Isolation and identification of Vibrio spp. with potential ability to produce polysaccharide monooxygenase from diseased Penaeus monodon

Ho Ta Giap, N. Vu, V. V. Van
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Abstract

Polysaccharide monooxygenases (PMOs) and Glycoside Hydrolases (GH) play a crucial role in breaking down chitin and cellulose. Research on PMOs has mainly focused on their role in the production of bioethanol from biomass. Still, the potential of PMOs as virulence factors in microorganisms is not been fully studied. Some pathogenic bacteria secrete PMOs, which have been identified as virulence factors. The ability of Vibrio species to metabolize chitin is well-known, but the question of which Vibrio species contains the PMO gene remains unanswered. Materials and methods: Black tiger shrimp Penaeus monodon was collected from a culture pond in Long An, Vietnam. Thiosulfate-citrate-bile salts-sucrose and CHROMagar Vibrio agar were used to isolate, Vibrio bacteria from the shrimp samples. Mixed bases primers were designed to detect PMOs gene from Vibrio strains. Results: Total of 35 different strains of bacteria were isolated from the guts and pancreas of the shrimp sample. Based on biochemical tests, there were a total of 20 different Vibrio strains identified among 35 isolated strains. The PCR results showed that six isolates could have the PMOs gene. The partial 16S rDNA fragments of six isolate Vibrio strains were sequenced. Comparing 16S rDNA sequences with sequences available in NCBI databases revealed that all the sequences are Vibrio spp. 16S rDNA sequences.
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从患病的单对虾中产生多糖单加氧酶的弧菌的分离与鉴定
多糖单加氧酶(PMOs)和糖苷水解酶(GH)在甲壳素和纤维素的分解中起着至关重要的作用。对PMOs的研究主要集中在它们在生物质生产生物乙醇中的作用。尽管如此,PMOs作为微生物毒力因子的潜力尚未得到充分研究。一些致病菌分泌PMOs,已被确定为毒力因子。几种弧菌代谢几丁质的能力是众所周知的,但是哪种弧菌含有PMO基因的问题仍然没有答案。材料与方法:采自越南龙安某养殖池的黑虎对虾。采用硫代硫酸盐-柠檬酸盐-胆汁盐-蔗糖和CHROMagar琼脂弧菌法从虾样品中分离弧菌。设计了混合碱基引物,用于检测弧菌PMOs基因。结果:从对虾的肠道和胰腺中共分离到35株不同的细菌。经生化检测,35株分离菌株中共鉴定出20株不同的弧菌。PCR结果显示,6株分离物可携带PMOs基因。对6株弧菌分离株的部分16S rDNA片段进行了测序。将16S rDNA序列与NCBI数据库中的序列进行比对,发现所有序列均为弧菌16S rDNA序列。
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