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Molecular Identification and Physiological Characteristics of Vibrio Alginolyticus-related Isolates

Author: GuoJing
Tutor: ZhangDeMin
School: Ningbo University
Course: Marine biology
Keywords: Vibrio alignolyticu 16S rRNA gene molecular identification physiological ecology
CLC: S943
Type: Master's thesis
Year: 2012
Downloads: 89
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Abstract


Vibrio is a large group of gram negative bacteria with short bending shape. They are widely existing inembouchure, bay, sea and animals,some species of them can cause Vibriosis. Identification Vibrios is soimportane to prevent Vibriosis. Vibrio alginolyticus´╝îa member of the core group of the genus Vibrio, alsocalled harveyi group, share high phenotypic and genotypic resemblance with the other species of this group.It is very difficult to absolutely identify novel strains to V. alginolyticus. In this study, we compared theresolution of16S rRNA gene, toxR gene, pyrH gene and rpoB gene on V. alginolyticus-related isolates.There are multiple copies of16S rRNA gene in the genome of Vibrio and the intragenomic heterogeneityamong different copies might be higher than intergenomic heterogeneity in some strains. Our study haveproved it with16S rRNA sequense analysis,16S rRNA gene of Vibrio have more PCR-DGGE bands andclone16S rRNA gene of strain418shows6different sequence.All these facts make the16S rRNA gene notsutible for discrimination of harveyi group species.Both of single-copy gene toxR and pyrH show a considerable gap between the maximal interspeciessimilarity and the minimal intraspecies similarity. The toxR gene similarity analyses of eighteen isolatesand five reference strains can clearly group them into four species. The phylogenetic tree based on toxRgene shows that strains of individual species clustered together independently, and there are two differentclusters among V. alginolyticus, indicating that they have different evolving path. The similarity andphylogenetic analyses based on pyrH gene come to the same result of toxR gene. As pyrH gene is moreconservative, its discriminatory power is sightly lower than that of toxR gene. The phylogenetic tree basedon rpoB gene shows mistakes in identification V. alginolyticus and lowest discriminatory power, also thedatabase does not include the type strain sequences of V. campbellii, V. rotiferianus, V. natriegens in thisgroup. We suggested that toxR gene can be the first method for fast identification of V. alginolyticus isolates,and pyrH gene may used for verification when necessary.The physiological and biochemical reactions between Vibrio alginolyticus and V. parahaemolyticus aremostly the same, mainly different are can growing at10%NaCl and using sucrose. But there are some V.parahaemolyticus isolates can growing slowly at10%NaCl and one of Vibrio alginolyticus isolate can notusing sucrose, this made biochemical identification confused and need more molecular method. The isolatealso have denitrification capabilities, which is one of reasons deterioration of water quality. The capabilityof producing extracellular enzyme is related with enviroment factors, and the Vibrios isolated fromaqucalture water or diseased crab are more able to producing extracellular enzyme than these from seawater.

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