Do the Escherichia coli European Union shellfish safety standards predict the presence of Arcobacter spp., a potential zoonotic pathogen?

The genus Arcobacter comprises Campylobacter-related species, considered zoonotic emergent pathogens, the presence of which in water has been associated with fecal pollution. Discharges of fecal polluted water into the sea have been considered as one of the main reasons for the presence of Arcobacter in shellfish, and this may represent a risk for public health. In this study, the European Union shellfish food safety criteria based on levels of Escherichia coli were studied in relation to their capacity to predict the presence of Arcobacter species. In addition, the accumulation factor (AF) that measures the concentration ratio between the microbes present in the shellfish and in the water, was also studied for both bacteria. The results show that the presence of E. coli correlated with the presence of the potentially pathogenic species A. butzleri and A. cryaerophilus. However, in 26.1% of the shellfish samples (corresponding to those taken during summer months) E. coli failed to predict the presence of, for instance A. butzleri and A. skirrowii, among other species. In the rest of the samples a significant correlation between the concentration of E. coli and Arcobacter spp. (mussels and oyster; R2=0.744) was found. This study indicates that the presence of E. coli can predict the presence of pathogenic Arcobacter species in shellfish samples harvested from water with temperatures lower than 26.2°C. Consumption of shellfish collected at higher temperatures which may not be permissive to the growth of E. coli but does allow growth of Arcobacter spp., may represent a risk for consumers.

Enhanced recovery of Arcobacter spp. using NaCl in culture media and re-assessment of the traits of Arcobacter marinus and Arcobacter halophilus isolated from marine water and shellfish.

The genus Arcobacter is a relatively poorly known group of bacteria, and the number of new species and sequences from non-culturable strains has increased considerably in recent years. This study investigates whether using media that contain NaCl might help to improve the recovery of Arcobacter spp. from marine environments. To this aim, 62 water and shellfish samples were analysed in parallel, with both a commonly used culture method (enrichment in Arcobacter-CAT broth followed by culture on Blood Agar) and a new one that supplements the Arcobacter-CAT enrichment broth with 2.5% NaCl (w/v) followed by culturing on Marine Agar. The new method yielded ca. 40% more positive samples and provided a higher diversity of known (11 vs. 7) and unknown (7 vs. 2) Arcobacter species. Among the 11 known species recovered, Arcobacter marinus and Arcobacter halophilus were isolated only by this new method. No more strains of these species have been isolated since their original descriptions, both of which were based only on a single strain. In view of that, the phenotypic characteristics of these species are re-evaluated in the present study, using the new strains. Strains of A. halophilus had the same phenotypic profile as the type strain. However, some strains of A. marinus differed from the type strain in that they did not hydrolyse indoxyl-acetate, becoming, therefore, the first Arcobacter species to show a varying ability to hydrolyse indoxyl-acetate. This study shows to what extent a simple variation to the culture media can have a big influence on positive samples and on the community of species recovered.

Aeromonas lusitana sp. nov., Isolated from Untreated Water and Vegetables.

During previous studies to evaluate the phylogenetic diversity of Aeromonas from untreated waters and vegetables intended for human consumption, a group of isolates formed a unique gyrB phylogenetic cluster, separated from those of all other species described so far. A subsequent extensive phenotypic characterization, DNA-DNA hybridization, 16S rRNA gene sequencing, multi-locus phylogenetic analysis of the concatenated sequence of seven housekeeping genes (gyrB, rpoD, recA, dnaJ, gyrA, dnaX, and atpD; 4705 bp), and ERIC-PCR, were performed in an attempt to ascertain the taxonomy position of these isolates. This polyphasic approach confirmed that they belonged to a novel species of the genus Aeromonas, for which the name Aeromonas lusitana sp. nov. is proposed, with strain A.11/6(T) (=DSMZ 24095(T), =CECT 7828(T)) as the type strain.

Aeromonas dhakensis pneumonia and sepsis in a neonate Risso's dolphin Grampus griseus from the Mediterranean Sea.

A neonate Risso's dolphin Grampus griseus was found stranded alive on a beach in Catalonia, Spain. Rehabilitation attempts were unsuccessful and it died 2 d later, showing pneumonia and sepsis. A pure bacterial culture was obtained from all tissues and blood and identified as Aeromonas hydrophila using the API 20NE. However, sequencing the rpoD gene showed that the strain in fact belongs to A. dhakensis, making this the first report of fatal haemorrhagic-necrotizing pneumonia and sepsis due to this species in a marine mammal. The A. dhakensis strain GMV-704 produced ß-haemolysis, possessed several virulence genes and showed sensitivity to several antimicrobials. This study provides a new potential host for A. dhakensis, and its potential virulence in dolphins and its presence in the marine environment may warrant considering this species a potential threat to marine mammals.

Multilocus phylogenetic analysis of the genus Aeromonas.

A broad multilocus phylogenetic analysis (MLPA) of the representative diversity of a genus offers the opportunity to incorporate concatenated inter-species phylogenies into bacterial systematics. Recent analyses based on single housekeeping genes have provided coherent phylogenies of Aeromonas. However, to date, a multi-gene phylogenetic analysis has never been tackled. In the present study, the intra- and inter-species phylogenetic relationships of 115 strains representing all Aeromonas species described to date were investigated by MLPA. The study included the independent analysis of seven single gene fragments (gyrB, rpoD, recA, dnaJ, gyrA, dnaX, and atpD), and the tree resulting from the concatenated 4705 bp sequence. The phylogenies obtained were consistent with each other, and clustering agreed with the Aeromonas taxonomy recognized to date. The highest clustering robustness was found for the concatenated tree (i.e. all Aeromonas species split into 100% bootstrap clusters). Both possible chronometric distortions and poor resolution encountered when using single-gene analysis were buffered in the concatenated MLPA tree. However, reliable phylogenetic species delineation required an MLPA including several "bona fide" strains representing all described species.

New perspectives in monitoring drinking water microbial quality.

The safety of drinking water is evaluated by the results obtained from faecal indicators during the stipulated controls fixed by the legislation. However, drinking-water related illness outbreaks are still occurring worldwide. The failures that lead to these outbreaks are relatively common and typically involve preceding heavy rain and inadequate disinfection processes. The role that classical faecal indicators have played in the protection of public health is reviewed and the turning points expected for the future explored. The legislation for protecting the quality of drinking water in Europe is under revision, and the planned modifications include an update of current indicators and methods as well as the introduction of Water Safety Plans (WSPs), in line with WHO recommendations. The principles of the WSP approach and the advances signified by the introduction of these preventive measures in the future improvement of drinking water quality are presented. The expected impact that climate change will have in the quality of drinking water is also critically evaluated.

Clinical relevance of the recently described species Aeromonas aquariorum.

Twenty-two human extraintestinal isolates (11 from blood) and three isolates recovered from patients with diarrhea were genetically characterized as Aeromonas aquariorum, a novel species known only from ornamental fish. The isolates proved to bear a considerable number of virulence genes, and all were resistant to amoxicillin (amoxicilline), cephalothin (cefalotin), and cefoxitin. Biochemical differentiation from the most relevant clinical species is provided.