qnrS Genes Research Articles

Emergence of blaOXA-181-bearing tigecycline-resistant Klebsiella aerogenes in China.

We report the isolation of a blaOXA-181-positive, tigecycline-resistant Klebsiella aerogenes strain KA04 from a Chinese inpatient's fecal sample. Species identification was performed using MALDI-TOF MS. The antibiotic susceptibilities were assessed via the broth microdilution method. To elucidate the transmission and genetic structure of the blaOXA-181 gene, conjugation assays and whole-genome sequencing (WGS) were performed. KA04 displayed resistance to carbapenems, quinolones, piperacillin/tazobactam and tigecycline. Through WGS and conjugation experiments, it was possible to confirm blaOXA-181 and qnrS1 genes causing antibiotic resistance were located on a 51-kb IncX3 type mobile plasmid, blaOXA-181 gene could be successfully transferred into E. coli EC600 at a conjugation frequency of 1.1 × 10- 4. tet(A) gene was located on both the chromosome and non-transmissible IncFIB(K) plasmid. This is a tigecycline-resistant K. aerogenes harboring blaOXA-181 isolate from human fecal sample, highlighting a significant public health concern. Further comprehensive surveillance is needed.

A Study on Extended Spectrum β-Lactamase Producing (ESBL) and Fluoroquinolone-Resistant Escherichia coli in the Japanese Quails

Antimicrobial resistance (AMR) is a growing public health threat worldwide, and members of the Enterobacteriaceae family are among the clinically important bacteria rapidly developing resistance to available antibacterial agents. The development of extended-spectrum β-lactamase (ESBL) enzymes is the primary cause of resistance to third-generation cephalosporins among Enterobacteriaceae. Quails also represent a source of animal protein deficiency in developing countries including India. This study aimed to detect the presence of extended-spectrum β-lactamases (ESBLs) and fluoroquinolone-resistant Escherichia coli from fecal samples of quails in Puducherry. A total of 36 fecal samples of quails from six different retail outlets were collected. E. coli could be isolated from 27 fecal samples and confirmed as E. coli by polymerase chain reaction by targeting the uspA gene. The isolates were preliminarily screened for ESBL production by indicator antimicrobial disc and phenotypic ESBL production was confirmed by combination disc method. The isolates were subjected to genotypic detection of ESBL (bla CTX-M, bla TEM, and bla SHV) by PCR. Out of 27 isolates, 16 isolates were positive for the presence of ESBL-producing genes, in which, 9 harbored bla CTX-M, 2 harbored bla TEM, and 5 harboured bla SHV genes. Similarly, out of 27 isolates 13 isolates were found positive for fluoroquinolone resistance in which 13 (48.18%) isolates harbored the qnrS gene, and 11 (40.74%) isolates harbored the qnrB gene. Antimicrobial susceptibility test showed that the isolates exhibited a high level of resistance against Cefpodoxime (90%), Amoxycillin/clavulanic acid (70%), Ceftriaxone (60%), Cefotaxime (55%), Ceftazidime (45%), Ciprofloxacin (40%) and Aztreonam (30%).

Genetic and in-silico approaches for investigating the mechanisms of ciprofloxacin resistance in Salmonella typhi: Mutations, extrusion, and antimicrobial resistance

Salmonella enterica serovar Typhi spreads typhoid infection in humans through the consumption of contaminated food and water. Poor sanitation plays a pivotal role in its dissemination. Over time, the bacterium has acquired resistance to many promising antibiotics, posing a growing global health concern and hindering the achievement of sustainable development goals. This study aims to elucidate the molecular complexity of fluoroquinolone resistance, a first-line treatment for typhoid infection. To achieve this aim, 80 clinical isolates were collected from various diagnostic laboratories. These isolates were confirmed based on morphological characteristics and biochemical tests. Multidrug-resistant (MDR) and extensively drug-resistant (XDR) isolates were identified using the Kirby-Bauer disc diffusion method. The mechanism of ciprofloxacin resistance was investigated by sequencing the quinolone resistance-determining region (QRDR) genes and identifying the presence of the qnrS1 gene. As a result of this study, 60 % of isolates showed resistance to ciprofloxacin. At the same time, the qnrS1 gene was present in all the selected strains while mutation analysis identified significant mutation in QRDR of DNA gyrase subunit A (gyrA) and Topoisomerase IV (parC) gene. The combinatorial effect was further investigated by downloading 286 draft genomes. The Mutation analysis reveals significant mutations at gyrA S83F, gyrA D87N, gyrA S83Y, gyrB S464F, parC S80I, and parE L416F. Additionally, docking analysis indicates reduced binding affinity and altered solvent accessibility, which show the structural changes at mutation sites. This study provides crucial insights that mutation reduces the binding affinity while qnrS1 acts as a transport channel to extrude the ciprofloxacin. In the future, further validation through experimental mutagenesis is recommended, for targeted therapeutic interventions against the mounting threat of antibiotic-resistant S. Typhi.

Whole-genome sequencing of three ciprofloxacin-resistant Salmonella Reading (ST93) strains, an emerging Salmonella serovar in the poultry sector of Pakistan.

In this study, we performed whole-genome sequencing of three ciprofloxacin-resistant Salmonella Reading strains isolated from poultry meat. Genomes of S. Reading strains contained an average of 4.81 Mbp size with 52.1% GC. The isolates exhibited blaOXA-10, aac [6']-Iaa, aadA1, cmlA1, qnrS1, and tetA resistance genes and IncX1 and IncX2 plasmids.

Evaluation of extended-spectrum β-lactamase production and plasmid-mediated quinolone resistance in Escherichia coli isolated from raw milk

This study aimed to determine the antibiotic resistance and extended-spectrum β-lactamase production profiles of Escherichia coli isolates obtained from raw milk samples. To this end, the study used a chromogenic-based culture method, MALDI-TOF-MS, 16S rRNA gene sequencing, disc diffusion, and combined disc techniques. PCR was also performed to detect ESBL (blaSHV, blaTEM, and blaCTX-M), quinolone (qnrA, qnrB, and qnrS) genes, and the presence of class I integrons. A total of 47 (78.33%) E. coli isolates were identified by morphological, biochemical, and molecular tests. All isolates exhibited susceptibility to gentamicin and tobramycin. Resistance rates to different antibiotics were observed as follows: ampicillin, 51.06%; nalidixic acid, 23.40%; trimethoprim/sulfamethoxazole, 17.02%; meropenem, 14.89%; ceftazidime, 8.51%; cefotaxime, 8.51%; piperacillin-tazobactam, 4.25%; aztreonam, 4.25%; imipenem, 4.25%; and levofloxacin, 2.12%. ESBL production was positive in 3 (6.38%) of the isolates. Bla genes were detected in 11 isolates (blaSHV = 9 [19.14%], blaTEM = 4 [8.5%], and blaCTX = 3 [6.38%]). The qnrB, qnrS, and qnrA genes were found in 5 (10.63%), 4 (8.51%), and 1 (2.12%) isolates, respectively. The intI1 gene was detected in 14 (29.78%) isolates.

Investigation of plasmid-mediated quinolone resistance among extended-spectrum β-lactamase isolates of E. coli and K. pneumoniae.

Escherichia coli and Klebsiella pneumoniae are important members of the Enterobacteriaceae family, involved in many infections. The increased resistance rate towards β-lactams and fluoroquinolones -which are the main therapeutic options- limits their treatment options. This study aimed to assess the local resistance patterns against different antimicrobials and to determine the extended-spectrum β-lactamase (ESBLs) producers. The study revealed that 36% of clinical isolates were ESBL producers, showing high resistance rates towards β-lactams and non-β-lactams, especially sulphamethoxazole-trimethoprim and fluoroquinolones. However, they were susceptible to chloramphenicol and doxycycline (33% and 20%; respectively). Also, the investigation aimed to screen the plasmid profile of quinolone-resistant ESBLs-producers and to detect the plasmid-mediated quinolone resistance genes including qnrA, qnrS, qnrB, qnrC, qnrD, and qnrVC. Moreover, the conjugative plasmid among the quinolone-resistant isolates was elucidated. The results showed that extracted plasmids of sizes ranging from ≈0.9 to 21.23 Kb, divided into 7 plasmid patterns were detected. A plasmid of approximately 21.23 Kb was found in all isolates and the QnrS gene was the most predominant gene. Moreover, the frequency of transconjugation within the same genus was higher than that recorded between different genera; where 68% of E. coli isolates transferred the resistance genes compared to Klebsiella isolates (36.6%). Plasmid profiles of transconjugants demonstrated great similarity, where 21.23 Kb plasmid was detected in all transconjugants. Since these transconjugants were quinolone-resistant ESBL producers, it has been suggested that quinolone resistance determinants might be carried on that plasmid.

Exploring Plasmid-Mediated Quinolone Resistance Gene Diversity among Ciprofloxacin-Resistant Proteus Species in a Tertiary Care Hospital of Bangladesh

Background: Increased resistance to fluoroquinolones has been observed in members of the Enterobacteriaceae, including Proteus species. The extensive use of fluoroquinolones and the presence of plasmid-mediated quinolone resistance (PMQR) genes are believed to contribute significantly to fluoroquinolone resistance. This study highlights the prevalence of ciprofloxacin-resistant Proteus species and the presence of specific PMQR genes among them, emphasizing their potential role in quinolone resistance. Method: The study included 30 ciprofloxacin-resistant Proteus isolates collected from wound swab, pus, urine, and blood samples. Identification of Proteus species was performed using culture and biochemical tests. Antibiotic susceptibility test was conducted using the Kirbey-Bauer disc-diffusion method. The minimum inhibitory concentration (MIC) of ciprofloxacin among ciprofloxacin-resistant Proteus species was determined using the agar dilution method. The study aimed to detect specific PMQR genes, namely aac(6´) lb-cr, qnrA, qnrB, qnrD, and qnrS, using polymerase chain reaction (PCR). Result: A total of 42 Proteus species were isolated from 310 various samples. Among the Proteus isolates, 71.43% were ciprofloxacin resistant. The study found that 30% of ciprofloxacin-resistant isolates were positive for aac(6´) lb-cr gene, 13.33% for qnrA and qnrS respectively and 10% for qnrD gene. In total, 60% of ciprofloxacin-resistant Proteus isolates were positive for at least one PMQR gene. The qnrB gene was not detected among ciprofloxacin-resistant Proteus species. The MIC of ciprofloxacin ranged from 8 μg/ml to 128 μg/ml among ciprofloxacin-resistant Proteus isolates. Conclusion: This study suggests aac(6´) lb-cr, qnrA, qnrS, and qnrD genes are emerging in Proteus species, potentially contributing to the development of quinolone resistance and has implications for clinical management and public health. J MEDICINE 2024; 25: 149-154

The effect of Astragalus polysaccharides on the repair of adverse effects in largemouth bass (Micropterus salmoides) under enrofloxacin stress

Antibiotics have important role in controlling the outbreak of bacterial diseases in fish farming, but long-term use or persistent existence of antibiotics in water can cause adverse effects on aquatic animals. In order to evaluate repairing effect of Astragalus polysaccharides (APS) on damaged intestine of fish under enrofloxacin (ENR) stress, ENR-pretreated (1.0 mg/L, 7 days) largemouth bass were randomly divided into two groups, and subsequently were fed basal diet and diet supplemented with 1.0 g/kg APS for 14 days. Intestine tissue and content were sampled on day 0 (D0), days 7 (D7), days 21 (D21_APS and D21) to detect the intestinal structure, intestinal microbiota structure, oxidative stress, non-specific immune and inflammatory response.Intestinal tissue section showed that the height, width of intestinal villi, and muscular thickness in D21_APS were significantly greater than those on D0, D7, and D21 (P < 0.05). APS-supplemented diet was significantly increased expression of tight junction protein related genes occludin, claudin-1, ZO-1 (P < 0.05). APS addition significantly improved the activities of catalase (CAT) and peroxidase (POD) and decreased superoxide dismutase (SOD) activity (P < 0.05). The activity of CAT and SOD in D21_APS and D0 were similar. The highest activities of acid phosphatase (ACP), lysozyme (LZM) and alkaline phosphatase (AKP) were found in D21_APS group. The expression of proinflammatory factors IL-1β and TNF-α in D21_APS and D21 were significantly down-regulated compared to that of D7, while anti-inflammatory factor IL-10 was up-regulated (P < 0.05). Compared with D7 group, quinolone resistance gene (qnrB) in D21 and D21_APS groups were significantly down-regulated, and in contrast to D21 group, qnrB and qnrS genes in D21_APS group were significantly down-regulated (P < 0.05).High-throughput 16SrRNA gene sequencing revealed proportion of Cyanobacteria in intestinal microbiota increased from 8.23% to 52.66%, while Firmicutes decreased from 28.54% to 2.03% after ENR treatment. In D21 group, Proteobacteria was notable decrease compared to D0, while Firmicutes increased. In D21_APS, the dominant bacteria were Proteobacteria, Fusobacteriota and Firmicutes, and Acinetobacter, Cetobacterium and Aeromonas were the predominant genera. Cluster analysis indicated that bacterial community structure in D21_APS has a similar level to D0 at phylum level. Feeding APS increased abundance of genes related to styrene, isoleucine and lysine degradation; propanoate, butanoate phenylalanine, and tryptophan metabolism; lipopolysaccharide biosynthesis; bacterial secretion system; cell motility and secretion at KEGG pathway Level 3 compared to the basal diet. Taken together, APS can repair the damage of intestinal structure and intestinal microbiota caused by ENR.

Removal of metals and emergent contaminants from liquid digestates in constructed wetlands for agricultural reuse.

Given the increasing pressure on water bodies, it is imperative to explore sustainable methodologies for wastewater treatment and reuse. The simultaneous presence of multiples contaminants in complex wastewater, such as the liquid effluents from biogas plants, can compromise biological treatment effectiveness for reclaiming water. Vertical subsurface flow constructed wetlands were established as low-cost decentralized wastewater treatment technologies to treat the liquid fraction of digestate from municipal organic waste with metals, antibiotics, and antibiotic resistance genes, to allow its reuse in irrigation. Twelve lab-scale planted constructed wetlands were assembled with gravel, light expanded clay aggregate and sand, testing four different treating conditions (liquid digestate spiked with oxytetracycline, sulfadiazine, or ofloxacin, at 100 μg/ L, or without dosing) during 3 months. Physicochemical parameters (pH, chemical oxygen demand (COD), nutrients, metals, and antibiotics), the microbial communities dynamics (through 16S high-throughput sequencing) and antibiotic resistance genes removal (qPCR) were monitored in influents and effluents. Systems removed 85.8%-96.9% of organic matter (as COD), over 98.1% of ammonium and phosphate ions, and 69.3%-99.4% of nitrate and nitrite ions, with no significant differences between the presence or absence of antibiotics. Removal of Fe, Mn, Zn, Cu, Pb and Cr exceeded 82% in all treatment cycles. The treatment also removed oxytetracycline, sulfadiazine and ofloxacin over 99%, and decreased intl1, tetA, tetW, sul1 and qnrS gene copies. Nonetheless, after 3 months of ofloxacin dosing, qnrS gene started being detected. Removal processes relied on high HRT (14 days) and various mechanisms including sorption, biodegradation, and precipitation. Microbial community diversity in liquid digestate changed significantly after treatment in constructed wetlands with a decrease in the initial Firmicutes dominance, but with no clear effect of antibiotics on the microbial community structure. Removals above 85% and 94% were observed for Streptococcus and Clostridium, respectively. Results suggest that vertical subsurface flow constructed wetlands were a suitable technology for treating the liquid digestate to reuse it in irrigation agricultural systems, contributing to the circular bioeconomy concept. However, a more profound understanding of effective wastewater treatment strategies is needed to avoid antibiotic resistance genes dissemination.

Emergence of extensive drug resistance and high prevalence of multidrug resistance among clinical Proteus mirabilis isolates in Egypt

BackgroundProteus mirabilis is an opportunistic pathogen that has been held responsible for numerous nosocomial and community-acquired infections which are difficult to be controlled because of its diverse antimicrobial resistance mechanisms.MethodsAntimicrobial susceptibility patterns of P. mirabilis isolates collected from different clinical sources in Mansoura University Hospitals, Egypt was determined. Moreover, the underlying resistance mechanisms and genetic relatedness between isolates were investigated.ResultsAntimicrobial susceptibility testing indicated elevated levels of resistance to different classes of antimicrobials among the tested P. mirabilis clinical isolates (n = 66). ERIC-PCR showed great diversity among the tested isolates. Six isolates (9.1%) were XDR while all the remaining isolates were MDR. ESBLs and AmpCs were detected in 57.6% and 21.2% of the isolates, respectively, where blaTEM, blaSHV, blaCTX−M, blaCIT−M and blaAmpC were detected. Carbapenemases and MBLs were detected in 10.6 and 9.1% of the isolates, respectively, where blaOXA−48 and blaNDM−1 genes were detected. Quinolone resistant isolates (75.8%) harbored acc(6')-Ib-cr, qnrD, qnrA, and qnrS genes. Resistance to aminoglycosides, trimethoprim-sulfamethoxazole and chloramphenicol exceeded 80%. Fosfomycin was the most active drug against the tested isolates as only 22.7% were resistant. Class I or II integrons were detected in 86.4% of the isolates. Among class I integron positive isolates, four different gene cassette arrays (dfrA17- aadA5, aadB-aadA2, aadA2-lnuF, and dfrA14-arr-3-blaOXA−10-aadA15) and two gene cassettes (dfrA7 and aadA1) were detected. While class II integron positive isolates carried four different gene cassette arrays (dfrA1-sat1-aadA1, estXVr-sat2-aadA1, lnuF- dfrA1-aadA1, and dfrA1-sat2).ConclusionP. Mirabilis ability to acquire resistance determinants via integrons may be held responsible for the elevated rates of antimicrobial resistance and emergence of XDR or even PDR strains limiting the available therapeutic options for management of infections caused by those strains.

Study of plasmid mediated quinolone resistance genes among Escherichia coli and Klebsiella pneumoniae isolated from pediatric patients with sepsis

The resistance to antibiotics in Gram-negative bacilli causing sepsis is a warning sign of failure of therapy. Klebsiella pneumoniae (K. pneumoniae) and Escherichia coli (E. coli) represent major Gram-negative bacilli associated with sepsis. Quinolone resistance is an emerging resistance among E. coli and K. pneumoniae. Therefore, the present study aimed to study the presence of plasmid-mediated quinolone resistance (PMQR) genes qnrA, qnrB, and qnrS by polymerase chain reaction (PCR) in E. coli and K. pneumoniae isolated from pediatric patients with sepsis. This was a retrospective cross-sectional study that included pediatric patients with healthcare-associated sepsis. The E. coli and K. pneumoniae isolates were identified by microbiological methods. PMQR genes namely qnrA, qnrB, and qnrS were detected in E. coli and K. pneumoniae isolates by PCR. The results were analyzed by SPPS24, and the qualitative data was analyzed as numbers and percentages and comparison was performed by Chi-square test, P was significant if < 0.05. The most prevalent gene detected by PCR was qnrA (75%), followed by qnrB (28.1%), and qnrS (25%). The most frequently detected qnr gene in E coli and K. pneumoniae was qnrA (28.8%, and 16.3% respectively). The present study highlights the high prevalence of ciprofloxacin resistance among E. coli and K. pneumoniae isolated from pediatric patients with healthcare-associated sepsis. There was a high frequency of PMQR genes in E. coli and K. pneumoniae isolated from pediatric patients. Therefore, it is important to monitor the spread of PMQR genes in clinical isolates to ensure efficient antibiotic use in those children. The finding denotes the importance of an antibiotics surveillance program.

Coexistence of plasmid-mediated quinolone resistance (PMQR) and extended-spectrum beta-lactamase (ESBL) genes among clinical Pseudomonas aeruginosa isolates in Egypt

BackgroundData about the prevalence of plasmid-mediated quinolone resistance (PMQR) and extended-spectrum beta-lactamase (ESBL) production in P. aeruginosa compared to the Enterobacteriaceae family is limited. The availability of limited therapeutic options raises alarming concerns about the treatment of multidrug-resistant P. aeruginosa. This study aimed to assess the presence of PMQR and ESBL genes among P. aeruginosa strains.MethodsFifty-six P. aeruginosa strains were isolated from 330 patients with different clinical infections. Phenotypically fluoroquinolone-resistant isolates were tested by PCR for the presence of six PMQR genes. Then, blaTEM, blaSHV, and blaCTX-M type ESBL genes were screened to study the co-existence of different resistance determinants.ResultsOverall, 22/56 (39.3%) of the studied P. aeruginosa isolates were phenotypically resistant to fluoroquinolones. PMQR-producing P. aeruginosa isolates were identified in 20 isolates (90.9%). The acc(6')-Ib-cr was the most prevalent PMQR gene (77.3%). The qnr genes occurred in 72.7%, with the predominance of the qnrA gene at 54.5%, followed by the qnrS gene at 27.3%, then qnrB and qnrC at 22.7%. The qepA was not detected in any isolate. The acc(6')-Ib-cr was associated with qnr genes in 65% of positive PMQR isolates. Significant differences between the fluoroquinolone-resistant and fluoroquinolone-susceptible isolates in terms of the antibiotic resistance rates of amikacin, imipenem, and cefepime (P value < 0.0001) were found. The ESBL genes were detected in 52% of cephalosporin-resistant P. aeruginosa isolates. The most frequent ESBL gene was blaCTX-M (76.9%), followed by blaTEM (46.2%). No isolates carried the blaSHV gene. The acc(6')-Ib-cr gene showed the highest association with ESBL genes, followed by the qnrA gene. The correlation matrix of the detected PMQR and ESBL genes indicated overall positive correlations. The strongest and most highly significant correlation was between qnrA and acc(6')-Ib-cr (r = 0.602) and between qnrA and blaCTX-M (r = 0.519).ConclusionA high prevalence of PMQR genes among the phenotypic fluoroquinolone-resistant P. aeruginosa isolates was detected, with the co-carriage of different PMQR genes. The most frequent PMQR was the acc(6')-Ib-cr gene. Co-existence between PMQR and ESBL genes was found, with 75% of PMQR-positive isolates carrying at least one ESBL gene. A high and significant correlation between the ESBL and PMQR genes was detected.

Enhanced removal of ciprofloxacin and associated antibiotic-resistant genes from wastewater using a biological aeration filters in combination with Fe3O4-modified zeolite.

Antibiotics release into the water environment through sewage discharge is a significant environmental concern. In the present study, we investigated the removal of ciprofloxacin (CIP) in simulated sewage by biological aeration filter (BAF) equipped with Fe3O4-modified zeolite (Fe3O4@ZF). Fe3O4@ZF were prepared with impregnation method, and the Fe3O4 particles were successfully deposited on the surface of ZF in an amorphous form according to the results of XPS and XRD analysis. The modification also increased the specific surface area (from 16.22 m²/g to 22 m²/g) and pore volume (from 0.0047 cm³/g to 0.0063 cm³/g), improving the adsorption efficiency of antibiotics. Fe3O4 modified ZF improved the treatment performance significantly, and the removal efficiency of CIP in BAF-Fe3O4@ZF was 79%±2.4%. At 10ml/L CIP, the BAF-Fe3O4@ZF reduced the relative abundances of antibiotics resistance genes (ARGs) int, mexA, qnrB and qnrS in the effluent by 57.16%, 39.59%, 60.22%, and 20.25%, respectively, which effectively mitigate the dissemination risk of ARGs. The modification of ZF increased CIP-degrading bacteria abundance, such as Rhizobium and Deinococcus-Thermus, and doubled bacterial ATP activity, promoting CIP degradation. This study offers a viable, efficient method to enhance antibiotic treatment and prevent leakage via sewage discharge.

Whole genome sequencing analysis on antibiotic-resistant Escherichia coli isolated from pig farms in Banten Province, Indonesia.

The emergence and rapid increase in the incidence of multidrug-resistant (MDR) bacteria in pig farms has become a serious concern and reduced the choice of effective antibiotics. This study analyzed the phylogenetics and diversity of antibiotic resistance genes (ARGs) and molecularly identified the source of ARGs in antibiotic-resistant Escherichia coli isolated from pig farms in Banten Province, Indonesia. Forty-four antibiotic-resistant E. coli isolates from fecal samples from 44 pig farms in Banten Province, Indonesia, were used as samples. The samples were categorized into 14 clusters. Sequencing was performed using the Oxford Nanopore Technologies MinION platform, with barcoding before sequencing with Nanopore Rapid sequencing gDNA-barcoding (SQK-RBK110.96) according to manufacturing procedures. ARG detection was conducted using ResFinder, and the plasmid replicon was determined using PlasmidFinder. Three phylogenetic leaves of E. coli were identified in the pig farming cluster in Banten Province. The E. coli isolates exhibited potential resistance to nine classes of antibiotics. Fifty-one ARGs were identified across all isolates, with each cluster carrying a minimum of 10 ARGs. The ant(3'')-Ia and qnrS1 genes were present in all isolates. ARGs in the E. coli pig farming cluster originated mainly from plasmids, accounting for an average of 89.4%. The elevated potential for MDR events, coupled with the dominance of ARGs originating from plasmids, increases the risk of ARG spread among bacterial populations in animals, humans, and the environment.

Prevalence of CTX-M types among ESBL-producing pathogenic Escherichia coli isolates from foodborne diarrheal patients in Gyeonggi-do, South Korea

Prevalence and characteristics of extended-spectrum β-lactamase (ESBL)-producing pathogenic Escherichia coli from foodborne diarrheal patients were studied. Analysis of 495 E. coli isolates revealed that 80 isolates were ESBL-producing pathogenic E. coli, and enteroaggregative E. coli and enterotoxigenic E. coli were two of the most prevalent pathotypes. In silico Clermont phylo-typing of the 80 ESBL-producing E. coli showed that phylogroup A (49/80) and D (22/80) were the predominant phylogroups. The average nucleotide identity analysis of ESBL-producing E. coli disclosed that they could be grouped into two phylogenetic groups; 25 A and 55 B groups. All strains, except one, harbored the blaCTX-M gene. All CTX-M-15 type ESBL-producing strains also carried qnrS, a plasmid-mediated quinolone resistance gene (PMQR). These results suggest that the diversity of ESBL-producing E. coli is high and that co-existence of blaCTX-M-15 and qnrS genes is widespread, highlighting their high risk of antibiotic-resistance spreading in infectious disease outbreaks.

High prevalence of plasmid-mediated quinolone resistance in escherichia coli strains producing extended-spectrum beta-lactamases isolated from faeces and urine of pregnant women with acute cystitis.

Escherichia coli is the most common etiological agent of urinary tract infections (UTIs). Meanwhile, plasmid-mediated quinolone resistance (PMQR) is reported in E. coli isolates producing extended-spectrum β-lactamases (ESBLs). Furthermore, the reservoirs and mechanisms of acquisition of uropathogenic Escherichia coli (UPEC) strains are poorly understood. On the other hand, UTIs are common in pregnant women and the treatment challenge is alarming. In the present study, 54 pregnant women with acute cystitis were included. A total of 108 E. coli isolates, 54 isolates from UTI and 54 isolates from faeces of pregnant women (same host) were collected. In the antimicrobial susceptibility test, the highest rate of antibiotic resistance was to nalidixic acid (77%, 83/108) and the lowest rate was to imipenem (9%, 10/108). Among the isolates, 44% (48/108) were ESBLs producers. A high frequency of PMQR genes was observed in the isolates. The frequency of PMQR genes qnrS, qnrB, aac(6')-Ib-cr, and qnrA was 58% (63/108), 21% (23/108), 9% (10/108), and 4% (4/108), respectively. Meanwhile, PMQR genes were not detected in 24% (20/85) of isolates resistant to nalidixic acid and/or fluoroquinolone, indicating that other mechanisms, i.e. chromosomal mutations, are involved in resistance to quinolones, which were not detected in the present study. In ESBL-producing isolates, the frequency of PMQR genes was higher than that of non-ESBL-producing isolates (81% vs. 53%). Meanwhile, UTI and faeces isolates mainly belonged to phylogenetic group B2 (36/54, 67% and 25/54, 46%, respectively) compared to other phylogenetic groups. In addition, virulence factors and multidrug-resistant (MDR) were mainly associated with phylogenetic group B2. However, predominant clones in faeces were not found in UTIs. Rep-PCR revealed the presence of 85 clones in patients. Among the clones, 40 clones were detected only in faeces (faeces-only), 35 clones only in UTI (UTI-only) and 10 clones in both faeces and UTI (faeces-UTI). We found that out of 10 faeces-UTI clones, 5 clones were present in the host's faeces flora. This study revealed a high rate of resistance to the quinolone nalidixic acid and a widespread distribution of PMQR genes in MDR E. coli strains producing ESBLs. The strains represented virulence factors and phylogenetic group B2 are closely associated with abundance in UTI and faeces. However, the predominant clones in faeces were not found in UTIs and it is possible that rep-PCR is not sufficiently discriminating clones.

Pathovars, occurrence, and characterization of plasmid-mediated quinolone resistance in diarrheal Escherichia coli isolated from farmers and farmed chickens in Tunisia and Nigeria.

The poultry industry is a very important agricultural and industrial sector in Tunisia and Nigeria, with little information about occurrence of diarrheagenic Escherichia coli in the farmers and chickens. This study aimed to detect the prevalence of diarrheal E. coli in humans and poultry and to investigate plasmid-mediated quinolone resistance (PMQR) genes in both countries. Seventy-four isolates of E. coli were studied; nine different virulence genes were screened by PCR. Serotyping was performed only for pathotypes as well as the determining of antibiotic resistance profiles against 21 antibiotics. PMQR genes were investigated by PCR. EAEC was the most abundant pathotype (37/74; 50%) in human and chicken isolates, whereas single EHEC and EPEC (1/74, 1.35%) pathotypes were detected in Tunisia and Nigeria, respectively. About 17 (45.95%) quinolones/fluoroquinolones-resistant isolates were detected, from which the following PMQR genes were detected: aac(6')-Ib-cr (8/17, 47.05%), qepA (6/17, 35.29%), qnrA+qnrB (2/17, 11.76%), and qnrS gene (1/17, 5.88%). Our findings highlight high occurrence of EAEC pathotype in Tunisia and Nigeria, more frequent than EPEC and EHEC. Additionally, all E. coli pathotypes isolated from different sources (humans, poultry) showed resistance to several antibiotics, which are in use as therapeutic choices in Tunisia and Nigeria.

Carbapenem and colistin-resistant hypervirulent Klebsiella pneumoniae: An emerging threat transcending the egyptian food chain

BackgroundCarbapenem-resistant Klebsiella pneumoniae (CRKP) is a great public health problem and is associated with many disease outbreaks and high mortality rates. Alarmingly, K. pneumoniae has been isolated from food in several recent studies. This study aimed to investigate the prevalence and characteristics of CRKP in food samples from Egypt. MethodsA total of 311 food samples (including 116 minced meat, 92 chicken meat, 75 diced meat, and 28 mutton) were collected from local markets in Egypt and were screened for CRKP with the determination of their antimicrobial resistance profiles. The whole genome sequence was done for 23 CRKP isolates to clarify the relationship between CRKP from food and human cases in Egypt using the SNP core genome. The conjugation probability of the blaNDM-5 harboring plasmid was identified using oriTfinder ResultsCRKP was isolated from 11% (35/311) of the samples, with 45.71% (16/35) of them showing resistance to colistin, one of the last-resort options for treating CRKP-mediated infections. In addition to the carbapenem and colistin resistance, the CRKP isolates frequently exhibited resistance to multiple antimicrobials including β-lactams, fluoroquinolones, aminoglycosides, tetracyclines, and chloramphenicol. In addition, most of the CRKP were potentially hypervirulent K. pneumoniae (HvKP) identified as phylogroup Kp1 and of high-risk groups as detected in STs reported in many human outbreaks globally, such as ST383 and ST147. The core-genome phylogeny showed similarities between the isolates from this study and those previously isolated from clinical human samples in Egypt. In addition, analysis of the plasmid on which blaNDM is encoded revealed that several antimicrobial resistance genes such as blaOXA-9, blaCTX-M-15, aac(6′)-Ib, qnrS1, and several virulence genes are encoded on the same plasmid. ConclusionsThis study is significant for food safety and public health and is important to further identify the change in the epidemiology of CRKP infections, especially the consumption of contaminated food products.

Plasmid-mediated quinolone resistance in Escherichia coli isolates from commercial broiler chickens in Semnan, Iran

Background and Objectives: Antibiotic resistance within the poultry sector presents a considerable health concern due to treatment inefficacy and resistance transmission to humans and the environment. The investigation of plasmid-mediated quinolone resistance (PMQR) in Escherichia coli, acknowledged for its role in advancing resistance, remains inadequately studied in Iranian poultry. This study aimed to evaluate PMQR gene prevalence as well as to determine correlation between resistance phenotype and genotype in E. coli obtained from poultry colibacillosis. Materials and Methods: A collection of 100 E. coli isolates from the viscera of broilers suspected to colibacillosis was assessed. Using the Kirby-Bauer disk diffusion method, antimicrobial susceptibility tests were conducted for ofloxacin, nalidixic acid, levofloxacin, ciprofloxacin, and ampicillin. Additionally, PCR was employed to screen for qnrS, qnrB, and aac(6)Ib-cr genes. Results: Among the analyzed E. coli isolates, 51% demonstrated resistance to at least one of the tested antibiotics, with 17% exhibiting resistance to four different antibiotics. Nalidixic acid displayed the highest resistance rate at 48%, while ampicillin had the lowest at 16%. PMQR genes were detected in 28% of the E. coli isolates, with aac(6′)-Ib-cr being the most prevalent at 14%, followed by qnrB in 13%, and qnrS in 7%. Conclusion: The study underscores the vital need for careful antibiotic usage in poultry to curb the emergence of antibi- otic-resistant bacteria. The results illuminate the prevalence of PMQR genes and their association with resistance trends in Iranian poultry, forming a pivotal basis for forthcoming approaches to combat antibiotic resistance within the poultry sector.

Analysis of Antimicrobial Resistance in Bacterial Pathogens Recovered from Food and Human Sources: Insights from 639,087 Bacterial Whole-Genome Sequences in the NCBI Pathogen Detection Database.

Understanding the role of foods in the emergence and spread of antimicrobial resistance necessitates the initial documentation of antibiotic resistance genes within bacterial species found in foods. Here, the NCBI Pathogen Detection database was used to query antimicrobial resistance gene prevalence in foodborne and human clinical bacterial isolates. Of the 1,843,630 sequence entries, 639,087 (34.7%) were assigned to foodborne or human clinical sources with 147,788 (23.14%) from food and 427,614 (76.88%) from humans. The majority of foodborne isolates were either Salmonella (47.88%), Campylobacter (23.03%), Escherichia (11.79%), or Listeria (11.3%), and the remaining 6% belonged to 20 other genera. Most foodborne isolates were from meat/poultry (95,251 or 64.45%), followed by multi-product mixed food sources (29,892 or 20.23%) and fish/seafood (6503 or 4.4%); however, the most prominent isolation source varied depending on the genus/species. Resistance gene carriage also varied depending on isolation source and genus/species. Of note, Klebsiella pneumoniae and Enterobacter spp. carried larger proportions of the quinolone resistance gene qnrS and some clinically relevant beta-lactam resistance genes in comparison to Salmonella and Escherichia coli. The prevalence of mec in S. aureus did not significantly differ between meat/poultry and multi-product sources relative to clinical sources, whereas this resistance was rare in isolates from dairy sources. The proportion of biocide resistance in Bacillus and Escherichia was significantly higher in clinical isolates compared to many foodborne sources but significantly lower in clinical Listeria compared to foodborne Listeria. This work exposes the gaps in current publicly available sequence data repositories, which are largely composed of clinical isolates and are biased towards specific highly abundant pathogenic species. We also highlight the importance of requiring and curating metadata on sequence submission to not only ensure correct information and data interpretation but also foster efficient analysis, sharing, and collaboration. To effectively monitor resistance carriage in food production, additional work on sequencing and characterizing AMR carriage in common commensal foodborne bacteria is critical.