Inactivation of Legionella pneumophila by combined systems of copper and silver ions and free chlorine.

The aim of this study was to evaluate the efficacy of copper and silver ions and of free chlorine in different combinations and concentrations (0.4 to 0.8-0.04 to 0.08 mg/l Cu(2+) Ag(+); 0.4 to 0.8-0.04 to 0.08-0.2 mg/l Cu(2+) Ag(+) Cl; 0.4 to 0.8-0.04 to 0.08-2 mg/l Cu(2+) Ag(+) Cl; 0.4 to 0.8-0.04 to 0.08-4 mg/l Cu(2+) Ag(+) Cl; 2-20 mg/l Cl), in inactivating Legionella pneumophila in drinking and distilled water after a contact time of 24-hours. Treatment with chlorine alone at 20 mg/l concentration was found to be the most effective treatment leading to complete killing of bacteria within 4 minutes in all water samples. On the other hand, at 2 mg/l concentration complete inactivation was obtained after 3 hours. The association of copper and silver ions at concentrations of 0.4-0.04 mg/l was found to be less effective and live bacteria could still be identified in all water samples after a 24 hour contact time. When testing copper and silver ions in combination, at concentrations of 0.8-0.08 mg/l and different combinations of the three disinfectants, results varied according to the various concentrations and type of water. The combination of copper and silver with 2 mg/l of chlorine was found to be more effective than 2mg/l of chlorine alone; a synergistic effect can therefore be hypothesized. The physical and chemical properties of drinking water, in particular its chlorine content, may have affected the water disinfection process when disinfecting agents were used in low concentrations. In conclusion, this study confirms the efficacy of shock hyperchlorination in the inactivation of Legionella pneumophila. However, the combination of free chlorine with metal (copper and silver) ions may represent a valid option for reducing the concentration of disinfectants to safer levels for human health and avoiding damage to water distribution systems especially in facilities such as hotels and hospitals.

Techniques for the recovery of enteric viruses from artificially contaminated marine sediments.

Viruses are an important component of aquatic microbial communities and marine sediments may represent an optimal means for their survival. The aim of this study was to evaluate different methods for virus recovery from marine sediments. Three methods were used for virus recovery from artificially contaminated sediments: elution and centrifugation technique, sonication technique, and mechanical disgregation followed by elution and centrifuge technique. The sonication technique obtained the highest virus recovery percentages (94,25%). Eluent 2 provided more efficient recovery of enteric viruses than eluent 1 presumably due to the presence, in eluent 2, of NANO3, a chaotropic agent that enhances the solubilization of hydrophobic compounds in water. Finally, the authors confirm the importance of searching for viruses in sediments, which protect them from inactivation by biological, chemical and physical factors and allow them to survive longer than in the overlaying water column.

Use of multiparameter analysis for Vibrio alginolyticus viable but nonculturable state determination.

BACKGROUND: Vibrio alginolyticus is known to enter into a viable but nonculturable (VBNC) state in response to environmental conditions unfavorable to the growth. Cells in VBNC condition pose a public health threat because they are potentially pathogenic. METHODS: We constructed a pathway for the identification of the most significant variables and the characterization of those variables able to discriminate the groups under investigation. Different parameters measured by the image processing software were chosen as the most representative of V. alginolyticus cell morphology (length index for dimension) and metabolic activity (density profile indexes). To detect relationships between the groups of treatment performed, we carried out a principal components analysis (PCA). RESULTS: The PCA analysis indicated that increasing coccoid shape transformation was related to both metabolic and dimension variations, delineating a well defined graph profile. Indeed, we discovered that specific morphological variations occur when cells in the culturable state pass into VBNC condition, namely comma-shaped culturable bacteria are converted into coccoid-shaped VBNC cells. The results were also supported by scanning electron microscopy analysis. CONCLUSIONS: This technique allows the analysis of a large number of vibrio samples in a short period of time. The obtained multiparameter information may complement genetic/molecular analyses facilitating, in an automatic fashion, further studies to evaluate the potential risk of this pathogen in the environment. It may also be a useful tool for large-scale cell biology studies and high content screening.

Campylobacter jejuni loss of culturability in aqueous microcosms and ability to resuscitate in a mouse model.

Water is known as one of the main transmission routes of Campylobacter and contributes to increase the number of sporadic infections and outbreaks. Campylobacter jejuni persists in the environment, especially in water, in a viable but non-culturable (VBNC) form that is thought to be a possible cause of water-borne outbreaks. In this study, we evaluated the loss of culturability and viability of 9 C. jejuni strains of clinical origin and one ATCC reference strain when kept at 4 degrees C in artificial sea water (ASW). Culturability was measured as colony-forming units while viability was evaluated by CTC-DAPI double staining and the combined CTC-specific fluorescent antibody technique (CTC-FA). When cultured on Columbia Agar plates, strains exhibited different growth profiles which allowed to classify them into three different groups. Both techniques used to monitor the viability of the bacterial cells showed that C. jejuni strains survived in the VBNC form in the microcosms through a period lasting from 138 to 152 days. The recovery of C. jejuni VBNC forms to culturability, as evidenced by cell division, was obtained by passage in the mouse intestine. Our results indicate that C. jejuni VBNC cells were able to remain in this state for a few months and regain their culturability after in vivo passage depending on their lasting in the VBNC state, which affects the number of respiring bacteria. In fact, the resuscitation was achieved when the number of respiring bacteria became higher than 10(4) cell/ml. Therefore, a relatively high microbial titer of respiring bacteria in the VBNC state seems to be important for the resuscitation and subsequent intestinal colonisation.