Molecular identification of pathogenic and nonpathogenic strains of Vibrio harveyi using PCR and RAPD.
Fifteen environmental samples of Vibrio spp. isolated from healthy and diseased shrimps were tested for pathogenicity to juvenile shrimps. Two isolates, strains Z2 and Z3, were observed to be pathogenic, causing 100% mortality of the target host compared to the control strain Vibrio harveyi ATCC 14126. Environmental and type strains were subjected to molecular characterization by restriction fragment length polymorphism (RFLP) and PCR using primers targeted to different virulence, transcriptional regulator, or quorum sensing genes from V. harveyi. Primers designed for luxN were specific and identified all the environmental strains as V. harveyi. The random amplified polymorphic DNA (RAPD) method was used to differentiate between pathogenic and nonpathogenic strains of V. harveyi. These methodologies allowed us to detect and distinguish strains virulent and avirulent to juvenile shrimp.
Molecular Microbiology Laboratory, Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE), Department of Marine Biotechnology, Ensenada, B.C. México.
G
G
Hernández
J
J
Olmos
D000820:Animal Diseases / Q000382:microbiology; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D003445:Crustacea / Q000382:microbiology; D016172:DNA Fingerprinting / Q000379:methods*; D004269:DNA, Bacterial / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D011494:Protein Kinases / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D019105:Random Amplified Polymorphic DNA Technique; D014157:Transcription Factors / Q000235:genetics; D014733:Vibrio / Q000145:classification* / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification / Q000472:pathogenicity*; D014735:Vibrio Infections / Q000382:microbiology; D014774:Virulence; D014871:Water Microbiology
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D014157:Transcription Factors; D011494:Protein Kinases; C084290:luxN protein, Vibrio harveyi