山东地区驴源沙门氏菌的分离鉴定及其生物学特性比较分析

Isolation,identification and comparative analysis of biological characteristics of Salmonella of donkey origin in Shandong Province

为了解山东地区驴源沙门氏菌的流行情况及其生物学特性,试验将采自山东多地的14份驴流产病料及137份驴粪便样本分别接种于沙门氏菌显色培养基中对沙门氏菌进行分离培养,提取细菌基因组DNA后采用双重PCR方法判定分离菌是否为沙门氏菌并进一步判定是否为马流产沙门氏菌,并对分离到的沙门氏菌进行血清型鉴定、生化试验、细菌运动力和生物膜形成能力检测及药敏试验,选取部分菌株进行致病性试验,并采用双层平板法检测分离菌对9株噬菌体A18028、C59102、F59102、F18024、F18038、E17016、DS2、DS4、5FS4的敏感性。结果表明:从驴流产病料中共分离到14株马流产沙门氏菌(A1~A14株);从驴粪便样本中共分离到22株其他血清型沙门氏菌(F1~F22株),其中F1株为山夫顿堡沙门氏菌,F2、F3、F4、F5、F7、F8、F9、F10、F12、F13、F15株为肯塔基沙门氏菌,F6、F11株为Give沙门氏菌,F14株为乙型副伤寒Java型沙门氏菌,F16、F20株为阿哥纳沙门氏菌;F17株为鼠伤寒沙门氏菌,F18、F19株为汤卜逊沙门氏菌,F21株为阿伯丁沙门氏菌,F22株未鉴定出血清型;36株分离菌均符合沙门氏菌属的生化特性,但马流产沙门氏菌与其他血清型沙门氏菌生化特性略有差异;14株马流产沙门氏菌的平均运动力较22株其他血清型沙门氏菌弱;36株沙门氏菌均能形成生物膜;36株沙门氏菌对链霉素、庆大霉素、哌拉西林的耐药率较高,对亚胺培南和美罗培南的耐药率为0;小鼠接种A2株14 d后的死亡率为100%,接种F14株和F20株14 d后死亡率分别为50.0%和62.5%,接种F1、F2、F6、F18、F21、F22株14 d后无死亡情况;14株马流产沙门氏菌对9株噬菌体均敏感,而22株其他血清型沙门氏菌对9株噬菌体的敏感性不同,其中沙门氏菌F1、F6、F11株仅对1株噬菌体敏感,F18、F19、F20株对9株噬菌体均不敏感。说明不同沙门氏菌的生物学特性存在差异,马流产沙门氏菌对噬菌体具有很高的敏感性。

In order to understand the prevalence status and biological characteristics of Salmonella in donkey farms from Shandong Province,14 samples of donkey abortion and 137 samples of donkey feces were inoculated with Salmonella chromogenic medium for isolation and culture.After extracting genomic DNA of bacteria,double PCR method was used to determine whether the isolated bacteria were Salmonella and further determine whether the isolated bacteria were Salmonella abortus equi.Serotype identification,biochemical test,bacterial motility test,biofilm formation ability detection,and drug susceptibility test were carried out on the isolated Salmonella.Some strains were selected for pathogenicity test,and the sensitivity of the isolates(A18028,C59102,F59102,F18024,F18038,E17016,DS2,DS4,5FS4)was detected by double-layer plate method.The results showed that a total of 14 strains of Salmonella abortus equi(A1-A14)were isolated from the donkey abortion samples.A total of 22 strains of other serotypes of Salmonella(F1-F22)were isolated from donkey feces,of which F1 was Salmonella Senftenberg,F2,F3,F4,F5,F7,F8,F9,F10,F12,F13 and F15 were Salmonella Kentucky;F6 and F11 were Salmonella Give;F14 was Salmonella paratyphoid B var.Java;F16 and F20 were Salmonella argonauticus;F17 was Salmonella typhimurium;F18 and F19 were Salmonella thompson;F21 was Salmonella aberdeen.The serotype of F22 was not identified.The 36 isolates were consistent with the biochemical characteristics of Salmonella,but the biochemical characteristics of Salmonella abortus equi were slightly different from those of other serotypes.The average motility of 14 strains of Salmonella abortus equi were weaker than that of 22 strains of other serotypes of Salmonella.All 36 strains of Salmonella could form biofilm.The resistance rate of 36 Salmonella strains to streptomycin,gentamicin and piperacillin was high,and the resistance rate to imipenem and meropenem was 0.The mortality rate of mice inoculated with A2 strain was 100%after 14 days,and the mortality rate of mice inoculated with F14 strain and F20 strain was 50.0%and 62.5%after 14 days,respectively.No death occurred after inoculation with strains F1,F2,F6,F18,F21 and F22.The 14 strains of Salmonella abortus equi were sensitive to 9 phages,while the sensitivity of 22 strains of other serotypes of Salmonella was different.Salmonella F1,F3,F6 and F11 were only sensitive to 1 phage,while Salmonella F18,F19 and F20 were not sensitive to 9 phages.These results indicated that the biological characteristics of different Salmonella bacteria were different,and Salmonella equi abortus had a high sensitivity to phages.