Mobile genetic elements facilitate the transmission of antibiotic resistance genes in multidrug-resistant Enterobacteriaceae from duck farms

Multidrug-resistant(MDR)Enterobacteriaceae critically threaten duck farming and public health.The phenotypes,genotypes,and associated mobile genetic elements(MGEs)of MDR Enterobacteriaceae isolated from 6 duck farms in Zhejiang Province,China,were investigated.A total of 215 isolates were identified as Escherichia coli(64.65%),Klebsiella pneumoniae(12.09%),Proteus mirabilis(10.23%),Salmonella(8.84%),and Enterobacter cloacae(4.19%).Meanwhile,all isolates were resistant to at least two antibiotics.Most isolates carried tet(A)(85.12%),blaTEM(78.60%)and sul1(67.44%)resistance genes.Gene co-occurrence analysis showed that the resistance genes were associated with IS26 and integrons.A conjugative IncFII plasmid pSDM004 containing all the above MGEs was detected in Proteus mirabilis isolate SDM004.This isolate was resistant to 18 antibiotics and carried the blaNDM-5 gene.MGEs,especially plasmids,are the primary antibiotic resistance gene transmission route in duck farms.These findings provide a theoretical basis for the rational use of antibiotics in farms which are substantial for evaluating public health and food safety.

Performance Parameters:Demobilization Antibiotic Resistant Bacteria(ARB)and Carrying Genes(ARG)in Wastewater Disinfection

The UV irradiation is used for removing Antibiotic Resistant Bacteria(ARB)and Antibiotic Resistance Genes(ARG)from wastewater treatment.Bacteriophages are viruses that infect within bacteria,are recognized for bacterial control.The influence of some parameters in quantification and performance influencing of pathogen demobilization could be considered in disinfection of wastewater.The comparison of Polyvalent phage(NE1)versus Coliphage(NE4)in suppressing a bacterium Escherichia coli(NDM-1:b-lactam-resistant)with UV irradiation was observed the efficacy in reduction of cells in the disinfection and parameter process.The results with the effect of UV-C irradiation on NDM-1 infected with 1%of NE4 showed a decrease of cells from 8×10^(6)to 2×10^(5)in 60 min with UV-C dose.The NDM1(E.coli)was infected with 1%of NE4(Polyvalent Phage)under magnetic stirring for 1 h,the cells count was 8×10^(6).After 1 h in UV-C e×posure,the cells number reached 3×10^(5).The NDM1 that was e×posed in 1 h of UV-C irradiation and then was infected with 1%of NE4.Cells counting were done 24 h after this procedure.These cells were e×posed in UV-C and showed a reduction in the number of cells from 1×10^(8)to 4×10^(5)after 60 min.The results indicate that bacteriophages can mitigate bacteria species,and combined the conventional water disinfection technologies that can support the microbial safety control strategies.

Trifunctional Cu-Mesh/Cu_(2)O@FeO Nanoarrays for Highly Efficient Degradation of Antibiotic, Inactivation of Antibiotic-Resistant Bacteria, and Damage of Antibiotics Resistance Genes

Trifunctional Cu-mesh/Cu_(2)O@FeO nanoarrays heterostructure is designed and fabricated by integrating CuCu_(2)O@FeO nanoarrays onto Cu-mesh(CM)via an in situ growth and phase transformation process.It is successfully applied to efficiently mitigate the antibiotic pollution,including degradation of antibiotics,inactivation of antibiotic-resistant bacteria(ARB),and damage of antibiotics resistance genes(ARGs).Under visible-light irradiation,CM/CuCu_(2)O@FeO nanoarrays exhibit a superior degradation efficiency on antibiotics(e.g.,up to 99%in 25 min for tetracycline hydrochloride,TC),due to the generated reactive oxygen species(ROS),especially the dominant·O^(2−).It can fully inactivate E.coli(HB101)with initial number of~108 CFU mL^(−1) in 10 min,which is mainly attributed to the synergistic effects of 1D nanostructure,dissolved metal ions,and generated ROS.Meanwhile,it is able to damage ARGs after 180 min of photodegradation,including tetA(vs TC)of 3.3 log 10,aphA(vs kanamycin sulfate,KAN)of 3.4 log 10,and tnpA(vs ampicillin,AMP)of 4.4 log 10,respectively.This work explores a green way for treating antibiotic pollution under visible light.

UV-Based Advanced Oxidation Processes for Antibiotic Resistance Control: Efficiency, Influencing Factors, and Energy Consumption

Antibiotic resistant bacteria(ARB)with antibiotic resistance genes(ARGs)can reduce or eliminate the effectiveness of antibiotics and thus threaten human health.The United Nations Environment Programme considers antibiotic resistance the first of six emerging issues of concern.Advanced oxidation processes(AOPs)that combine ultraviolet(UV)irradiation and chemical oxidation(primarily chlorine,hydrogen peroxide,and persulfate)have attracted increasing interest as advanced water and wastewater treatment technologies.These integrated technologies have been reported to significantly elevate the efficiencies of ARB inactivation and ARG degradation compared with direct UV irradiation or chemical oxidation alone due to the generation of multiple reactive species.In this study,the performance and underlying mechanisms of UV/chlorine,UV/hydrogen peroxide,and UV/persulfate processes for controlling ARB and ARGs were reviewed based on recent studies.Factors affecting the process-specific efficiency in controlling ARB and ARGs were discussed,including biotic factors,oxidant dose,UV fluence,pH,and water matrix properties.In addition,the cost-effectiveness of the UV-based AOPs was evaluated using the concept of electrical energy per order.The UV/chlorine process exhibited a higher efficiency with lower energy consumption than other UV-based AOPs in the wastewater matrix,indicating its potential for ARB inactivation and ARG degradation in wastewater treatment.Further studies are required to address the trade-off between toxic byproduct formation and the energy efficiency of the UV/chlorine process in real wastewater to facilitate its optimization and application in the control of ARB and ARGs.

Fate and Behavior of Tetracycline Resistance Genes in Activated Carbon Adsorption

The accessibility of tetracycline resistance gene (tetG) into the pores of activated carbon (AC), as well as the impact of the pore size distribution (PSD) of AC on the uptake capacity of tetG, were investigated using eight types of AC (four coal-based and four wood-based). AC showed the capability to admit tetG and the average reduction of tetG for coal-based and wood-based ACs at the AC dose of 1 g·L-1 was 3.12 log and 3.65 log, respectively. The uptake kinetic analysis showed that the uptake of the gene followed the pseudo-second-order kinetics reaction, and the uptake rate constant for the coal-based and wood-based ACs was in the range of 5.97 × 10-12 - 4.64 × 10-9 and 7.02 × 10-11 - 1.59 × 10-8 copies·mg-1·min-1, respectively. The uptake capacity analysis by fitting the obtained experiment data with the Freundlich isotherm model indicated that the uptake constant (KF) values were 1.71 × 103 - 8.00 × 109 (copies·g-1)1-1/n for coal-based ACs and 7.00 × 108 - 3.00 × 1010 (copies·g-1)1-1/n for wood-based ones. In addition, the correlation analysis between KF values and pore volume as well as pore surface at different pore size regions of ACs showed that relatively higher positive correlation was found for pores of 50 - 100 Å, suggesting ACs with more pores in this size region can uptake more tetG. The findings of this study are valuable as reference for optimizing the adsorption process regarding antibiotic resistance-related concerns in drinking water treatment.

Dissemination of Resistance Integrons and Genes Coding for Blse and Cabapenemases in the Urban Drainage Network in Cote d’Ivoire

Antibiotic resistance has become a major threat to human health worldwide. Environment, particularly the water environment, has long been overlooked as a player in the antibiotic resistance cycle, although its role remains unclear. These can provide an ideal setting for the acquisition and dissemination of antibiotic resistance, as they are frequently affected by anthropogenic activities. The objective of this study was to establish a diffusion map of resistance integrons used as genetic markers of resistance associated with antibiotic resistance conferring genes (ARGs). Total DNA extracts from non-cultivable bacterial communities were used for the analyses. These communities were obtained from wastewater samples from 14 sites upstream and downstream of drainage channels or effluents in the cities of Abidjan, Bouaké, and Yamoussoukro. The results obtained correspond to the number of positives among the treated samples (n = 39). Among the genetic markers of dissemination, class 1 integrons were the most evident in 94.8% of samples in Abidjan (93.3%), Bouaké (100%) and Yamoussoukro (91.6%). Class 2 integrons and class 3 integrons were found respectively in 41% and 51% of all samples. Genes coding for β-lactamases and blaTEM was identified in almost all samples at a rate of 97.4%. A co-presence of the three genes blaTEM, blaSHV and blaCTX-M is also remarkable in the sites of the city of Yamoussoukro. Among the genes coding for carbapenemases, only blaKPC 17.94%, blaNDM 30.76% and blaOXA48 38.46% were detected in the samples.

The Role of Wastewater Treatment Plants in the Environmental Dissemination of Antibiotic Resistant Bacteria (ARB) and Resistance Genes (ARG)

This study was conducted to evaluate the influence of wastewater treatment processes on the prevalence of antibiotic resistance fecal coliform (FC) and antibiotic resistance genes (ARGs) of FC. In addition, the occurrence of antibiotic resistant bacteria (ARB) and antibiotic resistant genes (ARGs) in surface waters receiving wastewater was evaluated. Greater resistance against penicillin (P), colisitin (CT) and ampicillin (AMP) were observed for FC isolated from effluent disinfected by chlorine (71%), than that disinfected by UV (45%). The greatest resistance against six antibiotics was recorded for FC isolates from effluent disinfected by chlorine. The prevalence of tetB and blaSHV was lowest in isolates from chlorine-disinfected effluents. The occurrence of ARG blaSHV was highest in FC isolated from effluent disinfected by UV. A significant correlation was recorded between FC levels in surface waters and the level of bacterial resistance to ampicillin (P SHV in effluents and in surface waters. TetA and tetC were highly prevalent in surface water compared to tetB. The results of the study demonstrate the widespread prevalence of ARB and ARG in wastewater and receiving water bodies. The result indicates that the source of ARB and ARG in surface waters originate from wastewater. Released ARB and ARG may serve as the source of ARG to pathogenic bacteria in surface waters. Disinfection processes may influence the selection of antibiotic resistant patterns of bacteria.

Analysis of antibiotic resistance genes and their effects on marine environment

The application of overgrowth antibiotic resistance genes in marine research was discussed.Using the China National Knowledge Infrastructure(CNKI)network database to retrieve papers on resistance genes(n=30627)and antibiotic resistance genes(n=3277),the published trends,subject headings and other aspects of bibliometrics were analyzed.Literature in this field has increased rapidly in the past 38 years.55.3%of the papers were published in the last ten years,31.2%of the papers were published in the last five years.The relevance of research topics has increased in recent years,the current research mainly focus on antibiotics,resistance gene,antibiotic resistance gene,and drug resistance.Literature analysis is helpful to understand the overall development trend of overgrowth antibiotic resistance genes in marine research.

Antibiotic resistance analysis and coping strategies of helicobacter pylori

Helicobacter pylori(H.pylori)is a Gram-negative bacterium that mainly colonizes the stomach and duodenum,and it can cause gastrointestinal diseases such as gastric inflammation,peptic ulcer and gastric cancer,and eradication of H.pylori can effectively stop the occurrence and development of gastrointestinal diseases.Antibiotics are one of the main drugs used to treat H.pylori.Due to the long-term application of antibiotics,the resistance rate of H.pylori to antibiotics increases year by year,which greatly reduces the eradication rate of H.pylori and increases the difficulty of re-treatment and the economic burden of patients.In this paper,we will review three aspects of H.pylori resistance status,resistance mechanism and treatment to provide reference for the progress of H.pylori resistance research and its treatment strategy.

Genetic Diversity,Antibiotic Resistance,and Pathogenicity of Aeromonas Species from Food Products in Shanghai,China

Objective Aeromonas has recently been recognized as an emerging human pathogen.Aeromonasassociated diarrhea is a phenomenon occurring worldwide.This study was designed to determine the prevalence,genetic diversity,antibiotic resistance,and pathogenicity of Aeromonas strains isolated from food products in Shanghai.Methods Aeromonas isolates(n=79)collected from food samples were analyzed using concatenated gyrB-cpn60 sequencing.The antibiotic resistance of these isolates was determined using antimicrobial susceptibility testing.Pathogenicity was assessed usingβ-hemolytic,extracellular protease,virulence gene detection,C.elegans liquid toxicity(LT),and cytotoxicity assays.Results Eight different species were identified among the 79 isolates.The most prevalent Aeromonas species were A.veronii[62(78.5%)],A.caviae[6(7.6%)],A.dhakensis[3(3.8%)],and A.salmonicida[3(3.8%)].The Aeromonas isolates were divided into 73 sequence types(STs),of which 65 were novel.The isolates were hemolytic(45.6%)and protease-positive(81.0%).The most prevalent virulence genes were act(73.4%),fla(69.6%),aexT(36.7%),and ascV(30.4%).The results of C.elegans LT and cytotoxicity assays revealed that A.dhakensis and A.hydrophila were more virulent than A.veronii,A.caviae,and A.bivalvium.Antibiotic resistance genes[tetE,blaTEM,tetA,qnrS,aac(6)-Ib,mcr-1,and mcr-3]were detected in the isolates.The multidrug-resistance rate of the Aeromonas isolates was 11.4%,and 93.7%of the Aeromonas isolates were resistant to cefazolin.Conclusion The taxonomy,antibiotic resistance,and pathogenicity of different Aeromonas species varied.The Aeromonas isolates A.dhakensis and A.hydrophila were highly pathogenic,indicating that food-derived Aeromonas isolates are potential risks for public health and food safety.The monitoring of food quality and safety will result in better prevention and treatment strategies to control diarrhea illnesses in China.

Enhanced removal of antibiotic resistance genes during chicken manure composting after combined inoculation of Bacillus subtilis with biochar

This study explored the combined effects of Bacillus subtilis inoculation with biochar on the evolution of bacterial communities,antibiotic resistance genes(ARGs),and mobile genetic elements(MGEs)during the composting of chicken manure.The results showed that B.subtilis inoculation combined with biochar increased bacterial abundance and diversity as well as prolonged the compost thermophilic period.Promoted organic matter biodegradation and facilitated the organic waste compost humification process,reduced the proliferation of ARGs by altering the bacterial composition.Firmicutes and Actinobacteriota were the main resistant bacteria related to ARGs and MGEs.The decrease in ARGs and MGEs was associated with the reduction in the abundance of related host bacteria.Compost inoculation with B.subtilis and the addition of biochar could promote nutrient transformation,reduce the increase in ARGs and MGEs,and increase the abundance of beneficial soil taxa.

微藻生物系统去除抗生素和ARGs的研究进展

抗生素与抗生素抗性基因(ARGs)污染严重危害人类健康,微藻生物技术在处理抗生素废水过程中表现出良好的效果。文章首先总结了微藻对于不同种类抗生素的去除机理,包括生物吸附、生物累积、生物降解、光生物降解和水解挥发5种。分析了微藻在去除抗生素过程中的重要影响因素,包括pH、水力停留时间、光照、抗生素的种类与浓度和微藻的种类。并总结了微藻对ARGs的去除机制,包括移动基因元件的去除及活性氧自由基的猝灭。最后,文章指出了微藻处理抗生素废水过程中仍然存在的问题,进一步明确了今后微藻生物技术处理抗生素废水的重点研究方向。

Distribution characteristics and removal rate of antibiotics and antibiotic resistance genes in different treatment processes of two drinking water plants

Emerging pollutants,such as antibiotics and antibiotic-resistance genes,are becoming increasingly important sources of safety and health concerns.Drinking water safety,which is closely related to human health,should receive more attention than natural water body safety.However,minimal research has been performed on the efficacy of existing treatment processes in water treatment plants for the removal of antibiotics and antibiotic resistance genes.To address this research gap,this study detected and analyzed six main antibiotics and nine antibiotic resistance genes in the treatment processes of two drinking water plants in Wuhan.Samples were collected over three months and then detected and analyzed using ultra-high-performance liquid chromatography-tandem mass spectrometry and fluorescence quantitation.The total concentrations of antibiotics and antibiotic resistance genes in the influent water of the two water plants were characterized as December>March>June.The precipitation and filtration processes of the Zou Maling Water Plant and Yu Shidun Water Plant successfully removed the antibiotics.The ozone-activated carbon process increased the removal rate of most antibiotics to 100%.However,a large amount of antibiotic resistance gene residues remained in the effluents of the two water plants.The experiments demonstrated that the existing ozone-activated carbon processes could not effectively remove antibiotic resistance genes.This study provides a reference for the optimization of drinking water treatment processes for antibiotics and antibiotic resistance gene removal.

Evaluating human exposure to antibiotic resistance genes

Antibiotic resistance is an escalating global concern,leading to millions of annual fatalities.Antibiotic resistance genes(ARGs)present in bacteria equip them to withstand the effects of antibiotics.Intra-and interspecific ARGs transmission through horizontal gene transfer further exacerbates resistance dissemination.The presence of ARGs in the environment heightens the probability of human exposure via direct inhalation,ingestion,or contact with polluted air,food,or water,posing substantial biosafety and health hazards.Consequently,ARGs represent a critical focal point in public health and environmental safety and are classified as emerging contaminants.This perspective underscores the necessity to assess ARG exposure within the One Health framework and to accord greater attention to the mitigation strategies and tactics associated with ARGs.

Health risk ranking of antibiotic resistance genes in the Yangtze River

Antibiotic resistance is an escalating global health concern,exacerbated by the pervasive presence of antibiotic resistance genes(ARGs)in natural environments.The Yangtze River,the world's third-longest river,traversing areas with intense human activities,presents a unique ecosystem for studying the impact of these genes on human health.Here,we explored ARGs in the Yangtze River,examining 204 samples from six distinct habitats of approximately 6000 km of the river,including free-living and particle-associated settings,surface and bottom sediments,and surface and bottom bank soils.Employing shotgun sequencing,we generated an average of 13.69 Gb reads per sample.Our findings revealed a significantly higher abundance and diversity of ARGs in water-borne bacteria compared to other habitats.A notable pattern of resistome coalescence was observed within similar habitat types.In addition,we developed a framework for ranking the risk of ARG and a corresponding method for calculating the risk index.Applying them,we identified water-borne bacteria as the highest contributors to health risks,and noted an increase in ARG risks in particle-associated bacteria correlating with heightened anthropogenic activities.Further analysis using a weighted ARG risk index pinpointed the ChengdueChongqing and Yangtze River Delta urban agglomerations as regions of elevated health risk.These insights provide a critical new perspective on ARG health risk assessment,highlighting the urgent need for strategies to mitigate the impact of ARGs on human health and to preserve the ecological and economic sustainability of the Yangtze River for future human use.

Occurrence and possible sources of antibiotic resistance genes in seawater of the South China Sea

Antibiotic resistance genes(ARGs)might have great effect on ecological security and human health.Oceans are important reservoirs that receive tremendous amounts of pollutants globally.However,information on the proliferation of ARGs in seawater is still limited.This study performed field sampling to investigate the occurrence and distribution of ARGs in seawater of the South China Sea,which is the deepest and largest sea in China.The results showed that the total absolute abundances of ARGs in seawater samples ranged from 2.1×10^(3)to 2.3×10^(4)copies/mL,with an of 5.0×10^(3)copies/mL and a range of 2.2×10^(3)–1.8×10^(4)copies/mL for those with mobile genetic elements(MGEs).Genes resistant to multidrug,aminoglycoside,tetracycline,and fluoroquinolone antibiotics accounted for 77.3%–88.6%of total ARGs in seawater.Proteobacteria and Cyanobacteria represented 32.1%–56.2%and 30.4%–49.5%of microbial community,respectively.Prochlorococcus_MIT9313 and Clade_la were the prevalent genera in seawater of the South China Sea.Complex co-occurrence relationship existed among ARGs,MGEs,and bacteria.Anthropogenic activities had critical influence on ARGs and MGEs.Hospital wastewater,wastewater treatment plant effluent,sewage,aquaculture tailwater,and runoff were determined as the important sources of ARGs in seawater of the South China Sea based on positive matrix factorization analysis.

Nanoplastics promote the dissemination of antibiotic resistance genes and diversify their bacterial hosts in soil

The wide application of plastics has led to the ubiquitous presence of nanoplastics and microplastics in terrestrial environments.However,few studies have focused on the mechanism underlying the effects of plastic particles on soil microbiomes and resistomes,especially the differences between nanoplastics and microplastics.This study investigated the microbiome and resistome in soil exposed to polystyrene microplastics(mPS)or nanoplastics(nPS)through 16S rRNA and shotgun metagenomic sequencing.Distinct microbial communities were observed between mPS and nPS exposure groups,and nPS exposure significantly changed the bacterial composition even at the lowest amended rate(0.01%,w/w).The abundance of antibiotic resistance genes(ARGs)in nPS exposure(1%)was 0.26 copies per cell,significantly higher than that in control(0.21 copies per cell)and mPS exposure groups(0.21 copies per cell).It was observed that nanoplastics,bacterial community,and mobile genetic elements(MGEs)directly affected the ARG abundance in nPS exposure groups,while in mPS exposure groups,only MGEs directly induced the change of ARGs.Streptomyces was the predominant host for multidrug in the control and mPS exposure,whereas the primary host was changed to Bacillus in nPS exposure.Additionally,exposure to nPS induced several bacterial hosts to exhibit possible multi-antibiotic resistance characteristics.Our results indicated that the effects of plastic particles on the soil microbial community were size-dependent,and nano-sized plastic particles exhibited more substantial impacts.Both microplastics and nanoplastics promoted ARG transfer and diversified their bacterial hosts.These findings bear implications for the regulation of plastic waste and ARGs.

环境中抗生素抗性基因(ARGs)形成和传播机制研究进展

从DNA损伤和错误修复角度阐述了环境中抗生素抗性基因(Antibiotics resistance genes,ARGs)的形成机制,并对DNA损伤和错误修复过程中涉及的相关核心功能基因片段进行归纳总结。也从细胞代谢角度阐述了环境中ARGs进行水平传播的三种主要机制,包括:菌毛架桥作用、信号传导和IV型细胞分泌系统。现有研究进一步加强对ARGs在环境中形成和传播机制的认识,为今后研究ARGs的削减提供理论依据。目前关于抗生素诱导ARGs形成和传播的内在机制仍相对缺乏,后续可从生物毒性和分子生物学层面进行深入研究。

沈阳典型粳稻种植区稻田土壤ARGs和MGEs污染分布特征分析

农田土壤中抗生素抗性基因(ARGs)与可移动遗传元件(MGEs)的污染问题,在学术界已引发广泛关注。为研究沈阳市粳稻种植区稻田土壤中ARGs与MGEs的存在和相关性,并探讨其对土壤-植物系统的潜在影响,通过采集沈阳市辽中区、新民市、沈北新区和苏家屯区的粳稻种植区域土壤样品,利用高通量qPCR技术,对285个ARGs、10个MG-Es和一个16S rRNA基因进行检测和定量分析。结果表明:在不同地区的稻田土壤中共检测到273种ARGs和MGEs,且种类和丰度存在显著差异。进一步的相关性分析发现,ARGs与MGEs之间存在一定相关性,并且这种相关性在不同地区间有所差异。多种ARGs与MGEs之间存在显著正相关(p<0.05),其中部分ARGs[tet(32)等]与MGEs(tnpA-01等)存在及极显著正相关(p<0.01),这表明在土壤中MGEs对ARGs迁移和扩散起到促进作用。4个采样区根际土中ARGs和MGEs的总相对丰度均高于非根际土壤,这进一步凸显根际环境在ARGs和MGEs分布中的重要性。此外,还发现在不同地区的稻田土壤中,抗生素失活机制在ARGs中占据主导地位。综上所述,试验结果为深入了解东北水稻种植区稻田土壤中ARGs的污染现状及其潜在影响提供了数据支持和理论基础。这些发现不仅有助于认识农田土壤中ARGs和MGEs的分布与迁移规律,还为制定针对性的土壤污染防控措施提供了科学依据。

Effect of biofilm formation by clinical isolates of Helicobacter pylori on the efflux-mediated resistance to commonly used antibiotics

To evaluate the role of biofilm formation on the resistance of Helicobacter pylori (H. pylori) to commonly prescribed antibiotics, the expression rates of resistance genes in biofilm-forming and planktonic cells were compared.METHODSA collection of 33 H. pylori isolates from children and adult patients with chronic infection were taken for the present study. The isolates were screened for biofilm formation ability, as well as for polymerase chain reaction (PCR) reaction with HP1165 and hp1165 efflux pump genes. Susceptibilities of the selected strains to antibiotic and differences between susceptibilities of planktonic and biofilm-forming cell populations were determined. Quantitative real-time PCR (qPCR) analysis was performed using 16S rRNA gene as a H. pylori-specific primer, and two efflux pumps-specific primers, hp1165 and hefA.RESULTSThe strains were resistant to amoxicillin, metronidazole, and erythromycin, except for one strain, but they were all susceptible to tetracycline. Minimum bactericidal concentrations of antibiotics in the biofilm-forming cells were significantly higher than those of planktonic cells. qPCR demonstrated that the expression of efflux pump genes was significantly higher in the biofilm-forming cells as compared to the planktonic ones.CONCLUSIONThe present work demonstrated an association between H. pylori biofilm formation and decreased susceptibility to all the antibiotics tested. This decreased susceptibility to antibiotics was associated with enhanced functional activity of two efflux pumps: hp1165 and hefA.