Annals of Microbiology

, Volume 67, Issue 12, pp 827–835 | Cite as

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

  • Luca Settanni
  • Raimondo GaglioEmail author
  • Carlo Stucchi
  • Simone De Martino
  • Nicola Francesca
  • Giancarlo Moschetti
Original Article


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.


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



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

Supplementary material

13213_2017_1311_MOESM1_ESM.docx (77 kb)
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)
13213_2017_1311_MOESM2_ESM.docx (72 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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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature and the University of Milan 2017

Authors and Affiliations

  1. 1.Dipartimento Scienze Agrarie, Alimentari e ForestaliUniversità di PalermoPalermoItaly
  2. 2.Istituto Nazionale Ghiaccio Alimentare (INGA)RomeItaly

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