摘要
为确定浙江省德清县朱鹮抢救保护基地一只死亡朱鹮的发病原因及病原特征,本研究采集其肝脏、心脏等组织样品,通过TSA平板划线培养,观察菌落、革兰氏染色、生化鉴定和16S rRNA基因的PCR扩增与序列分析。结果显示,从朱鹮体内分离的细菌为红斑丹毒丝菌(E.rhusiopathiae),并命名为E.rhusiopathiae-716株。采用纸片扩散(K-B)法检测该菌对8类10种药物的敏感性;采用微量结晶紫染色法鉴定分离菌的生物被膜(BF)形成能力,结果显示该菌对青霉素类的氨苄西林和青霉素、喹诺酮类的诺氟沙星等5类6种抗生素敏感,对四环素类的四环素、大环内酯类的红霉素、氨基糖苷类的庆大霉素和阿米卡星等3类4种抗生素耐药,且其BF形成能力适中。采用Illumina二代测序平台对分离菌的全基因组测序,并采用SPAdes拼接序列;利用Prokka和PATRIC 3.6.9分析该菌基因组的分子特征。利用CARD数据库预测分离菌的耐药基因;采用VFDB数据库预测分离菌的毒力基因。采用ParSNP软件绘制该分离菌与GenBank中不同国家不同宿主来源的红斑丹毒丝菌全基因组的系统发育树。全基因组的分子特征分析结果显示,该分离菌基因组全长1.78×10^(3)kb,GC含量36.5%,含1694个蛋白编码基因、55个tRNA、6个rRNA、1个tmRNA,无质粒、无前噬菌体。经软件预测结果显示,该分离菌携带3个耐药基因,分别是耐四环素类药物的tetM、耐大环内酯类药物的mefA及兼具大环内酯类、林可酰胺类以及原霉素B类的msrD基因,这与其对四环素、红霉素的耐药表型相符,但未携带庆大霉素和阿米卡星耐药相关基因;其携带76个与菌毛、荚膜、铁摄取、溶胞素、分泌、转运等毒力相关基因;系统发育树结果显示分离株E.rhusiopathiae-716与韩国海豚源红斑丹毒丝菌分离株KC-Sb-R1处于同一分支,进一步表明分离菌为红斑丹毒丝菌。将不同剂量的E.rhusiopathiae-716株分别经腹
To determine the etiology and pathogenic characteristics of a dead crested ibis(nipponia nippon)in the rescue and protection base of crested ibis in Deqing County,Zhejiang Province,the liver,heart and other tissue were sampled aseptically.Bacterial colonies were observed through streak culture on TSA plates,followed by Gram staining,biochemical identification,and PCR amplification sequencing and analysis of the 16S rRNA gene.The results showed that one Erysipelothrix rhusiopathiae(E.rhusiopathiae)isolate were obtained from the crested ibis,which were named as E.rhusiopathiae-716.The sensitivity of the isolate to 10 antibiotics in 8 categories was detected by the universal Kirby Bauer(K-B)test,and its biofilm formation(BF)ability was detected by the microplate crystal violet staining method.The results showed that the strain was sensitive to 5 categories and 6 antibiotics such as ampicillin and penicillin of penicillins and norfloxacin of quinolones,and resistant to 3 categories and 4 antibiotics such as tetracycline of tetracyclines,erythromycin of macrolides,gentamicin of aminoglycosides and amikacin,and it had moderate BF ability.The whole genome of the isolate was sequenced using Illumina second-generation sequencing platform,and the sequence was assembled using SPAdes.Prokka and PATRIC 3.6.9 were used to analyze the functional characteristics of the genome,and the CARD database and VFDB database were used to predict the antibiotics resistance genes and the virulence genes of the isolate,respectively.A phylogenetic tree of the whole genome of the isolate and E.rhusiopathiae from different countries and host sources in GenBank was constructed using ParSNP.The functional annotation results of the whole genome showed that the genome of the isolate was 1.78×10^(3)kb in length,with a GC content of 36.5%,containing 1694 protein-coding gene sequences,55 tRNAs,6 rRNAs,1 tmRNA,and no plasmid,no prophagosome.The results of software prediction showed that the isolate carried three antibiotics resistance genes,namely tet
作者
邵乐乐
杨永春
马武林
白洪青
李颖
徐浩瑜
吴燕芬
宋厚辉
邱国强
SHAO Le-le;YANG Yong-chun;MA Wu-lin;BAI Hong-qing;LI Ying;XU Hao-yu;WU Yan-fen;SONG Hou-hui;QIU Guo-qiang(Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province,College of Animal Science and Technology&College of Veterinary Medicine,Zhejiang A&F University,Hangzhou 311300,China;Deqing County Ecological Forestry Comprehensive Service Center,Deqing 313200,China;Deqing Natural Resources and Planning Bureau of Zhejiang,Deqing 313200,China;Huzhou Ecological Forestry Protection Research Center,Deqing 313000,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2023年第12期1245-1252,共8页
Chinese Journal of Preventive Veterinary Medicine
基金
国家级大学生创新创业训练计划项目(202210341047)
国家自然科学基金(32102671)
湖州市科技计划项目(2022GZ40)。
关键词
红斑丹毒丝菌
朱鹮
全基因组测序
生物学特性
E.rhusiopathiae
crested ibis
whole genome sequence
biological characteristics
