两株不同血清型鱼源无乳链球菌生物学特性的比较研究
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摘要
为分析裂腹鱼源无乳链球菌(Streptococcus agalactiae)血清型Ia型和Ⅲ型的生物学特性的差异,本研究对该菌Ia型(S07)与Ⅲ型(S03)两株菌的生长曲线、耐药性、粘附和侵入能力、致病性,以及环境因子(pH、温度和盐度)对两株菌生长的影响等特性进行比较研究。结果显示,Ⅲ型S03株生长速度显著快于Ia型S07株。S03株生长受pH、温度和盐度的影响比S07株小。S03株和S07株在蔗糖、亮氨酸芳胺酶、β-溶血和VP试验等40种生化反应中结果一致,但S03株在葡萄糖、麦芽糖、α-葡萄糖苷酶和磷酸盐等11种生化反应中均呈阳性,而S07株呈阴性。S03株的耐药性明显强于S07株,且S03株表现出7重耐药,而S07株对所测抗生素均敏感或中度敏感。S07株黏附能力显著强于S03株。S07株对齐口裂腹鱼和斑马鱼的毒力(LD_(50)分别为8.9×10~3 cfu/mL和1.88×10~3cfu/mL)显著强于S03株(LD_(50)分别为5.27×10~8 cfu/mL和7.09×10~7 cfu/mL)。研究表明裂腹鱼源Ia型和Ⅲ型无乳链球菌在多种生物学特性上存在明显差异。本研究为阐明不同血清型无乳链球菌的致病机理及该菌引起的疾病的防治提供了实验依据。
To analyze the differences of the biological characteristics of serotype Ia strain S07 and serotype Ⅲ strain S03 of Streptococcus agalactiae from Schizothoraxs, we analyzed and compared diverse characteristics and capabilities of serotype Ia strain S07 and serotype Ⅲ strain S03, such as growth curve, adhesive capacity and invasion, antimicrobial resistance, pathogenicity and effects of environmental factors(pH curve, cultivating temperature curve and tolerance to salt). The result indicated that the growth rate of S03 was significantly faster than S07. S07 showed poor tolerance while S03 performed a relatively strong tolerance in different temperature, pH and salinity conditions. S03 and S07 were identical in the 40 kinds of biochemical test including sucrose, leucine arylamidase, beta-hemolysis and VP test, etc. But S03 was positive in 11 kinds of biochemical test including glucose, maltose, alpha-glucosidase and phosphate, etc. while S07 was negative.S03 showed resistant to seven kinds of antibiotics,however S07 was sensitive or intermediate to all antibiotics. The adhesion ability of S07 was much stronger than S03. In the pathogenicity test of Schizothorax prenanti and Danio rerio, S07 showed significantly more virulent with the LD_(50) reaching 8.9×10~3cfu/mL and 1.88×10~3 cfu/mL, compared to S03 with the LD_(50) reaching 5.27 ×10~8 cfu/mL and 7.09 ×10~7 cfu/mL, respectively. The results showed that there were significant differences in the biological characteristics between serotype Ia and serotype Ⅲ from Schizothoraxs, which provided a basis for the further study on the pathogenesis of S.agalactiae.
To analyze the differences of the biological characteristics of serotype Ia strain S07 and serotype Ⅲ strain S03 of Streptococcus agalactiae from Schizothoraxs, we analyzed and compared diverse characteristics and capabilities of serotype Ia strain S07 and serotype Ⅲ strain S03, such as growth curve, adhesive capacity and invasion, antimicrobial resistance, pathogenicity and effects of environmental factors(pH curve, cultivating temperature curve and tolerance to salt). The result indicated that the growth rate of S03 was significantly faster than S07. S07 showed poor tolerance while S03 performed a relatively strong tolerance in different temperature, pH and salinity conditions. S03 and S07 were identical in the 40 kinds of biochemical test including sucrose, leucine arylamidase, beta-hemolysis and VP test, etc. But S03 was positive in 11 kinds of biochemical test including glucose, maltose, alpha-glucosidase and phosphate, etc. while S07 was negative.S03 showed resistant to seven kinds of antibiotics,however S07 was sensitive or intermediate to all antibiotics. The adhesion ability of S07 was much stronger than S03. In the pathogenicity test of Schizothorax prenanti and Danio rerio, S07 showed significantly more virulent with the LD_(50) reaching 8.9×10~3cfu/mL and 1.88×10~3 cfu/mL, compared to S03 with the LD_(50) reaching 5.27 ×10~8 cfu/mL and 7.09 ×10~7 cfu/mL, respectively. The results showed that there were significant differences in the biological characteristics between serotype Ia and serotype Ⅲ from Schizothoraxs, which provided a basis for the further study on the pathogenesis of S.agalactiae.
引文
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[15]刘志刚,可小丽,卢迈新,等.温度对尼罗罗非鱼无乳链球菌毒力的影响[J].水产学报,2013,37(11):1733-1741.
[16] Xi Hong-bo, Yang Qi,Shang Hai-tao, et al. Biodegradation characteristics and enzymatic reaction kinetics of phenanthrenedegrading bacteria[J]. Earth Sci Front, 2008, 6(01):17-19.
[17]黄瑞,吴淑燕,闻玉梅.细菌接合性耐药质粒与毒力关系的研究[J].苏州大学学报:医学版,2000,20(1):13-17.
[2] Zhao Zuo-tao, Kong Fan-rong, Gabriela M,et al. Molecular serotype identification of Streptococcus agalactiae of bovine origin by multiplex PCR-based reverse line blot(mPCR/RLB)hybridization assay[J]. Fems Microbiol Lett, 2006, 263(2):236-239.
[3] Ren Si-yu, Geng Yi, Wang Kai-yu, et al. Streptococcus agalactiae infection in domestic rabbits, Oryctolagus cuniculus[J].Transbound Emerg Dis, 2014, 61(6):92-95.
[4] Liu L, Li Y W, He R Z, et al. Outbreak of Streptococcus agalactiae infection in barcoo grunter, Scortum barcoo(McCulloch&Waite), in an intensive fish farm in China[J]. J Fish Dis,2014, 37(12):1067-1072.
[5] Geng Yi, Wang Kai-yu, Huang Xiao-li, et al. Streptococcus aga-lactiae, an emerging pathogen for cultured ya-fish, Schizothorax prenanti, in China[J]. Transbound Emerg Dis, 2012, 59(4):369-375.
[6] Imperi M, Pataracchia M, Alfarone G, et al. A multiplex PCR assay for the direct identification of the capsular type(la to IX)of Streptococcus agalactiae[J]. J Microbiol Meth, 2010, 80(2):212-214.
[7] Pereira U, Mian G, Oliveira I, et al. Genotyping of Streptococcus agalactiae strains isolated from fish, human and cattle and their virulence potential in Nile tilapia[J]. Vet Microbiol, 2010,140(1-2):186-192.
[8] Wayne P. Clinical and laboratory standards institute:Performance standards for antimicrobial susceptibility testing[M].USA:2011.
[9] Wang Ya-min, Wang Qi-yao, Yang Wei-zheng, et al. Functional characterization of Edwardsiella tarda twin-arginine translocation system and its potential use as biological containment in live attenuated vaccine of marine fish[J]. Appl Microbiol Biot, 2013,97(8):3545-3557.
[10] Robinson J, Meyer F. Streptococcal fish pathogen[J]. J Bacteriol, 1966, 92(92):512-514.
[11]李莉萍,王瑞,黄婷,等.2007-2012年中国罗非鱼无乳链球菌流行菌株血清型分析[J].大连海洋大学学报,2014, 29(5):469-475.
[12]祝璟琳,杨弘.鱼源无乳链球菌致病机理研究进展[J].广东海洋大学学报,2013, 33(6):92-96.
[13] Isabelle R, Elisabeth S, Barbara M, et al. Reductive evolution in Streptococcus agalactiae and the emergence of a host adapted lineage[J]. Bmc Genomics, 2013, 14(1):252-255.
[14] Rodkhum C, Kayansamruaj P, Pirarat N. Effect of water temperature on susceptibility to Streptococcus agalactiae serotype la infection in Nile tilapia(Oreochromis niloticus)[J]. Thai J Vet Med, 2011, 41(3):309-314.
[15]刘志刚,可小丽,卢迈新,等.温度对尼罗罗非鱼无乳链球菌毒力的影响[J].水产学报,2013,37(11):1733-1741.
[16] Xi Hong-bo, Yang Qi,Shang Hai-tao, et al. Biodegradation characteristics and enzymatic reaction kinetics of phenanthrenedegrading bacteria[J]. Earth Sci Front, 2008, 6(01):17-19.
[17]黄瑞,吴淑燕,闻玉梅.细菌接合性耐药质粒与毒力关系的研究[J].苏州大学学报:医学版,2000,20(1):13-17.