Characterization of Aeromonas sobria hemolysin by use of monoclonal antibodies against Aeromonas hydrophila hemolysins

Kozaki, Shunji; Asao, Tsutomu; Kamata, Y; Sakaguchi, Genji

doi:10.1128/jcm.27.8.1782-1786.1989

Cited by 31 publications

(16 citation statements)

References 24 publications

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“…These results correlated with data from studies of A. sobria and A. hydrophila hemolysins, which also required trypsinization before the hemolysin could be activated (Kozaki et al, 1989). To demonstrate that this hemolytic activity was not associated with Act, the culture supernatant of wild-type A. hydrophila SSU strain as well as from the act-minus mutant was subjected to neutralization with Act-specific antibodies.…”

Section: Hemolytic Activity and Neutralization Testsupporting

confidence: 75%

“…The HlyA protein did not exhibit any homology with other hemolysins or the aerolysin and act genes that were cloned and characterized earlier from several Aeromonas spp. (Kozaki et al, 1989;Wong et al, 1998;Wang et al, 2003;Yu et al, 2005;Galindo et al, 2006b). However, the hlyA gene is present on the genome of A. hydrophila ATCC 7966 strain that was recently sequenced (GenBank accession number CP000462) (Seshadri et al, 2006).…”

Section: Resultsmentioning

confidence: 99%

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Identification of a new hemolysin from diarrheal isolate SSU ofAeromonas hydrophila

Erova

Sha

Horneman

et al. 2007

FEMS Microbiology Letters

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A clinical strain SSU of Aeromonas hydrophila produces a potent cytotoxic enterotoxin (Act) with cytotoxic, enterotoxic, and hemolytic activities. A new gene, which encoded a hemolysin of 439-amino acid residues with a molecular mass of 49 kDa, was identified. This hemolysin (HlyA) was detected based on the observation that the act gene minus mutant of A. hydrophila SSU still had residual hemolytic activity. The new hemolysin gene (hlyA) was cloned, sequenced, and overexpressed in Escherichia coli. The hlyA gene exhibited 96% identity with its homolog found in a recently annotated genome sequence of an environmental isolate, namely the type strain ATCC 7966 of A. hydrophila subspecies hydrophila. The hlyA gene did not exhibit any homology with other known hemolysins and aerolysin genes detected in Aeromonas isolates. However, this hemolysin exhibited significant homology with hemolysin of Vibrio vulnificus as well as with the cystathionine beta synthase domain protein of Shewanella oneidensis. The HlyA protein was activated only after treatment with trypsin and the resulting hemolytic activity was not neutralizable with antibodies to Act. The presence of the hlyA gene in clinical and water Aeromonas isolates was investigated and DNA fingerprint analysis was performed to demonstrate its possible role in Aeromonas virulence.

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Section: Hemolytic Activity and Neutralization Testsupporting

confidence: 75%

Section: Resultsmentioning

confidence: 99%

Identification of a new hemolysin from diarrheal isolate SSU ofAeromonas hydrophila

Erova

Sha

Horneman

et al. 2007

FEMS Microbiology Letters

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“…Kozaki et al reported that a monoclonal antibody against the hemolysin of A. hydrophila neutralizes the enterotoxic activity of the culture supernatant of A. sobria, indicating that the hemolysin of the latter also possesses enterotoxic activity (18). Subsequently, Millership et al examined the effects of A. sobria culture filtrates containing defined levels of hemolysin and CT immunoreactivity on water and ion flux in the gut by means of a modified rat jejunal perfusion assay.…”

mentioning

confidence: 99%

Purification and Characterization of Enterotoxin Produced by Aeromonas sobria

Fujii

Nomura

Kanzawa

et al. 1998

Microbiology and Immunology

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We purified the toxin of Aeromonas sobria capable of inducing a positive response in the mouse intestinal loop assay. The purified toxin showed a positive response not only in the loop assay but also in a hemolytic assay. Subsequently, we cloned the toxin gene and demonstrated that the product of this gene possessed both hemolytic and enterotoxic activities. These results showed that the enterotoxin of A. sobria possesses hemolytic activity. Nucleotide sequence determination of the toxin gene and amino acid sequence analysis of the purified toxin revealed that it is synthesized as a precursor composed of 488 amino acid residues, and that the 24 amino-terminal amino acid residues of the precursor is removed in the mature toxin. As antiserum against the purified toxin neutralized the fluid accumulation induced by living cells not only of A. sobria but also of A. hydrophila, this and antigenically related toxin(s) are thought to play an essential role in the induction of diarrhea by these organisms. The toxin-injured Chinese hamster ovary (CHO) cells induced the release of intracellular lactose dehydrogenase (LDH). The release of LDH from CHO cells and the lysis of erythrocytes by the toxin were repressed by the addition of dextran to the reaction solution, indicating that the toxin forms pores in the membranes and that the cells were injured by the osmotic gradient developed due to pore formation. However, the histopathological examination of intestinal cells exposed to the toxin showed that it caused fluid accumulation in the mouse intestinal loop without causing cellular damage.

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“…hydrophila. Kozaki et al (17) proved that hemolysin produced by A. sobria caused fluid accumulation in infant mouse intestine. More recently, A. schubertii was isolated from human wounds and characterized as causing primary cellulitis (6) .…”

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confidence: 99%

Distribution and Characterization of Hemolytic, and Enteropathogenic Motile Aeromonas in Aquatic Environment

Nakano

Kameyama

Venkateswaran

et al. 1990

Microbiology and Immunology

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A year-long survey on the distribution of motile Aeromonas species in the surface waters of riverine and marine environments was conducted. The filtered membranes were directly placed onto the modified Pril-xylose-ampicillin agar for the enumeration of Aeromonas species. High counts of motile aromonads were found in riverine stations and this bacterial population was also observed in significant quantities in polluted marine samples. In the identification of 2,444 isolates, three species of motile Aeromonas were observed. A. caviae (43%) was prevalent followed by A. sobria (35%) and A. hydrophila (20%). A. hydrophila was high in clean riverine samples, A. sobria was predominantly isolated from a stagnant water sampling area, and A. caviae was distributed more in marine samples. Statistical analyses suggested that the densities of Aeromonas were related to the cumulative effect of various physicochemical parameters rather than to a single factor. Among the species of Aeromonas, A. hydrophila, and A. sobria were highly hemolytic whereas only 11% of A. caviae were observed to lyse sheep erythrocytes. Suckling-mouse assay was performed to elucidate the enterotoxicity of motile aeromonads and 21% of the tested strains (one A. caviae strain) were found to produce enterotoxin.

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Characterization of Aeromonas sobria hemolysin by use of monoclonal antibodies against Aeromonas hydrophila hemolysins

Cited by 31 publications

References 24 publications

Identification of a new hemolysin from diarrheal isolate SSU ofAeromonas hydrophila

Identification of a new hemolysin from diarrheal isolate SSU ofAeromonas hydrophila

Purification and Characterization of Enterotoxin Produced by Aeromonas sobria

Distribution and Characterization of Hemolytic, and Enteropathogenic Motile Aeromonas in Aquatic Environment

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