海参、海胆养殖水体中多抗性细菌抗性基因的初步研究

抗药性细菌对生态环境、养殖业以及人类健康构成严重威胁。以大连海参、海胆养殖场多抗性细菌筛选为基础,采用分子生物学手段对40株抗性细菌进行了氯霉素、土霉素及氟氯霉素抗性基因检测,探讨抗药性分子遗传机理,从而为控制抗药性细菌的产生和传播提供研究基础。     

发光细菌法检测水产品中氯霉素体系的建立

从青岛近海分离的多株发光细菌中筛选出1株对氯霉素敏感的鳆发光杆菌Photobacterium leiognathi YL,接种单菌落快速培养,制备出适宜检测的菌悬液,添加不同浓度的氯霉素溶液,研究氯霉素浓度与细菌发光强度抑制率之间的关系.通过控制菌体起始发光强度、菌液与氯霉素作用时间,建立发光细菌检测水产品中氯霉素体系.结果表明,当菌体的起始发光强度控制在(2.0～4.0)×105、菌液与氯霉素的最佳作用时间为30 min时,氯霉素浓度与细菌发光强度抑制率呈良好的线性关系.该方法的线性范围为0.1～1.0 ng/mL,相关系数R2为0.989,该体系对氯霉素的检测灵敏度可以达到0.1 ng/mL,可以作为水产品中痕量氯霉素残留的1种快速、灵敏的检测方法.在水产品的氯霉素加标回收试验中,鱼肉中添加浓度为0.1～1.0 ng/g时,回收率在40.34%～114.26%之间.     

Acclimatization and screening of a chloramphenicol-degrading strain

Chloramphenicol （CAP） is an antibiotic commonly used in aquaculture as a prophylactic or disinfectant to prevent diseases, or as a chemotherapeutic agent to control diseases. This antibiotic is either spread directly in the aquatic environment or administered through medicated feeds. It may be directly exposed to or enter the environment by leaching from uneaten feeds or from the aquatic animals＇ excrements. The antibiotic also directly enters the environment through pharmaceutical wastewater. Toxicity of CAP to human bone marrow has been linked to fatal aplastic anemia. Antimicrobial residues enter the environment, which may also establish a selective pressure in favour of antimicrobial-resistant bacteria. The residues of antibiotics may pose potential risk to public health or increase the occurrence of antibiotic-resistant bacteria in the aquatic environment. Due to these perceived risk, treatment of CAP in farm animals intended for human consumption has been restricted. However, residue of CAP in the environment is still a concern. There have been some reports on the residual effects of CAP on aquaculture and on microbial resistance to CAP in certain environments. The biotransformation of CAP in aquaculture pond sediments has been reported, which indicated that salinity is one of the many environmental factors that affect the degradation of CAP. However, acclimatization and screening of dominant bacteria for biodegrading CAP have been rarely studied. This paper studied the microbial degradation of CAP. In order to screen the dominant bacteria for degrading, the source of bacteria was sampled from the aquaculture pond sediments of Baiyun Borough, Guangzhou, China. 60 mg/L CAP was used as a sole carbon source to screen CAP-degrading bacteria and the dominant bacteria were acclimated after 2 months.     

氯霉素单克隆抗体的研制及其特性分析

为了研制氯霉素单克隆抗体,建立氯霉素的简便有效的检测方法.采用碳二亚胺(EDC)方法制备氯霉素免疫抗原和包被抗原,经SDS-PAGE凝胶电泳,三硝基苯磺酸(TNBS)法鉴定表明偶联成功.采用制备的抗原CAP-HS-BSA((CAP-HS,氯霉素琥珀酸酯;BSA,牛血清白蛋白)免疫Balb/c小鼠,通过杂交瘤技术获得了2株抗氯霉素的杂交瘤细胞株,细胞株体外传代和冻存复苏后抗体分泌稳定.经体内诱生法产生腹水,ELISA检测纯化腹水的效价为1:10~6.抗体的亲和常数分别为:1.13×10~(10) L/mol,1.41×10~(10) L/mol.ELISA检测显示这2株单克隆抗体与氯霉素琥珀酸交叉反应率为300%,与其他抗生素及结构类似物的交叉反应小,表明该单抗可满足建立免疫学检测方法的需要和开发应用的要求.     

土霉素、氯霉素对湛江等鞭金藻细胞增殖和光合色素含量的影响

在室温下研究了不同浓度的土霉素和氯霉素对湛江等鞭金藻的细胞数量、个体大小 ,以及叶绿素、类胡萝卜素等的含量的影响。结果表明 :土霉素在 0～ 10mg/L范围内浓度越大 ,金藻细胞数量在增长期内增幅越小 在衰退期内衰退越快 ,叶绿素a越少 ,类胡萝卜素越多 ;氯霉素在 0～ 5mg/L范围内浓度越大 ,金藻细胞数量在增长中后期增幅越小且低于正常水平 ,衰退期内衰退越快 ,叶绿素类越少 ,类胡萝卜素越多。土霉素、氯霉素对金藻细胞个体大小的影响均不明显。     

条斑紫菜叶状体细胞的抗生素敏感性研究

为了进行条斑紫菜基因工程研究和培育转基因紫菜 ,首先必须确定适合的阳性选择标记基因。条斑紫菜对 8种抗生素的敏感性试验表明 ,它对各种抗生素的敏感性不同 ,其中 zeocin、腐草霉素和氯霉素半致死剂量都很低。因此 Cat(氯霉素乙酰转移酶 )基因和 Sh.ble(sh.bleomycin)基因均有望成为条斑紫菜基因工程研究中理想的选择标记基因。