一株从红藻中分离出的产芳基硫酸酯酶的海单胞菌FW-1的全基因组测序及结果分析（英文）

【目的】海单胞菌Marinomonas sp. FW-1是1株经验证可以获得高活性芳基硫酸酯酶的菌株。为深入研究FW-1菌株产芳基硫酸酯酶机制,进一步筛选高活性的芳基硫酸酯酶基因片段,有必要解析FW-1菌株的全基因组序列信息。【方法】本研究采用高通量测序技术对FW-1进行全基因组测序,使用相关软件对测序数据进行基因组装、基因预测与功能注释、COG聚类分析等。结合异源表达的方法对其不同基因片段所产生的芳基硫酸酯酶活性进行分析。【结果】全基因组测序结果表明该基因组大小为3964876 bp,GC含量为44.03%,编码3590个蛋白基因,含有78个tRNA和25个rRNA操纵子。从全基因组测序结果中找到22个可能具有芳基硫酸酯酶活性的基因,对其中4个进一步异源表达后发现FW-1中至少含有的3个具有芳基硫酸酯酶活性的基因,其均含有芳基硫酸酯酶的特异性氨基酸基团C-X-P-X-R基团。【结论】本研究首次报道了1株含有多个芳基硫酸酯酶基因序列的菌株FW-1的全基因组序列,分析了基因组的基本特征,为芳基硫酸酯酶的进一步应用提供了思路。     

一株从红藻中分离出的产芳基硫酸酯酶的海单胞菌FW-1的全基因组测序及结果分析

【目的】海单胞菌Marinomonas sp. FW-1是1株经验证可以获得高活性芳基硫酸酯酶的菌株。为深入研究FW-1菌株产芳基硫酸酯酶机制,进一步筛选高活性的芳基硫酸酯酶基因片段,有必要解析FW-1菌株的全基因组序列信息。【方法】本研究采用高通量测序技术对FW-1进行全基因组测序,使用相关软件对测序数据进行基因组装、基因预测与功能注释、COG聚类分析等。结合异源表达的方法对其不同基因片段所产生的芳基硫酸酯酶活性进行分析。【结果】全基因组测序结果表明该基因组大小为3964876 bp,GC含量为44.03%,编码3590个蛋白基因,含有78个tRNA和25个rRNA操纵子。从全基因组测序结果中找到22个可能具有芳基硫酸酯酶活性的基因,对其中4个进一步异源表达后发现FW-1中至少含有的3个具有芳基硫酸酯酶活性的基因,其均含有芳基硫酸酯酶的特异性氨基酸基团C-X-P-X-R基团。【结论】本研究首次报道了1株含有多个芳基硫酸酯酶基因序列的菌株FW-1的全基因组序列,分析了基因组的基本特征,为芳基硫酸酯酶的进一步应用提供了思路。     

全基因组外显子测序及其应用

近年来,众多研究小组开展了大量的全基因组关联研究(Genome-wide association studies,GWAS),发现并鉴定了许多与复杂疾病/性状相关联的遗传变异,为复杂疾病发病机制的研究提供了重要线索。由于GWAS的结果存在假阳性、假阴性、检测到的单核苷酸多态性很少位于功能区以及对稀有变异和结构变异不敏感等问题,导致了其应用的局限性。而新一代测序技术的进步,促进了全基因组测序和全基因组外显子测序的快速发展,为解决上述问题提供了契机。全基因组外显子测序是利用序列捕获技术将全基因组外显子区域DNA捕捉并富集后进行高通量测序的基因组分析方法。由于其具有对常见和罕见变异高灵敏度,能发现外显子区绝大部分疾病相关变异以及仅需要对约1%的基因组进行测序等优点,促使全基因组外显子测序成为鉴定孟德尔疾病的致病基因最有效的策略,也被运用于复杂疾病易感基因的研究和临床诊断中。     

基于比较基因组学解析耐酸乳杆菌G10的多碳源利用特征

【背景】耐酸乳杆菌（Lactobacillus acetotolerans）是白酒发酵过程中的优势乳酸菌,对白酒发酵具有重要作用。L. acetotolerans G10是分离自芝麻香型白酒发酵酒醅的一株能够利用多种碳源的菌株。【目的】基于全基因组测序,解析菌株G10多碳源利用机制。【方法】通过三代测序平台Oxford Nanopore完成菌株G10全基因组测序,分别利用Circlator和Prodigal对测序数据进行组装和基因预测;通过细菌基因组分析工具（bacterial pan genome analysis tool,BPGA）进行泛基因组分析。【结果】G10能够利用22种糖类及糖类衍生物,其全基因组大小为1 627 828 bp,含有1 878个编码基因;基于Koyto Encyclopedia of Genes and Genomes （KEGG）数据库注释获得292个碳源代谢相关基因,基于Carbohydrate-Active Enzymes （CAZy）数据库注释获得44个CAZy家族的编码基因。与其他发酵食品来源的耐酸乳杆菌相比,G10基因组最小,但其总基因数量以及...     

药用美洲大蠊全基因组测序分析

以美洲大蠊Periplaneta americana为原料生产的康复新液等药品临床疗效显著,得到了广泛应用。本文以四川好医生攀西药业有限责任公司饲养的药用美洲大蠊为材料,首次采用Illumina Hi Seq 2000和Pac Bio SMRT测序平台开展了全基因组测序,并进行基因组组装、注释和分析。原始测序数据经过滤后得到1.4 Tb的二代测序数据和33.81 Gb的三代测序数据。组装结果表明,美洲大蠊基因组大小为3.26 Gb,这在已报道的昆虫基因组中仅次于东亚飞蝗Locusta migratoria。基因组重复序列含量为62.38%,杂合度为0.635%,表明其为复杂基因组。组装的Contig N50和scaffold N50长度分别为28.2 kb、315 kb,单拷贝基因完整性为88.1%,小片段文库测序数据平均比对率为99.8%,测序和组装质量满足后续分析要求。采用De novo预测、同源预测和基于转录本预测3种方法共注释到14 568个基因,其中92.4%的基因获得了功能注释。本研究首次完成了美洲大蠊的全基因组测序,也是大蠊属Periplaneta昆虫的第一个基因组,为美洲大蠊遗传进化分析和药用基因资源挖掘打下了重要基础。     

雄烯二酮高产菌新金分枝杆菌MN4的全基因组测序及序列分析北大核心CSCD

【目的】新金分枝杆菌(Mycobacterium neoaurum)MN4是1株经诱变育种获得的高产雄烯二酮,并且能够耐受高浓度底物植物甾醇的突变菌株。为深入研究MN4菌株耐底物的机制及雄烯二酮的生物合成途径,有必要解析MN4菌株的全基因组序列信息。【方法】本研究采用高通量测序技术对MN4进行全基因组测序,然后使用相关软件对测序数据进行基因组装、基因预测与功能注释、COG聚类分析以及次级代谢产物合成基因簇预测等。【结果】新金分枝杆菌MN4基因组装获得33个Contigs,整个基因组大小为5.39 Mb,GC含量为66.9%,编码4920个蛋白基因,序列提交至Gen Bank数据库,登录号为JXYZ00000000。【结论】本研究首次报道了1株高产雄烯二酮菌株MN4的全基因组序列,分析了基因组的基本特征,初步解析了该菌株降解植物甾醇生产雄烯二酮的关键基因,将为MN4的功能基因组学研究及相关次级代谢产物的生物合成途径与异源表达研究提供基础。     

桑氏链霉菌KJ40全基因组测序及分析

【背景】桑氏链霉菌(Streptomyces sampsonii)KJ40是一株具有防病、促生多重功能的放线菌,有作为生物农药的潜力。目前还没有相关研究报道S.sampsonii全基因组序列,这限制了其功能基因、代谢产物合成途径及比较基因组学等研究。【目的】解析S.sampsonii KJ40的基因组序列信息,以深入研究该菌株防病促生机制及挖掘次级代谢产物基因资源。【方法】利用Illumina HiSeq高通量测序平台对KJ40菌株进行全基因组测序,使用相关软件对测序数据进行基因组组装、基因预测和功能注释、预测次级代谢产物合成基因簇、共线性分析等。【结果】基因组最后得到9个Scaffolds和578个Contigs,总长度为7 261 502 bp,G+C%含量平均为73.41%,预测到6 605个基因、1 260个串联重复序列、804个小卫星序列、67个微卫星序列、90个tRNA、9个rRNA和19个sRNA。其中,2 429、3 765、2 890、6 063和1 911个基因分别能够在COG、GO、KEGG、NR和Swiss-Prot数据库提取到注释信息。同时,还预测得到21个次级代谢产物合成基因簇。基因组测序数据提交至NCBI获得Gen Bank登录号:LORI00000000。S.sampsonii KJ40与Streptomyces coelicolor A3(2)、Streptomyces griseus subsp.griseus NBRC 13350三株链霉菌基因组存在翻转、易位等基因组重排,3个基因组共有1 711个蛋白聚类簇。【结论】研究为从基因组层面上解析KJ40菌株具有良好促生防病效果的内在原因提供基础数据,为深入了解链霉菌次级代谢合成途径提供参考信息,对S.sampsonii后续相关研究具有重要意义。     

基于金针菇全基因组的赖氨酸合成途径关键酶分析

【目的】以金针菇全基因组测序数据为基础研究其L-赖氨酸的从头合成途径及其关键基因。【方法】采用Illumina Hiseq2000和Roche454 FLX+两种方法完成金针菇单孢菌株Dan3的基因组测序,基于全基因组序列筛选金针菇中赖氨酸生物合成的关键基因,并分析这些基因编码的蛋白质基本理化性质;预测其亚细胞定位情况及蛋白的二级结构。【结果】金针菇Dan3全基因组序列长度为34.17 Mb,预测到8个参与α-氨基己二酸途径的关键基因,这些基因都含有多个内含子和外显子,二级结构主要由α-螺旋和无规则卷曲组成,有3个蛋白定位于线粒体。【结论】金针菇是通过α-氨基己二酸途径合成赖氨酸,基因组中预测到与该途径相关的几乎所有的酶。     

1株表现为鞭毛抗原3相的韦太夫雷登沙门氏菌的鉴定和基因组序列分析

利用高通量测序获取1株抗原式为3,10∶a,r,z6的沙门氏菌(Salmonella)GX150603的全基因组序列.根据鞭毛抗原序列、致病性和抗性基因预测GX150603的血清型,利用分子生物学软件分析基因组岛和前噬菌体,并与其他菌株进行全基因组系统发育分析.经鉴定GX150603为韦太夫雷登沙门氏菌(Salmone...     

冰冷杆菌PG-2的基因组测序及生物信息学分析

冰冷杆菌PG-2(Gelidibacter sp. PG-2)是一株能产类胡萝卜素的菌株,为深入研究其产类胡萝卜素的机制,对该菌株进行了全基因组测序。从PG-2中提取类胡萝卜素后通过LC-MS/MS进行定性分析,通过液体紫外全波长扫描计算其含量,基于全基因组测序结果对该菌株产类胡萝卜素的代谢通路进行了预测分析。结果表明,PG-2所产类胡萝卜素为玉米黄质,含量为185.81μg/g菌体干重,PG-2全基因组大小为3 850 413 bp,GC含量为37.91%,rRNA共计5个,tRNA共计37个,sRNA共计19个,在COG、GO、KEGG数据库分别注释到基因3 401、1 797、1 495个。菌株PG-2含有3个与产类胡萝卜素相关的核心基因crtI、crtZ、lcyB,并对其代谢通路进行了预测。基因组框架测序数据提交至NCBI获得GenBank登录号为JAMJTV000000000。上述结果表明,菌株PG-2具有产类胡萝卜素的能力,推测它产类胡萝卜素与基因crtI、crtZ和lcyB有关。     

The detection of <Emphasis Type="Italic">hip</Emphasis>O gene by real-time PCR in thermophilic <Emphasis Type="Italic">Campylobacter</Emphasis> spp. with very weak and negative reaction of hippurate hydrolysis

A total of 190 Campylobacter spp. isolates, of which 34 gave the result of very weak activity, and 156 gave the negative activity in the test for hippurate hydrolysis were characterized. The genomic DNA was isolated from a fresh culture of each isolate and the real-time PCR, targeting the hipO gene, was used to confirm the species distribution of Campylobacter isolates. The hipO gene was detected in 17 isolates (11%) within the total of 156 negative isolates for hippurate hydrolysis. Out of 34 isolates with very weak activity, 19 isolates (56%) were also found to be positive for hipO gene and characterized as C. jejuni. The real-time PCR assay used in this study could be employed for more accurate diagnosis of Campylobacter infections at species level after the biochemical characterization based on hippuricase activity of the isolates. This could also provide important data for the epidemiology of infections associated with these zoonotic pathogens.     

A Bayesian modelling framework to estimate Campylobacter prevalence and culture methods sensitivity: application to a chicken meat survey in Belgium

Aims: To estimate the true prevalence of Campylobacter and the diagnostic sensitivity of routine detection methods by applying a Bayesian modelling approach. Methods and Results: Results from a Belgium‐wide survey of Campylobacter contamination in chicken meat preparations (n = 656 samples) showed that Campylobacter was detected in 24·2% of the samples by enrichment, compared with 41% detected by direct plating. Combining positive results from both methods increased the apparent prevalence to 48·02%. Bayesian model was set up in WinBUGS software, the model estimates Campylobacter prevalence as 60% (95% Credibility interval (CI): 47–82%), and the sensitivity of enrichment culture and direct plating as 41% (95% CI: 31–52%) and 69% (95% CI: 50–85%), respectively. Conclusions: The parallel use of direct plating and enrichment culture adds value for Campylobacter detection from chicken meat preparations, but the false‐negative results from each culture method must be taken into account. Significance and Impact of the Study: Monitoring data could be strongly biased by the microbiological techniques used to generate it. To circumvent this bias, we describe an applied Bayesian framework for better interpretation of Campylobacter survey data in view of the imperfect test characteristics of routine culture methods.     

Local genes for local bacteria: Evidence of allopatry in the genomes of transatlantic Campylobacter populations

The genetic structure of bacterial populations can be related to geographical locations of isolation. In some species, there is a strong correlation between geographical distance and genetic distance, which can be caused by different evolutionary mechanisms. Patterns of ancient admixture in Helicobacter pylori can be reconstructed in concordance with past human migration, whereas in Mycobacterium tuberculosis it is the lack of recombination that causes allopatric clusters. In Campylobacter, analyses of genomic data and molecular typing have been successful in determining the reservoir host species, but not geographical origin. We investigated biogeographical variation in highly recombining genes to determine the extent of clustering between genomes from geographically distinct Campylobacter populations. Whole‐genome sequences from 294 Campylobacter isolates from North America and the UK were analysed. Isolates from within the same country shared more recently recombined DNA than isolates from different countries. Using 15 UK/American closely matched pairs of isolates that shared ancestors, we identify regions that have frequently and recently recombined to test their correlation with geographical origin. The seven genes that demonstrated the greatest clustering by geography were used in an attribution model to infer geographical origin which was tested using a further 383 UK clinical isolates to detect signatures of recent foreign travel. Patient records indicated that in 46 cases, travel abroad had occurred <2 weeks prior to sampling, and genomic analysis identified that 34 (74%) of these isolates were of a non‐UK origin. Identification of biogeographical markers in Campylobacter genomes will contribute to improved source attribution of clinical Campylobacter infection and inform intervention strategies to reduce campylobacteriosis.     

不同宿主源弯曲菌黏附侵袭肠上皮细胞及在鸡肠道内定殖的比较

【背景】弯曲菌(Campylobacter)是重要的人畜共患病原菌,可在多种动物肠道定殖,但不同宿主源弯曲菌对肠上皮细胞的黏附侵袭特征及在鸡肠道内的定殖能力并不明确。【目的】探究不同宿主源弯曲菌对不同宿主肠上皮细胞黏附侵袭及在鸡肠道内定殖能力的差异性。【方法】利用 5株来自不同宿主源弯曲菌,包括人源、鸡源、鸭源和牛源空肠弯曲菌(Campylobacter jejuni)及猪源结肠弯曲菌(Campylobacter coli),在对菌株PCR鉴定、运动力及生物膜形成能力测定的基础上,分别测定各菌株对人源肠上皮细胞Caco-2、猪源肠上皮细胞IPEC-J2和大鼠源肠上皮细胞IEC-6的黏附能力,通过庆大霉素保护试验测定菌株对肠上皮细胞的侵袭能力,比较黏附量和侵袭量的差异;将5株弯曲菌分别口服攻毒鸡,于攻毒后不同日龄(different days post inoculation,DPI)采集肠道样品测定弯曲菌的菌落数,比较不同弯曲菌在鸡肠道内定殖的差异。【结果】人源弯曲菌运动力显著高于其他4株动物源弯曲菌,而牛源和猪源弯曲菌生物膜形成能力显著高于其他菌株。黏附侵袭测定结果显示,人源弯曲菌对Caco-2细胞的黏附能力显著高于动物源弯曲菌,但侵袭能力显著低于动物源弯曲菌;鸭源和牛源弯曲菌对IPEC-J2细胞的黏附能力显著低于其他菌株,而且鸭源弯曲菌的侵袭能力显著低于其他菌株;不同菌株对IEC-6细胞的黏附能力无显著差异,但鸡源弯曲菌侵袭能力显著低于其他菌株。不同弯曲菌口服攻毒鸡后1、3和6d动物源弯曲菌定殖水平显著高于人源,在攻毒后10d和15d仅牛源弯曲菌显著高于人源,于攻毒后15d所有菌株达到约8-10Log₁₀(CFU/g)的稳定定殖水平。【结论】来源于不同宿主的弯曲菌对不同宿主肠上皮细胞均具有黏附侵袭能力,同时可在鸡肠道内稳定定殖,提示弯曲菌在不同动物间传播和适应性定殖的特征,对开展弯曲菌针对性防控措施具有一定的借鉴意义。     

Comparison of swabbing and destructive methods for microbiological pig carcass sampling

Aims: To compare the Belgian swabbing sampling method for pig carcasses with the reference destructive method with regard to Escherichia coli and aerobic plate counts, Salmonella and Campylobacter prevalence and their relationship. Methods and Results: Recovery was significantly lower for the swabbing method and corresponded to a recovery of 36% for E. coli counts and 81% for aerobic plate counts in comparison with the destructive method. There was no significant difference between the swabbing and destructive sampling methods for the prevalence of Salmonella or Campylobacter. A higher median for E. coli counts was detected for samples where Salmonella or Campylobacter were detected. The same association was also observed between the median for aerobic plate counts and the presence of Campylobacter. Conclusions: The method of swabbing used, covering 600 cm² on each half‐pig carcass, is efficient for the sampling of pig carcasses in comparison with the reference destructive method. Significance and Impact of the Study: This study describes an efficient method for microbiological pig carcass sampling. The Belgian swabbing method should continue to be used to allow the follow up of bacterial contamination in the Belgian meat production chain.     

Utilization of hydrogen and formate by Campylobacter spec. under aerobic and anaerobic conditions

A free-living aspartate-fermenting Campylobacter spec. was shown to utilize hydrogen produced in mixed culture by Clostridium cochlearium from glutamate. Resting cells of Campylobacter were shown to reduce aspartate, fumarate and malate as well as nitrate, nitrite, hydroxylamine, sulphite, thiosulphate and elemental sulphur with molecular hydrogen. Growth of Campylobacter spec. was demonstrated with formate as electron donor and nitrate, thiosulphate, elemental sulphur or oxygen as electron acceptor in the presence of acetate as carbon source.     

The occurrence of Campylobacter in river water and waterfowl within a watershed in southern Ontario,Canada

Aims: Quantitative PCR and a culture method were used to investigate Campylobacter occurrence over 3 years in a watershed located in southern Ontario, Canada that is used as a source of drinking water. Methods and Results: Direct DNA extraction from river water followed by quantitative PCR analysis detected thermophilic campylobacters at low concentrations (<130 cells 100 ml^?1) in 57–79% of samples taken from five locations. By comparison, a culture‐based method detected Campylobacter in 0–23% of samples. Water quality parameters such as total Escherichia coli were not highly correlated with Campylobacter levels, although higher pathogen concentrations were observed at colder water temperatures (<10°C). Strains isolated from river water were primarily nalidixic acid‐susceptible Campylobacter lari, and selected isolates were identified as Campylobacter lari ssp. concheus. Campylobacter from wild birds (seagulls, ducks and geese) were detected at a similar rate using PCR (32%) and culture‐based (29%) methods, and although Campylobacter jejuni was isolated most frequently, C. lari ssp. concheus was also detected. Conclusions: Campylobacter were frequently detected at low concentrations in the watershed. Higher prevalence rates using quantitative PCR was likely because of the formation of viable but nonculturable cells and low recovery of the culture method. In addition to animal and human waste, waterfowl can be an important contributor of Campylobacter in the environment. Significance and Impact of the Study: Results of this study show that Campylobacter in surface water can be an important vector for human disease transmission and that method selection is important in determining pathogen occurrence in a water environment.     

Repeated therapeutic dosing selects macrolide‐resistant Campylobacter spp. in a turkey facility

Aims: This study assessed the effects of the therapeutic use of Tylan^® in a large‐scale turkey production facility on the selection of macrolide‐resistant Campylobacter. Methods and Results: A flock of production turkeys (c. 30 000 birds) was followed from brooding to slaughter, and the effects of macrolide application was assessed in one half of the flock from finishing stage to final product and compared against the control barn where no macrolide was used. Overall, Campylobacter prevalence in turkeys was almost 100% by 4 weeks of age. When Campylobacter prevalence was assessed in relation to treatment, high levels of macrolide resistance were evident in this group following treatment, with Campylobacter coli becoming the dominant strain type. Over time, and in the absence of a selection agent, the population of resistant strains decreased suggesting that there was a fitness cost associated with macrolide resistance carriage and persistence. Macrolide resistance was detected in the control barn at a very low level (four isolates recovered during the study), suggesting that the creation or selection of macrolide‐resistant Campylobacter was correlated with the treatment regime used. Molecular analysis of a selection of macrolide‐resistant Campylobacter recovered was assessed using PCR, RFLP and sequence analysis of the 23S rRNA. The majority of isolates displaying high‐level macrolide resistance (>256 μg ml^?1) possessed an A2075G transition mutation in the 23S rRNA and the CmeABC efflux pump. Conclusions: These studies suggest that macrolide resistance can be promoted through the application of treatment during the grow‐out phase and once established in a production facility has the potential to persist and be transferred to final product. Significance and Impact of the Study: The study highlights the prudent use of antimicrobials in treatment of disease in poultry. Of significance is the presence of macrolide‐resistant Campylobacter in poultry production and finished product as a consequence of macrolide usage.     

Bacteriocin-like inhibitory substances from <Emphasis Type="Italic">Campylobacter</Emphasis> spp.

Twenty-five Campylobacter isolates were screened for production of antimicrobial substances using a deferred antagonism assay. Sixteen isolates showed activity against either Staphylococcus aureus, Salmonella enterica serovar Enteritidis or Candida albicans. The inhibitory activity was sensitive to treatment with pronase E, trypsin and pepsin, suggesting that the antimicrobial compound(s) are proteinaceous. Activity spectra of isolates included S. aureus, Micrococcus luteus, Streptococcus sp., Bacillus subtilis, a drug-resistant clinical isolate of S. aureus and one isolate of C. albicans. Producing isolates showed cross-immunity and inhibitory activity was only observed on solid media. The findings of this study suggest that Campylobacter produces proteinaceous inhibitory substances.     

Comprehensive Genomic Characterization of Campylobacter Genus Reveals Some Underlying Mechanisms for its Genomic Diversification

Campylobacter species.are phenotypically diverse in many aspects including host habitats and pathogenicities, which demands comprehensive characterization of the entire Campylobacter genus to study their underlying genetic diversification. Up to now, 34 Campylobacter strains have been sequenced and published in public databases, providing good opportunity to systemically analyze their genomic diversities. In this study, we first conducted genomic characterization, which includes genome-wide alignments, pan-genome analysis, and phylogenetic identification, to depict the genetic diversity of Campylobacter genus. Afterward, we improved the tetranucleotide usage pattern-based naïve Bayesian classifier to identify the abnormal composition fragments (ACFs, fragments with significantly different tetranucleotide frequency profiles from its genomic tetranucleotide frequency profiles) including horizontal gene transfers (HGTs) to explore the mechanisms for the genetic diversity of this organism. Finally, we analyzed the HGTs transferred via bacteriophage transductions. To our knowledge, this study is the first to use single nucleotide polymorphism information to construct liable microevolution phylogeny of 21 Campylobacter jejuni strains. Combined with the phylogeny of all the collected Campylobacter species based on genome-wide core gene information, comprehensive phylogenetic inference of all 34 Campylobacter organisms was determined. It was found that C. jejuni harbors a high fraction of ACFs possibly through intraspecies recombination, whereas other Campylobacter members possess numerous ACFs possibly via intragenus recombination. Furthermore, some Campylobacter strains have undergone significant ancient viral integration during their evolution process. The improved method is a powerful tool for bacterial genomic analysis. Moreover, the findings would provide useful information for future research on Campylobacter genus.