The Isolation And Identification Of Vibrio Parahaemolyticus In Aquatic Products, And Drug-resistant Test Of The Part Strains

Vibrio parahaemolyticus (Vibrio parahaemolyticus, called V.p), also known as halophilic bacteria belonging to Vibrionaceae, Vibrio genus.At present, Vibrio parahaemolyticus food poisoning caused by bacterial food poisoning in the proportion of the higher, its harm degree is only inferior to salmonella, E. coli, Staphylococcus aureus and Clostridium botulinum,the bacteria often exist in the near shore waters ,the seabed sediment, seafood and pickled foods,the main cause of food poisoning and acute diarrhea,especially in Japan, Southeast Asia, the United States and China's in the Taipei area more common, but also our country's mainland coastal area food poisoning in the most common pathogenic bacteria.Along with the improvement of living standards,The world pay more attention to the safety of aquatic products, especially in recent years, due to popular seafood in the mainland cities, the inland aquatic product security incident also repeatedly occurrence,hazardous are becoming increasingly serious. This study collected Vibrio parahaemolyticus- positive patients suspected of 30 copies of materials, A total of 30 suspected strains isolated,As well as suspected strains isolated from the conventional morphological observation, biochemical identification, serotyping, tolerance test and the local strain of experimental drug susceptibility and carried out amplification of 16S rRNA sequencing to explore the molecular level. With a view to further study the drug-resistant Vibrio parahaemolyticus,Vibrio parahaemolyticus situation and clinical treatment of infections of public health significance,and provide a reference,while the production of reality prevention and treatment of Vibrio parahaemolyticus has guiding significance.The main research topic content:(A) Collection of Vibrio parahaemolyticus-positive patients suspected of 30 copies of materials: a small sample of 16 lobsters, fish samples were 10, 4 copies of shellfish samples (these samples come from different seafood markets in Zhengzhou region derived from random sampling). By bacteria prior to training, and use TCBS medium and Kema Jia Vibrio selective chromogenic medium for isolation and culture, a total of 30 suspected isolated strains.Suspected of 30 strains of morphological features, culture characteristics such as observation and identification of bacteria using automatic analyzer (VITEK-32) for biochemical identification, a preliminary determination were 30 suspected bacteria Vibrio parahaemolyticus.(B) bacterial lipopolysaccharide O antigen and capsular K antigen-based study found that any bacteria are present a variety of serotypes.The same kind of bacteria exist more extensive serum homology, serotyping of bacteria not only have a certain Sero-epidemiological significance, and in the right bacterial diseases diagnosis and prevention of specific immunity also has a certain reference value. In order to detect isolates of serotypes and antibiotic drug sensitivity, reference GB/T 4789 17-2003 with the standard serum for Vibrio parahaemolyticus serological test,The results indicate that the test 30 suspected Vibrio parahaemolyticus main points belonging to two serogroups, O1 group were 60% (18/30), 05 group accounted for 40% (12/30). The 30 test strains is Divided into 6 serotypes: O1: K3, O1: K4, O1: K6, O1: K7, O5: K9, O5: K4.(C) 16S rRNA gene sequence analysis has become a bacterial species identification and classification of the standard method, this study was one of five representative strains (small lobster disease material isolated 2, fish disease material isolated from 2 strains, shellfish -like illness material isolated from a), which conducted a 16S rRNA gene sequence analysis, design the upstream primer (5'-GACTCAACACAAGAAGAGATGC-3'), downstream primer (5'-ATGCGTGACATCCCAGAAC-3'), according to bacterial phylogeny Research reference to relevant literature, using DNA-Star in the MEGA version software ,relying on Kimura2-Pameter Distance analysis methods have been published on a suspected strains and strains analyzed and compared with the Neighbor-Joining build iaolates phylogenetic tree showed that the measured sequence and has been published in the standard strains of Vibrio parahaemolyticus 16S rRNA sequences of up to 99% similarity. Contrast to Vibrio-related strains of 16S rRNA sequences and isolates of the phylogenetic tree analysis show that the evolution of characteristics of isolates with the published sequence of Vibrio parahaemolyticus strains closest to the final to determine the 5 vice hemolytic bacteria Vibrio.(D) drug-min paper method of ceftazidime, ampicillin, gentamicin, polymyxin, levofloxacin, cefazolin, cefuroxime sodium, streptomycin, different serotypes 6 strain Vibrio parahaemolyticus susceptibility test. From the results we can see 6 strains of resistance to essentially the same, the vast majority are sensitive to ampicillin, gentamicin, ceftazidime, streptomycin sensitive; but on polymyxin, levofloxacin, Cefazolin, cefuroxime sodium has the emergence of drug-resistant strains, especially polymyxin antibiotic resistance rate of 83.3%.