Isolation and identification of cadmium- and lead-resistant lactic acid bacteria for application as metal removing probiotic

  • J. N. Bhakta
  • Y. Munekage
  • K. Ohnishi
  • B. B. Jana
Original Paper

Abstract

The purpose of the present study was to isolate and identify the metal-resistant lactic acid bacteria from sediments of coastal aquaculture habitats for removal of cadmium and lead from ambience. Collected sediment samples were used to isolate the cadmium- and lead-resistant bacterial colonies by spread plate technique using agar media (De Man, Rogosa and Sharpe) supplemented with cadmium or lead at 50 mg/l. Isolates were identified by bacterial colony polymerase chain reaction and sequencing of 16S ribosomal deoxyribonucleic acid. Metal removing probiotic was determined by characterizing the lactic acid yield in culture media, viability in fish intestine, metal-resistant and metal-removal efficiencies. 16S ribosomal deoxyribonucleic acid sequencing data of five (Cd10, Cd11, Pb9, Pb12 and Pb18) and other all isolates clearly showed 99 % similarities to Enterococcus faecium and Bacillus cereus, respectively. The Pb12 exhibited higher lactic acid yield (180 mmol) than that of the remaining E. faecium strains and excellent viability without pathogenicity; therefore, further study was carried out using Pb12 strain. The selected Pb12 strain showed elevated metal resistant (minimum inhibitory concentrations 120 and 800 mg/l for cadmium and lead, respectively) and removal efficiencies [Cadmium 0.0377 mg/h/g and lead 0.0460 mg/h/g of cells (wet weight)]. From the viability and metal removal points of view, it can be concluded that isolated metal-resistant E. faecium Pb12 strains might be used as potential probiotic strains for removing heavy metals from fish intestinal milieu to control the progressive bioaccumulation of heavy metals in the fish.

Keywords

Bioaccumulation Enterococcus faecium Heavy metal Removal efficiency Sediment 

References

  1. Aksu Z (2005) Application of biosorption for the removal of organic pollutants: a review. Process Biochem 40(3–4):997–1026CrossRefGoogle Scholar
  2. Amundsen PA, Staldvik FJ, Lukin AA, Kashulin NA, Popova OA, Reshetnikov YS (1997) Heavy metal contamination in freshwater fish from the border region between Norway and Russia. Sci Total Environ 201(3):211–224CrossRefGoogle Scholar
  3. Anadon A, Martynes-Larranaga MR, Martynes MA (2006) Probiotic for animal nutrition in the European Union. Regulation and safety assessment. Regul Toxicol Pharmacol 45(1):91–95CrossRefGoogle Scholar
  4. Ashraf MA, Maah MJ, Yusoff I (2011) Heavy metals accumulation in plants growing on former tin mining catchment. Int J Environ Sci Technol 8(2):401–416Google Scholar
  5. Belimov AA, Ontzeas N, Safronova VI (2005) Cadmium-tolerant plant growth-promoting bacteria associated with the roots of Indian mustard (Brassica juncea L. Czern.). Soil Biol Biochem 37(2):241–250CrossRefGoogle Scholar
  6. Brown MJ, Lester JN (1982) Role of bacterial extracellular polymers in metal uptake in pure bacterial culture and activated sludge-1. Effect of metal concentration. Water Res 16(11):1539–1548CrossRefGoogle Scholar
  7. Bunting SW, Pretty J, Edwards P (2010) Wastewater-fed aquaculture in the East Kolkata Wetlands, India: anachronism or archetype for resilient ecocultures? Rev Aquac 2(3):138–153CrossRefGoogle Scholar
  8. Campbell PGC (2006) Cadmium—a priority pollutant. Environ Chem 3(6):387–388CrossRefGoogle Scholar
  9. Edwards P (2005a) Demise of periurban wastewater-fed aquaculture? Urban Agric Mag 14:27–29Google Scholar
  10. Edwards P (2005b) Development status of, and prospects for, wastewater-fed aquaculture in urban environments. In: Costa-Pierce B, Desbonnet A, Edwards P, Baker D (eds) Urban aquaculture. CABI Publishing, Wallingford, pp 45–59CrossRefGoogle Scholar
  11. Gadd GM (1990) Heavy metal accumulation by bacteria and other microorganisms. Experientia 46(8):834–840CrossRefGoogle Scholar
  12. Giller K, Witter E, McGrath SP (1998) Toxicity of heavy metals to microorganisms and microbial processes in agricultural soils: a review. Soil B Biochem 30(10–11):1389–1414CrossRefGoogle Scholar
  13. Gupta UC, Gupta SC (1998) Trace element toxicity relationships to crop production and livestock and human health: implications for management. Commun Soil Sci Plant Anal 29(11 & 14):1491–1522CrossRefGoogle Scholar
  14. Herranz C, Casaus P, Mukhopadhyay S, Martínez JM, Rodríguez JM, Nes IF, Hernández PE, Cintas LM (2001) Enterococcus faecium P21: a strain occurring naturally in dry-fermented sausages producing the class II bacteriocins enterocin A and enterocin B. Food Microbiol 18(2):115–131CrossRefGoogle Scholar
  15. Hollis L, Hogstrand C, Wood CM (2001) Tissue-specific cadmium accumulation, metallothionein induction, and tissue zinc and copper levels during chronic sublethal cadmium exposure in juvenile rainbow trout. Arch Environ Contam Toxicol 41(4):468–474CrossRefGoogle Scholar
  16. Hrudey SE, Chen W, Rousseaux CG (1996) Bioavailability in environmental risk assessment. Lewis Publications, Boca RatonGoogle Scholar
  17. Idris R, Trifonova R, Puschenreiter M (2004) Bacterial communities associated with flowering plants of the Ni hyperaccumulator Thaspi goesingense. Appl Environ Microbiol 70(5):2667–2677CrossRefGoogle Scholar
  18. Jana BB (1998) Sweage-fed aquaculture: the Calcutta model1. Ecol Eng 11(1–4):73–85CrossRefGoogle Scholar
  19. Jarup L (2003) Hazards of heavy metal contamination. Br Med Bull 68(1):167–182CrossRefGoogle Scholar
  20. Kalay M, Aly O, Canil M (1999) Heavy metal concentrations in fish tissues from the Northeast Mediterranean sea. Bull Environ Contam Toxicol 63(5):673–681CrossRefGoogle Scholar
  21. Kara Y (2005) Bioaccumulation of Cu, Zn and Ni from the wastewater by treated Nasturtium officinale. Int J Environ Sci Technol 2(1):63–67Google Scholar
  22. Kratochvil D, Volesky D (1998) Advances in the biosorption of heavy metals. Tybtech 16(7):291–300CrossRefGoogle Scholar
  23. Ling T, Jun R, Fangke Y (2011) Effect of cadmium supply levels to cadmium accumulation by Salix. Int J Environ Sci Technol 8(3):493–500Google Scholar
  24. Long A, Wang W (2005) Assimilation and bioconcentration of Ag and Cd by the marine black bream after waterborne and dietary metal exposure. Environ Toxicol Chem 24(3):709–716CrossRefGoogle Scholar
  25. Macey BM, Coyne VE (2005) Improved growth rate and disease resistance of farmed Haliotis midae through probiotic treatment. Aquaculture 245(1–4):249–261CrossRefGoogle Scholar
  26. Macha M, Taras D, Vahjen W (2004) Specific enumeration of the probiotic strain Enterococcus faecium NCIMB 10415 in the intestinal tract and in faeces of piglets and sows. Arch Anim Nutr 58(6):443–452CrossRefGoogle Scholar
  27. Marcussen H, Holm PE, Ha LT, Dalsgaard A (2007) Food safety aspects of toxic element accumulation in fish from wastewater-fed ponds in Hanoi, Vietnam. Trop Med Int Health 12(2):34–39CrossRefGoogle Scholar
  28. Min-sheng H, Jing P, Le-ping Z (2001) Removal of heavy metals from aqueous solutions using bacteria. J Shanghai Univ 5(3):253–259CrossRefGoogle Scholar
  29. Mohideen MMAK, Selva TM, Mohamed SP, Hussain MIZ (2010) Effect of probiotic bacteria on the growth rate of freshwater fish, Catla catla. Int J Biol Technol 1(2):113–117Google Scholar
  30. Musikasang H, Tani A, H-kittikun A, Maneerat S (2009) Probiotic potential of lactic acid bacteria isolated from chicken gastrointestinal digestive tract. World J Microbiol Biotechnol 25(8):337–1345CrossRefGoogle Scholar
  31. Nwachukwu MA, Feng H, Alinnor J (2010) Assessment of heavy metal pollution in soil and their implications within and around mechanic villages. Int J Environ Sci Technol 7(2):347–358Google Scholar
  32. Panigrahi A, Kiron V, Puangkaew J, Kobayashi T, Satoh S, Sugita H (2005) The viability of probiotic bacteria as a factor influencing the immune response in rainbow trout Oncorhynchus mykiss. Aquaculture 243(1–4):241–254CrossRefGoogle Scholar
  33. Pazirandeh M, Wells BM, Ryan RL (1998) Development of bacterium-based heavy metal biosorbents: enhanced uptake of cadmium and mercury by Escherichia coli expressing a metal binding motif. Appl Environ Microbiol 64(10):4068–4072Google Scholar
  34. Phuong NTD, Tuan PA (2005) Current status of periurban aquatic production in Hanoi. Urban Agric Mag 14:10–12Google Scholar
  35. Qing H, Min-Na D, Hong-Yan Q, Xiang-Ming X, Guo-Qiang Z, Min Y (2007) Detection, isolation, and identification of cadmium-resistant bacteria based on PCR-DGGE. J Environ Sci 19(9):1114–1119CrossRefGoogle Scholar
  36. Rengpipat S, Rueangruklikhit T, Piyatiratitivorakul S (2008) Evaluations of lactic acid bacteria as probiotics for juvenile seabass Lates calcarifer. Aquac Res 39(2):134–143CrossRefGoogle Scholar
  37. Ruangsomboon S, Wongrat L (2006) Bioaccumulation of cadmium in an experimental aquatic food chain involving phytoplankton (Chlorella vulgaris), zooplankton (Moina macrocopa), and the predatory catfish Clarias macrocephalus and C. gariepinus. Aquat Toxicol 78(1):15–20CrossRefGoogle Scholar
  38. Ruiz-Barba JL, Maldonado A, Jiménez-Díaz R (2005) Small-scale total DNA extraction from bacteria and yeast for PCR applications. Anal Biochem 347(2):333–335CrossRefGoogle Scholar
  39. Scharek L, Guth J, Reiter K, Weyrauch KD, Taras D, Schwerk P, Schierack P, Schmidt MFG, Wieler LH, Tedin K (2005) Influence of a probiotic Enterococcus faecium strain on development of the immune system of sows and piglets. Vet Immunol Immunopathol 105(1–2):151–161CrossRefGoogle Scholar
  40. Sugita H, Ishigaki T, Iwai D, Suzuki Y, Okano R, Matsuura S, Asfie M, Aono E, Deguchi Y (1998) Antibacterial abilities of intestinal bacteria from three coastal fishes. Suisan Zoshoku 46(4):563–568Google Scholar
  41. Taras D, Vahjen W, Macha M (2006) Performance, diarrhea incidence, and occurrence of Escherichia coli virulence genes during long-term administration of a probiotic Enterococcus faecium strain to sows and piglets. Anim Sci 84(3):608–617Google Scholar
  42. USEPA (1992) Common chemicals found at superfund sites. U.S. Gov Print Office, Washington, DC, Miner Eng 14:317–340Google Scholar
  43. Vahjen W, Jadamus A, Simon O (2002) Influence of a probiotic Enterococcus faecium stain on selected bacterial groups in the small intestine of growing turkey poults. Arch Anim Nutr 56(6):419–429Google Scholar
  44. Wong CK, Wong PPK, Chu LM (2001) Heavy metal concentrations in marine fishes collected from fish culture sites in Hong Kong. Arch Environ Contam Toxicol 40(1):60–69CrossRefGoogle Scholar

Copyright information

© CEERS, IAU 2012

Authors and Affiliations

  • J. N. Bhakta
    • 1
  • Y. Munekage
    • 2
  • K. Ohnishi
    • 1
  • B. B. Jana
    • 3
  1. 1.Research Institute of Molecular Genetics, Faculty of AgricultureKochi UniversityNankokuJapan
  2. 2.Department of Environmental Engineering, Faculty of AgricultureKochi UniversityNankokuJapan
  3. 3.International Centre of Ecological EngineeringUniversity of KalyaniKalyaniIndia

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