사람 및 가축 유래 분변 미생물 군집과 항생제 내성 유전자 간 상관 관계에 대한 연구

정지원,반다리 아프라지타,운노 타쯔야 한국환경농학회 2022 한국환경농학회지 Vol.41 No.4

BACKGROUND: Antibiotics used in animal husbandry for disease prevention and treatment have resulted in the rapid progression of antibiotic resistant bacteria which can be introduced into the environment through livestock feces/manure, disseminating antibiotic resistant genes (ARGs). In this study, fecal samples were collected from the livestock farms located in Jeju Island to investigate the relationship between microbial communities and ARGs. METHODS AND RESULTS: Illumina MiSeq sequencing was applied to characterize microbial communities within each fecal sample. Using quantitative PCR (qPCR), ten ARGs encoding tetracycline resistance (tetB, tetM), sulfonamide resistance (sul1, sul2), fluoroquinolone resistance (qnrD, qnrS), fluoroquinolone and aminoglycoside resistance (aac(6')-Ib), beta-lactam resistance (blaTEM, blaCTX-M), macrolide resistance (ermC), a class 1 integronsintegrase gene (intI1), and a class 2 integrons-integrase gene (intI2) were quantified. The results showed that Firmicutes and Bacteroidetes were dominant in human, cow, horse, and pig groups, while Firmicutes and Actinobacteria were dominant in chicken group. Among ARGs, tetM was detected with the highest number of copies, followed by sul1 and sul2. Most of the genera belonging to Firmicutes showed positive correlations with ARGs and integron genes. There were 97, 34, 31, 25, and 22 genera in chicken, cow, pig, human, and horse respectively which showed positive correlations with ARGs and integron genes. In network analysis, we identified diversity of microbial communities which correlated with ARGs and integron genes. CONCLUSION(S): In this study, antibiotic resistance patterns in human and livestock fecal samples were identified. The abundance of ARGs and integron genes detected in the samples were associated with the amount of antibiotics commonly used for human and livestocks. We found diverse microbial communities associated with antibiotics resistance genes in different hosts, suggesting that antibiotics resistance can disseminate across environments through various routes. Identifying the routes of ARG dissemination in the environment would be the first step to overcome the challenge of antibiotic resistance in the future.

Tetracycline‑resistant bacteria and ribosomal protection protein genes in soils from selected agricultural fields and livestock farms

Nogrado Kathyleen,Unno Tatsuya,Hur Hor-Gil,Lee Ji-Hoon 한국응용생명화학회 2021 Applied Biological Chemistry (Appl Biol Chem) Vol.64 No.3

Antibiotic resistance in soil environment has eminently been compared and studied between agricultural and pristine soils, and the role of concentrated animal feeding operations has markedly been recognized as one of the major sources of antibiotic resistance. This study described the tetracycline resistance in small-scale farms in pursuit of presenting its possible role and contribution to the persistence of antibiotic resistance in the environment. Results of the study would render additional information on the occurrence of the ribosomal protection protein (RPP) tet genes among the isolated bacteria from the selected agricultural soils. Four tetracycline resistance and RPP genes were determined in two different agricultural soil settings. Both the culture and molecular method were used to determine and measure tetracycline resistance in soils from arable land and animal house. Results revealed a significantly higher number of culturable antibiotic-resistant bacteria in animal houses than arable lands which was suggestive of higher antibiotic resistance in areas where there was direct administration of the antibiotics. However, quantification of the gene copy numbers in the agricultural soils indicated a different result. Higher gene copy number of tetO was determined in one animal house (IAH-3), while the two other tet genes tetQ and tetW were found to be higher in arable lands. Of the total 110 bacterial isolates, tetW gene was frequently detected, while tetO gene was absent in any of the culturable bacterial isolates. Principal component analysis of occurrence and gene copy number of RPP tet genes tetO, tetQ, and tetW also revealed highest abundance of RPP tet genes in the manure and arable soils. Another important highlight of this study was the similarity of the RPP tet genes detected in the isolated bacteria from the agricultural soils to the identified RPP tet genes among pathogenic bacteria. Some of the tetracycline-resistant bacterial isolates were also multidrug resistant as it displayed resistance to tetracycline, erythromycin, and streptomycin using disk diffusion testing.

Comparison of Harboring the Resistance Gene and Disc Diffusion Susceptibility Test Result in Staphylococcus pseudintermedius from the Bacterial Dermatitis

장혜진,손형원,강효민,한재익,나기정 한국임상수의학회 2015 한국임상수의학회지 Vol.32 No.2

Bacterial dermatitis is common disease that is necessary to treat with antibiotics. In recent, antibiotic-resistantbacteria is being increased in worldwide. The purpose of the present study was to evaluate the prevalence of resistantgenes in Staphylococcus (S.) pseudintermedius isolated from dogs, and to compare the resistant gene profile with theresult of antibiotic disc diffusion test. A total of seven S. pseudintermedius was included in the study. Bacterialidentification was performed by 16S ribosomal RNA gene sequence analysis. S. pseudintermedius isolates had morethan one antibiotic resistant gene (mecA, blaZ and aac(6')/aph(2"). While all isolates were PCR positive to blaZ gene,only two isolates were resistant to amoxicillin/clavulanate. Among five isolates harboring gentamicin resistance, oneisolate was negative to aac(6')/aph(2")-targeted PCR. Taken together, the results suggest that resistant gene-targetedPCR and disc diffusion test are complementary to detect antibiotic resistance. Bacterial dermatitis is common disease that is necessary to treat with antibiotics. In recent, antibiotic-resistant bacteria is being increased in worldwide. The purpose of the present study was to evaluate the prevalence of resistant genes in Staphylococcus (S.) pseudintermedius isolated from dogs, and to compare the resistant gene profile with the result of antibiotic disc diffusion test. A total of seven S. pseudintermedius was included in the study. Bacterial identification was performed by 16S ribosomal RNA gene sequence analysis. S. pseudintermedius isolates had more than one antibiotic resistant gene (mecA, blaZ and aac(6')/aph(2"). While all isolates were PCR positive to blaZ gene, only two isolates were resistant to amoxicillin/clavulanate. Among five isolates harboring gentamicin resistance, one isolate was negative to aac(6')/aph(2")-targeted PCR. Taken together, the results suggest that resistant gene-targeted PCR and disc diffusion test are complementary to detect antibiotic resistance.

Antibiotic resistance fate in the full-scale drinking water and municipal wastewater treatment processes: A review

Rui Gao,Minghao Sui 대한환경공학회 2021 Environmental Engineering Research Vol.26 No.4

Antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) have been widely identified as emerging pollutants in various aquatic compartments. Concerns have been raised that the antibiotic resistance determinants may occur in treated drinking water and wastewater, weakening the therapeutic efficacy of antibiotics and so posing threat to public health. Most often, laboratory trials are conducted to assess the removal efficiency of ARB and ARGs in individual treatment processes (e.g., disinfection). However, the random variations of microbial distribution and chemical compositions in real-world environments cannot be entirely simulated, presumably leading to false-positive results as a consequence. It is therefore useful to provide a summary of recent advancements regarding the antibiotic resistance attenuation during full-scale water/wastewater treatments, which has not been adequately evaluated so far. In this review, the prevalence, proliferation and transmission of ARB and ARGs in urban water cycle, coupled with corresponding detection methods are presented as a short overview. The fate of ARB and ARGs in the sequential drinking water and wastewater processing units is critically summarized, the aim of which is to provide guidance for improving the current water treatment facilities to further reduce the antibiotic resistance in finished water.

Removal of antibiotics from wastewater and its problematic effects on microbial communities by bioelectrochemical Technology: Current knowledge and future perspectives

Mahdi Hassan,Guangcan Zhu,Yong-ze LU,Ali Hamoud AL-Falahi,Yuan LU,Shan Huang,Ziren Wan 대한환경공학회 2021 Environmental Engineering Research Vol.26 No.1

In this review, antibiotics are considered an emerging pollutant that has drawn worldwide attention in recent years. Therefore, the effective removal of antibiotic contaminants has become a hot issue in the field of environmental research. Most antibiotics applied to humans eventually enter municipal Wastewater Treatment Plants (WWTPs), because there are no appropriate commercially available pretreatment techniques. However, increasing anthropogenic activities, the high demand for animal-protein in developing countries as a nutritional alternative, and the extensive usage of antibiotics are mainly responsible for the persistence of antibiotic pollutants. One of the serious concerns regarding the presence of antibiotics in water and their potential role in exacerbating the emergence of antibiotics-resistance bacteria (ARB) and antibiotics-resistance genes (ARGs). In recent years, bioelectrochemical technologies are found promising for suppressing antibiotic contaminants through microbial metabolism and electrochemical redox reactions. Therefore, this review provides up-to-date insight research on bioelectrochemical systems (BESs), which improves the removal of the antibiotic in an efficient way. The focus of this review has been on the environmental sources of antibiotics, their health effects and possible degradation pathways, bacterial-antibiotics resistance mechanisms, and treatment of antibiotic-contained water using BES technology.

Metagenomic analysis reveals the prevalence and persistence of antibiotic- and heavy metal-resistance genes in wastewater treatment plant

Sachin Kumar Gupta,신한섭,한덕기,허호길,Tatsuya Unno 한국미생물학회 2018 The journal of microbiology Vol.56 No.6

The increased antibiotic resistance among microorganisms has resulted into growing interest for investigating the wastewater treatment plants (WWTPs) as they are reported to be the major source in the dissemination of antibiotic resistance genes (ARGs) and heavy metal resistance genes (HMRGs) in the environment. In this study, we investigated the prevalence and persistence of ARGs and HMRGs as well as bacterial diversity and mobile genetic elements (MGEs) in influent and effluent at the WWTP in Gwangju, South Korea, using high-throughput sequencing based metagenomic approach. A good number of broad-spectrum of resistance genes (both ARG and HMRG) were prevalent and likely persistent, although large portion of them were successfully removed at the wastewater treatment process. The relative abundance of ARGs and MGEs was higher in effluent as compared to that of influent. Our results suggest that the resistance genes with high abundance and bacteria harbouring ARGs and MGEs are likely to persist more through the treatment process. On analyzing the microbial community, the phylum Proteobacteria, especially potentially pathogenic species belonging to the genus Acinetobacter, dominated in WWTP. Overall, our study demonstrates that many ARGs and HMRGs may persist the treatment processes in WWTPs and their association to MGEs may contribute to the dissemination of resistance genes among microorganisms in the environment.

Prevalence of multidrug-resistant (MDR) Escherichia coli in untreated effluents from a wastewater treatment plant (WWTP) in Dhaka, Bangladesh

Md Naeem Hossain,Asim kumar Roy,Humayra Habib,Zenat Zebin Hossain,Humaira Akhter,Anowara Begum 한국미생물학회 2022 미생물학회지 Vol.58 No.3

The widespread use of antibiotics in human and animal husbandry results in the potential release of antibiotics into surrounding environments and leads to the emergence of antibiotic-resistant bacteria and antibiotic resistance genes. With this concern, we aimed to study the molecular mechanisms associated with multidrug resistance (MDR) and biofilm formation capacity in Escherichia coli isolates from Dhaka City’s wastewater treatment plant. A total of 37 antimicrobialresistant E. coli isolates were analyzed for the presence of genes associated with antibiotic resistance and biofilm formation using several phenotypic and polymerase chain reaction (PCR) assays and curing plasmids through the SDS method. Among the 37 isolates, most prevalent gene detected was qnrS (32.43%), followed by blaTEM (29.73%), blaCTX-M-15 (24.32%), dfrA17 (21.62%), sul2 (8.1%), qnrB (5.41%), sul1 (5.41%), and dfrA1 (2.70%). Twelve E. coli isolates (32.43%) possessed only class 1 integrons. A comparison of results prior to and after plasmid curing revealed most E. coli had plasmid-mediated antibiotic resistance. Among the 37 isolates, nine strains showed weak biofilm production and one showed strong biofilm production. All of the isolates were curli producers; fimH and csgA genes were present in 72.9% and 64.9% of the isolates, respectively. The dynamic antibiotic resistance diversity revealed in this study may pose concern regarding the potential development of drug-resistant bacteria and their dissemination into the surrounding environment via WWTPs.

Gain and loss of antibiotic resistant genes in multidrug resistant bacteria: One Health perspective

Kim Misung,Park Jaeeun,Kang Mingyeong,Yang Jihye,Park Woojun 한국미생물학회 2021 The journal of microbiology Vol.59 No.6

The emergence of multidrug resistance (MDR) has become a global health threat due to the increasing unnecessary use of antibiotics. Multidrug resistant bacteria occur mainly by accumulating resistance genes on mobile genetic elements (MGEs), made possible by horizontal gene transfer (HGT). Humans and animal guts along with natural and engineered environments such as wastewater treatment plants and manured soils have proven to be the major reservoirs and hotspots of spreading antibiotic resistance genes (ARGs). As those environments support the dissemination of MGEs through the complex interactions that take place at the human-animalenvironment interfaces, a growing One Health challenge is for multiple sectors to communicate and work together to prevent the emergence and spread of MDR bacteria. However, maintenance of ARGs in a bacterial chromosome and/or plasmids in the environments might place energy burdens on bacterial fitness in the absence of antibiotics, and those unnecessary ARGs could eventually be lost. This review highlights and summarizes the current investigations into the gain and loss of ARG genes in MDR bacteria among human-animal- environment interfaces. We also suggest alternative treatments such as combinatory therapies or sequential use of different classes of antibiotics/adjuvants, treatment with enzymeinhibitors, and phage therapy with antibiotics to solve the MDR problem from the perspective of One Health issues.

Genomic characterization of extensively drug-resistant <i>Acinetobacter baumannii</i> strain, KAB03 belonging to ST451 from Korea

Lee, Sang-Yeop,Oh, Man Hwan,Yun, Sung Ho,Choi, Chi-Won,Park, Edmond Changkyun,Song, Hyun Seok,Lee, Hayoung,Yi, Yoon-Sun,Shin, Juhyun,Chung, Chaeuk,Moon, Jae Young,Lee, Je Chul,Kim, Gun-Hwa,Kim, Seung Elsevier 2018 INFECTION GENETICS AND EVOLUTION Vol.65 No.-

<P><B>Abstract</B></P> <P>Extensively drug-resistant (XDR) <I>Acinetobacter baumannii</I> strains have emerged rapidly worldwide. The antibiotic resistance characteristics of XDR <I>A. baumannii</I> strains show regional differences; therefore, it is necessary to analyze both genomic and proteomic characteristics of emerging XDR <I>A. baumannii</I> clinical strains isolated in Korea to elucidate their multidrug resistance. Here, we isolated new sequence type of XDR <I>A. baumannii</I> clinical strain (KAB03) from Korean hospitals and performed comprehensive genome analyses. The strain belongs to new sequence type, ST451. Single nucleotide polymorphism (SNP) analysis with other types of <I>A. baumannii</I> strains revealed that KAB03 has unique SNP pattern in the regions of <I>gyrB</I> and <I>gpi</I> of MLST profiles. <I>A. baumannii</I> KAB03 harbours three antibiotic resistance islands (AbGRI1, 2, and 3). AbGRI1 harbours two copies of Tn<I>2006</I> containing <I>bla</I> <SUB>OXA-23</SUB>, which play an important role in antibiotic resistance. AbGRI2 possesses aminoglycoside resistant gene <I>aph(3′)-Ic</I> and class A β-lactamase <I>bla</I> <SUB>TEM</SUB>. AbGIR3 has macrolide resistant genes and aminoglycoside resistant gene <I>armA</I>. <I>A. baumannii</I> KAB03 harbours mutations in <I>pmrB</I> and <I>pmrC</I>, which are believed to confer colistin resistance. In addition, proteomic and transcriptional analysis of KAB03 confirmed that β-lactamases (ADC-73 and OXA-23), Ade efflux pumps (AdeIJK), outer membrane proteins (OmpA and OmpW), and colistin resistance genes (PmrCAB) were major proteins responsible for antibiotic resistance. Our proteogenomic results provide valuable information for multi-drug resistance in emerging XDR <I>A. baumannii</I> strains belonging to ST451.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Whole genome of an XDR <I>A. baumannii</I> KAB03 belonging to ST451, isolated in South Korea, was analyzed </LI> <LI> <I>A.s baumannii</I> strains belonging to ST451 have unique SNP pattern in the regions of <I>gyrB</I> and <I>gpi</I> of MLST profiles </LI> <LI> Antibiotic resistance proteins of <I>A. baumannii</I> KAB03 were suggested by proteomic and transcriptomic analysis </LI> </UL> </P>

Survey for Correlation between Biofilm Formation and Virulence Determinants in a Collection of Pathogenic and Fecal Enterococcus faecalis Isolates

Fereshteh Saffari,Mohammad Sadegh Dalfardi,Shahla Mansouri,Roya Ahmadrajabi 대한감염학회 2017 Infection and Chemotherapy Vol.49 No.3

Background: Enterococcus faecalis is an opportunistic pathogen that causes most of the enterococcal infections. Among the different factors implicated in the pathogenesis of these organisms, biofilm formation and antibiotic resistance are the most important. The ability for biofilm formation has been attributed to the presence of some virulence genes. However, no definite correlation has been found. This study aimed to detect biofilm formation and antibiotic resistance patterns in E. faecalis isolates collected from clinical and fecal samples, and to investigate possible correlation between some virulence genes (esp, cyl, gelE) and biofilm formation. Materials and Methods: A collection of 123 E. faecalis isolates were investigated for antibiotic resistance and production of hemolysin, gelatinase, and biofilm using phenotypic methods. The esp, gelE and cyl genes were detected using polymerase chain reaction. Results: Thirty-eight pathogenic isolates (37%) were positive for biofilm formation. Additionally, the gelE, esp, and cyl genes were detected in 74 (71.8%), 79 (76.7%) and 42 (40.8%) isolates, respectively. In the fecal samples, 18 (90%) isolates were biofilm producers and 11 (55%), 17 (85%) and 8 (40%) isolates were positive for gelE, esp, and cyl, respectively. There were significant differences in biofilm production between pathogenic and fecal isolates (P <0.001). Multidrug resistance (MDR) was found among 32% (n = 33) and 15% (n = 3) of the clinical and fecal isolates, respectively. However, no significant difference was seen between MDR and biofilm formation. Five pathogenic and two fecal isolates were negative for all investigated genes while they were they were biofilm producers. In contrast, 22 pathogenic isolates and 1 fecal isolate were positive for the tested genes, but did not form any biofilm. No significant differences were observed between biofilm formation and the presence of the esp, gelE and cyl genes in the pathogenic and fecal isolates (P ˃0.05). Conclusion: The presence of the esp, gelE and cyl genes might not be determining factors for biofilm formation in enterococci and other mechanisms might be involved in this process.