Occurrence of Vibrio vulnificus in mussel farms from the Varano lagoon environment.

AIMS: Monitoring the occurrence of the human pathogen Vibrio vulnificus in a mussel farm located in the lagoon of Varano (Italy). METHODS AND RESULTS: A total of 72 samples of mussel, water and sediment, collected from two locations of Varano lagoon in the Gargano peninsula, during a 7-month survey, were analysed. Isolation and PCR characterization of six V. vulnificus environmental genotype strains revealed that this pathogen was isolated when with T was above 22 °C and salinity ranged between 22.7 and 26.4‰. No significant correlation of the occurrence of V. vulnificus with water pH or salinity was observed. Moreover, 8% of mussel samples were found to be contaminated by V. vulnificus. All of that positive mussel samples originated from the same sampling station. CONCLUSION: It is suggested that warmer season are risky to eat raw or undercooked bivalve molluscs in the local area. SIGNIFICANCE AND IMPACT OF THE STUDY: To increase knowledge about environmental conditions that may affect the occurrence of waterborne pathogen Vibrio vulnificus in seafood.

Occurrence of Giardia and Cryptosporidium in Italian water supplies.

A total of 21 samples: raw water (RW) samples; water samples after coagulation with aluminium sulfate (clarified water: CW); and water after chlorination (treated water: TW) from a water purification plant that treats river surface water from the neighbourhood of Foggia (Italy), were analysed for the presence of Giardia cysts and Cryptosporidium oocysts. Bacteriological indicator of faecal contamination (total and faecal coliforms, faecal streptococci,), total bacterial count at 22 and 36 degrees C and physicochemical parameters (turbidity, temperature, pH) were evaluated. Cryptosporidium oocysts were not found in any samples examined, while Giardia cysts were found only in RW samples, with the maximal concentration of 8 cysts/100 l. A positive correlation was found between the Giardia densities and quality parameters such as TC, FC and TBC at 22 degrees C. Giardia levels in raw water samples correlated (p < 0.05) with TC, FC and with temperature. No other water quality parameters was consistently correlated with cysts level.

Cloning, molecular characterization and expression analysis of two small heat shock genes isolated from wine Lactobacillus plantarum.

AIM: Understanding the molecular response to stress tolerance of wine Lactobacillus plantarum. METHODS AND RESULTS: Two genes codifying for heat shock proteins were cloned from wine L. plantarum. The coding regions of the two heat shock genes are 420 and 444 nucleotides long, and started with an ATG codon suggesting that they were translated. The protein sequences deduced from the isolated genes have a molecular mass of 18.483 and 19.282 kDa, respectively, and were therefore named hsp18.5 and hsp19.3. The expression of small heat shock genes was analysed by RT-PCR analysis. Moreover, the 5' and 3' noncoding regions were cloned and sequenced. CONCLUSIONS: The expression of the heat shock genes was strongly induced by heat, cold and ethanol stress. Analysis of the 5' and 3' flanking regions of hsp18.5 and hsp19.3 genes, revealed the presence of an inverted repeat sequence (TTAGCACTC-N(9)-GAGTGCTAA) homologue to the CIRCE elements found to the upstream regulatory region of heat shock operons, and an inverted sequence that could form a stem and loop structure that it is likely to function as a transcriptional terminator. Based on their structures, the genes were classified as belonging to Class I of heat shock genes according to the B. subtilis nomenclature of heat response genes. SIGNIFICANCE AND IMPACT OF THE STUDY: Small heat shock genes isolated from wine L. plantarum might have a role in preventing damage by cold stress.

Molecular characterization of lactic acid populations associated with wine spoilage.

We have investigated the prevalence of spoilage lactic acid bacteria (LAB) in table wines produced in the Apulia region. The occurrence of LAB was evaluated in wines produced with low sulphur dioxide doses and not supplemented with selected malolactic starters such as Oenococcus oeni. About 150 strains were isolated from wine must and a molecular characterization was performed using PCR-based techniques. Most of the strains analysed belonged to Lactobacillus plantarum species. However, some of the strains were identified as Pediococcus damnosus and Leuconostoc sp. The amplified fragments of Pediococcus damnosus were cloned and sequenced. The coding sequence was highly homologous to that of the ropy plasmid confirming that the isolated strain was a ropy(+) Pediococcus damnosus. In all the samples analysed, the final must pH value reached was relatively high (from 3.78 to 3.90). The high pH values had probably negatively influenced (counteracted) the activity of sulphur dioxide added, allowing proliferation of spoilage wine microorganisms.