Isolation and identification of Micrococcus luteusin rice field eel(Monopterus albus) in Sichuan Province
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    Abstract:

    This experiment was conducted to analyse the species and drug resistance of hemorrhagic pathogens from Monopterus albus. One hemorrhagic disease pathogens(HM1) of M. albus from Mingshan in Sichuan was isolated using bacteria classification and identification technique, and was identified according to the morphological, physiological and biological characteristics and the phylogenetic analysis. On pathogenicity test of animals, the experimental Ictalurus nebulosus demonstrated weak pathogenic symptom, but not any pahthogenic symptom in mice and Misgurnus anguillicaudatus. Morphological and physiological biochemical character homophyly of HM1 strain was in accordance with Micrococcus luteus, which presented gram positive coccus, nonmotility, arranged in couples or quadruple or clumps, nonchainlike conformation, nonsporulation; catalasepositive, gelatin liquefactionpositive, lactosenegative, mannitolnegative, glucosenegative, aesculin hydrolysisnegative, nitrate reductionnegative, arginine hydrolytic enzymenegative and so on. The 16S rDNA gene of HM1 was amplified by PCR using the universal primers. Then, the 16S rDNA gene was cloned and sequenced. A sequence of 1 382 base pair (bp) was obtained from HM1 (GenBank accession number: HM044913). The analysis using online Classifer software form the Ribosomal Database Project (RDP) database showed that the strain HM1 was classified in Micrococcus. The nucleotide sequences homology of 16S rDNA from HM1 strain had 99% identity with 7 strains M. luteus in GenBank. Phylogenetic trees reconstructed based on genes 16S rDNA indicate that the strains of HM1 and other M. luteus in GenBank are clustered in the same clade. The results suggest that the isolated strain (HM1) was classified in M. luteus. Antibiotic susceptibility tests showed that the isolated strain (HM1) was sensitive to 18 kinds of drugs inculding norfloxacin, cefazolin sodium and carbenicillin, etc; and was medium sensitive to ceftazidime; and insusceptible to 5 kind of drugs such as nalidixic acid, gatifloxacin and sulfafurazole, etc.

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彭 彬,杨光友,陈晓利,占爱思,何明莉.黄鳝藤黄微球菌的分离鉴定及药敏试验[J].上海海洋大学学报,2011,20(3):405-411.
PENG Bin,YANG Guang-you, CHEN Xiao-li, ZHAN Ai-si, HE Ming-li. Isolation and identification of Micrococcus luteusin rice field eel(Monopterus albus) in Sichuan Province[J]. Journal of Shanghai Ocean University,2011,20(3):405-411.

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