Effect of dilution factor on the isolation of Helicobacter pylori from municipal wastewater using culture technique
-
Mehri Solaimany Aminabad
,
Mahdi Hadi
,
Seyedeh Zohreh Mirbagheri
,
Alireza Mesdaghinia
,
Ronak Bakhtiari
,
Masoud Alebouyeh
,
Shahrokh Nazmara
Abstract
Background and Objectives: Isolating Helicobacter pylori (H. pylori) from wastewater and culturing it using a conventional method has always been a controversial issue because the bacterium converts into a coccoid form when exposed to an unfavourable environment like wastewater. To clarify the cultivability behaviour of the bacterium in fresh wastewater samples, the effect of municipal wastewater dilation on the cultivation of the bacterium using a conventional method was examined.
Materials and Methods: Several dilutions of wastewater samples were inoculated with fresh H. pylori suspension (with McFarland's dilution 0.5) to examine the dilution effect of wastewater on the bacterium isolation.
Results: The H. pylori growth was found to be possible for a dilution factor from 1/106 to 1/107 of raw wastewater. In higher dilution factors the growth of fungi was dominant and could prevent the isolation of the bacterium.
Conclusion: The optimized technique could be applied in future studies for increasing the chance of H. pylori isolation from fresh wastewater environments.
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2. Mungazi SG, Chihaka OB, Muguti GI. Prevalence of Helicobacter pylori in asymptomatic patients at surgical outpatient department: Harare hospitals. Ann Med Surg (Lond) 2018; 35: 153-157.
3. Berendonk TU, Manaia CM, Merlin C, Fatta-Kassinos D, Cytryn E, Walsh F, et al. Tackling antibiotic resistance: the environmental framework. Nat Rev Microbiol 2015; 13: 310-317.
4. Hooi JKY, Lai WY, Ng WK, Suen MMY, Underwood FE, Tanyingoh D, et al. Global prevalence of Helicobacter pylori infection: systematic review and meta-analysis. Gastroenterology 2017; 153: 420-429.
5. Dube C, Tanih NF, Ndip RN. Helicobacter pylori in water sources: a global environmental health concern. Rev Environ Health 2009; 24: 1-14.
6. Queralt N, Bartolome R, Araujo R. Detection of Helicobacter pylori DNA in human faeces and water with different levels of faecal pollution in the north‐east of Spain. J Appl Microbiol 2005; 98: 889-895.
7. Goh K-L, Chan W-K, Shiota S, Yamaoka Y. Epidemiology of Helicobacter pylori infection and public health implications. Helicobacter 2011; 16 Suppl 1 (0 1): 1-9.
8. Bai X, Xi C, Wu J. Survival of Helicobacter pylori in the wastewater treatment process and the receiving river in Michigan, USA. J Water Health 2016; 14: 692-698.
9. López-Serna R, Postigo C, Blanco J, Pérez S, Ginebreda A, De Alda ML, et al. Assessing the effects of tertiary treated wastewater reuse on the presence emerging contaminants in a Mediterranean river (Llobregat, NE Spain). Environ Sci Pollut Res Int 2012; 19: 1000-1012.
10. Lemarchand K, Berthiaume F, Maynard C, Harel J, Payment P, Bayardelle P, et al. Optimization of microbial DNA extraction and purification from raw wastewater samples for downstream pathogen detection by microarrays. J Microbiol Methods 2005; 63: 115-126.
11. Lu Y, Redlinger TE, Avitia R, Galindo A, Goodman K. Isolation and genotyping of Helicobacter pylori from untreated municipal wastewater. Appl Environ Microbiol 2002; 68: 1436-1439.
12. Shannon KE, Lee D-Y, Trevors JT, Beaudette LA. Application of real-time quantitative PCR for the detection of selected bacterial pathogens during municipal wastewater treatment. Sci Total Environ 2007; 382: 121-129.
13. Straub TM, Chandler DP. Towards a unified system for detecting waterborne pathogens. J Microbiol Methods 2003; 53: 185-197.
14. Hadi M, Shokoohi R, Namvar AE, Karimi M, Aminabad MS. Antibiotic resistance of isolated bacteria from urban and hospital wastewaters in Hamadan City. Iran J Health Environ 2011; 4: 105-114.
15. Rizzo L, Manaia C, Merlin C, Schwartz T, Dagot C, Ploy MC, et al. Urban wastewater treatment plants as hotspots for antibiotic resistant bacteria and genes spread into the environment: a review. Sci Total Environ 2013; 447: 345-360.
16. Nguyen AQ, Vu HP, Nguyen LN, Wang Q, Djordjevic SP, Donner E, et al. Monitoring antibiotic resistance genes in wastewater treatment: Current strategies and future challenges. Sci Total Environ 2021; 783: 146964.
17. Lagier J-C, Edouard S, Pagnier I, Mediannikov O, Drancourt M, Raoult D. Current and past strategies for bacterial culture in clinical microbiology. Clin Microbiol Rev 2015; 28: 208-236.
18. Al-Awadhi H, Dashti N, Khanafer M, Al-Mailem D, Ali N, Radwan S. Bias problems in culture-independent analysis of environmental bacterial communities: a representative study on hydrocarbonoclastic bacteria. Springerplus 2013; 2: 369.
19. Al-Dhabaan FA. Morphological, biochemical and molecular identification of petroleum hydrocarbons biodegradation bacteria isolated from oil polluted soil in Dhahran, Saud Arabia. Saudi J Biol Sci 2019; 26: 1247-1252.
20. Hortelano I, Moreno Y, Vesga FJ, Ferrús MA. Evaluation of different culture media for detection and quantification of H. pylori in environmental and clinical samples. Int Microbiol 2020; 23: 481-487.
21. Ayrapetyan M, Williams TC, Oliver JD. Bridging the gap between viable but non-culturable and antibiotic persistent bacteria. Trends Microbiol 2015; 23: 7-13.
22. Li L, Mendis N, Trigui H, Oliver JD, Faucher SP. The importance of the viable but non-culturable state in human bacterial pathogens. Front Microbiol 2014; 5: 258.
23. Zeng B, Zhao G, Cao X, Yang Z, Wang C, Hou L. Formation and resuscitation of viable but nonculturable Salmonella typhi. Biomed Res Int 2013; 2013: 907170.
24. Frenck Jr RW, Clemens J. Helicobacter in the developing world. Microbes Infect 2003; 5: 705-713.
25. Adams BL, Bates TC, Oliver JD. Survival of Helicobacter pylori in a natural freshwater environment. Appl Environ Microbiol 2003; 69: 7462-7466.
26. Rizvi F, Hannan A. Evaluation of different transport and enrichment media for the isolation of Helicobacter pylori. JAMC 2000; 12: 31-33.
27. Castillo-Rojas G, Mazarí-Hiriart M, López-Vidal Y. Helicobacter pylori: focus on CagA and VacA major virulence factors. Salud Publica Mex 2004; 46: 538-548.
28. Miehlke S, Kirsch C, Agha-Amiri K, Günther T, Lehn N, Malfertheiner P, et al. The Helicobacter pylori vacA s1, m1 genotype and cagA is associated with gastric carcinoma in Germany. Int J Cancer 2000; 87: 322-327.
29. Mirbagheri SZ, Bakhtiari R, Fakhre Yaseri H, Rahimi Foroushani A, Eshraghi SS, Alebouyeh M. Transcriptional alteration of genes linked to gastritis concerning Helicobacter pylori infection status and its virulence factors. Mol Biol Rep 2021; 48: 6481-6489.
30. Saberi S, Douraghi M, Azadmanesh K, Shokrgozar MA, Zeraati H, Hosseini ME, et al. A potential association between Helicobacter pylori CagA EPIYA and multimerization motifs with cytokeratin 18 cleavage rate during early apoptosis. Helicobacter 2012; 17: 350-357.
31. Moreno Y, Ferrús MA. Specific detection of cultivable Helicobacter pylori cells from wastewater treatment plants. Helicobacter 2012; 17: 327-332.
32. Andersen LP, Wadstrom T. Basic bacteriology and culture. In: Mobley HL, Mendz GL, Hazell SL, editors. Helicobacter Pylori: Physiology and Genetics. Washington, DC: ASM Press; 2001. p. 27-38.
33. Robben C, Fister S, Witte AK, Schoder D, Rossmanith P, Mester P. Induction of the viable but non-culturable state in bacterial pathogens by household cleaners and inorganic salts. Sci Rep 2018; 8: 15132.
34. Jiang X, Doyle MP. Effect of environmental and substrate factors on survival and growth of Helicobacter pylori. J Food Prot 1998; 61: 929-933.
35. Saito N, Konishi K, Sato F, Kato M, Takeda H, Sugiyama T, et al. Plural transformation-processes from spiral to coccoid Helicobacter pylori and its viability. J Infect 2003; 46: 49-55.
36. Cellini L, Del Vecchio A, Di Candia M, Di Campli E, Favaro M, Donelli G. Detection of free and plankton‐associated Helicobacter pylori in seawater. J Appl Microbiol 2004; 97: 285-292.
37. Azevedo NF, Pacheco AP, Keevil CW, Vieira MJ. Nutrient shock and incubation atmosphere influence recovery of culturable Helicobacter pylori from water. Appl Environ Microbiol 2004; 70: 490-493.
38. Queralt N, Araujo R. Analysis of the survival of H. pylori within a laboratory-based aquatic model system using molecular and classical techniques. Microb Ecol 2007; 54: 771-777.
39. Konishi K, Saito N, Shoji E, Takeda H, Kato M, Asaka M, et al. Helicobacter pylori: longer survival in deep ground water and sea water than in a nutrient‐rich environment. APMIS 2007; 115: 1285-1291.
40. Azevedo NF, Almeida C, Cerqueira L, Dias S, Keevil CW, Vieira MJ. Coccoid form of Helicobacter pylori as a morphological manifestation of cell adaptation to the environment. Appl Environ Microbiol 2007; 73: 3423-3427.
41. Degnan AJ, Sonzogni WC, Standridge JH. Development of a plating medium for selection of Helicobacter pylori from water samples. Appl Environ Microbiol 2003; 69: 2914-2918.
42. Moreno Y, Piqueres P, Alonso JL, Jiménez A, González A, Ferrús MA. Survival and viability of Helicobacter pylori after inoculation into chlorinated drinking water. Water Res 2007; 41: 3490-3496.
43. Jiang X, Doyle MP. Optimizing enrichment culture conditions for detecting Helicobacter pylori in foods. J Food Prot 2002; 65: 1949-1954.
44. Janzon A, Sjöling Å, Lothigius Å, Ahmed D, Qadri F, Svennerholm A-M. Failure to detect Helicobacter pylori DNA in drinking and environmental water in Dhaka, Bangladesh, using highly sensitive real-time PCR assays. Appl Environ Microbiol 2009; 75: 3039-3044.
45. Piqueres P, Moreno Y, Alonso JL, Ferrús MA. A combination of direct viable count and fluorescent in situ hybridization for estimating Helicobacter pylori cell viability. Res Microbiol 2006; 157: 345-349.
46. Bellack NR, Koehoorn MW, MacNab YC, Morshed MG. A conceptual model of water's role as a reservoir in Helicobacter pylori transmission: a review of the evidence. Epidemiol Infect 2006; 134: 439-449.
2. Mungazi SG, Chihaka OB, Muguti GI. Prevalence of Helicobacter pylori in asymptomatic patients at surgical outpatient department: Harare hospitals. Ann Med Surg (Lond) 2018; 35: 153-157.
3. Berendonk TU, Manaia CM, Merlin C, Fatta-Kassinos D, Cytryn E, Walsh F, et al. Tackling antibiotic resistance: the environmental framework. Nat Rev Microbiol 2015; 13: 310-317.
4. Hooi JKY, Lai WY, Ng WK, Suen MMY, Underwood FE, Tanyingoh D, et al. Global prevalence of Helicobacter pylori infection: systematic review and meta-analysis. Gastroenterology 2017; 153: 420-429.
5. Dube C, Tanih NF, Ndip RN. Helicobacter pylori in water sources: a global environmental health concern. Rev Environ Health 2009; 24: 1-14.
6. Queralt N, Bartolome R, Araujo R. Detection of Helicobacter pylori DNA in human faeces and water with different levels of faecal pollution in the north‐east of Spain. J Appl Microbiol 2005; 98: 889-895.
7. Goh K-L, Chan W-K, Shiota S, Yamaoka Y. Epidemiology of Helicobacter pylori infection and public health implications. Helicobacter 2011; 16 Suppl 1 (0 1): 1-9.
8. Bai X, Xi C, Wu J. Survival of Helicobacter pylori in the wastewater treatment process and the receiving river in Michigan, USA. J Water Health 2016; 14: 692-698.
9. López-Serna R, Postigo C, Blanco J, Pérez S, Ginebreda A, De Alda ML, et al. Assessing the effects of tertiary treated wastewater reuse on the presence emerging contaminants in a Mediterranean river (Llobregat, NE Spain). Environ Sci Pollut Res Int 2012; 19: 1000-1012.
10. Lemarchand K, Berthiaume F, Maynard C, Harel J, Payment P, Bayardelle P, et al. Optimization of microbial DNA extraction and purification from raw wastewater samples for downstream pathogen detection by microarrays. J Microbiol Methods 2005; 63: 115-126.
11. Lu Y, Redlinger TE, Avitia R, Galindo A, Goodman K. Isolation and genotyping of Helicobacter pylori from untreated municipal wastewater. Appl Environ Microbiol 2002; 68: 1436-1439.
12. Shannon KE, Lee D-Y, Trevors JT, Beaudette LA. Application of real-time quantitative PCR for the detection of selected bacterial pathogens during municipal wastewater treatment. Sci Total Environ 2007; 382: 121-129.
13. Straub TM, Chandler DP. Towards a unified system for detecting waterborne pathogens. J Microbiol Methods 2003; 53: 185-197.
14. Hadi M, Shokoohi R, Namvar AE, Karimi M, Aminabad MS. Antibiotic resistance of isolated bacteria from urban and hospital wastewaters in Hamadan City. Iran J Health Environ 2011; 4: 105-114.
15. Rizzo L, Manaia C, Merlin C, Schwartz T, Dagot C, Ploy MC, et al. Urban wastewater treatment plants as hotspots for antibiotic resistant bacteria and genes spread into the environment: a review. Sci Total Environ 2013; 447: 345-360.
16. Nguyen AQ, Vu HP, Nguyen LN, Wang Q, Djordjevic SP, Donner E, et al. Monitoring antibiotic resistance genes in wastewater treatment: Current strategies and future challenges. Sci Total Environ 2021; 783: 146964.
17. Lagier J-C, Edouard S, Pagnier I, Mediannikov O, Drancourt M, Raoult D. Current and past strategies for bacterial culture in clinical microbiology. Clin Microbiol Rev 2015; 28: 208-236.
18. Al-Awadhi H, Dashti N, Khanafer M, Al-Mailem D, Ali N, Radwan S. Bias problems in culture-independent analysis of environmental bacterial communities: a representative study on hydrocarbonoclastic bacteria. Springerplus 2013; 2: 369.
19. Al-Dhabaan FA. Morphological, biochemical and molecular identification of petroleum hydrocarbons biodegradation bacteria isolated from oil polluted soil in Dhahran, Saud Arabia. Saudi J Biol Sci 2019; 26: 1247-1252.
20. Hortelano I, Moreno Y, Vesga FJ, Ferrús MA. Evaluation of different culture media for detection and quantification of H. pylori in environmental and clinical samples. Int Microbiol 2020; 23: 481-487.
21. Ayrapetyan M, Williams TC, Oliver JD. Bridging the gap between viable but non-culturable and antibiotic persistent bacteria. Trends Microbiol 2015; 23: 7-13.
22. Li L, Mendis N, Trigui H, Oliver JD, Faucher SP. The importance of the viable but non-culturable state in human bacterial pathogens. Front Microbiol 2014; 5: 258.
23. Zeng B, Zhao G, Cao X, Yang Z, Wang C, Hou L. Formation and resuscitation of viable but nonculturable Salmonella typhi. Biomed Res Int 2013; 2013: 907170.
24. Frenck Jr RW, Clemens J. Helicobacter in the developing world. Microbes Infect 2003; 5: 705-713.
25. Adams BL, Bates TC, Oliver JD. Survival of Helicobacter pylori in a natural freshwater environment. Appl Environ Microbiol 2003; 69: 7462-7466.
26. Rizvi F, Hannan A. Evaluation of different transport and enrichment media for the isolation of Helicobacter pylori. JAMC 2000; 12: 31-33.
27. Castillo-Rojas G, Mazarí-Hiriart M, López-Vidal Y. Helicobacter pylori: focus on CagA and VacA major virulence factors. Salud Publica Mex 2004; 46: 538-548.
28. Miehlke S, Kirsch C, Agha-Amiri K, Günther T, Lehn N, Malfertheiner P, et al. The Helicobacter pylori vacA s1, m1 genotype and cagA is associated with gastric carcinoma in Germany. Int J Cancer 2000; 87: 322-327.
29. Mirbagheri SZ, Bakhtiari R, Fakhre Yaseri H, Rahimi Foroushani A, Eshraghi SS, Alebouyeh M. Transcriptional alteration of genes linked to gastritis concerning Helicobacter pylori infection status and its virulence factors. Mol Biol Rep 2021; 48: 6481-6489.
30. Saberi S, Douraghi M, Azadmanesh K, Shokrgozar MA, Zeraati H, Hosseini ME, et al. A potential association between Helicobacter pylori CagA EPIYA and multimerization motifs with cytokeratin 18 cleavage rate during early apoptosis. Helicobacter 2012; 17: 350-357.
31. Moreno Y, Ferrús MA. Specific detection of cultivable Helicobacter pylori cells from wastewater treatment plants. Helicobacter 2012; 17: 327-332.
32. Andersen LP, Wadstrom T. Basic bacteriology and culture. In: Mobley HL, Mendz GL, Hazell SL, editors. Helicobacter Pylori: Physiology and Genetics. Washington, DC: ASM Press; 2001. p. 27-38.
33. Robben C, Fister S, Witte AK, Schoder D, Rossmanith P, Mester P. Induction of the viable but non-culturable state in bacterial pathogens by household cleaners and inorganic salts. Sci Rep 2018; 8: 15132.
34. Jiang X, Doyle MP. Effect of environmental and substrate factors on survival and growth of Helicobacter pylori. J Food Prot 1998; 61: 929-933.
35. Saito N, Konishi K, Sato F, Kato M, Takeda H, Sugiyama T, et al. Plural transformation-processes from spiral to coccoid Helicobacter pylori and its viability. J Infect 2003; 46: 49-55.
36. Cellini L, Del Vecchio A, Di Candia M, Di Campli E, Favaro M, Donelli G. Detection of free and plankton‐associated Helicobacter pylori in seawater. J Appl Microbiol 2004; 97: 285-292.
37. Azevedo NF, Pacheco AP, Keevil CW, Vieira MJ. Nutrient shock and incubation atmosphere influence recovery of culturable Helicobacter pylori from water. Appl Environ Microbiol 2004; 70: 490-493.
38. Queralt N, Araujo R. Analysis of the survival of H. pylori within a laboratory-based aquatic model system using molecular and classical techniques. Microb Ecol 2007; 54: 771-777.
39. Konishi K, Saito N, Shoji E, Takeda H, Kato M, Asaka M, et al. Helicobacter pylori: longer survival in deep ground water and sea water than in a nutrient‐rich environment. APMIS 2007; 115: 1285-1291.
40. Azevedo NF, Almeida C, Cerqueira L, Dias S, Keevil CW, Vieira MJ. Coccoid form of Helicobacter pylori as a morphological manifestation of cell adaptation to the environment. Appl Environ Microbiol 2007; 73: 3423-3427.
41. Degnan AJ, Sonzogni WC, Standridge JH. Development of a plating medium for selection of Helicobacter pylori from water samples. Appl Environ Microbiol 2003; 69: 2914-2918.
42. Moreno Y, Piqueres P, Alonso JL, Jiménez A, González A, Ferrús MA. Survival and viability of Helicobacter pylori after inoculation into chlorinated drinking water. Water Res 2007; 41: 3490-3496.
43. Jiang X, Doyle MP. Optimizing enrichment culture conditions for detecting Helicobacter pylori in foods. J Food Prot 2002; 65: 1949-1954.
44. Janzon A, Sjöling Å, Lothigius Å, Ahmed D, Qadri F, Svennerholm A-M. Failure to detect Helicobacter pylori DNA in drinking and environmental water in Dhaka, Bangladesh, using highly sensitive real-time PCR assays. Appl Environ Microbiol 2009; 75: 3039-3044.
45. Piqueres P, Moreno Y, Alonso JL, Ferrús MA. A combination of direct viable count and fluorescent in situ hybridization for estimating Helicobacter pylori cell viability. Res Microbiol 2006; 157: 345-349.
46. Bellack NR, Koehoorn MW, MacNab YC, Morshed MG. A conceptual model of water's role as a reservoir in Helicobacter pylori transmission: a review of the evidence. Epidemiol Infect 2006; 134: 439-449.
| Files | ||
| Issue | Vol 14 No 6 (2022) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v14i6.11264 | |
| Keywords | ||
| Helicobacter pylori; Wastewater; Culture techniques | ||
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How to Cite
1.
Solaimany Aminabad M, Hadi M, Mirbagheri SZ, Mesdaghinia A, Bakhtiari R, Alebouyeh M, Nazmara S. Effect of dilution factor on the isolation of Helicobacter pylori from municipal wastewater using culture technique. Iran J Microbiol. 2022;14(6):891-900.
