Virulence Factor Genes Research Articles

Environmentally persistent microbial contamination in agricultural soils: High risk of pathogenicity and antibiotic resistance

Persistent microbial contamination commonly occurs in the environment. However, the characteristics and associated risks remain largely unknown. The coexistence of virulence factor genes (VFGs) and “last-resort” antibiotic resistance genes (LARGs) on human bacterial pathogens (HBPs) are notorious, creating ecological concerns and health risks. Herein, we explored the pathogenicity and antibiotic resistance levels of LARG-harboring HBPs in agricultural soils. Our findings revealed a high distribution level of VFGs and LARGs in soils (an absolute abundance up to 4.7 × 107 gene copies/g soil) by quantitative PCR (qPCR). Furthermore, most isolated LARG-harboring HBPs exhibited a 100 % lethality rate to Galleria mellonella. LARG-carrying plasmids had a low fitness cost to their host bacteria, implying the high adaptation of these plasmids within the HBPs. Most importantly, multiple LARG and VFG plasmid fusion and core genetic arrangements suggested that these LARG/VFG-linked plasmids endowed the stable and persistent horizontal spread of these genes in and/or cross the species and environments. This study not only unveiled high risk, multisource, compliance and stability aspects of environmentally persistent microbial contamination but also illuminated the importance of linking the phenotype-genotype-niche colonization of environmental microbial contamination within “One Health” framework.

Draft genome sequence of multidrug-resistant Kurthia gibsonii strain Hakim RU_BHWE isolated from sewage water in Bangladesh.

We have sequenced the genome of Kurthia gibsonii strain Hakim RU_BHWE, isolated from sewage water. The assembled genome consists of 2.891 Mb with 58.6883× coverage, presenting an average GC content of 36.60%. This genome includes 8 CRISPR arrays, 3 prophages, 3 antibiotic resistance genes, and 12 virulence factor genes.

Travertine deposition rather than tourism activity is the primary contributor to the microplastic risks in alpine karst lakes

Microplastics (MPs) are emerging as anthropogenic vectors to form plastisphere, facilitating microbiome colonization and pathogenic dissemination, thus contributing to environmental and health crises across various ecosystems. However, a knowledge gap persists regarding MPs risks and their driving factors in certain unique and vulnerable ecosystems, such as Karst travertine lakes, some of which are renowned World Natural Heritage Sites under ever-increasing tourism pressure. We hypothesized that tourism activities serve as the most important factor of MPs pollution, whereas intrinsic features, including travertine deposition can exacerbate potential environmental risks. Thus, metagenomic approaches were employed to investigate the geographical distribution of the microbiome, antibiotic resistance genes (ARGs), virulence factor genes (VFGs), and their combined environmental risks in Jiuzhaigou and Huanglong, two famous tourism destinations in Southwest China. The plastisphere risks were higher in Huanglong, contradicting our hypothesis that Jiuzhaigou would face more crucial antibiotic risks due to its higher tourist activities. Specifically, the levels of Lipopolysaccharide Lewis and fosD increased by sevenfold and 20-fold, respectively, from upstream to downstream in Huanglong, whereas in Jiuzhaigou, no significant accrual was observed. Structural equation modeling results showed that travertine deposition was the primary contributor to MPs risks in alpine karstic lakes. Our findings suggest that tourism has low impact on MPs risks, possibly because of proper management, and that travertine deposition might act as an MPs hotspot, emphasizing the importance of considering the unique aspects of travertine lakes in mitigating MPs pollution and promoting the sustainable development of World Natural Heritage Sites.

Unveiling potential roles of earthworms in mitigating the presence of virulence factor genes in terrestrial ecosystems

Earthworms can redistribute soil microbiota, and thus might affect the profile of virulence factor genes (VFGs) which are carried by pathogens in soils. Nevertheless, the knowledge of VFG profile in the earthworm guts and its interaction with earthworm gut microbiome is still lacking. Herein, we characterized earthworm gut and soil microbiome and VFG profiles in natural and agricultural ecosystems at a national scale using metagenomics. VFG profiles in the earthworm guts significantly differed from those in the surrounding soils, which was mainly driven by variations of bacterial communities. Furthermore, the total abundance of different types of VFGs in the earthworm guts was about 20-fold lower than that in the soils due to the dramatic decline (also by approximately 20-fold) of VFG-carrying bacterial pathogens in the earthworm guts. Additionally, five VFGs related to nutritional/metabolic factors and stress survival were identified as keystones merely in the microbe–VFG network in the earthworm guts, implying their pivotal roles in facilitating pathogen colonization in earthworm gut microhabitats. These findings suggest the potential roles of earthworms in reducing risks related to the presence of VFGs in soils, providing novel insights into earthworm-based bioremediation of VFG contamination in terrestrial ecosystems.

SYNERGISTIC EFFECTS OF NEEM OIL AND GENTAMICIN ON PSEUDOMONAS AERUGINOSA VIA PHZM GENE DOWNREGULATION: A COMPREHENSIVE REVIEW

Exploring the antibacterial potential of neem oil (Azadirachta indica) in combination with gentamicin (GEN) against pathogenic molds, especially Pseudomonas aeruginosa, has drawn concern due to the quest for natural treatment options against incurable diseases. Prospective research directions include looking for natural cures for many of the currently incurable diseases available now. microbial identification system, were used to identify the isolates. The research utilized a range of methods, such as the diffusion agar well (AWD) assays, TEM (transmission electron microscopy) analysis, minimum inhibitory concentration (MIC) assays, and real-time PCR (RT-qPCR) to analyze bacterial expression and the antibacterial action of neem oil (Azadirachta indica) combined with gentamicin (GEN) against the pathogenic bacteria Pseudomonas aeruginosa between others. The combined effects of neem oil and GEN on P. aeruginosa are extensively investigated in this study, with particular attention paid to the downregulation of the virulent factor gene phzM, inhibition of biofilm formation, morphological changes, and gene expression, in that proportion. Results show that neem oil has strong antibacterial activity against P. aeruginosa and S. aureus, preventing the formation of biofilms and causing morphological alterations. In addition, the synergistic effects of neem oil and GEN are demonstrated, together in order with the downregulation of the phzM gene in P. aeruginosa. These results highlight the increased effectiveness of neem oil when combined with GEN and point to the potential of the oil as a stand-alone antibacterial agent. They also highlight the need for more study to overcome antibiotic resistance in harmful bacteria. Overall, our results point to a possibility that neem oil extract, either alone or in combination with GEN, can suppress the development of pathogenic microbes. This synergistic impact is related to the downregulation of a virulence factor gene. To address the drug resistance linked to pathogenic microbes, greater study is required. Novelty of study addresses the important issue of antibiotic resistance by exploring the potential synergistic effects of neem oil and gentamicin on pathogenic bacteria, particularly Pseudomonas aeruginosa. This topic is highly relevant in the context of the global challenge of antimicrobial resistance.Novelty and relevanc the important issue of antibiotic resistance by exploring the potential synergistic effects of neem oil and gentamicin on pathogenic bacteria, particularly Pseudomonas aeruginosa. This topic is highly relevant in the context of the global challenge of antimicrobial resistance.

Differential Effects of Bacillus Species-Fermented Products on Antibiotic Resistome and Virulence Factor Gene Composition in the Cecal Digesta of Broilers

Abstract This study investigated the differential effects of Bacillus subtilis- and Bacillus licheniformis-fermented products (defined as SFP and LFPs, respectively) on microbial antibiotic resistance gene (ARG) and virulence factor gene (VFG) composition in the cecal digesta of 35-day-old broilers by using metagenomic sequencing. First, 160 1-day-old unsexed Arbor Acres broiler chicks were randomly allocated to four treatment groups: basal diet (CON group); basal diet + enramycin (10 mg/kg, ENM group); basal diet + SFPs (108 colony-forming units (CFU) of B. subtilis spores/kg, SFP group); and basal diet + LFPs (108 CFU of B. licheniformis spores/kg, LFP group). Principal coordinate analysis of ARG and VFG composition indicated distinct clustering among the cecal samples of the groups. At the antibiotic resistance class level, LFP treatment increased the expression of peptide resistance genes and decreased the expression of aminocoumarin resistance genes compared with the other groups. Compared with the other groups, LFP treatment promoted bcrA, ugd, and efrB expression but suppressed parY expression, whereas SFP treatment inhibited efrA expression. The abundance of the peptide resistance gene bcrA in Lachnoclostridium species was higher in the CON and LFP groups than in the ENM and SFP groups, whereas the abundance of the peptide resistance gene rpoB2 in Bacteroides species was lower in the ENM and LFP groups than in the SFP group. No specific VFGs were regulated only by SFPs or LFPs. SFP and LFP treatment inhibited clpC expression compared with the other groups. clpC abundance in Bacteroides species was lower in the LFP group than in the CON group, whereas its abundance in Faecalibacterium species was lower in the SFP group than in the CON and ENM groups. These results demonstrated that SFPs and LFPs differentially regulate microbial ARG and VFG composition in the cecal digesta of broilers. LFP supplementation modulated more antibiotic resistance classes and ARGs than did SFP supplementation.

Molecular Detection of the mecA and some Virulence Determinants in Methicillin-Resistant Staphylococcus aureus

Background: Methicillin Resistant Staphylococcus aureus (MRSA) is globally acknowledged as a prominent contributor to both hospital-acquired and community infections. Understanding key virulence factors including coagulase production, hemolysis ability and biofilm formation, is crucial. Objective: The study aimed to establish a molecular characterization of mecA gene and virulence factors genes (hla, icaA, and coa) in clinical isolates of MRSA obtained from two hospitals in Baghdad. Materials and Methods: A hundred and five isolates were obtained from clinical sources from November 2022 to March 2023 and their antibiotic sensitivity was assessed using the agar diffusion test against seven different antibiotics (Azithromycin, Ciprofloxacin, Nitrofurantoin, Rifampin, Trimethoprim, Ofloxacin and Oxacillin), through Conventional Polymerase Chain Reaction, the presence of virulence factor genes including mecA, hla, icaA, and coa, was determined in MRSA isolates. Results: All MRSA isolates (100%) harbored the mecA, hla, and icaA genes while the coa gene was recognized in 50% of the isolates. Regarding antibiotic susceptibility, all MRSA isolates (100%) demonstrated sensitivity to Nitrofurantoin. Additionally, 96.8% of the isolates were sensitive to Oxacillin. Conclusion: Molecular detection of methicillin resistance genes and virulence genes can be used to diagnose MRSA isolates in hospitals. The presence of these genes may affect their pattern of sensitivity to antibiotics.

Effect of phenylalanine arginyl β-naphthylamide on the imipenem resistance, elastase production, and the expression of quorum sensing and virulence factor genes in Pseudomonas aeruginosa clinical isolates.

Pseudomonas aeruginosa is one of the most important nosocomial pathogens that possess the ability to produce multiple antibiotic resistance and virulence factors. Elastase B (LasB) is the major factor implicated in tissue invasion and damage during P. aeruginosa infections, whose synthesis is regulated by the quorum sensing (QS) system. Anti-virulence approach is now considered as potential therapeutic alternative and/or adjuvant to current antibiotics' failure. The aim of this study is primarily to find out the impact of the efflux pump inhibitor (EPI) phenylalanine arginyl β-naphthylamide (PAβN) on the production of elastase B and the gene expression of lasI quorum sensing and lasB virulence factor in clinical isolates of P. aeruginosa. Five P. aeruginosa isolates recovered from patients with respiratory tract infections were examined in this study. Antimicrobial susceptibility of isolates was performed by the disk agar diffusion method. Effect of the PAβN on imipenem susceptibility, bacterial viability, and elastase production was evaluated. The expression of lasB and lasI genes was measured by quantitative real-time PCR in the presence of PAβN. All isolates were identified as multidrug-resistant (MDR) and showed resistance to carbapenem (MIC = 64-256µg/mL). Susceptibility of isolates to imipenem was highly increased in the presence of efflux inhibitor. PAβN significantly reduced elastase activity in three isolates tested without affecting bacterial growth. In addition, the relative expression of both lasB and lasI genes was diminished in all isolates in the presence of inhibitor. Efflux inhibition by using the EPI PAβN could be a potential target for controlling the P. aeruginosa virulence and pathogenesis. Furthermore, impairment of drug efflux by PAβN indicates its capability to be used as antimicrobial adjuvant that can decrease the resistance and lower the effective doses of current drugs.

Molecular Characterization and Antibacterial Resistance Determination of Escherichia coli Isolated from Fresh Raw Mussels and Ready-to-Eat Stuffed Mussels: A Major Public Health Concern.

Our study focused exclusively on analyzing Escherichia coli (E. coli) contamination in fresh raw mussels and ready-to-eat (RTE) stuffed mussels obtained from authorized and regulated facilities. However, it is critical to recognize that such contamination represents a significant public health threat in regions where unauthorized harvesting and sales practices are prevalent. This study aimed to comprehensively assess the prevalence, molecular characteristics, and antibacterial resistance profiles of E. coli in fresh raw mussels and RTE stuffed mussels. E. coli counts in fresh raw mussel samples ranged from 1 to 2.89 log CFU/g before cooking, with a significant reduction observed post-cooking. RTE stuffed mussel samples predominantly exhibited negligible E. coli presence (<1 log CFU/g). A phylogenetic analysis revealed a dominance of phylogroup A, with variations in the distribution observed across different sampling months. Antibacterial resistance was prevalent among the E. coli isolates, notably showing resistance to ampicillin, streptomycin, and cefotaxime. Extended-spectrum β-lactamase (ESβL) production was rare, with only one positive isolate detected. A variety of antibacterial resistance genes, including tetB and sul1, were identified among the isolates. Notably, virulence factor genes associated with pathogenicity were absent. In light of these findings, it is imperative to maintain rigorous compliance with quality and safety standards at all stages of the mussel production process, encompassing harvesting, processing, cooking, and consumption. Continuous monitoring, implementation of rigorous hygiene protocols, and responsible antibacterial drug use are crucial measures in mitigating food safety risks and combating antibacterial resistance. Stakeholders, including seafood industry players, regulatory agencies, and healthcare professionals, are essential to ensure effective risk mitigation and safeguard public health in the context of seafood consumption.

Multi-omics strategy reveals potential role of antimicrobial resistance and virulence factor genes responsible for Simmental diarrheic calves caused by Escherichia coli.

Escherichia coli (E. coli) is reported to be an important pathogen associated with calf diarrhea. Antibiotic resistance genes (ARGs) and virulence factor genes (VFGs) pose a considerable threat to both animal and human health. However, little is known about the characterization of ARGs and VFGs presented in the gut microbiota of diarrheic calves caused by E. coli. In this study, we used multi-omics strategy to analyze the ARG and VFG profiles of Simmental calves with diarrhea caused by E. coli K99. We found that gut bacterial composition and their microbiome metabolic functions varied greatly in diarrheic calves compared to healthy calves. In total, 175 ARGs were identified, and diarrheal calves showed a significantly higher diversity and abundance of ARGs than healthy calves. Simmental calves with diarrhea showed higher association of VFGs with pili function, curli assembly, and ferrienterobactin transport of E. coli. Co-occurrence patterns based on Pearson correlation analysis revealed that E. coli had a highly significant (P < 0.0001) correlation coefficient (>0.8) with 16 ARGs and 7 VFGs. Metabolomics analysis showed that differentially expressed metabolites in Simmental calves with diarrhea displayed a high correlation with the aforementioned ARGs and VFGs. Phylotype analysis of E. coli genomes showed that the predominant phylogroup B1 in diarrheic Simmental calves was associated with 10 ARGs and 3 VFGs. These findings provide an overview of the diversity and abundance of the gut microbiota in diarrheic calves caused by E. coli and pave the way for further studies on the mechanisms of antibiotic resistance and virulence in the calves affected with diarrhea.IMPORTANCESimmental is a well-recognized beef cattle breed worldwide. They also suffer significant economic losses due to diarrhea. In this study, fecal metagenomic analysis was applied to characterize the antibiotic resistance gene (ARG) and virulence factor gene (VFG) profiles of diarrheic Simmental calves. We identified key ARGs and VFGs correlated with Escherichia coli isolated from Simmental calves. Additionally, metabolomics analysis showed that differentially expressed metabolites in Simmental calves with diarrhea displayed a high correlation with the aforementioned ARGs and VFGs. Our findings provide an insight into the diversity and abundance of the gut microbiota in diarrheic calves caused by Escherichia coli and pave the way for further studies on the mechanisms of antibiotic resistance and virulence in the diarrheal calves from cattle hosts.

Detection and Characterization of Virulent Pseudomonas aeruginosa genes in Fresh Fish at Al-Diwaniya City Market, Iraq.

Fish meat is more susceptible to pathogen contamination than other protein-rich foodstuffs due to its high deterioration. Pseudomonas aeruginosa is considered one of the most common microbial strains responsible for the deterioration of fish and seafood products. the study's goals are to detecting virulence variables Pseudomonas aeruginosa and estimating the level of contamination in a fish sample, In this research, 120 fresh fish samples, gathered from a the local Diwaniyah city market local fish meat sample. including (60 marine fish and 60 common carp (Cyprinus carpio). The isolates were subjected to numerous biochemical, microbiological, and molecular analyses, and the results showed that about 26 samples were contaminated with Pseudomonas spp. performed on 21.6% of the total sample. Polymerase chain reaction (PCR) was used to amplify 539 bp, 511 bp, and 495 bp, for 16S DNA, Tox A, and Tox S respectively. A total of 21 Tox A genes were identified in 26 P. aeruginosa isolates, whereas the ToxS gene was detected in 17 of the same isolates. According to the results Pseudomonas aeruginosa isolated from fish samples contains multiple virulence factor genes, indicating high pathogenicity, which may affect consumer health because these variants exhibit the capability to increase the pathogenesis of the microorganisms with the severity of the infection and cause economic losses

WGS of intrauterine E. coli from cows with early postpartum uterine infection reveals a non-uterine specific genotype and virulence factors.

Escherichia coli has been attributed to playing a major role in a cascade of events that affect the prevalence and severity of uterine disease in cattle. The objectives of this project were to (i) define the association between the prevalence of specific antimicrobial resistance and virulence factor genes in E. coli with the clinical status related to uterine infection, (ii) identify the genetic relationship between E. coli isolates from cows with diarrhea, with mastitis, and with and without metritis, and (iii) determine the association between the phenotypic and genotypic antimicrobial resistance identified on the E. coli isolated from postpartum cattle. Bacterial isolates (n = 148) were obtained from a larger cross-sectional study. Cows were categorized into one of three clinical groups before enrollment: metritis, cows with purulent discharge, and control cows. For genomic comparison, public genomes (n = 130) from cows with diarrhea, mastitis, and metritis were included in a genome-wide association study, to evaluate differences between the drug classes or the virulence factor category among clinical groups. A distinct E. coli genotype associated with metritis could not be identified. Instead, a high genetic diversity among the isolates from uterine sources was present. A virulence factor previously associated with metritis (fimH) using PCR was not associated with metritis. There was moderate accuracy for whole-genome sequencing to predict phenotypic resistance, which varied depending on the antimicrobial tested. Findings from this study contradict the traditional pathotype classification and the unique intrauterine E. coli genotype associated with metritis in dairy cows.IMPORTANCEMetritis is a common infectious disease in dairy cattle and the second most common reason for treating a cow with antimicrobials. The pathophysiology of the disease is complex and is not completely understood. Specific endometrial pathogenic Escherichia coli have been reported to be adapted to the endometrium and sometimes lead to uterine disease. Unfortunately, the specific genomic details of the endometrial-adapted isolates have not been investigated using enough genomes to represent the genomic diversity of this organism to identify specific virulence genes that are consistently associated with disease development and severity. Results from this study provide key microbial ecological advances by elucidating and challenging accepted concepts for the role of Intrauterine E. coli in metritis in dairy cattle, especially contradicting the existence of a unique intrauterine E. coli genotype associated with metritis in dairy cows, which was not found in our study.

Draft genome sequence of carbapenems-resistant Acinetobacter baumannii Hakim RU_CBWP strain isolated from a pond surface water in Bangladesh.

We have revealed the genomic sequence of Acinetobacter baumannii strain Hakim RU_CBWP isolated from pond surface water. Our assembled genome covers 3.787 Mb with 45.5629× coverage, showcasing an average GC content of 38.60%. This genome contains two CRISPR arrays, 17 prophages, 22 antibiotic resistance genes, and 20 virulence factor genes.

Neglected risks of enhanced antimicrobial resistance and pathogenicity in anaerobic digestion during transition from thermophilic to mesophilic

Minimization of antibiotic resistance genes (ARGs) and potential pathogenic antibiotic-resistant bacteria (PARB) during anaerobic digestion (AD) is significantly impacted by temperature. However, knowledge on how ARGs and PARB respond to temperature transition from thermophilic to mesophilic is limited. Here, we combined metagenomic-based with culture-based approaches and revealed the risks of antimicrobial resistance and pathogenicity during transition from 55 °C to 35 °C for AD, with strategies of sharp (ST, one-step by 20 °C/d) and mild (MT, step-wise by 1 °C/d). Results indicated a lower decrease in methane production with MT (by 38.9%) than ST (by 88.8%). Phenotypic assays characterized a significant propagation of multi-resistant lactose-fermenting Enterobacteriaceae and indicator pathogens after both transitions, especially via ST. Further genomic evidence indicated a significant increase of ARGs (29.4-fold), virulence factor genes (1.8-fold) and PARB (65.3-fold) after ST, while slight enrichment via MT. Bacterial succession and enhanced horizontal transfer mediated by mobile genetic elements promoted ARG propagation in AD during transition, which was synchronously exacerbated through horizontal transfer mechanisms mediated by cellular physiological responses (oxidative stress, membrane permeability, bacterial conjugation and transformation) and co-selection mechanisms of biomethanation metabolic functions (acidogenesis and acetogenesis). This study reveals temperature-dependent resistome and pathogenicity development in AD, facilitating microbial risk control.

Draft genome sequence of Escherichia coli MTR_GS_S1457 strain isolated from a soil sample of a vegetable garden in Gazipur, Bangladesh.

We announce the sequence of the Escherichia coli MTR_GS_S1457 strain isolated from a soil sample of a vegetable gardening system for the first time in Bangladesh. With a length of 4,918,647 bp, this strain contained one plasmid, two CRISPR arrays, 54 predicted antibiotic resistance genes, and 81 predicted virulence factor genes.

Genomic characterization of Staphylococcus aureus isolated from patients admitted to intensive care units of a tertiary care hospital: epidemiological risk of nasal carriage of virulent clone during admission.

We conducted a molecular epidemiological study of Staphylococcus aureus using whole-genome sequence data and clinical data of isolates from nasal swabs of patients admitted to the intensive care unit (ICU) of Hiroshima University hospital. The relationship between isolate genotypes and virulence factors, particularly for isolates that caused infectious diseases during ICU admission was compared with those that did not. The nasal carriage rates of methicillin-resistant S. aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) in patients admitted to the ICU were 7.0% and 20.1%, respectively. The carriage rate of community-acquired (CA)-MRSA was 2.3%, accounting for 32.8% of all MRSA isolates. Whole-genome sequencing analysis of the MRSA isolates indicated that most, including CA-MRSA and healthcare-associated (HA)-MRSA, belonged to clonal complex (CC) 8 [sequence type (ST) 8] and SCCmec type IV. Furthermore, results for three disease foci (pneumonia, skin and soft tissue infection, and deep abscess) and the assessment of virulence factor genes associated with disease conditions [bacteremia, acute respiratory distress syndrome (ARDS), disseminated intravascular coagulopathy (DIC), and septic shock] suggested that nasal colonization of S. aureus clones could represent a risk for patients within the ICU. Particularly, MRSA/J and MSSA/J may be more likely to cause deep abscess infection; ST764 may cause ventilation-associated pneumonia, hospital-acquired pneumonia and subsequent bacteremia, and ARDS, and tst-1-positive isolates may cause DIC onset.IMPORTANCENasal colonization of MRSA in patients admitted to the intensive care unit (ICU) may predict the development of MRSA infections. However, no bacteriological data are available to perform risk assessments for Staphylococcus aureus infection onset. In this single-center 2-year genomic surveillance study, we analyzed all S. aureus isolates from nasal swabs of patients admitted to the ICU and those from the blood or lesions of in-patients who developed infectious diseases in the ICU. Furthermore, we identified the virulent clones responsible for causing infectious diseases in the ICU. Herein, we report several virulent clones present in the nares that are predictive of invasive infections. This information may facilitate the design of preemptive strategies to identify and eradicate virulent MRSA strains, reducing nosocomial infections within the ICU.

Molecular detection of Virulence Factor Genes in Candida albicans Isolated from the Oral Cavity of Local Chicken and Antifungal Susceptibility in Mosul Province

Molecular detection of Virulence Factor Genes in Candida albicans Isolated from the Oral Cavity of Local Chicken and Antifungal Susceptibility in Mosul Province

Prevalence and Molecular Detection of Virulence Genes among Multidrug-Resistant Escherichia coli from Human Clinical Samples and Poultry in Duhok City, Iraq

Abstract Background: Extended-spectrum beta-lactamase (ESBL)-producing, carbapenem-resistant Escherichia coli has increased virulence and multidrug resistance (MDR). Objectives: This study was designed to ascertain the frequency of some virulence factor genes, antibiotic susceptibility patterns, ESBLs, and MDR, focused on colistin-resistant E. coli strains of human and animal origin in Duhok city, Iraq. Materials and Methods: Between December 2020 and April 2021, a total of 150 E. coli isolates (110 from human clinical specimens and 40 from poultry cloacal swabs) were included in this study. The isolates underwent screening for antibiotic susceptibility, MDR, ESBL, and molecular detection of four virulence genes (fimA, cnfL, crL, and hlyA) was conducted using the polymerase chain reaction. Results: Urine specimens (77.2%) compared to blood, wound, vaginal swab, sputum, and semen from outpatients (71.8%). All strains from humans and poultry showed high resistance to ampicillin (86%–97%), ceftriaxone (74%–47%), tetracycline (72%–85%), ciprofloxacin (48%–97%), and colistin (17%–12%). The lowest levels of resistance were found for carbapenems (4%–4%), and the MDR for the isolates was 63%–93%. Apart from carbapenems, colistin-resistant isolates, especially those from poultry, exhibited significant resistance to other antibiotics, and 57% of these isolates being ESBL producers. Three virulence genes (fimA, cnfL, and crL) were highly prevalent (92%) in human isolates, with the crL gene being predominant (100%). Among poultry isolates, fimA was more prevalent (94%) while crL was less common (6%). Conclusion: The predominance of isolates of colistin-resistant poultry origin and the virulence of isolates of human E. coli origin indicate that both strains are currently experiencing an increase in antibiotic resistance.

Community coalescence under variable hydrochemical conditions of the Chesapeake Bay shaped bacterial diversity and functional traits

Community coalescence related to bacterial mixing events regulates community characteristics and affects the health of estuary ecosystems. At present, bacterial coalescence and its driving factors are still unclear. The present study used a dataset from the Chesapeake Bay (2017) to address how bacterial community coalescence in response to variable hydrochemistry in estuarine ecosystems. We determined that variable hydrochemistry promoted the deterioration of water quality. Temperature, orthophosphate, dissolved oxygen, chlorophyll a, Secchi disk depth, and dissolved organic phosphorus were the key environmental factors driving community coalescence. Bacteria with high tolerance to environmental change were the primary taxa accumulated in community coalescence, and the significance of deterministic processes to communities was revealed. Community coalescence was significantly correlated with the pathways of metabolism and organismal systems, and promoted the co-occurrence of antibiotic resistance and virulence factor genes. Briefly, community coalescence under variable hydrochemical conditions shaped bacterial diversity and functional traits, to optimise strategies for energy acquisition and lay the foundation for alleviating environmental pressures. However, potential pathogenic bacteria in community coalescence may be harmful to human health and environmental safety. The present study provides a scientific reference for ecological management of estuaries.

Removal of intracellular and extracellular antibiotic resistance genes and virulence factor genes using electricity-intensified constructed wetlands

Constructed wetland (CW) is considered a promising technology for the removal of emerging contaminants. However, its removal performance for antibiotic resistance genes (ARGs) is not efficient and influence of virulence factor genes (VFGs) have not been elucidated. Here, removal of intracellular and extracellular ARGs as well as VFGs by electricity-intensified CWs was comprehensively evaluated. The two electrolysis-intensified CWs can improve the removal of intracellular ARGs and MGEs to 0.96- and 0.85-logs, respectively. But cell-free extracellular ARGs (CF-eARGs) were significantly enriched with 1.8-logs in the electrolysis-intensified CW. Interestingly, adding Fe-C microelectrolysis to the electrolysis-intensified CW is conducive to the reduction of CF-eARGs. However, the detected number and relative abundances of intracellular and extracellular VFGs were increased in all of the three CWs. The biofilms attached onto the substrates and rhizosphere are also hotspots of both intracellular and particle-associated extracellular ARGs and VFGs. Structural equation models and correlation analysis indicated that ARGs and VFGs were significantly cooccurred, suggesting that VFGs may affect the dynamics of ARGs. The phenotypes of VFGs, such as biofilm, may act as protective matrix for ARGs, hindering the removal of resistance genes. Our results provide novel insights into the ecological remediation technologies to enhance the removal of ARGs.