中国水产科学
中國水產科學
중국수산과학
Journal of Fishery Sciences of China
2013年
5期
1023-1031
,共9页
林桂芳%鄢庆枇%陈文博%苏永全%覃映雪
林桂芳%鄢慶枇%陳文博%囌永全%覃映雪
림계방%언경비%진문박%소영전%담영설
嗜水气单胞菌%黏附%间接ELISA%鳗鲡%表皮黏液
嗜水氣單胞菌%黏附%間接ELISA%鰻鱺%錶皮黏液
기수기단포균%점부%간접ELISA%만려%표피점액
Aeromonas hydrophila%adhesion%indirect ELISA%Anguilla anguilla%skin mucus
采用间接 ELISA 法研究菌浓度、孵育时间、温度、pH、阳离子及碳源等因子对嗜水气单胞菌(Aeromonas hydrophila)黏附鳗鲡(Anguilla anguilla)表皮黏液的影响。结果表明,改良后的间接 ELISA 法的检测灵敏度约为9.92×104 CFU,细菌的黏附量随菌浓度的升高而逐渐增大并符合饱和黏附动力学方程: y=0.135ln(x)-0.936(R2=0.986);嗜水气单胞菌黏附鳗鲡表皮黏液的最佳条件为:温度20~28℃, pH 6.2~6.6, NaCl、MgCl2质量浓度分别为15~25 g/L和3 g/L,孵育时间为150 min。碳源对嗜水气单胞菌的黏附作用有不同程度的影响,葡萄糖和麦芽糖能显著提高嗜水气单胞菌的黏附量(P<0.05),果糖则显著降低嗜水气单胞菌的黏附量(P<0.05)。以上结果说明,嗜水气单胞菌对鳗鲡表皮黏液具有较强的黏附作用,且其黏附作用具有可控性。
採用間接 ELISA 法研究菌濃度、孵育時間、溫度、pH、暘離子及碳源等因子對嗜水氣單胞菌(Aeromonas hydrophila)黏附鰻鱺(Anguilla anguilla)錶皮黏液的影響。結果錶明,改良後的間接 ELISA 法的檢測靈敏度約為9.92×104 CFU,細菌的黏附量隨菌濃度的升高而逐漸增大併符閤飽和黏附動力學方程: y=0.135ln(x)-0.936(R2=0.986);嗜水氣單胞菌黏附鰻鱺錶皮黏液的最佳條件為:溫度20~28℃, pH 6.2~6.6, NaCl、MgCl2質量濃度分彆為15~25 g/L和3 g/L,孵育時間為150 min。碳源對嗜水氣單胞菌的黏附作用有不同程度的影響,葡萄糖和麥芽糖能顯著提高嗜水氣單胞菌的黏附量(P<0.05),果糖則顯著降低嗜水氣單胞菌的黏附量(P<0.05)。以上結果說明,嗜水氣單胞菌對鰻鱺錶皮黏液具有較彊的黏附作用,且其黏附作用具有可控性。
채용간접 ELISA 법연구균농도、부육시간、온도、pH、양리자급탄원등인자대기수기단포균(Aeromonas hydrophila)점부만려(Anguilla anguilla)표피점액적영향。결과표명,개량후적간접 ELISA 법적검측령민도약위9.92×104 CFU,세균적점부량수균농도적승고이축점증대병부합포화점부동역학방정: y=0.135ln(x)-0.936(R2=0.986);기수기단포균점부만려표피점액적최가조건위:온도20~28℃, pH 6.2~6.6, NaCl、MgCl2질량농도분별위15~25 g/L화3 g/L,부육시간위150 min。탄원대기수기단포균적점부작용유불동정도적영향,포도당화맥아당능현저제고기수기단포균적점부량(P<0.05),과당칙현저강저기수기단포균적점부량(P<0.05)。이상결과설명,기수기단포균대만려표피점액구유교강적점부작용,차기점부작용구유가공성。
Adhesion to the host mucus, which is closely related to the virulence of pathogenic bacteria, is the first crucial step in most infections. In this paper, the adhesion characteristics of Aeromonas hydrophila to eel(Anguilla anguilla) skin mucus was investigated by indirect ELISA. First, we confirmed that OD492 can be used as an index to reflect the number of adhering bacteria. Second, we confirmed that the indirect ELISA method can detect bac-teria with a sensitivity of 9.92×104CFU. The number of adhering A. hydrophila on eel mucus rose with increased bacterial density;a saturation kinetics equation y=0.135 n(x)?0.936(R2=0.986) was obtained by plotting the number of adhering bacteria against cell density. The factors affecting bacterial adhesion were then investigated. The re-sults revealed that the number of adhering A. hydrophila increased with incubation time and reached a peak after 150 min. Temperature was one of the major factors affecting bacterial adhesion, with peak adhesion occurring at 20?28℃. pH also influenced bacterial adhesion. In a weak acid with a pH of 6.2?6.6, the adhesion ability of A. hydrophila was remarkably enhanced. The optimum concentrations of Na+and Mg2+ for A. hydrophila adhesion were 1.5?2.5 g/L and 3 g/L, respectively. In summary, the optimum conditions for the adhesion of A. hydrophila to eel skin mucuswere 20?28℃, pH 6.2?6.6, 1.5?2.5 g/L NaCl, 0.3 g/L MgCl2, and 150 min incubation time. In ad-dition, we found that bacterial adhesion was influenced by carbohydrates. Seven kinds of carbohydrates were investigated and the results indicate that glucose and maltose significantly enhance A. hydrophila adhesion (P<0.05), while fructose inhibited it (P<0.05). Understanding the mechanisms of A. hydrophila adhesion will help to further elucidate bacterial pathogenicity and help control fish disease.