高浓度柴油降解菌的筛选、鉴定及降解特性研究

从某石油化工退役场地石油烃污染土壤中筛选得到一株高效柴油降解菌株,经过形态学观察和16S rRNA基因序列分析,鉴定该菌株属于鲍特氏菌属(Bordetella),命名其为Bordetella sp.T3(简称T3)。T3适宜在中温、弱碱性的环境条件下生长,在柴油体积分数为1%、接种量(φ)为2%、温度为30℃、pH为8.0的条件下降解柴油,第7天可达89.79%的降解率。T3能够耐受柴油体积分数为10%的高浓度环境,在最适培养条件下培养7 d,其对柴油的降解率为38.57%。降解前后柴油的气相色谱对比结果表明,T3可以有效降解柴油的主要组分(C_(10)~C_(22)),具有良好的石油烃污染物修复应用潜力。

标准菌株CMCC(B)40001的分类学研究

通过形态学观察、16S rRNA序列分析、MALDI-TOF质谱分析、VITEK 2 COMPACT全自动微生物分析、API试剂条方法,对GB 4789.28-2013《食品安全国家标准食品微生物学检验培养基和试剂的质量要求》中指定使用的培养基质控菌株CMCC(B)40001进行多相复核鉴定,确定其分类学地位。结果表明:CMCC(B)40001为支气管炎鲍特氏菌,与GB 4789.28-2013收录的名称粪产碱杆菌不一致。

Biological Characteristics and Pathogenicities of Shewanella algae and Shewanella abalone from Babylonia

[Objective] The biological characteristics and pathogenicities of Shewanella algae and Shewanella abalone from Babylonia were studied in this paper. [Method]The hemolytic bacteria were isolated from the hepatopancreas of Babylonia suffered from proboscis edema with blood agar plate. The dominant bacterial community in the ill Babylonia was identified by 16 S r DNA sequence analysis, and the bacterial cultural and biochemical characteristics and pathogenicities were studied. [Result]The Shewanella bacteria, including Shewanella algae and Shewanella abalone, are the dominant bacterial community in Babylonia suffered from proboscis edema.The colony characteristics of Shewanella algae in nutrient agar medium, TCBS agar medium and CHROMagar vibrio colored medium were similar to those of Shewanella abalone. Shewanella algae possessed β-hemolysis and Shewanella abalone possessed α-hemolysis in the blood agar plate. The biochemical reaction of Shewanella algae and Shewanella abalone was all of non-fermentation type. The results of artificial infection test showed that half lethal dose（LD50） of the test strains of Shewanella algae was 10-5.50/0.1 ml. The test strains of Shewanella algae have strong toxicity, and could cause mice and chickens to die of sepsis with mortality of100%. The mortality of Babylonia infected with Shewanella algae was 10%; while the survived Babylonia lost the ability of moving and intaking for a long time, but they were not suffered from proboscis edema. There was no death in mice or chicks infected with Shewanella abalone, but their livers and spleens were slightly hyperemic and swelling. There was also no death in Babylonia infected with Shewanella abalone, but their intaking and moving ability was lost for a short time.[Conclusion] Although Shewanella algae and Shewanella abalone were the dominant bacteria in Babylonia suffered from proboscis edema, they were not the main pathogenic bacteria for proboscis edema. Shewanella algae had strong pathogenicity to mice, chicks and Babylonia, while Shewanella abalone showed no marked pathogenicity to those experimental animals in this study.

新生儿百日咳临床特点和致病菌耐药性分析

目的：总结新生儿百日咳的临床特点和致病株的耐药性。方法分析7例新生儿百日咳患儿的临床特点，采用Etest方法和纸片扩散法检测分离株对红霉素等抗生素的敏感性。结果7例患儿中，6例患儿的母亲或祖辈有持续10d以上的咳嗽，其中4例患儿的家庭成员中，≥3人咳嗽。患儿早期均有鼻塞、轻咳症状，5例在病程4-7d出现典型痉挛性咳嗽；5例口唇青紫，4例有呼吸暂停，3例有屏气发作，仅2例有发热表现。可见程度不一的鼻扇、三凹征等呼吸困难表现。分离株中5株在红霉素纸片周围没有形成抑菌环，Etest检查显示红霉素、阿奇霉素、克拉霉素和克林霉素对其的最小抑菌浓度（MIC）均〉256mg/L。结论对于有呼吸道症状，且曾与呼吸道感染患者密切接触的新生儿应警惕百日咳的可能，应详问流行病学史，并及时进行病原学检查；大环内酯类抗生素耐药菌株在百日咳病原菌中已很常见。

百日咳鲍特菌铁离子相关分泌蛋白研究

目的探讨儿童呼吸道致病菌之一百日咳鲍特菌不同铁离子培养条件下分泌蛋白质组的差异以及与其致病的关系。方法百日咳鲍特菌分别在加铁、加铁螯合剂和加铁加镁的包-姜氏培养基中培养,于对数生长期(培养48h)收集细菌,分别用三氯乙酸-丙酮沉淀法制备分泌蛋白,制备的分泌蛋白做MALDI-TOF MS质谱分析,并进行分泌蛋白的细菌亚细胞定位和分子信号肽预测。结果加铁培养从47h开始细菌生长进入平台期,加铁离子螯合剂组细菌随铁离子螯合剂的增加而趋于不生长状态,加铁加镁组细菌在培养至47h时缓慢生长;百日咳鲍特菌在3种培养条件下的蛋白亚细胞定位分别为:加铁组胞质蛋白326个,内膜蛋白61个,分泌蛋白3个,外膜蛋白13个,周质蛋白33个,未知蛋白201个;加铁螯合剂组胞质蛋白466个,外膜蛋白21个,周质蛋白63个,内膜蛋白66个,分泌蛋白3个,未知蛋白160个;加铁加镁组胞质蛋白601个,内膜蛋白97个,分泌蛋白5个,外膜蛋白26个,周质蛋白7个,未知蛋白185个。结论通过亚细胞定位和分子信号肽预测,锁定百日咳鲍特菌6个分泌蛋白,其中3个分泌蛋白核糖核苷二磷酸激酶(NDPK)、假定输出蛋白和血清三型菌毛亚单位是本研究首次发现的百日咳鲍特菌分泌蛋白。