2011; 178:293–308 [View Article][PubMed]
     [Google Scholar]
  52. Almuzara M, Radice M, de Gárate N, Kossman A, Cuirolo A et al. VIM-2–producing Pseudomonas putida, Buenos Aires. Emerg Infect Dis 2007; 13:668–669 [View Article]
     [Google Scholar]
  53. Bennett JW, Herrera ML, Lewis JS, Wickes BW, Jorgensen JH. Kpc-2-Producing Enterobacter cloacae and Pseudomonas putida coinfection in a liver transplant recipient. Antimicrob Agents Chemother 2009; 53:292–294 [View Article][PubMed]
     [Google Scholar]
  54. Bogaerts P, Huang T-D, Rodriguez-Villalobos H, Bauraing C, Deplano A et al. Nosocomial infections caused by multidrug-resistant Pseudomonas putida isolates producing VIM-2 and VIM-4 metallo-beta-lactamases. J Antimicrob Chemother 2008; 61:749–751 [View Article][PubMed]
     [Google Scholar]
  55. Molina L, Udaondo Z, Duque E, Fernández M, Molina-Santiago C et al. Antibiotic resistance determinants in a Pseudomonas putida strain isolated from a hospital. PLoS One 2014; 9:e81604 [View Article][PubMed]
     [Google Scholar]
  56. Yoshino Y, Kitazawa T, Kamimura M, Tatsuno K, Ota Y et al. Pseudomonas putida bacteremia in adult patients: five case reports and a review of the literature. J Infect Chemother 2011; 17:278–282 [View Article][PubMed]
     [Google Scholar]
  57. Durve A, Naphade S, Bhot M, Varghese J, Chandra S. Plasmid curing and protein profiling of heavy metal tolerating bacterial isolates. Arch Appl Sci Res 2013; 5:46–54
     [Google Scholar]
  58. LG L, Xia Y, Zhang T. Co-Occurrence of antibiotic and metal resistance genes revealed in complete genome collection. Internat Soc Microb Ecol 2017; 11:651–662
     [Google Scholar]
  59. Di Cesare A, Eckert EM, D'Urso S, Bertoni R, Gillan DC et al. Co-Occurrence of integrase 1, antibiotic and heavy metal resistance genes in municipal wastewater treatment plants. Water Res 2016; 94:208–214 [View Article][PubMed]
     [Google Scholar]
  60. Seiler C, Berendonk TU. Heavy metal driven co-selection of antibiotic resistance in soil and water bodies impacted by agriculture and aquaculture. Front Microbiol 2012; 3:399 [View Article][PubMed]
     [Google Scholar]
  61. Stepanauskas R, Glenn TC, Jagoe CH, Tuckfield RC, Lindell AH et al. Elevated microbial tolerance to metals and antibiotics in metal-contaminated industrial environments. Environ Sci Technol 2005; 39:3671–3678 [View Article][PubMed]
     [Google Scholar]
  62. Chatterjee S, Das J, Chatterjee S, Choudhuri P, Sarkar A. Isolation, characterization and protein profiling of lead resistant bacteria. BMRJ 2014; 4:116–131 [View Article]
     [Google Scholar]
  63. Lemire JA, Harrison JJ, Turner RJ. Antimicrobial activity of metals: mechanisms, molecular targets and applications. Nat Rev Microbiol 2013; 11:371–384 [View Article][PubMed]
     [Google Scholar]
  64. Wright MS, Loeffler Peltier G, Stepanauskas R, McArthur JV. Bacterial tolerances to metals and antibiotics in metal-contaminated and reference streams. FEMS Microbiol Ecol 2006; 58:293–302 [View Article][PubMed]
     [Google Scholar]
  65. Luo G, Li B, Li L-G, Zhang T, Angelidaki I. Antibiotic resistance genes and correlations with microbial community and metal resistance genes in full-scale biogas reactors as revealed by metagenomic analysis. Environ Sci Technol 2017; 51:4069–4080 [View Article][PubMed]
     [Google Scholar]
  66. Tuckfield RC, McArthur JV. Spatial analysis of antibiotic resistance along metal contaminated streams. Microb Ecol 2008; 55:595–607 [View Article][PubMed]
     [Google Scholar]
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Principal component analysis exploring the association between antibiotic resistance and heavy metal tolerance of plasmid-bearing sewage wastewater bacteria of clinical relevance
Access Microbiology 2, e000095 (2020); https://doi.org/10.1099/acmi.0.000095
/content/journal/acmi/10.1099/acmi.0.000095
/content/journal/acmi/10.1099/acmi.0.000095
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