Internalization and cytotoxicity are important virulence mechanisms in vibrio-f ishepithelial cell interactions

Wang, X. H.; Oon, H. L.; Ho, G. W. P.; Wong, W.S. Fred; Lim, Tit Meng; Leung, Ka Yin

doi:10.1099/00221287-144-11-2987

Cited by 46 publications

(31 citation statements)

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“…Adhesion assays were performed as described previously with minor modifications (64). Briefly, 72-h-old epithelioma papillosum of carp (Cyprinus carpio) (EPC) cells in 24-well tissue culture plates were washed with Hanks' balanced salt solution (HBSS) and then incubated for an additional 30 min in tissue culture medium containing different concentrations (0.1 to 0.6 g/ml) of O-antigen and capsular polysaccharides.…”

Section: Methodsmentioning

confidence: 99%

“…Naïve tilapia serum was used to perform the serum resistance assay. Bacteria were prepared and treated with 50% tilapia serum as described previously (64). Different concentrations (0.1 to 0.6 g/ml) of A. hydrophila PPD134/91 O-antigen and capsular polysaccharides were added to the serum in different tubes, followed by addition of washed A. hydrophila PPD35/85 (serum-sensitive strain) cells.…”

Section: Methodsmentioning

confidence: 99%

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NovelAeromonas hydrophilaPPD134/91 Genes Involved in O-Antigen and Capsule Biosynthesis

Zhang¹,

Arakawa

Leung

2002

Infect Immun

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The sequences of the O-antigen and capsule gene clusters of the virulent Aeromonas hydrophila strain PPD134/91 were determined. The O-antigen gene cluster is 17,296 bp long and comprises 17 genes. Seven pathway genes for the synthesis of rhamnose and mannose, six transferase genes, one O unit flippase gene, and one O-antigen chain length determinant gene were identified by amino acid sequence similarity. PCR and Southern blot analysis were performed to survey the distribution of these 17 genes among 11 A. hydrophila strains of different serotypes. A. hydrophila PPD134/91 might belong to serotype O:18, as represented by JCM3980; it contained all the same O-antigen genes as JCM3980 (97 to 100% similarity at the DNA and amino acid levels). The capsule gene cluster of A. hydrophila PPD134/91 is 17,562 bp long and includes 13 genes, which were assembled into three distinct regions similar to those of the group II capsule gene cluster of Escherichia coli and other bacteria. Regions I and III contained four and two capsule transport genes, respectively. Region II had five genes which were highly similar to capsule synthesis pathway genes found in other bacteria. Both the purified O-antigen and capsular polysaccharides increased the ability of the avirulent A. hydrophila strain PPD35/85 to survive in naïve tilapia serum. However, the purified surface polysaccharides had no inhibitory effect on the adhesion of A. hydrophila PPD134/91 to carp epithelial cells.Surface polysaccharides, such as O-antigen and capsule, are important bacterial cell surface components. The O-antigen polysaccharide is covalently ligated to the lipid A-core complex and extends outward from the cell surface. The capsule is an extracellular polysaccharide enclosing the bacterium while remaining attached to the cell. Both the O-antigen polysaccharide and the capsule are composed of repeating oligosaccharide units (44). They act as prominent antigens and play important roles in the pathogenicity of many bacterial pathogens, such as protecting bacterial cells from complement-mediated serum killing (20, 30), acting as adhesion factors (31), protecting the bacteria from the effects of desiccation (38), and aiding survival in phagocytes (56). The serogrouping of bacterial strains within a genus is determined by the structural variability of surface polysaccharides. For example, Escherichia coli strains are divided into more than 160 serogroups based on the different surface polysaccharides (67). Klebsiella species have been classified into 72 serogroups based on the structural variability of their capsular polysaccharides (39).Aeromonas hydrophila is an important pathogen of a wide variety of aquatic and terrestrial animals, especially fish (4). In fish, it causes hemorrhagic septicemia, which often results in high mortalities in commercial aquaculture. Some strains of A. hydrophila are also reported to cause infections in humans. The clinical symptoms include septicemia (17), meningitis (25), peritonitis (35), pneumonia (32), myonecrosis (34), and diarrh...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

NovelAeromonas hydrophilaPPD134/91 Genes Involved in O-Antigen and Capsule Biosynthesis

Zhang¹,

Arakawa

Leung

2002

Infect Immun

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“…infections (Toranzo & Barja 1993). In addition, the ECP fractions from pathogenic Vibrio appear to play a major role during disease and the enzymatic components of this fraction have been implicated as important factors necessary for invasion, survival and proliferation in host fish (Balebona et al 1998, Wang et al 1998, Wang & Leung 2000. Although ECP is an important factor in pathogenesis, a positive correlation between the pathogenicity of a particular strain of Vibrio and the virulence of the ECP from the same strain cannot be assumed as cytotoxicity of ECP prepared from nonpathogenic Vibrio strains can also occur (Toranzo et al 1983).…”

Section: Discussionmentioning

confidence: 99%

Modulated heat shock protein expression during pathogenic Vibrio alginolyticus stress of sea bream

Deane

Li²,

Woo³

2004

Dis. Aquat. Org.

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The effects of infective Vibrio alginolyticus and its extracellular product (ECP) on host fish function are not well understood. In this study a partial biochemical characterization of the ECP from an infective strain of V. alginolyticus isolated from diseased silver sea bream Sparus sarba was achieved and the effects of live V. alginolyticus and ECP on hepatic heat shock protein (hsp) expression was compared. The ECP fraction was found to contain several hydrolytic enzymes including both haemolytic and proteolytic activities. Intramuscular administration of ECP to sea bream resulted in vibriosis with similar pathological signs as those observed with live V. alginolyticus administration. Using quantitative immunoassays we assessed the levels of the major hsp families, hsp90, hsp70 and hsp60, in hepatic tissue of diseased sea bream between 12 and 48 h post-infection. Throughout the infective period, live V. alginolyticus did not alter hsp90 whereas ECP significantly reduced hepatic hsp90 during the late stages of acute infection. The levels of hsp70 were found to be rapidly and drastically increased with both live V. alginolyticus and ECP. The transcript levels of both gene members of the hsp70 family (hsc70 and hsp70) were significantly increased with both live V. alginolyticus and ECP. The levels of hsp60 remained unchanged with both live V. alginolyticus and ECP. The data presented in this study is the first report describing an effect of both live V. alginolytus and ECP on hsp expression in diseased fish.KEY WORDS: Fish · Disease · Vibrio · Heat shock protein · hsp90 · hsp70 · hsp60 Resale or republication not permitted without written consent of the publisherDis Aquat Org 62: [205][206][207][208][209][210][211][212][213][214][215] 2004 When normal cellular processes are adversely affected the synthesis of a group of proteins belonging to the heat shock protein (hsp) families (hsp90, hsp70 and hsp60) are rapidly increased. Members of these different hsp families play different roles in the cell; for example, the hsp90 family is involved in steroid receptor formation and protein folding (Smith & Toft 1993, Pratt 1997, the hsp70 family is necessary for protein synthesis, translocation and protein folding (Gething & Sambrook 1992), and mitochon-drial bound hsp60 is defined as being involved in protein stability and folding (Cheng et al. 1989, Ostermann et al. 1989, Martin et al. 1992. Indeed all of these groups of hsps have been demonstrated to be upregulated in fish during abiotic stress (Iwama et al. 1998). Evidence from mammalian studies has suggested that hsps are important during disease as they may play critical roles in immune function and protection (Jacquier-Sarlin et al. 1994), antigen presentation (DeNagel & Pierce 1993) and non-specific immune responses (Guzik et al. 1999). Although strong evidence exists for an important role of hsps in disease responses for mammals, much less is known about pathogenic stress and its effect on hsp90, 70 and 60 expression in fish. A better understanding...

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“…Cytotoxicity assays were basically carried out as described by Wang et al (1998) on mammalian and fish cell lines. Two different mammalian cell lines were employed: NIH/3T3, a murine fibroblastic cell line, and HT-1080, a human epithelial cell line.…”

Section: Methodsmentioning

confidence: 99%

Toxicity of Photobacterium damselae subsp. damselae strains isolated from new cultured marine fish

Labella¹,

Sanchez-Montes²,

Berbel³

et al. 2010

Dis. Aquat. Org.

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The in vivo and in vitro toxicity of bacterial cells and their extracellular products (ECPs) from 16 strains of Photobacterium damselae subsp. damselae isolated from 7 epizootic outbreaks were evaluated. On the basis of their 50% lethal dose (LD 50 ) values (about 1 × 10 5 CFU), these strains may be considered as moderately virulent. However, their ECPs were strongly lethal for redbanded seabream Pagrus auriga causing fish death within 2 h post-inoculation (protein concentration ranged between 2.1 and 6.41 µg g -1 fish). The bacterial ECPs tested exhibited several enzymatic activities, such as amylase, lipase, phospholipase, alkaline phosphatase, esterase-lipase, acid phosphatase, and β-glucosaminidase. These ECPs displayed a strong cytotoxic effect on 4 fish and 2 mammalian cell lines, although this activity disappeared when ECPs were heated at 100°C. The virulence of the strains tested could not be related to the hemolytic activity or to the production of the toxin damselysin. Therefore, another unknown type of toxin could play an important role in the virulence mechanisms of this bacterial pathogen.KEY WORDS: Toxicity · ECP · Photobacterium damselae subsp. damselae · Cultured marine fish Resale or republication not permitted without written consent of the publisherDis Aquat Org 92: [31][32][33][34][35][36][37][38][39][40] 2010 brane, has been described (Kreger 1984, Kothary & Kreger 1985, Kreger et al. 1987. In addition, a relationship between the degree of virulence and the hemolytic activity has been demonstrated in P. damselae subsp. damselae strains isolated from fish (Fouz et al. 1993). However, Cutter & Kreger (1990) found that not all the P. damselae subsp. damselae strains presented the damselysin gene (dly), but only those strains showing intense hemolytic activity. A further study demonstrated that the presence of this gene was not correlated to the virulence in mice and fish of 17 P. damselae subsp. damselae strains isolated from different sources (Osorio et al. 2000).The extracellular products (ECPs) are produced by bacterial pathogens to facilitate the uptake of nutrients from the surrounding environment, and/or for the successful penetration and survival of pathogens inside the host (Bakopoulos et al. 2003). However, the role of ECPs in the pathogenesis of Photobacterium damselae subsp. damselae in fish is poorly known, which is a considerable disadvantage for the development of vaccines and vaccine strategies, since it has been suggested that the ECP components are major antigenic compounds of several vaccine formulations (Collado et al. 2000, Bakopoulos et al. 2004.The aim of this study was to determine the toxicity of different Photobacterium damselae subsp. damselae strains isolated from cultured diseased fish. For this purpose, we performed in vivo and in vitro assays using bacterial cultures and their ECPs. In addition, the enzymatic activities of the ECPs and their cytotoxicity in fish and mammalian cell lines were compared. MATERIALS AND METHODSBacterial strains. In this...

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Internalization and cytotoxicity are important virulence mechanisms in vibrio-f ishepithelial cell interactions

Cited by 46 publications

References 47 publications

NovelAeromonas hydrophilaPPD134/91 Genes Involved in O-Antigen and Capsule Biosynthesis

NovelAeromonas hydrophilaPPD134/91 Genes Involved in O-Antigen and Capsule Biosynthesis

Modulated heat shock protein expression during pathogenic Vibrio alginolyticus stress of sea bream

Toxicity of Photobacterium damselae subsp. damselae strains isolated from new cultured marine fish

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