Integron Gene Research Articles

Dissemination of sulfonamide resistance genes in digester microbiome during anaerobic digestion of food waste leachate

The preeminence of sulfonamide drug resistance genes in food waste (FW) and the increased utilization of high-strength organic FW in anaerobic digestion (AD) to enhance methane production have raised severe public health concerns in wastewater treatment plants worldwide. In this regard, the dissemination patterns of different sulfonamide resistance genes (sul1 and sul2) and their impact on the digester core microbiota during AD of FW leachate (FWL) were evaluated. The presence of various sulfonamide antibiotics (SAs) in FWL digesters improved the final methane yield by 37 % during AD compared with FWL digesters without SAs. Microbial population shifts towards hydrolytic, acidogenic, and acetogenic bacteria in the phyla Actinobacteriota, Bacteroidota, Chloroflexi, Firmicutes, Proteobacteria, and Synergistota occurred due to SA induced substrate digestion and absorption through active transport; butanoate, propanoate, and pyruvate metabolism; glycolysis; gluconeogenesis; the citrate cycle; and pentose phosphate pathway. The initial dominance of Methanosaeta (89–96 %) declined to 47–53 % as AD progressed and shifted towards Methanosarcina (40 %) in digesters with the highest SA concentrations at the end of AD. Dissemination of sul1 depended on class 1 integron gene (intl1)-based horizontal gene transfer to pathogenic members of Chloroflexi, Firmicutes, and Patescibacteria, whereas sul2 was transmitted to Synergistota independent of intl1. Low susceptibility and ability to utilize SAs during methanogenesis shielded methanogenic archaea against selection pressure, thus preventing them from interacting with sul or intl1 genes, thereby minimizing the risk of antibiotic resistance development. The observed emergence of cationic antimicrobial peptide, vancomycin, and β-lactam resistance in the core microbiota during AD of FWL in the presence of SAs suggests that multidrug resistance caused by bacterial transformation could lead to an increase in the environmental resistome through wastewater sludge treatment.

Insight into the spatiotemporal distribution of antibiotic resistance genes in estuarine sediments during long-term ecological restoration

In this study, we aimed to investigate the long-term spatiotemporal changes in hydrodynamics, antibiotics, nine typical subtypes of antibiotic resistance genes (ARGs), class 1 integron gene (intI1), and microbial communities in the sediments of a semi-enclosed estuary during ecological restoration with four treatment stages (influent (#1), effluent of the biological treatment area (#2), oxic area (#3), and plant treatment area (#4)). Ecological restoration of the estuary reduced common pollutants (nitrogen and phosphorus) in the water, whereas variations in ARGs showed noticeable seasonal and spatial features. The absolute abundance of ARGs at sampling site #2 considerably increased in autumn and winter, while it significantly increased at sampling site #3 in spring and summer. The strong intervention of biological treatment (from #1 to #2) and aerators (from #2 to #3) in the estuary substantially affected the distribution of ARGs and dominant antibiotic-resistant bacteria (ARB). The dominant ARB (Thiobacillus) in estuarine sediments may have low abundance but important dissemination roles. Meanwhile, redundancy and network analysis revealed that the microbial communities and intl1 were key factors related to ARG dissemination, which was affected by spatial and seasonal ecological restoration. A positive correlation between low flow velocity and certain ARGs (tetM, tetW, tetA, sul2, and ermC) was observed, implying that flow optimization should also be considered in future ecological restoration to remediate ARGs. Furthermore, the absolute abundance of ARGs can be utilized as an index to evaluate the removal capacity of ARGs by estuarine restoration.

Microbial source tracking and land use associations for antibiotic resistance genes in private wells influenced by human and livestock fecal sources.

Antimicrobial resistance is a growing public health problem that requires an integrated approach among human, agricultural, and environmental sectors. However, few studies address all three components simultaneously. We investigated the occurrence of five antibiotic resistance genes (ARGs) and the class 1 integron gene (intI1) in private wells drawing water from a vulnerable aquifer influenced by residential septic systems and land-applied dairy manure. Samples (n=138) were collected across four seasons from a randomized sample of private wells in Kewaunee County, Wisconsin. Measurements of ARGs and intI1 were related to microbial source tracking (MST) markers specific to human and bovine feces; they were also related to 54 risk factors for contamination representing land use, rainfall, hydrogeology, and well construction. ARGs and intI1 occurred in 5%-40% of samples depending on target. Detection frequencies for ARGs and intI1 were lowest in the absence of human and bovine MST markers (1%-30%), highest when co-occurring with human and bovine markers together (11%-78%), and intermediate when co-occurring with just one type of MST marker (4%-46%). Gene targets were associated with septic system density more often than agricultural land, potentially because of the variable presence of manure on the landscape. Determining ARG prevalence in a rural setting with mixed land use allowed an assessment of the relative contribution of human and bovine fecal sources. Because fecal sources co-occurred with ARGs at similar rates, interventions intended to reduce ARG occurrence may be most effective if both sources are considered.

The fate of sulfonamide resistance genes and anthropogenic pollution marker intI1 after discharge of wastewater into a pristine river stream.

Currently there are sparse regulations regarding the discharge of antibiotics from wastewater treatment plants (WWTP) into river systems, making surface waters a latent reservoir for antibiotics and antibiotic resistance genes (ARGs). To better understand factors that influence the fate of ARGs in the environment and to foster surveillance of antibiotic resistance spreading in such habitats, several indicator genes have been proposed, including the integrase gene intI1 and the sulfonamide resistance genes sul1 and sul2. Here we used quantitative PCR and long-read nanopore sequencing to monitor the abundance of these indicator genes and ARGs present as class 1 integron gene cassettes in a river system from pristine source to WWTP-impacted water. ARG abundance was compared with the dynamics of the microbial communities determined via 16S rRNA gene amplicon sequencing, conventional water parameters and the concentration of sulfamethoxazole (SMX), sulfamethazine (SMZ) and sulfadiazine (SDZ). Our results show that WWTP effluent was the principal source of all three sulfonamides with highest concentrations for SMX (median 8.6 ng/l), and of the indicator genes sul1, sul2 and intI1 with median relative abundance to 16S rRNA gene of 0.55, 0.77 and 0.65%, respectively. Downstream from the WWTP, water quality improved constantly, including lower sulfonamide concentrations, decreasing abundances of sul1 and sul2 and lower numbers and diversity of ARGs in the class 1 integron. The riverine microbial community partially recovered after receiving WWTP effluent, which was consolidated by a microbiome recovery model. Surprisingly, the relative abundance of intI1 increased 3-fold over 13 km of the river stretch, suggesting an internal gene multiplication. We found no evidence that low amounts of sulfonamides in the aquatic environment stimulate the maintenance or even spread of corresponding ARGs. Nevertheless, class 1 integrons carrying various ARGs were still present 13 km downstream from the WWTP. Therefore, limiting the release of ARG-harboring microorganisms may be more crucial for restricting the environmental spread of antimicrobial resistance than attenuating ng/L concentrations of antibiotics.

Class 1 Integron gene harbouring multidrug resistant Pseudomonas isolates from clinical specimen

Gram-negative Pseudomonas spp. particularly P. aeruginosa is an important pathogen in hospitalized patients, contributing to the morbidity and mortality due to its multiple resistance mechanisms. The present study was undertaken to determine the characteristics of Pseudomonas spp. isolated from pus of patients admitted in a local diagnostic center, Dhaka, Bangladesh. A total of 14 clinical isolates were obtained. Detailed biochemical tests, antibiotic resistance pattern and class 1 integron gene profiling were successfully performed. All isolates (100%) showed resistance against amoxicillin-clavulanate, ampicillin, ciprofloxacin, cephalexin, ceftriaxone, kanamycin, nitrofurantoin and rifampicin. About 35% of the isolates (n=5) showed resistance against Amoxicillin-clavulanate, ampicillin, azithromycin, amikacin, ciprofloxacin, cephalexin, co-trimoxazole, ceftriaxone, imipenem, kanamycin, nitrofurantoin and rifampicin. Only 28.5% isolates (n=4) were sensitive to Imipenem. Plasmid was observed in none of the isolates, whereas 50% of the isolates carried Class 1 Integron gene. The presence of mobile genetic element is an added threat as this enables transfer of resistance genes to sensitive isolates. The findings of this study are significant from a public health viewpoint as therapeutic options become restricted.
 Bangladesh J Microbiol, Volume 39, Number 1, June 2022, pp 15-19

Effect of bioaugmentation on tetracyclines influenced chicken manure composting and antibiotics resistance

Antibiotic residue in husbandry waste has become a serious concern. In this study, contaminated chicken manure composting was conducted to reveal the bioaugmentation effect on tetracyclines residue and antibiotics resistance genes (ARGs). The bioaugmented composting removed most of the antibiotics in 7 days. Under bioaugmentation, 96.88 % of tetracycline and 92.31 % of oxytetracycline were removed, 6.32 % and 20.93 % higher than the control (P < 0.05). The high-temperature period was the most effective phase for eliminating antibiotics. The treatment showed a long high-temperature period (7 days), while no high-temperature period was in control. After composting, the treatment showed 13.87 % higher TN (26.51 g/kg) and 13.42 % higher NO3−-N (2.45 g/kg) than control (23.28 and 2.16 g/kg, respectively) but 12.72 % lower C/N, indicating fast decomposition and less nutrient loss. Exogenous microorganisms from bioaugmentation significantly reshaped the microbial community structure and facilitated the enrichment of genera such as Truepera and Fermentimonas, whose abundance increased by 71.10 % and 75.37 % than the control, respectively. Remarkably, ARGs, including tetC, tetG, and tetW, were enhanced by 198.77 %, 846.77 %, and 62.63 % compared with the control, while the integron gene (intl1) was elevated by 700.26 %, indicating horizontal gene transfer of ARGs. Eventually, bioaugmentation was efficient in regulating microbial metabolism, relieving antibiotic stress, and eliminating antibiotics in composting. However, the ability to remove ARGs should be further investigated. Such an approach should be further considered for treating pollutants-influenced organic waste to eliminate environmental concerns.

Assessment of Beta-lactamases and Integrons Genes among Bacteria Isolated From Bladder Cancer Patients with Urinary Tract Infections

Assessment of Beta-lactamases and Integrons Genes among Bacteria Isolated From Bladder Cancer Patients with Urinary Tract Infections

The Prevalence of bla VIM, bla KPC, bla NDM, bla IMP, bla SHV, bla TEM, bla CTX-M, and class I and II integrons Genes in Aeromonas hydrophila Isolated from Clinical Specimens of Qom, Iran.

Aeromonas hydrophila is an opportunistic gram-negative bacillus that causes diseases such as gastroenteritis, muscle infections, soft tissue, sepsis, and skin diseases in humans. Today, the prevalence of antibiotic resistance in bacteria has led to treatment failure and prolonged treatment. Therefore, the aim of this study was to evaluate the level of antibiotic resistance in isolates carrying bla VIM, bla KPC, bla NDM, bla IMP, bla SHV, bla TEM, bla CTX-M and class I and II integrons in Aeromonas hydrophila. In this cross-sectional study, Aeromonas hydrophila were collected from different clinical specimens in Hazrat Masoumeh Hospital, Qom Province, Iran, from 2018 to 2020. The collected isolates were identified by standard biochemical tests. Then, using specific primers bla VIM, bla KPC, bla NDM, bla IMP, bla SHV, bla TEM, bla CTX-M genes, and class I and II integrons were evaluated by PCR method. Then, data were analyzed using SPSS software and chi-squared tests, and the significance level was determined as p ≤ 0.05. During the sample collection period, 100 Aeromonas hydrophila were collected. Based on the results of the antibiotic resistance pattern, the highest and lowest rate of antibiotic resistance to ampicillin (92%) and azithromycin (4%) were determined for both. Among the 100 isolates, 60 isolates produced broad-spectrum beta-lactamase (ESBL) and 50 isolates produced carbapenemase. Among the studied beta-lactamase genes, the highest and lowest frequencies were related to bla CTX-M (58%) and bla TEM (1%), respectively. The frequency of class I and II integron genes was 27% and 15%, respectively. The results of the study of antibiotic resistance, beta-lactamase, and carbapenemase genes showed high resistance in Aeromonas hydrophila, which raises concerns with regard to controlling infection in medical centers. Also, the study of antibiotic resistance in the presence of beta-lactamase genes showed that there was only a significant relationship between the presence of bla CTX-M gene and resistance to imipenem (p = 0.037).

Occurrence of Class 1, 2, and 3 Integrons among Multidrug-resistant Pseudomonas aeruginosa in Babylon Province, Iraq

Background: Clinical management of bacterial infections has faced significant difficulties in recent years due to the advent and spread of multiple drug-resistant (MDR) bacteria. Worldwide, nosocomial infections are brought on by Pseudomonas aeruginosa, a clinically significant Pseudomonas species. Objectives: This research aimed to identify class 1, 2, and 3 integrons in P. aeruginosa in Babylon, Iraq. Materials and Methods: From February 2022 to October 2022, 131 isolates from various sites including (burn, wound, and urine) were collected from different hospitals in Babylon Province for both genders and ages. These isolates were identified using traditional techniques as well as the Vitek 2 system (bioMerieux, France). Pseudomonas aeruginosa isolates were subjected to disc diffusion antimicrobial susceptibility testing. Class 1, 2, and 3 integron-specific primers were used in the polymerase chain reaction technique for the molecular identification of integron genes. Results: Pseudomonas aeruginosa isolates that were 131 (100%) had integron class 1. On the contrary, only five (3.81%) contained a class 2 integron. There was no presence of class 3 integron in any isolate. Conclusion: The MDR P. aeruginosa was highly prevalent (100%) and this suggested that the availability of class 1 integrons in our area was alarmingly high, showing the need for epidemiological monitoring.

Genomic and Evolutionary Insights into Australian Toxigenic Vibrio cholerae O1 Strains.

Vibrio cholerae O1 is the causative agent of cholera, a severe diarrheal disease which can cause death if left untreated. In this study, a collection of clinical and environmental V. cholerae serogroup O1 isolates from Australia (1977 to 1987) (from local cases and cases acquired through international travel) and publicly available international isolates were characterized for genotypic features (virulence genes, mobile genetic elements [MGEs], and antimicrobial resistance gene profiles). Whole-genome sequencing (WGS) was used to investigate and compare the genetic relatedness between the 44 Australian and nine travel-associated isolates and the 60 publicly available international V. cholerae sequences representing pre-seventh-pandemic (pre-7PET) isolates and different waves of 7PET isolates. In this study, 36 (81%) Australian clinical and aquatic isolates harbored the cholera toxin-producing genes located in the CTX bacteriophage region. All the Australian environmental and clinical isolates lacked the seventh-pandemic virulence-associated genomic islands (VSP-I and -II). In silico multilocus sequence typing (MLST) classified all nine internationally acquired isolates as sequence type 69 (ST69), 36 clinical and aquatic isolates as ST70, and eight isolates from Australia as ST71. Most of the nontoxigenic clinical and aquatic isolates of ST71 had diverse genetic variations compared to ST70 Australian strains. The antimicrobial resistance-associated genes gyrA, parC, and parE had no mutations in all the environmental and clinical isolates from Australia. The SXT genetic element and class 1 integron gene sequences were not detected in Australian strains. Moreover, in this study, a Bayesian evolutionary study suggests that two distinct lineages of ST71 (new set of strains) and ST70 strains were prevalent around similar times in Australia, in ~1973 and 1969. IMPORTANCE Australia has its own indigenous V. cholerae strains, both toxigenic and nontoxigenic, that are associated with disease. Exotic strains are also detected in Australian patients returning from overseas travel. The clinical and aquatic V. cholerae O1 toxin gene-positive isolates from Australia responsible for cases in 1977 to 1987 were linked to acquisition from Queensland waterways but until now had not been characterized genetically. It is important to determine the genetic relatedness of Australian strains to international strains to assist in understanding their origin. This is the first extensive study to provide sequences and genomic analysis focused on toxigenic O1 V. cholerae clinical and environmental strains from Australia and its possible evolutionary relationship with other publicly available pre-7PET and 7PET V. cholerae strains. It is important to understand the population genetics of Australian V. cholerae from a public health perspective to assist in devising control measures and management plans for reducing V. cholerae exposure in Australia, given previous Australian disease clusters.

Bacillus subtilis reduces antibiotic resistance genes of animal sludge in vermicomposting by improving heat stress tolerance of Eisenia foetida and bacterial community adjustment

Antibiotic resistance genes (ARGs) in livestock industry have been recognized as a kind of pollutant. The effect of Bacillus subtilis (B. subtilis) as an additive for the reduction of ARGs in animal sludge from livestock and poultry wastewater treatment plant during vermicomposting was investigated. We also evaluated the oxidative stress level and growth of earthworms, Eisenia foetida, bacterial community succession, and the quality of the end products. Two treatments were conducted using B. subtilis, one at 18 °C and another at 28 °C. Controls were setup without the bacteria. The results showed that inoculation of B. subtilis promoted the degradation of organics at 28 °C and increased the germination index to 236%. The increased activities of the superoxide dismutase (1.69 U/mg pr) and catalase (8.05 U/mg pr) and the decreased activity of malondialdehyde (0.02 nmol/mg pr) by B. subtilis at 28 °C showed that the earthworms were relieved of heat stress. The addition of B. subtilis reduced the abundance of 32 target ARGs, including integron (intI-1), transposase (IS613) and resistant genes, such as sulfonamide (sul2), quinolone (oprJ), macrolide-lincosamide-streptogramin group B (ermF, ermB), tetracycline (tetL-02, tetX), β-lactama (blaOXA10-01) and aminoglycoside [strB, aac(6′)-Ib(aka aacA4)-01, aac(6′)-Ib(aka aacA4)-02]. Organic matter degrading Membranicola, Paludisphaera, Sphingorhabdus and uncultured bacterium belonging to the order Chitinophagales, nitrifying and nitrogen-fixing Singulisphaera and Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, soil remediating Achromobacter, and plant growth promoting Kaistia, Galbibacter and Ilumatobacter were increased significantly (P < 0.05). However, the growth of harmful bacteria such as Burkholderiaceae was inhibited in the vermicompost. In earthworm guts, the probiotic Mesorhizobium was promoted, while the pathogenic uncultured bacterium belonging to the family Enterobacteriaceae was reduced. Besides, B. subtilis enhanced the host relationships between bacteria and ARGs. These findings might be helpful in the removal of ARGs in animal wastes and in understanding the synergy between earthworms and microorganisms.

Multidrug resistance-encoding gene in Citrobacter freundii isolated from healthy laying chicken in Blitar District, Indonesia

Background and Aim: The increasing prevalence of resistance (MDR) of Enterobacteriaceae in Indonesia has caused concern regarding human health. Citrobacter freundii reportedly targets the gastrointestinal tract of animals and is a common cause of foodborne diseases associated with diarrhea, peritonitis, meningitis, brain abscess, bacteremia, and urinary tract infection. This study aimed to estimate the prevalence of MDR and the presence of Class 1 integron-encoding genes in C. freundii isolates obtained from cloacal swabs of healthy laying chickens in Blitar district, Indonesia. Materials and Methods: One hundred and sixty-five cloacal swab samples were collected from 33 farms in Blitar over a period of 4 months. Standard microbiological techniques such as bacterial culture in MacConkey agar, Simmons citrate agar, and triple sugar iron agar and biochemical tests such as the indole test were performed to identify the isolates. The antibiotic sensitivity patterns of C. freundii isolates were determined by the disk diffusion method, and MDR-encoding genes (Class 1 integron) were detected by polymerase chain reaction (PCR). Results: Out of 165 cloacal swab samples, 7 (4.24%) were positive for C. freundii. Citrobacter freundii was highly resistant to erythromycin (71.43%) and moderately to streptomycin, tetracycline, and trimethoprim-sulfamethoxazole (all 42.86%); however, it showed low resistance to ampicillin (28.57%). All isolates were found to exhibit MDR. Only 1 (14.29%) of the seven C. freundii isolates harbored a Class 1 integron gene. This study revealed that Class 1 integron-encoding genes have a low prevalence in C. freundii isolated from healthy laying chickens in Blitar, Indonesia. Conclusion: Poultry animals can play a role in the transmission of resistance genes to humans due to the MDR of Enterobacteriaceae, including C. freundii in the intestines.

Detection of ESBL and AmpC producing Klebsiella pneumoniae ST11 and ST147 from urinary tract infections in Iran.

In the present study a total of 200 Klebsiella pneumoniae isolates were collected from patients with urinary tract infections (UTIs) in Tehran, Iran. Antibiotic resistance was determined by disk diffusion and broth dilution methods. Detection of extended-spectrum β-lactamases (ESBLs) and AmpCs was performed using phenotypic tests. Polymerase chain reaction (PCR) was applied to detect the ESBL, AmpC, and integron genes. Analysis of AmpC and cassette arrays of integron genes was performed using DNA sequencing. Plasmids were analyzed by PCR-based replicon typing and conjugation. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were applied to explore the genomic relatedness among the isolates. The highest levels of resistance were observed against ampicillin (100%), followed by piperacillin (57.5%), ceftazidime (46%), trimethoprim/sulfamethoxazole (44%), ciprofloxacin (32.5%), and imipenem (19%). Approximately, 66.5% of isolates harbored at least one of the beta-lactamase genes (blaTEM, blaSHV, blaCTX-M, and blaOXA-1). In addition, 22.5% of isolates carried at least one of the AmpC genes including blaDHA and blaCIT. Integron class I was the most prevalent integron among resistant isolates. According to the results of replicon typing, IncFII, IncL/M, and IncA/C were the most frequent replicons, respectively. All selected isolates were able to transfer blaCTX-M, also two isolates transferred the blaDHA-1 gene to Escherichia coli K12 through conjugation. Finally, 21 isolates were categorized into 4 pulsotypes and 11 unique clusters in PFGE. MLST identified ST147 and ST11 sequence types but ST147 was the most prevalent in the current study.

Novel aadA5 and dfrA17 variants of class 1 integron in multidrug-resistant Escherichia coli causing bovine mastitis.

Mobile genetic elements (MGEs) are associated with the emergence of multidrug resistance in extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae. This study explores the role of class 1 integrons and IS26 elements in breaching taxonomic barriers. A total of 110 E. coli bacteria were isolated from 300 clinical mastitis milk samples. The 98% E. coli isolates were extended-spectrum beta-lactamase- producers. About 83% of these isolates carried co-resistance for fluoroquinolones. The co-existence of (extended-spectrum beta-lactamase + quinolone resistance determining region mutations) and (extended-spectrum beta-lactamase + plasmid-mediated quinolone resistance genes) was found in 76% and 44% of isolates, respectively. The MGEs were detected in 88% of isolates with IS26 in 82% and class 1 integrase in 40% of isolates. The types of class 1 integron gene cassettes detected includes dfrA7, (dfrA17 + aadA5), and (dfrA1 + aadA1). We discovered 2 and 4 novel variants of the dfrA17 and aadA5 genes, respectively. We report a variant of aadA5 with mutation E235G in the Indian subcontinent earlier reported only in a human clinical isolate from Belgium. About 19 isolates carried IS26 linked to integrase gene intI1 with an internal deletion of 265bp in the 5`CS of integrase gene intI1, earlier reported only in E. coli ST131 isolates from human clinical, wastewater samples. This study suggests intercontinental dissemination of antibiotic resistant genes (ARGs) across different microbiomes via mobile genetic elements. KEY POINTS: • The role of mobile genetic elements in the emergence of multidrug-resistant E. coli in bovine mastitis. • Novel variants of the aadA5 (aminoglycoside adenyl transferase) and dfrA17 (dihydrofolate reductase) genes were identified in pathogenic E. coli isolated from bovine mastitis in class 1 integron gene cassette. • Sequence analysis of mobile genetic components revealed the physical connection between IS26 and intI1 genes with an internal deletion in 5'CS of class 1 integrase.

Distribution and drivers of antibiotic resistance genes in brackish water aquaculture sediment

Brackish water aquaculture has brought numerous economic benefits, whereas anthropogenic activities in aquaculture may cause the dissemination of antibiotic resistance genes (ARGs) in brackish water sediments. The intricate relationships between environmental factors and microbial communities as well as their role in ARGs dissemination in brackish water aquaculture remain unclear. This study applied PCR and 16S sequencing to identify the variations in ARGs, class 1 integron gene (intI1) and microbial communities in brackish water aquaculture sediment. The distribution of ARGs in brackish water aquaculture sediment was similar to that in freshwater aquaculture, and the sulfonamide resistance gene sul1 was the indicator of ARGs. Proteobacteria and Firmicutes were the dominant phyla, and Paenisporosarcina (p_ Firmicutes) was the dominant genus. The results of correlation, network and redundancy analysis indicated that the microbial community in the brackish water aquaculture sediment was function-driven. The neutral model and variation partitioning analysis were used to verify the ecological processes of the bacterial community. The normalized stochasticity ratio showed that pond bacteria community was dominated by determinacy, which was affected by aquaculture activities. The total nitrogen and organic matter influenced the abundance of ARGs, while Proteobacteria and Thiobacillus (p_Proteobacteria) were the key antibiotic-resistant hosts. Our study provides insight into the prevalence of ARGs in brackish water aquaculture sediments, and indicates that brackish water aquaculture is a reservoir of ARGs.

Removal of antibiotic resistance genes and pathogenicity in effluent from municipal wastewater treatment plant by plasma oxidation

Municipal wastewater treatment plants (MWTP) are the main gathering places for antibiotic resistance genes (ARGs) and pathogenic bacteria. In this study, plasma oxidation was applied to remove the ARGs and inhibit pathogenicity in the effluent. The experimental results demonstrated that approximately 1.32 log, 1.34 log, 2.39 log, and 2.80 log of aac(3)-II, blaTEM-1, TetW, and TetC were removed within 25 min of the plasma treatment, respectively. OH, 1O2, O3, H2O2, NO2−, and NO3− all contributed to ARG removal. Coexisting dissolved organic matter was also oxidized, which was positively related to ARG removal. Plasma oxidation led to a decrease in the diversity and richness of microbial communities; Parcubacteria and Proteobacteria were the main potential hosts of ARGs and mobile genetic elements. Integron gene IntI1 decreased by 2.46 log after plasma treatment, which was positively related to ARG removal. Gene expression related to ATP binding, DNA repair, gene transfer, and the cell membrane was reduced after plasma treatment, resulting in effective ARG removal. Furthermore, pathogenic genes related to lumpy skin disease, Niemann-Pick disease, polycystic kidney and hepatic disease were all down-regulated by plasma treatment. Overall, the plasma treatment inhibited the transmission risks of ARGs and pathogenicity in the effluent of MWTP.

Class 1 integron and related antimicrobial resistance gene dynamics along a complex freshwater system affected by different anthropogenic pressures

The risk for human health posed by polluted aquatic environments, and especially those carrying antibiotic resistance genes (ARGs) of clinical interest, is still debated. This is because of our limited knowledge of the dynamics of antimicrobial resistance in the environment, the selection mechanisms underlying the spread of ARGs, and the ecological factors potentially favoring their return to humans. The Class 1 integron is one of the most effective platforms for the dissemination of ARGs. In this study we investigated a freshwater system consisting of a lake-river-lake continuum, determining the abundance of class 1 integrons and their associated ARGs by a modulated metagenomic approach. Bacterial abundance and community composition were used to identify the potential carriers of class 1 integrons and their associated ARGs over a period of six months. Class 1 integrons and their ARG cargoes were significantly more abundant in riverine sampling sites receiving treated wastewater. Further, class 1 integrons carried ARGs ranked at the highest risk for human health (e.g., catB genes), in particular, genes encoding resistance to aminoglycosides. Genera of potential pathogens, such as Pseudomonas and Escherichia-Shigella, were correlated with class 1 integrons. The lake-river-lake system demonstrated a clear relationship between the integrase gene of class 1 integrons (intI1) and anthropogenic impact, but also a strong environmental filtering that favored the elimination of intI1 once the human derived stressors were reduced. Overall, the results of this study underline the role class 1 integrons as proxy of anthropogenic pollution and suggest this genetic platform as an important driver of aminoglycoside resistance genes, including high risk ARGs, of potential concern for human health.

Molecular Study of Integrase Gene I and Integrase Gene II in Clinical Isolates of Pseudomonas aeruginosa.

The presence of the class I integron gene is associated with the emergence of multiple drug resistance (MDR) phenotype in Pseudomonas aeruginosa (P. aeruginosa) isolates. The objectives of this research were to study the prevalence of integrase genes I (Intel I) and integrase genes II (Intel II) in clinical isolates of P. aeruginosa and its association with antibiotic resistance in these isolates. The study was a retrograde cross-sectional study that was carried out on 150 clinical isolates of P. aeruginosa isolated from patients with healthcare-associated infections. The isolates were subjected to biochemical identification and antibiotic sensitivity study by discs diffusion test. Intel I & Intel II genes were detected by polymerase chain reaction (PCR). Intel I gene was present in 48% of the isolates, and Intel II was present in 1.3% of the isolates. Intel I gene was detected at a statistically significant high rate in MDR- P. aeruginosa (76.9%, P=0.001) compared to non-MDR- P. aeruginosa (3.4%), while intel II had a statistically insignificant increase in MDR- P. aeruginosa (1.1%, P=1.00) compared to non-MDR-P. aeruginosa (1.7%). Both Intl I/Intl II genes were detected in 2.2% of MDR-P. aeruginosa isolates and were absent in non- MDR-P. aeruginosa isolates with statistically insignificant difference (P=1.00). P. aeruginosa isolates with Intel I gene had an increase in antibiotic resistance pattern to the used antibiotics discs. However, this increase had statistically significant rates only for gentamicin (63.9%, P≤0.001), meropenem (47.2%, P=0.009), trimethoprim/sulfamethoxazole (37.5%, P=0.013) and imipenem (44.4%, P=0.025). The present study highlights the high prevalence of the Intel I gene in clinical isolates of P. aeruginosa, while the Intel II gene was less prevalent in these isolates. There was a significant association between the prevalence of the Intel I gene and the MDR phenotype of P. aeruginosa and resistance to gentamicin, meropenem, trimethoprim/sulfamethoxazole, and imipenem. These findings need future evaluation in a higher number of clinical isolates of P. aeruginosa.

Distribution and co-occurrence patterns of antibiotic resistance genes in black soils in Northeast China.

Black soils (Mollisols) are among the most important soil resources for crop production and food security. In China, they are mainly distributed in the northeastern region. To investigate soil antibiotic resistance distribution patterns and monitor soil quality, we randomly chose nine corn fields in Northeast China and analyzed the antibiotic resistance gene (ARG) distribution and co-occurrence patterns on the basis of high-throughput approaches and network analyses. High genetic diversity (136 unique genes) and low ARG abundances (10-5-10-2 copies/16S rRNA gene copy) were detected, with relatively few interactions among ARGs. Type I integron genes were prevalent in the soil and were positively correlated with ARGs, which may increase the risk of ARG transmission. Most ARGs were strongly associated with microorganisms. Moreover, several ARGs were significantly correlated with antibiotics, nutrients, and metal elements. The generation and dissemination of ARGs, which were most likely mediated by mobile genetic elements (MGEs) and bacteria, were affected by environmental conditions. These results provide insights into the widespread co-occurrence patterns in soil resistomes.

Fate of antibiotic resistance genes and class 1 integrons during sludge treatment using pilot-scale anaerobic digestion with thermal hydrolysis pretreatment

This work evaluated the fate of antibiotic resistance genes (ARGs) and class 1 integron gene in sewage sludge before and after pilot-scale thermal hydrolysis pretreatment (THP) and subsequent mesophilic anaerobic digestion (AD) treatment. Variables investigated include THP temperatures, feedstock types, and AD solids retention times. Real-time polymerase chain reaction was performed to quantify the ARGs in feedstocks, THP and AD effluent. Results show that THP significantly (t test, p < 0.05) reduced the absolute abundances of most ARGs, with the reduction ranging from 0.03 to 3.09 log units. Rebound effects of ARGs in the subsequent AD were observed and were relevant with tested variables; shorter solids retention time (10 days) and higher THP temperature (165 ℃) can significantly reduce ARGs in AD effluent. These findings provide references about the effects of the THP and AD on the control of ARG spread from sewage sludge to environments.