Genetic and Phenotypic Variation of Campylobacter jejuni NCTC11168 Caused by flhA Mutation during Laboratory Passage

Objective Campylobacter jejuni NCTC11168 is commonly used as a standard strain for flagellar biosynthesis research.In this report,two distinguished phenotypic isolates(CJ1Z,flhA mutant strain,lawn;CJ2S,flhA complemented strain,normal colony)appeared during laboratory passages for NCTC11168.Methods Phenotypic assessments,including motility plates,transmission electron microscopy,biofilm formation assay,autoagglutination assay,and genome re-sequencing for these two isolates(CJ1Z,flhA mutant strain;CJ2S,flhA complemented strain)were carried out in this study.Results Transmission electron microscopy revealed that the flagellum was lost in CJ1Z.Phenotypic assessments and genome sequencing of the two isolates were performed in this study.The capacity for biofilm formation,colony auto-agglutination,and isolate motility was reduced in the mutant CJ1Z.Comparative genomic analysis indicated a unique native nucleotide insertion in flhA(nt,2154)that caused the I719Y and I720Y mutations and early truncation in flhA.Conclusion FlhA has been found to influence the expression of flagella in C.jejuni.To the best of our knowledge,this is the first study to describe the function of the C-terminal of this protein.

2016-2022年北京市人源空肠弯曲菌多重耐药及流行特征分析

目的 对2016-2022年北京地区人源空肠弯曲菌流行特征和多重耐药进行分析。方法 对2016-2022年肠道门诊的感染性腹泻患者粪便样本进行滤膜驱动法分离培养空肠弯曲菌并采用琼脂稀释法(MIC)进行耐药实验。结果 2016-2022年共检出人源空肠弯曲菌614株,检出月份以4-10月为主,占87.46%(537/614),发病年龄集中在20~<40岁,占51.63%(317/614),男、女性别比为1.52∶1。耐药结果显示,614株菌以萘啶酸耐药率最高,为78.99%(485/614),红霉素耐药率最低,为4.89%(30/614)。614株菌有116种耐药谱型,耐三重及以上耐药菌株占65.64%(403/614),耐药谱以耐萘啶酸、环丙沙星、四环素谱型最多,占23.62%(145/614)。结论 空肠弯曲菌引起的腹泻逐年增多,多重耐药严重且谱型复杂。

大肠杆菌肠菌素对空肠弯曲菌的促生长作用研究

[目的]调查鸡肠道内大肠杆菌分泌肠菌素的水平,以及其对空肠弯曲菌的促生长作用。[方法]利用间接竞争ELISA(ic-ELISA)方法检测临床分离的30株鸡肠道大肠杆菌分离株的肠菌素产量;利用λRed同源重组系统构建大肠杆菌entB基因缺失突变株,并用ic-ELISA方法测定大肠杆菌亲本株和突变株的肠菌素产量;检测亲本株和突变株在不同限铁培养基中的生长曲线;将大肠杆菌亲本株和突变株培养上清接种空肠弯曲菌,探究大肠杆菌肠菌素对空肠弯曲菌生长的影响。[结果]30株鸡肠道大肠杆菌分离株分泌肠菌素水平介于11~126μmol/L之间。利用λRed同源重组系统成功构建2株大肠杆菌entB基因缺失突变株。定性结果显示,大肠杆菌亲本株在CAS培养基中产生黄色晕圈,而entB基因缺失株不产生晕圈。ic-ELISA检测结果显示,亲本株分泌肠菌素浓度为100~150μmol/L,而基因缺失株未检测出肠菌素。大肠杆菌亲本株和entB基因缺失株在富铁培养基中生长无显著差异(P>0.05),在限铁培养基中达到平台期的D 600 nm值分别为0.6和0.3。促生长试验结果显示,大肠杆菌亲本株培养上清接种空肠弯曲菌24和36 h,菌落数分别为7.8和8.2 lg CFU/mL,而entB基因缺失株培养上清接种后菌落数分别为7.0和7.3 lg CFU/mL。[结论]大肠杆菌具有分泌肠菌素的能力,且其在限铁环境下能促进空肠弯曲菌的生长,试验结果为深入研究肠菌素介导的肠道细菌之间的相互作用奠定基础。

空肠弯曲菌双分子荧光互补系统构建及其在蛋白质互作研究中的应用

为研究空肠弯曲菌中蛋白质的相互作用,建立适合于该菌稳定高效表达的双分子荧光互补(bimolecular fluorescence complementation,BiFC)系统,通过构建基于基因启动子、荧光蛋白以及基因回补方式等不同条件的重组载体,分别导入空肠弯曲菌进行荧光数值比较分析,确定荧光表达最优的参数,建立以porA启动子和Venus荧光蛋白的质粒回补的BiFC体系。用该体系探究空肠弯曲菌鞭毛表达蛋白的互作。结果表明:已知flhF与flhG互作的BiFC-flhF-flhG载体在菌体内表达的荧光值显著高于阴性对照载体,而BiFC-flhF-fliM和BiFC-flhF-fliG与阴性对照无差异,说明BiFC体系建立成功,且flhF与fliG、fliM不存在互作关系。综上,本研究建立了适用于空肠弯曲菌的BiFC体系,可为该菌蛋白质互作研究提供高效便捷的方法。

苏州市空肠弯曲菌耐药与分子特征分析

目的了解苏州市空肠弯曲菌的耐药情况和分子特征。方法对2018年-2022年苏州市分离的空肠弯曲菌进行全基因组测序分析了解毒力基因、耐药基因、多位点序列分型(Multi-locus sequence typing,MLST)等分子特征,通过琼脂稀释法了解其耐药表型。结果61株空肠弯曲菌被分为42个ST型别,以ST464占比最高(9.8%),包含12个CC群,以CC21占比最高(19.7%)。61株菌共检出18种耐药基因,97种毒力基因。药敏试验耐药率排前3位的抗生素分别是萘啶酸,四环素和环丙沙星,耐药率分别为90.2%,88.5%和73.8%,对红霉素100.0%敏感,多重耐药率达到41.0%。结论苏州市空肠弯曲菌多重耐药率高,携带耐药基因和毒力基因较多,克隆群以CC21为主,但ST型别较为分散,呈现较高的遗传多样性。

微酸性电解水对空肠弯曲菌的杀灭特性及机制

探究微酸性电解水(slightly acidic electrolyzed water,SAEW)对空肠弯曲菌(Campylobacter jejuni)的杀灭效果和杀灭机制。通过使用不同有效氯质量浓度(available chlorine concentration,ACC)的SAEW对C. jejuni进行不同时间的处理,考察ACC和处理时间对杀菌效果的影响;通过加入蛋白质,考察蛋白质对SAEW杀菌效果的影响;此外,分析了SAEW对多代培养菌的杀灭效果。通过对细胞膜完整性、形态以及细胞内容物含量等变化分析,探究SAEW杀灭C. jejuni的机制。结果表明:SAEW对C. jejuni的杀灭效果与ACC和处理时间呈正相关;蛋白质会减弱SAEW的杀菌效果;C. jejuni在10代内未对SAEW产生抗性反应;SAEW处理可破坏细胞膜完整性,菌体表面出现凹陷褶皱、破损,胞内核酸、蛋白质和ATP泄漏等变化,表明细胞膜受损可能是SAEW杀灭C. jejuni的主要原因。

婴幼儿血流感染空肠弯曲菌1例并文献复习

空肠弯曲菌(Campylobacter jejuni)广泛存在于家禽肠道,是全球范围内广受重视的人畜共患食物源性病原菌,主要引起人类和牲畜急性胃肠道感染。空肠弯曲菌导致的肠外感染较为少见,本资料报道1例空肠弯曲菌导致婴幼儿血流感染病例,并加以相关文献复习,以提高临床对空肠弯曲菌的认识。

Development and application of a real-time polymerase chain reaction method for Campylobacter jejuni detection

AIM:To develop a real-time polymerase chain reaction(PCR) method to detect and quantify Campylobacter jejuni(C.jejuni) from stool specimens.METHODS:Primers and a probe for real-time PCR were designed based on the specific DNA sequence of the hipO gene in C.jejuni.The specificity of the primers and probe were tested against a set of Campylobacter spp.and other enteric pathogens.The optimal PCR conditions were determined by testing a series of conditions with standard a C.jejuni template.The detection limits were obtained using purified DNA from bacterial culture and extracted DNA from the stool specimen.Two hundred and forty-two specimens were analyzed for the presence of C.jejuni by direct bacterial culture and real-time PCR.RESULTS:The optimal PCR system was determined using reference DNA templates,1 × uracil-DNA glycosylase,3.5 mmol/L MgCl 2,1.25 U platinum Taq polymerase,0.4 mmol/L PCR nucleotide mix,0.48 μmol/L of each primer,0.2 μmol/L of probe and 2 μL of DNA template in a final volume of 25 μL.The PCR reaction was carried as follows:95 ℃ for 4 min,followed by 45 cycles of 10 s at 95 ℃ and 30 s at 59 ℃.The detection limit was 4.3 CFU/mL using purified DNA from bacterial culture and 10 3 CFU/g using DNA from stool specimens.Twenty(8.3%,20/242) C.jejuni strains were isolated from bacterial culture,while 41(16.9%,41/242) samples were found to be positive by realtime PCR.DNA sequencing of the PCR product indicated the presence of C.jejuni in the specimen.One mixed infection of C.jejuni and Salmonella was detected in one specimen and the PCR test for this specimen was positive.CONCLUSION:The sensitivity of detection of C.jejuni from stool specimens was much higher using this PCR assay than using the direct culture method.

Genetic and Antibiotic Resistance Characteristics ofCampylobacterjejuni Isolated from Diarrheal Patients,Poultry and Cattle in Shenzhen

Objective To investigate genetic and antibiotic resistance characteristics of Campylobacter jejuni(C. jejuni) isolated from Shenzhen. Methods Multilocs sequence typing and agar dilution methods were used to define the genotype and antibiotic resistance of C. jejuni, respectively. Results In total, 126 C. jejuni strains were isolated. The prevalence of C. jejuni was 5.3% in diarrheal patients. The prevalence in poultry meat(36.5%) was higher than that in cattle meat(1.1%). However, the prevalence in poultry cloacal swabs(27.0%) was lower than that in cattle stool(57.3%). Sixty-two sequence types were obtained, among which 27 of the STs and 10 alleles were previously unreported. The most frequently observed clonal complexes were ST-21(11.9%), ST-22(10.3%), and ST-403(7.1%). ST-21, ST-45, ST-354, ST-403, and ST-443 complexes overlapped between isolates from patients and cattle, whereas ST-45 and ST-574 complexes overlapped between isolates from patients and poultry. All C. jejuni were resistant to at least one antibiotic. The highest resistance rate was toward ciprofloxacin(89.7%), followed by tetracycline(74.6%), and nalidixic acid(69.0%). Conclusion This is the first report of the genotypes and antibiotic resistance of C. jejuni in Shenzhen. Overlapping clonal complexes were found between isolates from patients and cattle, and between patients and poultry.

Reaction of antibodies to Campylobacter jejuni and cytolethal distending toxin B with tissues and food antigens

BACKGROUND The bacteria Campylobacter jejuni(C. jejuni) is commonly associated with GuillaneBarré syndrome(GBS) and irritable bowel syndrome(IBS), but studies have also linked it with Miller Fisher syndrome, reactive arthritis and other disorders, some of which are autoimmune. It is possible that C. jejuni and its toxins may be crossreactive with some human tissues and food antigens, potentially leading to autoimmune responses.AIM To measure the immune reactivity of C. jejuni and C. jejuni cytolethal distending toxin(Cdt) antibodies with tissue and food antigens to examine their role in autoimmunities.METHODS Using enzyme-linked immunosorbent assay(ELISA) methodology, specific antibodies made against C. jejuni and C. jejuni Cdt were applied to a variety of microwell plates coated with 45 tissues and 180 food antigens. The resulting immunoreactivities were compared to reactions with control wells coated with human serum albumin(HSA) which were used as negative controls and with wells coated with C. jejuni lysate or C. jejuni Cdt which served as positive controls.RESULTS At 3 SD above the mean of control wells coated with HSA or 0.41 OD, the mouse monoclonal antibody made against C. jejuni showed moderate to high reactions with zonulin, somatotropin, acetylcholine receptor, β-amyloid and presenilin.This immune reaction was low with an additional 25 tissue antigens including asialoganglioside, and the same antibody did not react at all with another 15 tissue antigens. Examining the reaction between C. jejuni antibody and 180 food antigens, we found insignificant reactions with 163 foods but low to high immune reactions with 17 food antigens. Similarly, we examined the reaction of C. jejuni Cdt with the same tissues and food antigens. The strongest reactions were observed with zonulin, intrinsic factor and somatotropin. The reaction was moderate with 9 different tissue antigens including thyroid peroxidase, and reaction was low with another 10 different antigens, including neuronal antigens.The reaction of C. jejuni Cdt antibody with an additional 23 tissue antigens was insignificant. Regarding the reaction of C. jejuni Cdt antibody with different food antigens, 160 out of 180 foods showed insignificant reactions, while 20 foods showed reactions ranging from low to high.CONCLUSION Our findings indicate that C. jejuni and its Cdt may play a role in inflammation and autoimmunities beyond the gut.

Polyphosphate and associated enzymes as global regulators of stress response and virulence in Campylobacter jejuni

Campylobacter jejuni(C. jejuni),a Gram-negative microaerophilic bacterium,is a predominant cause of bacterial foodborne gastroenteritis in humans worldwide. Despite its importance as a major foodborne pathogen,our understanding of the molecular mechanisms underlying C. jejuni stress survival and pathogenesis is limited. Inorganic polyphosphate(poly P) has been shown to play significant roles in bacterial resistance to stress and virulence in many pathogenic bacteria. C. jejuni contains the complete repertoire of enzymes required for poly P metabolism. Recent work in our laboratory and others have demonstrated that poly P controls a plethora of C. jejuni properties that impact its ability to survive in the environment as well as to colonize/infect mammalian hosts. This review article summarizes the current literature on the role of poly P in C. jejuni stress survival and virulence and discusses on how poly P-related enzymes can be exploited for therapeutic/prevention purposes. Additionally,the review article identifies potential areas for future investigation that would enhance our understanding of the role of poly P in C. jejuni and other bacteria,which ultimately would facilitate design of effective therapeutic/preventive strategies to reduce not only the burden of C. jejuni-caused foodborne infections but also of other bacterial infections in humans.

Antibiotic susceptibility profiling and virulence potential of Campylobacter jejuni isolates from different sources in Pakistan

Objective:To determine antibiotic resistance patterns and virulence potential of Campylobacter jejuni(C.jejuni) isolates from clinical human diarrheal infections,cattle and healthy broilers.Methods:Antibiotic sensitivity patterns of C.jejuni isolates were determined by Kirby Bauer Disc Diffusion assay.These isolates were then subjected to virulence profiling for the detection of map A(membrane-associated protein).cadF(fibronectin binding protein).wlaN(beta-1.3-galaclosyltransferase) and neu AB(sialic acid biosynthesis gene).Further C.jejuni isolates were grouped by random amplification of polymorphic DNA(RAPD) profiling.Results:A total of436 samples from poultry(n=88).cattle(n=216) and humans(n=132) from different locations were collected.Results revealed percentage of C.jejuni isolates were 35.2%(31/88).25.0%(54/216) and 11.3%(15/132) among poultry,cattle and clinical human samples respectively.Antibiotic susceptibility results showed that similar resistance patterns to cephalothin was ie.87.0%,87.1%and 89%among humans,poultry and cattle respectively,followed by sulfamethoxazolc+trimcthoprim 40.0%,38.7%and 31.0%in humans,poultry and cattle and Ampicillin 40%,32%and 20%in humans,poultry and cattle respectively.Beta-lactamase activity was detected in 40.00%humans.20.37%cattle and 32.25%in poultry C.jejuni isolates.CadF and mapA were present in all poultry,cattle and human C jejuni isolates.wlauN was not detected in any isolate and neu AB was found in 9/31(36%) poultry isolates.RAPD profiling results suggested high diversity of C.jejuni isolates.Conclusions:Detection of multidrug resistant C.jejuni strains from poultry and cattle is alarming as they can be potential hazard to humans.Moreover,predominant association of virulence factors,cadF and map A(100%each) in C.jejuni isolates from all sources and neuAB(36%) with poultry isolates suggest the potential source of transmission of diverse types of C.jejuni to humans.

In Vitro Protein Expression Profile of Campylobacter jejuni Strain NCTC11168 by Two-dimensional Gel Electrophoresis and Mass Spectrometry

Objective To investigate the protein expression profiles of the major food‐borne pathogen Campylobacter jejuni NCTC11168.Methods Membrane and soluble cellular proteins were extracted from the genome‐sequenced C.jejuni strain NCTC11168.Protein expression profiles were determined using two‐dimensional gel electrophoresis（2‐DE）.All the detected spots on the 2‐DE map were subjected to matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry（MALDI‐TOF/TOF） analysis.Results A total of 537 and 333 spots were detected from the whole cell and membrane‐associated proteins of C.jejuni NCTC11168 cultured on Columbia agar medium at 42 ℃ by 2‐DE and Coomassie Brilliant Blue staining,respectively.Analyses of whole cell and membrane‐associated proteins included 399 and 133 spots,respectively,which included 182 and 53 functional proteins identified by MALDI‐TOF/TOF analysis.Conclusion The comprehensive expression protein profiles of C.jeuni NCTC11168 obtained in this study will be useful for elucidating the roles of these proteins in further pathogenesis investigation.

Can Campylobacter jejuni play a role in development of celiac disease? A hypothesis

Celiac disease （CD） is an entropathy with malabsortive condition in which an allergic reaction to the cereal grain-protein （gluten） causes small intestine rnucosal injury. CD is a multifactorial disorder in which both genetic and environmental factors contribute to the disease development. Mechanisms have been described to explain the pathology of CD. T cells specific for multiple gluten peptides are found in virtually all patients. Generation of such a broad T cell response may be a prerequisite for disease development. CD is associated with multiple extraintestinal presentations, including neurological deficits. Recent studies have shown a significant correlation between anti-ganglioside antibodies and neurological disorders in patients with underlying CD. Gangliosides are glycosphingolipids which are abundant in nervous system and in other tissues including gastrointestinal tract. It is not known what triggers the release of anti-ganglioside antibodies in people with gluten sensitivity. But, the mechanism is likely to involve the intestinal immune system response to ingested gliadin, a component of wheat gluten. Studies showed that mechanisms different from gluten exposure may be implicated in antibody formation, and other environmental factors may also exist. In addition, considering the fact that genetic predisposition dysregulating mucosal immune responses in the presence of certain environmental triggers like gastrointestinal infections may be strong etiological factors for developing chronic intestinal inflammation including CD, the hypothesis raised in our mind that antiganglioside antibody formation in CD may play a role not only in development of neurological complications in celiac patients, but also in development of CD itself. As presence of Campylobacter jejuni in other diseases with antigangliosides antibody formation has been established, we propose the possible role of Campylobacter jejuni in development of CD in association with other genetic and environmental factors by the mechanism that molecular mimicry of gangliosides-like epitopes common to both lipo-polysacharide coats of certain strains of Campylobacter jejuni and gangliosides in cell structure of gastrointestinal mucosa may cause an autoimmune response and consequently lead to atrophy and degeneration of mucosa possibly by apoptosis.

Disruption of colonic barrier function and induction of mediator release by strains of Campylobacter jejuni that invade epithelial cells

AIM: To study the mechanisms by which Campylobacter jejuni (C. jejuni) causes inflammation and diarrhea. In particular, direct interactions with intestinal epithelial cells and effects on barrier function are poorly under- stood. METHODS: To model the initial pathogenic effects of C. jejuni on intestinal epithelium, polarized human colonic HCA-7 monolayers were grown on permeabilized filters and infected apically with clinical isolates of C. jejuni. Integrity of the monolayer was monitored by changes in monolayer resistance, release of lactate dehydrogenase, mannitol fluxes and electron microscopy. Invasion of HCA-7 cells was assessed by a modified gentamicin protection assay, translocation by counting colony forming units in the basal chamber, stimulation of mediator release by immunoassays and secretory responses in monolayers stimulated by bradykinin in an Ussing chamber. RESULTS: All strains translocated across monolayers but only a minority invaded HCA-7 cells. Strains that invaded HCA-7 cells destroyed monolayer resistance over 6 h, accompanied by increased release of lactate dehydrogenase, a four-fold increase in permeability to [3H] mannitol, and ultrastructural disruption of tight junctions, with rounding and lifting of cells off the filter membrane. Synthesis of interleukin (IL)-8 and prostaglandin E2 was increased with strains that invaded the monolayer but not with those that did not. CONCLUSION: These data demonstrate two distinct effects of C. jejuni on colonic epithelial cells and provide an informative model for further investigation of initial host cell responses to C. jejuni.

Genetic Characteristics of Lipooligosaccharide and Capsular Polysaccharide of Campylobacter jejuni from Different Sources in China

Objective To determine the distribution of two important virulence factors[lipooligosaccharide(LOS)and capsular polysaccharide(CPS)]in Campylobacter jejuni(C.jejuni)isolated from different sources in China and to develop a rapid screening method for Guillain–Barrésyndrome(GBS)-associated strains.Methods Whole-genome sequencing was carried out for 494 C.jejuni strains.The Ortho MCL software was used to define the LOS/CPS gene clusters.CPS genotyping was performed with serotype-specific sequence alignment using the BLAST software.Real-time Polymerase chain reaction(PCR)was developed with the unique sequences of specific CPS types.Results Nine novel and 29 previously confirmed LOS classes were identified.LOS classes A,B,and C were the most common(48.2%,238/494)among the 494 strains.Twenty-six capsular types were identified in 448 strains.HS2,HS4c,HS5/31,HS19,and HS8/17 were the most frequent CPS genotypes(58.7%,263/448).Strains of 17 CPS genotypes(strain number>5)had one or two prevalent LOS classes(P<0.05).Multiplex real-time PCR for rapid identification of HS2,HS19,and HS41 was developed and validated with strains of known serotypes.Conclusion Our results describe the genetic characteristics of the important virulence factors in C.jejuni strains in China.The multiplex real-time PCR developed in this study will facilitate enhanced surveillance of GBS-associated strains in China.

Use of reducing agents for the aerobic growth of Campylobacter jejuni

Objective:To produce a technique for the growth of Campylobacter jejuni in aerobic condition Methods:Different combinations of reducing agents were tested in brucella broth and the growth turbidity was compared with tubes containing normal broth only.Microaerophilic environment was also provided in a petri plate seeded with Campylobacter culture by pouring 3 different concentrations（10%,5%and 2.5%） of five reducing agents along with bacto-agar in the lid which was used to cover and seal the culture plate.Six reducing agents were also added in broth in concentration of 0.25 mg/mL of each with different combinations.Results:In lid agar technique,Campylobacter jejuni growth appeared in all three concentrations of reducing agents, that is 10%,5%and 2.5%after 24 hours of incubation but the best results were observed in 10% concentration.The colonial and morphological characters were not affected when the organisms were grown by this technique.Conclusions:It was found that reducing agents enhance the growth of C.jejuni/coli.In combination of FeSO4,Na2CO3 with H3BO3 worked as ideal mixture for the aerobic growth of Campylobacter.This technique is more economical as compared to commercially available media in the market and can be used for the oral facultative and microaerophilic bacterial growth in laboratory.

Antibiotic resistance profile and RAPD analysis of Campylobacter jejuni isolated from vegetables farms and retail markets

Objective:To investigate antibiotic resistance profile and characterize Campylobacter jejuni(C.jejuni) isolates using random amplified polymorphic DNA(RAPD) analysis.Methods:Ninety eight C.jejuni isolates from farms and retail outlets were screened against 10 antibiotics commonly used clinically and agriculturally by using disk diffusion method.RAPD analysis was done to characterize 98 C.jejuni isolates.Results:Fifty-one percent of the isolates had multiple antibiotic resistance index 0.2 and below.This indicated that the isolates in the vegetables were not from the high risk environment or extensive farming practices.C.jejuni isolates found resistant towards penicillin G(93%),vancomycin(86%),ampicillin(35%),erythromycin(28%),gentamycin(4%),amikacin(3%),enrofloxacin(1%),norfloxacin(1%) and no resistance towards ciprofloxacin.RAPD clustering analysis showed that the contamination of C.jejuni in vegetables was likely due to cross contamination at retail markets.Conclusions:C.jejuni contamination in vegetables at retail markets was due to cross contamination.Current finding proved that C.jejuni in small scale vegetables production was less expose towards antibiotic abuse.

胶东地区不同家禽弯曲菌流行分布及耐药特征分析

目的通过了解胶东地区不同禽源弯曲菌的耐药现状及分子流行分布情况,为有效防控禽源弯曲菌对家禽产品和人类健康带来的风险提供依据。方法采用细菌常规分离培养、质谱鉴定、微量肉汤稀释法和多位点序列分型(Multilocus Sequence Typing,MLST)技术,对2021年8月至10月期间,在胶东地区采集的565份泄殖腔样品进行弯曲菌分离鉴定,并别对131株代表性菌株(67株空肠弯曲菌、64株结肠弯曲菌)进行了耐药性及分子分型研究。结果分离菌株对环丙沙星、萘啶酸、四环素显示出高水平耐药,耐药率分别为96.18%、96.18%、94.66%,除2株结肠弯曲菌外,其余62株结肠弯曲菌对这3种药物均为完全耐药(100%);131株菌株中共65株多重耐药,总体多重耐药率为49.62%,其中11株空肠弯曲菌(16.42%)分布在3~5耐,54株结肠弯曲菌(84.38%)分布在3~6耐;在不同禽源分离菌株中,水禽分离菌株耐药程度最严重,其次是肉鸡。MLST分型结果表明,67株空肠弯曲菌共得到72个等位基因、35个序列型,分布较分散,无优势ST型和同源复合体;64株结肠弯曲菌共得到27个等位基因、19个序列型,59.38%(38/64)的菌株为同源复合体CC-828,其中ST-1586序列型最多,其次是ST-825;肉鸡源结肠弯曲菌主要以ST-1586、ST-9944、ST-3735为主,水禽以ST-825、ST-1586居多。结论胶东地区不同家禽中空肠弯曲菌和结肠弯曲菌的携带存在差异,蛋鸡携带两种菌的现象较肉鸡、水禽较为普遍;胶东地区禽源空肠弯曲菌对环丙沙星、萘啶酸、四环素耐药严重,对其它药物敏感性好,结肠弯曲菌对多种抗菌药物耐药严重,且多重耐药普遍;空肠弯曲菌ST型分散,该菌具有较高的遗传多样性,结肠弯曲菌在肉鸡和水禽以ST-1586为主的克隆传播;家禽携带能引起人类严重疾病的空肠弯曲菌,应加强禽源弯曲菌的动态监测。

Effect of Stress-Adaptation on Antibiotic Sensitivity Profiles of <i>Campylobacter jejuni</i>

Campylobacter jejuni is one of the leading causes of human gastroenteritis. Campylobacter jejuni requires special conditions and media in the laboratory for its growth. In nature, however, this organism is able to survive in very diverse and hostile environments and produce disease in humans and animals. The different mechanisms by which C. jejuni survives stressful conditions in the environment still remain unclear. Stress-adaptation may be one of the factors helping this organism to survive stresses. Some C. jejuni strains have been found to have increased antibiotic resistance in last several years. To determine the effect of acid adaptation on the antibiotic sensitivity profile of C. jejuni, 4 different isolates of C. jejuni (a human isolate and 3 poultry isolates) were exposed to an acid pH of 5.5 and then rechallenged with different stresses. The antibiotic sensitivity profiles of C. jejuni after stress-adaptation were compared with antibiotic sensitivity profiles of non-stressed C. jejuni using the Kirby Bauer agar disc diffusion assay. The antibiotic sensitivity profiles of the C. jejuni isolates used in this study were found to change when the acidadapted bacteria were subjected to further stresses such as an acidic pH of 4.5, aerobic atmosphere and starvation. In the majority of the cases, antibiotic-resistant C. jejuni isolates were found to be more sensitive to antibiotics after stress-adaptation, but in a few cases C. jejuni showed increased resistance. These results indicate that increasing various stresses in a sequential pattern may, in some cases, reduce antibiotic resistance of C. jejuni isolates.