Presence of pathogenic bacteria in ice cubes and evaluation of their survival in different systems

Abstract

In this study, 60 samples of ice cubes produced at different levels (domestic, restaurant and industrial facilities), within a restricted geographical area, were investigated for their general microbiological characteristics through the analysis of populations other than enteric bacteria. Total mesophilic bacteria were in the range 1.01 × 102–9.55 × 103, 3.12 × 102–6.31 × 103 and 1.30 × 102–3.99 × 103 CFU/100 mL of thawed ice from domestic freezer (DF), stock boxes (SB) for self-production performed with ice machines in bars and pubs, and from sales packages (SP) of industrial productions, respectively. Some DF and SP samples were negative for the presence of total psychrotrophic bacteria, showing that there are no specific microbial groups associated with ice. Pseudomonads were found in the majority of ice samples analyzed. The levels of contamination of the ice samples were significantly different among the three ice cube production levels. The samples produced at domestic level and those collected from bars and pubs were characterised by the highest cell densities. The colonies representative for the different bacterial morphologies were randomly picked up from plates, purified to homogeneity and subjected to the phenotypic and genotypic characterisation. Fifty-two strains representing 31 species of eight bacterial genera were identified, with the most numerous groups included in Pseudomonas, Staphylococcus, Bacillus and Acinetobacter. A consistent percentage of the microorganisms identified from ice are known agents of human infections, and their presence indicate an environmental contamination. In order to evaluate the effectiveness of the ice cubes to transfer pathogenic agents to consumers, a bar consumption was simulated with different drink systems added with ice cubes artificially contaminated with the strains found at dominant levels (Acinetobacter lwoffii ICE100, Bacillus cereus ICE170, Pseudomonas putida ICE224 and Staphylococcus haemolyticus ICE182), and the results showed a consistent reduction of bacterial risk due to alcohol, CO2, pH and antibacterial ingredients of vodka, whisky, Martini, peach tea, tonic water and coke.

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References

  1. Altinok I, Kayis S, Capkin E (2006) Pseudomonas putida infection in rainbow trout. Aquaculture 261:850–855. https://doi.org/10.1016/j.aquaculture.2006.09.009

    Article  Google Scholar 

  2. Awuor L, Thompson S, Thompson B, Liberda EN, Meldrum R (2016) Microbiological quality and handling practices of ice served in selected downtown Toronto food premises. Environ Health Rev 59:83–87. https://doi.org/10.5864/d2016-017

    Article  Google Scholar 

  3. Chavasit V, Sirilaksanamanon K, Phithaksantayothin P, Norapoompipat Y, Parinyasiri T (2011) Measures for controlling safety of crushed ice and tube ice in developing country. Food Control 22:118–123. https://doi.org/10.1016/j.foodcont.2010.05.016

    Article  Google Scholar 

  4. Commission Regulation (1998) No 98/83/EC of 3 November 1998 on the quality of water intended for human consumption. Off J Eur Union 330:32–54

    Google Scholar 

  5. Dabboussi F, Hamze M, Elomari M, Verhille S, Baida N, Izard D, Leclerc H (1999) Pseudomonas libanensis sp. nov., a new specie isolated from Lebanese spring waters. Int J Syst Bacteriol 49:1091–1101. https://doi.org/10.1099/00207713-49-3-1091

    CAS  Article  PubMed  Google Scholar 

  6. Economou V, Gousia P, Kemenetzi D, Sakkas H, Papadopoulou C (2017) Microbial quality and histamine producing microflora analysis of the ice used for fish preservation. J Food Saf 37:12285. https://doi.org/10.1111/jfs.12285

    Article  Google Scholar 

  7. Falcao JP, Dias AMG, Correa EF, Falcao DP (2002) Microbiological quality of ice used to refrigerate foods. Food Microbiol 19:269–276. https://doi.org/10.1006/fmic.2002.0490

    Article  Google Scholar 

  8. Falcao JP, Falcao DP, Gomes TAT (2004) Ice as a vehicle for diarrheagenic Escherichia coli. Int J Food Microbiol 91:99–103. https://doi.org/10.1016/S0168-1605(03)00327-1

    Article  PubMed  Google Scholar 

  9. Fredheim EGA, Klingenberg C, Rohde H, Frankenberger S, Gaustad P, Flægstad T, Sollid JE (2009) Biofilm formation by Staphylococcus haemolyticus. J Clin Microbiol 47:1172–1180. https://doi.org/10.1128/JCM.01891-08

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  10. Gaglio R, Francesca N, Di Gerlando R, Mahony J, De Martino S, Stucchi C, Moschetti G, Settanni L (2017) Enteric bacteria of food ice and their survival in alcoholic beverages and soft drinks. Food Microbiol 67:17–22. https://doi.org/10.1016/j.fm.2017.04.020

    CAS  Article  PubMed  Google Scholar 

  11. Gerokomou V, Voidarou C, Vatopoulos A, Velonakis E, Rozos G, Alexopoulos A, Plessas S, Stavropoulou E, Bezirtzoglou E, Demertzis PG, Akrida-Demertzi K (2011) Physical, chemical and microbiological quality of ice used to cool drinks and foods in Greece and its public health implications. Anaerobe 17:351–353. https://doi.org/10.1016/j.anaerobe.2011.06.005

    CAS  Article  PubMed  Google Scholar 

  12. Graman PS, Quinlan GA, Rank JA (1997) Nosocomial legionellosis traced to a contaminated ice machine. Infect Control Hosp Epidemiol 18:637–640

    CAS  Article  PubMed  Google Scholar 

  13. Gregersen T (1978) Rapid method for distinction of gram-negative from gram-positive bacteria. Appl Microbiol Biotechnol 5:123–127. https://doi.org/10.1007/BF00498806

    Article  Google Scholar 

  14. Hampikyan H, Bingol EB, Cetin O, Colak H (2017) Microbiological quality of ice and ice machines used in food establishments. J Water Health 15:410–417. https://doi.org/10.2166/wh.2017.159.

    Article  PubMed  Google Scholar 

  15. Istituto Nazionale Ghiaccio Alimentare (2015) Manuale di corretta prassi operativa per la produzione di ghiaccio alimentare. In: http://www.ghiaccioalimentare.it/il-manuale/. Accessed 13 May 2017

  16. Kamath U, Singer C, Isenberg HD (1992) Clinical significance of Staphylococcus warneri bacteremia. J Clin Microbiol 30:261–264

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Kharal SA, Hussain Q, Ali S (2009) Quinine is bactericidal. J Pak Med Assoc 59:208–212

    PubMed  Google Scholar 

  18. Kotiranta A, Lounatmaa K, Haapasalo M (2000) Epidemiology and pathogenesis of Bacillus cereus infections. Microbes Infect 2:189–198. https://doi.org/10.1016/S1286-4579(00)00269-0

    CAS  Article  PubMed  Google Scholar 

  19. Lateef A, Oloke JK, Kana EBG, Pacheco E (2006) The microbiological quality of ice used to cool drinks and foods in Ogbomoso metropolis, southwest, Nigeria. Internet J Food Saf 8:39–43

    Google Scholar 

  20. Lee KH, Ab Samad LS, Lwin PM, Riedel SF, Magin A, Bashir M, Vaishampayan PA, Lin WJ (2017) On the rocks: microbiological quality and microbial diversity of packaged ice in Southern California. J Food Prot 80:1041–1049. https://doi.org/10.4315/0362-028X.JFP-16-295

    Article  PubMed  Google Scholar 

  21. López JR, Diéguez AL, Doce A, De la Roca E, De la Herran R, Navas JI, Toranzo AE, Romalde JL (2012) Pseudomonas baetica sp. nov., a fish pathogen isolated from wedge sole, Dicologlossa cuneata (Moreau). Int J Syst Evol Microbiol 62:874–882. https://doi.org/10.1099/ijs.0.030601-0.

    Article  PubMed  Google Scholar 

  22. Nichols G, Gillespie I, de Louvois J (2000) The microbiological quality of ice used to cool drinks and ready-to-eat from retail and catering premises in the United Kingdom. J Food Prot 63:78–82. https://doi.org/10.4315/0362-028X-63.1.78

    CAS  Article  PubMed  Google Scholar 

  23. Noor Izani NJ, Zulaikha AR, Mohamad Noor MR, Amri MA, Mahat NA (2012) Contamination of faecal coliforms in ice cubes sampled from food outlets in Kubang Kerian, Kelantan. Trop Biomed 29:71–76

    CAS  PubMed  Google Scholar 

  24. Northcutt JK, Smith D (2010) Microbiological and chemical analyses of ice collected from a commercial poultry processing establishment. Poult Sci 89:145–149. https://doi.org/10.3382/ps.2009-00205

    CAS  Article  PubMed  Google Scholar 

  25. Palmisano MM, Nakamura LK, Duncan KE, Istock CA, Cohan FM (2001) Bacillus sonorensis sp. nov., a close relative of Bacillus licheniformis, isolated from soil in the Sonoran Desert, Arizona. Int J Syst Evol Microbiol 51:1671–1679. https://doi.org/10.1099/00207713-51-5-1671

    CAS  Article  PubMed  Google Scholar 

  26. Pawsey RK, Howard P (2001) Drinking ice as a vector for gastrointestinal disease. Brit Food J 103:253–263. https://doi.org/10.1108/00070700110391380

    Article  Google Scholar 

  27. Settanni L, Di Grigoli A, Tornambé G, Bellina V, Francesca N, Moschetti G, Bonanno A (2012) Persistence of wild Streptococcus thermophilus strains on wooden vat and during the manufacture of a Caciocavallo type cheese. Int J Food Microbiol 155:73–81. https://doi.org/10.1016/j.ijfoodmicro.2012.01.022

    CAS  Article  PubMed  Google Scholar 

  28. Shivaji S, Chaturvedi P, Suresh K, Reddy GSN, Dutt CBS, Wainwright M, Narlikar JV, Bhargava PM (2006) Bacillus aerius sp. nov., Bacillus aerophilus sp. nov., Bacillus stratosphericus sp. nov. and Bacillus altitudinis sp. nov., isolated from cryogenic tubes used for collecting air samples from high altitudes. Int J Syst Evol Microbiol 56:1465–1473. https://doi.org/10.1099/ijs.0.64029-0

    CAS  Article  PubMed  Google Scholar 

  29. Shivaji S, Chaturvedi P, Begum Z, Pindi PK, Manorama R, Padmanaban DA, Shouche YS, Pawar S, Vaishampayan P, Dutt CB, Datta GN, Manchanda RK, Rao UR, Bhargava PM, Narlikar JV (2009) Janibacter hoylei sp. nov., Bacillus isronensis sp. nov. and Bacillus aryabhattai sp. nov., isolated from cryotubes used for collecting air from the upper atmosphere. Int J Syst Evol Microbiol 59:2977–2986. https://doi.org/10.1099/ijs.0.002527-0

    CAS  Article  PubMed  Google Scholar 

  30. Ukwo SP, Ndaeyo NU, Udoh EJ (2011) Microbiological quality and safety evaluation of fresh juices and edible ice sold in Uyo metropolis, south-south, Nigeria. Internet J Food Saf 13:374–378

    Google Scholar 

  31. Venkateswaran K, Kempf M, Chen F, Satomi M, Nicholson W, Kern R (2003) Bacillus nealsoni sp. nov., isolated from a spacecraft-assembly facility, whose spores are γ-radiation resistant. Int J Syst Evol Microbiol 53:165–172. https://doi.org/10.1099/ijs.0.02311-0

    CAS  Article  PubMed  Google Scholar 

  32. Verhille S, Baida N, Dabboussi F, Izard D, Leclerc H (1999) Taxonomic study of bacteria isolated from natural mineral waters: proposal of Pseudomonas jessenii sp. nov. and Pseudomonas mandelii sp. nov. Syst Appl Microbiol 22:45–58

  33. Visca P, Seifert H, Towner KJ (2011) Acinetobacter infection—an emerging threat to human health. IUBMB Life 63:1048–1054. https://doi.org/10.1002/iub.534

    CAS  Article  PubMed  Google Scholar 

  34. Vuong C, Otto M (2002) Staphylococcus epidermidis infections. Microbes Infect 4:481–489

    Article  PubMed  Google Scholar 

  35. Wilson IG, Hogg GM, Barr JG (1997) Microbiological quality of ice in hospital and community. J Hosp Infect 36:171–180

    CAS  Article  PubMed  Google Scholar 

  36. Wisplinghoff H, Bischoff T, Tallent SM, Seifert H, Wenzel RP, Edmond MB (2004) Nosocomial bloodstream infections in US hospitals: analysis of 24,179 cases from a prospective nationwide surveillance study. Clin Infect Dis 39:309–317. https://doi.org/10.1086/421946

    Article  PubMed  Google Scholar 

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Acknowledgement

This work was financed by the National Institute of Food Ice (INGA), Rome, Italy.

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Correspondence to Raimondo Gaglio.

Electronic supplementary material

ESM 1

Supplementary Fig. S1. Dendrogram obtained with combined RAPD-PCR patterns generated with three primers (M17, AB111, and AB106) for Gram positive bacterial strains isolated from ice cubes. The line at the top indicates percentages of similarity. Abbreviations: B., Bacillus; Br., Brevibacterium; L., Lysinobacillus; M., Microbacterium; S., Staphylococcus (DOCX 76.5 kb)

ESM 2

Supplementary Fig. S2. Dendrogram obtained with combined RAPD-PCR patterns generated with three primers (M17, AB111, and AB106) for Gram negative bacterial strains isolated from ice cubes. The line at the top indicates percentages of similarity. Abbreviations: A., Acinetobacter; P., Pseudomonas; Px., Pseudoxanthomonas (DOCX 72.4 kb)

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Settanni, L., Gaglio, R., Stucchi, C. et al. Presence of pathogenic bacteria in ice cubes and evaluation of their survival in different systems. Ann Microbiol 67, 827–835 (2017). https://doi.org/10.1007/s13213-017-1311-1

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Keywords

  • Cross-contamination
  • Genetic identification
  • Human infections
  • Hygiene
  • Ice cubes
  • Microbial survival