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The interaction of Vibrio cholerae with chitin exemplifies for microbial ecology a successful bacteria-substrate interaction with complex and significant influence on the lifestyle of the bacterium. Chitin is one of the most abundant polymers on earth and possibly the most abundant in the aquatic environment, where its association with V. cholerae has provided the microorganism with a number of advantages, including food availability, adaptation to environmental nutrient gradients, tolerance to stress and protection from predators. Emergent properties of V. cholerae-chitin interactions occur at multiple hierarchical levels in the environment and include cell metabolic and physiological responses e.g. chemotaxis, cell multiplication, induction of competence, biofilm formation, commensal and symbiotic relationship with higher organisms, cycling of nutrients, and pathogenicity for humans and aquatic animals. As factors mediating virulence of V. cholerae for humans and aquatic animals derive from mechanisms of adaptation to its environment, at different levels of hierarchical scale, V. cholerae interactions with chitin represent a useful model for examination of the role of primary habitat selection in the development of traits that have been identified as virulence factors in human disease.

Forty-one Tnpho A mutants of Vibrio cholerae O1 classical strain CD81 were analyzed for their ability to interact with chitin particles, Tigriopus fulvus copepods and the Intestine 407 cell line compared to the parent strain. Thirteen mutants were less adhesive than CD81; in particular, T21, T33 and T87 were less adhesive towards all substrates and insensitive to inhibition by N-acetyl glucosamine (GlcNAc). By SDS-PAGE analysis of sarkosyl-insoluble membrane proteins (siMPs) isolated from mutants and parent, it was found that a 53 kDa siMP is missing in T21, T33 and T87 mutants. It is hypothesized that this protein might have the function to mediate adherence to GlcNAc-containing substrates both in the aquatic environment and in human intestine.

The Vibrio cholerae N-acetyl glucosamine-binding protein A (GbpA) is a chitin-binding protein involved in V. cholerae attachment to environmental chitin surfaces and human intestinal cells. We previously investigated the distribution and genetic variations of gbpA in a large collection of V. cholerae strains and found that the gene is consistently present and highly conserved in this species. Primers and probe were designed from the gbpA sequence of V. cholerae and a new Taq-based qPCR protocol was developed for diagnostic detection and quantification of the bacterium in environmental and stool samples. In addition, the positions of primers targeting the gbpA gene region were selected to obtain a short amplified fragment of 206 bp and the protocol was optimized for the analysis of formalin-fixed samples, such as historical Continuous Plankton Recorder (CPR) samples. Overall, the method is sensitive (50 gene copies), highly specific for V. cholerae and failed to amplify strains of the closely-related species Vibrio mimicus. The sensitivity of the assay applied to environmental and stool samples spiked with V. cholerae ATCC 39315 was comparable to that of pure cultures and was of 102 genomic units/l for drinking and seawater samples, 101 genomic units/g for sediment and 102 genomic units/g for bivalve and stool samples. The method also performs well when tested on artificially formalin-fixed and degraded genomic samples and was able to amplify V. cholerae DNA in historical CPR samples, the earliest of which date back to August 1966. The detection of V. cholerae in CPR samples collected in cholera endemic areas such as the Benguela Current Large Marine Ecosystem (BCLME) is of particular significance and represents a proof of concept for the possible use of the CPR technology and the developed qPCR assay in cholera studies. [ABSTRACT FROM AUTHOR]

Summary In the aquatic environment, Vibrio cholerae has been reported to be associated with a variety of living organisms, including animals with an exoskeleton of chitin, aquatic plants, protozoa, bivalves, waterbirds, as well as abiotic substrates (e.g. sediments). Most of these are well-known or putative environmental reservoirs for the bacterium, defined as places where the pathogen lives over time, with the potential to be released and to cause human infection. Environmental reservoirs also serve as V. cholerae disseminators and vectors. They can be responsible for the start of an epidemic, may be critical to cholera endemicity, and affect the evolution of pathogen virulence. To date, in addition to the generally recognized role of zooplankton as the largest environmental reservoir for V. cholerae, other environmental reservoirs play some role in cholera epidemiology by favouring persistence of the pathogen during inter-epidemic periods. Little is known about the ecological factors affecting V. cholerae survival in association with aquatic substrates. Studies aimed at these aspects, i.e. understanding how environmental reservoirs interact, are affected by climate, and contribute to disease epidemiology, will be useful for understanding global implications of V. cholerae and the disease cholera. [ABSTRACT FROM AUTHOR]

Summary Vibrio cholerae O1 El Tor attachment to chitin and biofilm formation on polyvinylchloride surfaces via the N-acetylglucosamine-binding protein A (GbpA) and the mannose-sensitive haemagglutinin (MSHA) were investigated under different temperature and salinity conditions simulating those found in the aquatic environment. In vitro tests showed that mshA and gbpA defective V. cholerae N16961 strains displayed a significant reduction ( P < 0.05) in attachment to chitin in comparison with the parent in all the environmental conditions tested. The lack of mshA, but not gbpA, resulted in a significant decrease ( P < 0.05) of V. cholerae N16961 strain ability to form biofilm. Wild-type attachment to chitin and biofilm formation increased from 15°C to 25°C as did gbpA and mshA expression. In situ data obtained analysing zooplankton and water samples collected in coastal waters of NW Mediterranean Sea over an annual cycle showed that the percentage of plankton-associated V. cholerae was positive correlated with sea surface temperature, and increased dramatically at temperature values above 22°C. It is suggested that temperature plays a major role in affecting persistence of V. cholerae in the aquatic environment by promoting colonization of environmental surfaces, via an enhanced expression of both mshA and gbpA. [ABSTRACT FROM AUTHOR]

Recent results for Vibrio cholerae interactions with bivalves and chitin-containing substrates are reviewed. Chitin, composed of β-1,4-linked N-acetylglucosamine residues, is one of the most abundant biopolymers in nature and the most abundant in the marine environment. V. cholerae connection to chitin is a well known phenomenon and one of the best documented examples of a successful bacteria-substrate interaction, affecting both the lifestyle of the microorganisms and natural system functioning. In sea water, vibrios can be entrapped by filter-feeding invertebrates that sieve suspended food particles from the aquatic environment. Persistence of bacteria in bivalves depends on different factors including their sensitivity to the bactericidal activity of hemolymph. Both V. cholerae survival inside bivalves and chitin colonization depend on cell surface properties, with contribution of different ligands (e.g. chitin-binding proteins, MSHA). The role of these ligands in interactions with human surfaces is discussed and evidence is presented supporting (i) the link between vibrio persistence in the environment (both inside bivalves and adhering to chitin-containing surfaces) and infection of the human host, and (ii) the hypothesis that virulence mechanisms of bacteria that have environmental reservoirs represent a consequence of adaptive mechanisms to the environment. [ABSTRACT FROM AUTHOR]

Presents a study that explored the role of surface proteins in Vibrio cholerae (v. cholerae) attachment to chitin particles in vitro. Effects of treatment of bacteria with pronase E, sodium m-periodate and sugars on V. cholerae attachment to chitin; Effects of treatment of chitin with membrane proteins (MP) and lectins on V. cholerae attachment to chitin; Involvement of surface proteins in interactions between V. cholerae 01 classical strains and chitin particles.

Investigates the ability of marine organism Vibrio alginolyticus to adhere to adhere to chitin surfaces by evolving specific systems to adapt to the substrate. Interaction of chitin particles and V. alginolyticus; Isolation of chitin-binding proteins from V. alginolyticus strains; Temperature of highest degree of attachment.

Vibrio cholerae exhibits two distinctive lifestyles, one inside the milieu of the human intestine and the other in the aquatic environment. Recently, the existence of V. cholerae ligands involved in colonization of both human intestine and environmental chitin surfaces via the same binding specificity has been shown. Such molecules, here named ‘dual role colonization factors (DRCFs)’, are example of a tight connection between the two V. cholerae''s lifestyles. It is suggested that DRCFs and, more generally, bacterial factors and pathways having roles in pathogenesis and in the out of the human body life may be promising targets for development of novel prophylactic or therapeutic interventions that may also affect V. cholerae fitness in its environmental reservoirs. [Copyright &y& Elsevier]