Sul1 Genes Research Articles

Distribution of bacterial community structures and spread of antibiotic resistome at industrially polluted sites of Mini River, Vadodara, Gujarat, India.

The influence of anthropogenic pollution on the distribution of bacterial diversity, antibiotic-resistant bacteria (ARBs), and antibiotic resistance genes (ARGs) was mapped at various geo-tagged sites of Mini River, Vadodara, Gujarat, India. The high-throughput 16S rRNA gene amplicon sequencing analysis revealed a higher relative abundance of Planctomycetota at the polluted sites, compared to the pristine site. Moreover, the relative abundance of Actinobacteriota increased, whereas Chloroflexi decreased in the water samples of polluted sites than the pristine site. The annotation of functional genes in the metagenome samples of Mini River sites indicated the presence of genes involved in the defence mechanisms against bacitracin, aminoglycosides, cephalosporins, chloramphenicol, streptogramin, streptomycin, methicillin, and colicin. The analysis of antibiotic resistome at the polluted sites of Mini River revealed the abundance of sulfonamide, beta-lactam, and aminoglycoside resistance. The presence of pathogens and ARB was significantly higher in water and sediment samples of polluted sites compared to the pristine site. The highest resistance of bacterial populations in the Mini River was recorded against sulfonamide (≥ 7.943 × 103CFU/mL) and ampicillin (≥ 8.128 × 103CFU/mL). The real-time PCR-based quantification of ARGs revealed the highest abundance of sulfonamide resistance genes sul1 and sul2 at the polluted sites of the Mini River. Additionally, the antimicrobial resistance genes aac(6')-Ib-Cr and blaTEM were also found abundantly at polluted sites of the Mini River. The findings provide insights into how anthropogenic pollution drives the ARG and ARB distribution in the riverine ecosystem, which may help with the development of antimicrobial resistance mitigation strategies.

Detection of Antimicrobial Resistance by Phenotypic and Genotypic Methods in E. coli Isolated from Diarrhea of Cattle Calves in Mhow, Indore

Background: Neonatal calf diarrhoea, a complex illness, continues to be the leading cause of neonatal calf deaths. The complex pathophysiology of infectious calf diarrhoea results in severe losses to the dairy sector, either directly via mortality or indirectly through the costs of prophylaxis and slowed growth in infected calves. Neonatal calf diarrhoea during the first month of life causes between 80 to 85 per cent of all deaths in India. Methods: A total 137 cattle calve diarrhea samples were collected from calves that show clinical signs of diarrhea. Swabs of the rectum were taken directly from diarrhoeic calves and collected in sterile test tubes. They were then immediately transported to the laboratory for the isolation of E. coli. Result: A total 64 E. coli isolates were detected and identified from samples via biochemical and molecular assays employing PCR targeting the E. coli specific 16s rRNA gene. The result of antibiotic sensitivity of 64 isolated strains of E. coli to 15 antimicrobial drugs shows highest sensitivity towards antibiotic gentamycin (68.75%) followed by sulfadiazine and co-trimoxazole (39.06), ciprofloxacin (32.81%) and highest resistance against ampicillin (87.5%) followed by ceftriaxone (82.813%), ceftazidime plus clav (81.25%). There is the variation in the expression of the AMR genes in the isolated E. coli strains. The detection rate of AMR gene blaTEM was highest 90% and that of aadA was the lowest (0%). The other AMR genes blaCTX-M, blaSHV, sul1 and tetA showing 50%, 20%, 50% and 30%.

Prevalence of diverse antimicrobial resistance genes and bacteria in sewage treatment plant-derived sludge environment.

Antimicrobial resistance (AMR) contamination in the environment is one of the most significant worldwide threats of the 21st century. Since sludge is heavily exposed to diverse contaminants, including pharmaceuticals, the inhabitant bacterial population is expected to exhibit resistance to antimicrobial agents. In this study, sewage treatment plant (STP) sludge samples were analyzed to assess the antibiotic-resistant bacterial population, abundance of AMR genes (ermF, qnrS, Sul1, blaGES, blaCTX-M, and blaNDM), and mobile genetic elements (intl1 and IS26). Out of 16, six bacterial isolates exhibited resistance to 13 antibiotics with a high multiple antibiotic resistance index (MARI) (0.93) and high metal tolerance. Quantitative polymerase chain reaction showed the abundance of target genes ranging from 6.6×103 to 6.5×108 copies g-1 sludge. The overall outcome reveals that STP sludge comprised varied multidrug-resistant bacterial populations. It will give insights into the functions of heavy metals and biofilm development in the selection and spread of AMR genes and the associated bacteria. Therefore, the application of sludge needs proper screening for AMR and metal contamination prior to its countless applications. This study will contribute immensely to the risk analysis of STP effluents on environmental health, including control of AMR transmission.

Revealing the response characteristics of periphyton biomass and community structure to sulfamethoxazole exposure in aquaculture water: The perspective of microbial network relationships

The widespread application of sulfonamide antibiotics in aquaculture has raised concerns about their adverse environmental impacts. Periphyton plays a crucial role in the aquatic ecosystem. In this study, we examined sulfamethoxazole (SMX) effects on the community structure and interactions of periphyton in simulated aquaculture water. Our findings indicated that the total biomass of periphyton decreased, while the biomass of periphytic algae and the secretion of extracellular polymeric substances (EPS) increased at 0.7 × 10−3 mg/L. Under higher SMX concentrations (5 mg/L and 10 mg/L), periphyton growth was severely inhibited, the microbial community structure of periphyton were sharply altered, characterized by the cyanobacteria growth suppression and decrease in the diversity index of community. Furthermore, elevated SMX concentrations (5 mg/L and 10 mg/L) increased the ratio of negative relationships from 45.4% to 49.4%, which suggested that high SMX concentrations promoted potential competition among microbes and disrupted the microbial food webs in periphyton. The absolute abundance of sul1 and sul2 genes in T2 and T3 groups were 2–3 orders of magnitude higher than those in control group after 30 days of SMX exposure, which elevated the risk of resistance gene enrichment and dissemination in the natural environment. The study contributes to our understanding of the detrimental effects of antibiotic pollution, which can induce changes in the structure and interaction relationship of microbial communities in aquaculture water.

Occurrence and fate of CECs (OMPs, ARGs and pathogens) during decentralised treatment of black water and grey water

Decentralised wastewater treatment is becoming a suitable strategy to reduce cost and environmental impact. In this research, the performance of two technologies treating black water (BW) and grey water (GW) fractions of urban sewage is carried out in a decentralised treatment of the wastewater produced in three office buildings. An Anaerobic Membrane Bioreactor (AnMBR) treating BW and a Hybrid preanoxic Membrane Bioreactor (H-MBR) containing small plastic carrier elements, treating GW were operated at pilot scale. Their potential on reducing the release of contaminants of emerging concern (CECs) such as Organic Micropollutants (OMPs), Antibiotic Resistance Genes (ARGs) and pathogens was studied. After 226 d of operation, a stable operation was achieved in both systems: the AnMBR removed 92.4 ± 2.5 % of influent COD, and H-MBR removed 89.7 ± 3.5 %. Regarding OMPs, the profile of compounds differed between BW and GW, being BW the matrix with more compounds detected at higher concentrations (up to μg L−1). For example, in the case of ibuprofen the concentrations in BW were 23.63 ± 3.97 μg L−1, 3 orders of magnitude higher than those detected in GW. The most abundant ARGs were sulfonamide resistant genes (sul1) and integron class 1 (intl1) in both BW and GW. Pathogenic bacteria counts were reduced between 1 and 3 log units in the AnMBR. Bacterial loads in GW were much lower than in BW, being no bacterial re-growth observed for the GW effluents after treatment in the H-MBR. None of the selected enteric viruses was detected in GW treatment line.

Template switching between the leading and lagging strands at replication forks generates inverted copy number variants through hairpin-capped extrachromosomal DNA.

Inherited and germ-line de novo copy number variants (CNVs) are increasingly found to be correlated with human developmental and cancerous phenotypes. Several models for template switching during replication have been proposed to explain the generation of these gross chromosomal rearrangements. We proposed a model of template switching (ODIRA-origin dependent inverted repeat amplification) in which simultaneous ligation of the leading and lagging strands at diverging replication forks could generate segmental inverted triplications through an extrachromosomal inverted circular intermediate. Here, we created a genetic assay using split-ura3 cassettes to trap the proposed inverted intermediate. However, instead of recovering circular inverted intermediates, we found inverted linear chromosomal fragments ending in native telomeres-suggesting that a template switch had occurred at the centromere-proximal fork of a replication bubble. As telomeric inverted hairpin fragments can also be created through double strand breaks we tested whether replication errors or repair of double stranded DNA breaks were the most likely initiating event. The results from CRISPR/Cas9 cleavage experiments and growth in the replication inhibitor hydroxyurea indicate that it is a replication error, not a double stranded break that creates the inverted junctions. Since inverted amplicons of the SUL1 gene occur during long-term growth in sulfate-limited chemostats, we sequenced evolved populations to look for evidence of linear intermediates formed by an error in replication. All of the data are compatible with a two-step version of the ODIRA model in which sequential template switching at short inverted repeats between the leading and lagging strands at a replication fork, followed by integration via homologous recombination, generates inverted interstitial triplications.

Clonal spread of trimethoprim-sulfamethoxazole-resistant Stenotrophomonas maltophilia isolates in a tertiary hospital.

The aims of this study were to: (i) determine antibiotic susceptibility of clinical Stenotrophomonas maltophilia isolates, (ii) investigate the presence of different classes of integrons and sul genes responsible for sulphonamide resistance, (iii) assess the molecular epidemiology of the isolates by determining their clonal relatedness, and (iv) investigate the potential sources of infection by collecting environmental samples when necessary. 99 S. maltophilia isolates from clinical specimens of hospitalized patients were screened by PCR for sul1, sul2, sul3 genes, and integron-associated integrase genes: intI1, intI2, and intI3. PFGE was used to determine the clonal relatedness of the isolates. Susceptibility rates for trimethoprim-sulfamethoxazole, levofloxacin, and ceftazidime were 90.9%, 91.9%, and 53.5% respectively. All trimethoprim-sulfamethoxazole-resistant isolates were positive for intI1 and sul1. PFGE analysis revealed that 24 of the isolates were clonally related, clustering in seven different clones. Five of the nine trimethoprim-sulfamethoxazole-resistant isolates were clonally related. The first isolate in this clone was from a wound sample of a patient in the infectious diseases clinic, and the other four were isolated from the bronchoalveolar lavage samples of patients in the thoracic surgery unit. The patient with the first isolate neither underwent bronchoscopy nor stayed in the thoracic surgery unit. Although clustering was observed in bronchoalveolar lavage samples, no S. maltophilia growth was detected in environmental samples. The findings demonstrated that the sul1 gene carried by class 1 integrons plays an important role in trimethoprim-sulfamethoxazole resistance in S. maltophilia isolates. PFGE analysis revealed a high degree of genetic diversity. However, detection of clonally related isolates suggests the acquisition from a common source and/or cross-transmission of this microorganism between the patients.

Genetic detection of Resistance Genes among Enterobacter cloacae and Citrobacter spp. isolates

Internationally, there is an increasing prevalence of diseases caused by germs that are resistant to many drugs, which is often a reason for worry. The objective of the present investigation was to identify specific genes linked to the resistance of beta-lactam, macrolide, and sulfa medications in Enterobacter cloacae and Citrobacter spp. isolates. A total of 364 patients were included in the study, with 132 (36.26%) being male and 232 (63.74%) being female. The results indicated that 135 samples, accounting for 37.08% of the total, exhibited positive bacterial growth. The findings indicated that Enterobacter cloacae was present in 25 (6.86%) of the patients with urinary tract infections, whereas Citrobacter spp. was found in only 2 (0.54%) cases (consisting of 1 (0.27%) C. freundii isolate and 1 (0.27%) C. farmer isolate). The PCR findings indicated that the sul-1 gene was present in 23 out of 25 (92%) E. cloacae isolates and in 2 out of 2 (100%) Citrobacter spp. isolates. On the other hand, the sul-2 gene was detected in 22 out of 25 (88%) E. cloacae isolates and in 2 out of 2 (100%) Citrobacter spp. isolates. The mph(A) gene was present in 80% (20/25) of E. cloacae isolates and in 100% (2/2) of Citrobacter spp. isolates. The findings showed that 96% of the E. cloacae isolates and 100% of the Citrobacter spp. isolates tested positive for both ctx-m and ctx-m-10 genes. The prevalence of the veb gene in E. cloacae isolates was 76% (19/25), but in Citrobacter spp. isolates it was 100% (2/2). The investigation did not detect the gene ctx-m-14 in any of the local isolates examined. Genes of sul-3 and ctx-m-14 were no recorded in this study.

Antibiotic Resistance in a Coastal River in Mississippi, USA – Potential Drivers

Wastewater treatment plants (WWTPs) are major sources of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in water bodies. Most studies on the impact of WWTPs on antibiotic resistance have focused on freshwater systems, with little information on coastal and estuarine waters with variable salinity. This study monitored seasonal levels of ARGs at the effluent and downstream of the Pascagoula— Moss Point WWTP in the lower Pascagoula River, a coastal river in southeastern Mississippi, USA. Surface water samples were collected seasonally at upstream, outflow, and 3 downstream sites from February to November 2016. Bacterial resistance to sulfamethazine, tetracycline, and ciprofloxacin was quantified using selective culture and qPCR. Mixed—effects models were developed to identify potential driving factors of ARG concentrations related to the WWTP and local environmental conditions (salinity, water temperature, and pH). The best model was selected based on the lowest Akaike Information Criterion (AIC) corrected for small sample size. The results show that the genes sul1, sul2, and intI1 were detected, with intI1 having the highest relative concentration. The qPCR analysis suggests a negative relation between ARG levels and temperature and salinity. ARG concentrations peaked immediately downstream of the WWTP and decreased gradually further downstream in some months, but the spatial pattern varied widely between sampling months. The study highlights the complex patterns of environmental ARGs and the importance of accounting for the impact of WWTPs, local environmental factors, and other anthropogenic influences to understand their potential drivers.

Multirresistência antimicrobiana e formação de biofilmes em isolados de Salmonella enterica da cadeia produtiva de frangos

ABSTRACT: Poultry and poultry products are considered the predominant sources of Salmonella enterica contamination and are important reservoirs of bacteria with antimicrobial resistance. The objective of this study was to identify Salmonella with multidrug resistance (MDR) phenotype, with the ability to form biofilms and elucidate the presence of genes that encode antimicrobial resistance in isolates from the broiler production chain in the state of Maranhão, Brazil. A total of 121 strains of S. enterica of different serovars were evaluated for antimicrobial susceptibility, and of these, 26 strains were used to detect the ability to form biofilms and identify resistance genes using PCR. Antimicrobial resistance was observed in 95 (78.5%) Salmonella isolates, and 57 (47.1%) showed MDR phenotype. The isolates showed greater resistance to the sulfonamide principles (58.7%), trimethoprim (48.8%), tetracycline (45.4%), nalidixic acid (44.6%), amoxicillin and ampicillin (26.4%), and cefazolin (22.3%). Salmonella Schwarzengrund (n=21/61.7%), Albany (n=15/62.5%), and Enteritidis (n=4/44.5%) showed the highest indices of MDR phenotype. The ability to form biofilms at 37°C was found in 13 of the 26 strains evaluated, which were considered poor producers. The resistance genes blaCTX-M, blaCTX-M2, blaSHV, sul1, sul2, tetA, tetB, tetC, tetE, dfrA12, and dfrA1 were observed in the serovars Schwarzengrund, Albany, Enteritidis, Heidelberg, and Typhimurium. The results showed a high occurrence of S. enterica, with multiple resistance to conventional antimicrobials and the ability to form biofilms in the poultry production chain.

Tracking antimicrobial resistance indicator genes in wild flatfish from the English Channel and the North Sea area: A one health concern

Antimicrobial resistance (AMR) is a burgeoning environmental concern demanding a comprehensive One Health investigation to thwart its transmission to animals and humans, ensuring food safety. Seafood, housing bacterial AMR, poses a direct threat to consumer health, amplifying the risk of hospitalization, invasive infections, and death due to compromised antimicrobial treatments. The associated antimicrobial resistance genes (ARGs) in diverse marine species can amass and transmit through various pathways, including surface contact, respiration, and feeding within food webs. Our research, focused on the English Channel and North Sea, pivotal economic areas, specifically explores the occurrence of four proposed AMR indicator genes (tet(A), blaTEM, sul1, and intI1) in a benthic food web. Analyzing 350 flatfish samples' skin, gills, and gut, our quantitative PCR (qPCR) results disclosed an overall prevalence of 71.4% for AMR indicator genes. Notably, sul1 and intI1 genes exhibited higher detection in fish skin, reaching a prevalence of 47.5%, compared to gills and gut samples. Proximity to major European ports (Le Havre, Dunkirk, Rotterdam) correlated with increased AMR gene frequencies in fish, suggesting these ports' potential role in AMR spread in marine environments. We observed a broad dispersion of indicator genes in the English Channel and the North Sea, influenced by sea currents, maritime traffic, and flatfish movements. In conclusion, sul1 and intI1 genes emerge as robust indicators of AMR contamination in the marine environment, evident in seawater and species representing a benthic food web. Further studies are imperative to delineate marine species' role in accumulating and transmitting AMR to humans via seafood consumption. This research sheds light on the urgent need for a concerted effort in comprehending and mitigating AMR risks in marine ecosystems within the context of One Health.

Characterization of an ST38 carbapenem-resistant and highly virulent Escherichia coli carrying conjugatively transferable ColV virulence-resistance and blaNDM-5-positive resistance plasmids.

To characterize an Escherichia coli strain causing bloodstream infection encoding both high-virulence and carbapenem-resistance phenotypes. Antimicrobial susceptibility testing, WGS and bioinformatics analysis were performed to characterize strain E1. The function of the ColV plasmid was investigated by the Galleria mellonella infection model, serum killing and macrophage killing assays. The fitness effect of the ColV plasmid was tested by growth curve, plasmid stability tests and the in vitro competition assay. The conjugation assay was performed to test the transferability of the ColV and blaNDM-5-carrying plasmids. E. coli E1 from bloodstream infection was MDR and highly virulent in the G. mellonella infection model. It belonged to phylogroup D, ST38 and serotype O7:H8. E1 carried a conjugatively transferable IncI1-type blaNDM-5-positive plasmid, which conferred carbapenem resistance, a conjugative IncFIB/FII-type ColV plasmid encoding an array of virulence-associated genes and antibiotic resistance genes blaTEM-1B, strAB and sul2, and seven other plasmids. Co-transfer of the ColV plasmid and the blaNDM-5-positive plasmid was observed. The ColV virulence-resistance hybrid plasmid contributed to the virulence, resistance to serum killing, and macrophage phagocytosis in E. coli E1. The carriage of this ColV plasmid did not constitute an in vitro fitness burden to strain E1 but caused fitness costs to E. coli strain EC600. The emergence of such a highly virulent and resistant strain with conjugative blaNDM-5-positive and ColV plasmids posed a significant threat to public health. Implementation of control measures is needed to prevent such strains from further disseminating in hospital settings and the community.

Molecular characterization of the multi-drug resistant Myroides odoratimimus isolates: a whole genome sequence-based study to confirm carbapenem resistance.

The bacteria belonging to the Myroides genus are opportunistic pathogens causing community or hospital-acquired infections that result in treatment failure due to antibiotic resistance. This study aimed to investigate molecular mechanisms of antibiotic resistance, clonal relatedness, and the biofilm forming capacity of the 51 multi-drug resistant Myroides odoratimimus. All isolates were screened for blaKPC, blaOXA, blaVIM, blaIMP, blaMUS, blaTUS, blaNDM, and blaB genes by using PCR amplification. Whole genome sequencing (WGS) was applied on three randomly selected isolates for further investigation of antibiotic resistance mechanisms. Clonal relatedness was analyzed by Pulsed-field gel electrophoresis (PFGE) and the microtiter plate method was used to demonstrate biofilm formation. All isolates were positive for biofilm formation. PCR analysis resulted in a positive for only the blaMUS-1 gene. WGS identified blaMUS-1, erm(F), ere(D), tet(X), and sul2 genes in all strains tested. Moreover, the genomic analyses of three strains revealed that genomes contained a large number of virulence factors (VFs). PFGE yielded a clustering rate of 96%. High clonal relatedness, biofilm formation, and multi-drug resistance properties may lead to the predominance of these opportunistic pathogens in hospital environments and make them cause nosocomial infections.

Quorum Quenching with a Diffusible Signal Factor Analog in Stenotrophomonas maltophilia.

Stenotrophomonas maltophilia is a multidrug-resistant Gram-negative bacillus associated with nosocomial infections in intensive care units, and nowadays, its acquired resistance to trimethoprim-sulfamethoxazole (SXT) by sul genes within class 1 integrons is a worldwide health problem. Biofilm and motility are two of the major virulence factors in this bacterium and are auto-induced by the diffusible signal factor (DSF). In recent studies, retinoids have been used to inhibit (Quorum Quenching) these virulence factors and for their antimicrobial effect. The aim was to reduce biofilm formation and motility with retinoic acid (RA) in S. maltophilia SXT-resistant strains. Eleven SXT-resistant strains and two SXT-susceptible strains were tested for biofilm formation/reduction and planktonic/sessile cell viability with RA and SXT-MIC50/RA; motility (twitching, swimming, swarming) was measured with/without RA; and MLST typing was determined. The biofilm formation of the strains was classified as follows: 15.38% (2/13) as low, 61.54% (8/13) as moderate, and 23.08% (3/13) as high. It was significantly reduced with RA and SXT-MIC50/RA (p < 0.05); cell viability was not significantly reduced with RA (p > 0.05), but it was with SXT-MIC50/RA (p < 0.05); and swimming (p < 0.05) and swarming (p < 0.05) decreased significantly. MLST typing showed the first and novel strains of Mexican S. maltophilia registered in PubMLST (ST479-485, ST497, ST23, ST122, ST175, ST212, and ST300). In conclusion, RA reduced biofilm formation and motility without affecting cell viability; furthermore, antimicrobial synergism with SXT-MIC50/RA in different and novel STs of S. maltophilia was observed.

Exogenous mobile genetic elements and their associated integrons drive the enrichment of antibiotic-resistant genes in the river of a valley basin city (Lanzhou, China).

River is a unique source of drinking water in valley-type cities, affecting local urban development and human lifestyles. However, the key driving factors for dissemination of antibiotic-resistant genes (ARGs) in valley-type urban environments remain unclear. This study aimed to investigate the distribution of ARGs in the Yellow River and to clarify the driving factors of ARGs in a typical valley basin city (Lanzhou, China). The seven selected ARGs with higher abundances including tetracycline resistance genes (tetM, tetX), macrolide resistance genes (ermB, ermF, ereA), and sulfonamide resistance genes (sul1, sul2) were detected. The results showed that the total absolute abundance of all the selected ARGs varied from 9.97 × 1012 to 1.04 × 1015 copies/L in the water body, with higher abundances in the wet season, relative to the dry season. Among these, sulfonamide resistance genes (sul1, sul2) displayed the highest absolute abundance in the river and soil. The ARGs and mobile genetic elements (MGEs) were significantly correlated with bacterial abundance, dissolved organic carbon (DOC), ammonia nitrogen (NH4+), and total nitrogen (TN) levels in the water environment (Mantel test, P < 0.01). Structural equation modeling revealed the direct input of point-source and nonpoint-source ARGs in this area contributed less to the overall level of the ARGs in the water. Among the multiple drivers, the MGEs derived from wastewater treatment plant and anthropogenic nonpoint area positively and directly affected the ARG profiles in water (P < 0.01), rather than the factors of bacterial abundance and physicochemical properties. According to this study, the exogenous MGEs from anthropogenic activities are the main driver for the enrichment of ARGs in the valley-type urban river environment.

Effect of pH on antibiotic resistance genes removal and bacterial nucleotides metabolism function in the wastewater by the combined ferrate and sulfite treatment

The variations of the antibiotic resistance genes removal and the bacterial nucleotide metabolism function in the wastewater by the combined ferrate and sulfite treatment at different pH levels were investigated. The total DNA as well as the genes sul2, sul3 and intI1 removal by the combined 10 mg/L ferrate and 28.5 mg/L sulfite treatment and the contributions of the sulfate radicals and the hydroxyl radicals produced by the reaction of ferrate with sulfite to the DNA removal decreased as the pH increase from 6.0 to 9.0. Bacterial nucleotide metabolism level changes affected the DNA removal, and the highest nucleotide degradation metabolism level and the lowest level of the nucleotide biosynthesis and salvage metabolism were found at pH 6.0. The reduction of the genes sul2, sul3 and intI1 host level contributed to the genes sul2, sul3 and intI1 removal. This study provides novel insights into the antibiotic resistance genes removal by the combined ferrate and sulfite treatment at different pH levels.

Removal of selected sulfonamides and sulfonamide resistance genes from wastewater in full-scale constructed wetlands

Sulfonamides are high-consumption antibiotics that reach the aquatic environment. The threat related to their presence in wastewater and the environment is not only associated with their antibacterial properties, but also with risk of the spread of drug resistance in bacteria. Therefore, the aim of this work was to evaluate the occurrence of eight commonly used sulfonamides, sulfonamide resistance genes (sul1–3) and integrase genes intI1–3 in five full-scale constructed wetlands (CWs) differing in design (including hybrid systems) and in the source of wastewater (agricultural drainage, domestic sewage/surface runoff, and animal runs runoff in a zoo). The CWs were located in low-urbanized areas in Poland and in Czechia. No sulfonamides were detected in the CW treating agricultural tile drainage water. In the other four systems, four sulfonamide compounds were detected. Sulfamethoxazole exhibited the highest concentration in those four CWs and its highest was 12,603.23 ± 1000.66 ng/L in a CW treating a mixture of domestic sewage and surface runoff. Despite the high removal efficiencies of sulfamethoxazole in the tested CWs (86 %–99 %), it was still detected in the treated wastewater. The sul1 genes occurred in all samples of raw and treated wastewater and their abundance did not change significantly after the treatment process and it was, predominantly, at the level 105 gene copies numbers/mL. Noteworthy, sul2 genes were only found in the influents, and sul3 were not detected. The sulfonamides can be removed in CWs, but their elimination is not complete. However, hybrid CWs treating sewage were superior in decreasing the relative abundance of genes and the concentration of SMX. CWs may play a role in the dissemination of sulfonamide resistance genes of the sul1 type and other determinants of drug resistance, such as the intI1 gene, in the environment, however, the magnitude of this phenomenon is a matter of further research.

Characterization of microbial contamination in agricultural soil: A public health perspective

Soil is widely recognized as a reservoir of microbial contaminants including antibiotic resistance genes (ARGs) and human bacterial pathogens (HBPs), which are major public health concerns. Although the risks associated with soil safety in different soil habitats have been studied, the results are not comprehensive. In this study, dryland soils used for vegetable, corn, and soybean planting, and submerged soils used for rice planting and crab farming were collected and subjected to metagenomic sequencing to characterize HBPs, ARGs, and virulence factor genes (VFGs). The results showed that submerged soils had a higher abundance of HBP than dryland soils. In addition, the submerged soil microbiome acquired significantly higher levels of high-risk ARGs than the dryland soil microbiome and these ARGs were mainly assigned to bacA, sul1, and aadA genes submerged. Network analysis revealed that 11 HBPs, including Yersinia enterocolitica, Vibrio cholerae, Escherichia coli, and Leptospira interrogans, were high-risk because of their close association with ARGs, VFGs, and mobile genetic elements (MGEs). Procrustes and network analyses showed that HBPs and ARGs were more closely linked in submerged soil. This study confirms that submerged field has higher ecological environment risk and human health risk than dryland soil.

Chloroxylenol positively affects the aerobic sequencing batch reactor performance and reshapes microbial communities and antibiotic resistance genes

The antibacterial ingredient chloroxylenol (PCMX) has been frequently detected in various environments, raising increasing concerns about its potential risks to organisms and ecosystems. Herein, the impacts of PCMX on the performance and microbial communities of aerobic sequencing batch reactors were investigated. The results demonstrated that 0.1 and 1.0 mg/L PCMX did not affect the removal performance of COD and NH4+-N, as well as the production of extracellular polymeric substances. However, a notable enhancement in these performances was observed when the concentration of PCMX reached 10 mg/L. Analysis of microbial community revealed that exposure to PCMX did not alter α diversity but significantly shifted the community structure. Spirosoma and Ferruginibacter were identified as dominant genera across all stages in the sludge, with Ferruginibacter being inhibited by 10 mg/L PCMX (from 20.3 % to 11.1 %). Network analysis revealed that PCMX had a diminishing effect on the interaction and complexity of microbial. High throughput qPCR analysis indicated that PCMX reshaped the antibiotic resistance genes (ARGs). The relative gene copy number increased from 0.23 to 0.29 at a concentration of 0.1 mg/L PCMX, while it decreased from 0.51 to 0.40 by 10 mg/L PCMX. Genes tetX and sul2 were identified as dominant genes unaffected by 0.1 and 1.0 mg/L but significantly inhibited by 10 mg/L PCMX. These findings suggested that environmental levels of PCMX might promote the enrichment of ARGs, while higher concentrations impeded gene propagation. This study provides new information regarding the roles, potential risks, and removal strategies of PCMX in wastewater treatment processes.

Isolation and identification of antimicrobial susceptibility, biofilm formation, efflux pump activity, and virulence determinants in multi-drug resistant Pseudomonas aeruginosa isolated from freshwater fishes.

The present study was undertaken to evaluate the prevalence, underlying resistance mechanism, and virulence involved in Pseudomonas aeruginosa (n = 35) isolated from freshwater fishes in Andhra Pradesh, India. Antibiogram studies revealed that 68.5, 62.8, 37.1, 11.4, 8.5, 57.1, 54.2, and 48.5% of isolates had resistance to oxytetracycline, co-trimoxazole, doxycycline, enrofloxacin, ciprofloxacin, cefotaxime, ceftazidime, and ampicillin, respectively. The resistant isolates harboured the tetA (85.7%), tetD (71.4%), tetM (91.4%), sul1 (80%), blaCTX-M (57.1%), blaTEM (42.8%), and blaSHV (48.5%) genes. In total, 50% of the isolates were altered as multi-drug resistant, and the multiple antibiotic resistance index was calculated as 0.4. Furthermore, 37.3, 48.5, and 14.2% of isolates were categorized as strong, moderate, and weak biofilm formers, possessing pslA (91.5%) and pslD (88.6%) biofilm encoding genes. In total, 82.8% of the isolates exhibited efflux pump activity and harboured the mexA (74.2%), mexB (77.1%), and oprM (37.1%) genes. Virulent genes oprL, toxA, exoS, and phzM were detected in 68.5, 68.5, 100, and 17.1% of isolates, respectively. The data suggested that P. aeruginosa harbours multiple resistance mechanisms and virulence factors that may contribute to antibiotic resistance and pathogenicity, and their distribution in fish culture facilities highlights the public health hazards of the food chain.