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探討俄羅斯遠東分離之耐冷蕈狀桿菌生長特性以及冷適應相關基因體分析

為了解決Alteration synonym的問題，作者潘氏荷 這樣論述：

Psychrophilic and psychrotolerant are the most abundant microbes that contributes to the extremophiles on the Earth with their habitat occupies almost 80% of the biosphere on earth. The psychrotolerant are much more diverse because of their cold-adaptation ability with optimal growth temperatures >

15°C and maximal growth temperatures >20°C. In additon, they have the ability to grow at low temperatures. Psychrotolerant organisms, therefore, provide more knowledge about the biology of the cold-adaptation lifestyle. In this study, Bacillus mycoides U53 (synonym of Bacillus weihenstephanensis) wa

s isolated from soil under a bamboo tree in Sakhalin Region (50°33′N, 143°00′E), Russia, where the annual average temperature is around 2.2°C. B. mycoides is a Gram-positive, soil-dwelling bacterium belonging to the Bacillus cereus group. Because of their abundant in soil and growth potential in foo

d products at low temperatures, the cold adaptation mechanism has been of concern in food poisoning. The strain U53 was shown to be able to grow at 6 to 35°C in the culture broth and 4-37°C on agar plate, therefore, demonstrated to be psychrotolerant. During the growth, the activity of extracellular

enzymes has also been screened. It shows that endoglucanase, amylase, xylanase, pectinase and protease produced in the range of 4-37°C on the Nutrient agar plates (in which endoglucanase and amylase activity at 20°C > 30°C; xylanase activity at 20°C > 10°C > 30°C; protease activity at 10°C > 4°C >

37°C) but the strain has no exoglucanase activity from the screening at 4-37°C. Observations of the activity of these enzyme groups are also consistent with the real living environment, soil under a tree, of strain U53 which proposes the role of cold-active enzyme in B. mycoides U53 nutritional acti

vity. Based on this study, the strain is considered a potential source of cold-active enzymes for applications. B. mycoides U53 genome was sequenced and submit to NCBI under accession number JABURZ000000000. The genome size of B. mycoides U53 is 5,710,017 bp, with an average G+C content of 35.4%. Th

e chromosome size is 5,601,634 bp (G+C content, 35.4%), the numbers of predicted coding sequences, rRNA operons, tRNAs, other RNA, and pseudogenes in the genome were 4,734; 42; 106; 5 and 836, respectively. The strain U53 contains a plasmid of 82,515 bp (G+C content, 32.2%) with 88 predicted coding

sequences. There are 2 contigs with the size of 15,970 bp (G+C content, 35.6%), 9,898 (G+C content, 32.4%) containing 24 and 12 predicted coding sequences, respectively. A total of 163 genes (4% of the total annotated genes by using PGAP) with known or predicted roles in cold adaptation in U53 geno

me including cold shock, stress and heat shock proteins (8 genes), DNA replication and repair (10 genes), membrane and peptidoglycan alteration (76 genes), polysaccharide capsule (14 genes), osmotic stress (11 genes), oxidative stress (11 genes), translation and transcription factors (22 genes), car

bon storage/ starvation (9 genes) have been listed. In the carbon storage/ starvation group, gene-related poly-beta-hydroxybutyrate (PHB)/polyhydroxyalkanoic acids (PHAs) production have been listed which are also common in other B. mycoides genome. The functions of PHB/ PHAs in cold protection, mai

ntenance of cellular oxidative balance, oxidative stress resistance and cellular motility have also been proposed in many psychrophiles (such as Sphingopyxis alaskensis, Pseudomonas extremaustralis, P. haloplanktis, Psychrobacter sp. PAMC 21119), its roles in Bacillus cold adaptation has not been co

nfirmed using U53 as a model. Corresponding to the action of the cold-active enzyme, we hypothesize that under cold temperatures, the cold-active enzyme will be released outside the cell, providing monomer for intracellular PHAs accumulation, helping the cell withstand the harsh conditions of the en

vironment. In addition, by the pan-genome analysis, compared to the six available genomes of B. mycoides, 20 of these 163 genes are specific in strain U53, in which, 13 and 4 genes belong to the group of glycosyltransferase involved in cell wall biosynthesis and capsular polysaccharide biosynthesis

protein, respectively. The strain U53 specific genes with predicted roles in cold adaptation are all related to the cell envelope proposed the novel strategies in cold tolerant of strain U53. In summary, in this study, a psychrotolerant, B. mycoides U53 has been isolated and characterized its growth

property. The potential mechanisms to cope with the cold condition of B. mycoides were proposed based on its genome analysis and extracellular enzyme screening, which have not been attention before. Cold-active enzymes, poly-beta-hydroxybutyrate (PHB)/polyhydroxyalkanoic acids (PHAs) production and

membrane, peptidoglycan alteration are potential responses to cope with the harsh from cold conditions of strain U53.

探討院內鮑氏不動桿菌之抗藥性、生物膜形成和抗藥基因/酶的交互作用

為了解決Alteration synonym的問題，作者蘇百薇 這樣論述：

抗藥菌問題已成為全球規模性的公眾議題，抗藥性的產生不僅限制了治療上的選擇，也造成致病率、死亡率和治療成本的增加。多重抗藥性鮑氏不動桿菌被認為是最難控制和治療的院內感染菌株，尤其近年來出現的多重抗藥性鮑氏不動桿菌 (Multidrug-resistant Acinetobacter baumannii, MDRAB)、碳青黴烯類抗藥性鮑氏不動桿菌(Carbapenem resistant Acinetobacter baumannii , CRAB)、超廣譜乙內醯胺酶格蘭氏陰性菌(Extended-spectrum β-lactamases producing gram-negative ba

cteria，ESBL producing GNB) ，高度抗藥性 (Extensively drug-resistant Acinetobacter baumannii, XDRAB)及泛抗藥性 (Pandrug-resistant Acinetobacter baumannii, PDRAB)等菌株。由於抗藥性機轉的不同，雖然許多醫療單位已著手管控抗生素的使用與關注抗藥性的議題，但唯有瞭解院內抗藥菌的傳播路徑與抗藥機制，方能有效即時掌控、制定預防之道。本研究收集自嘉義基督醫院所分離的臨床菌株，並應用統計方法進行抗藥性、特定抗藥基因組合與生物膜生成的交互作用分析。利用紙錠擴散法和最低抑菌濃度

進行抗生素敏感性測試，包括11種抗生素，分別為Cephalexin, Ceftazidime, Imipenem, Gentamycin, Amikacin, Streptomycin, Piperacillin, Ticarcillin, Carbenicillin, Tetracyline, Sulfamethoxazole-Triethoprim。 基因分型探討計分析9個與抗藥性有關的基因，包括: blaOXA-51-like, blaOXA-23-like, blaOXA-58-like, blaOXA-24-like, blaOXA-143-like, tnpA, ISAba1, b

laNDM和blaADC。此外，利用Carba NP test快速檢測抗藥菌所攜帶的基因，及分析碳青黴烯類抗藥性鮑氏不動桿菌其抗藥基因的關聯性。生物膜生成利用分光光度測量法及掃描式電子顯微鏡觀察，並分析生物膜生成相關的基因bap, blaPER, ompA和csuE。本研究結果顯示大部分測試菌株具有多重抗藥性，且多數的碳青黴烯類抗藥性鮑氏不動桿菌 (93%) 攜帶ISAba1基因，其中高達68%的菌株同時攜帶tnpA和ISAba1基因。在生物膜生成分析，發現具有抗藥基因的菌株其生物膜形成量明顯較高，且多數具有多重抗藥性。本研究證實測試菌株的抗藥性、生物膜的生成和攜帶的抗藥基因型具有關聯性，分析

其抗藥機轉有助於臨床抗藥菌的篩選、鑑定、診斷、治療與管控，可提供預防菌株產生抗藥性的重要參考指標。