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Metagenomic analysis of microbial community structure and distribution of resistance genes in Daihai Lake, China
2022
Du, Caili | Yang, Fang | Li, Xiaoguang | Liao, Haiqing | Li, Zhonghong | Gao, Jiayue | Zhang, Lieyu
The emergence of resistance genes is a global phenomenon that poses a significant threat to both animals and humans. Lakes are important reservoirs of genes that confer resistant to antibiotics and metals. In this study, we investigated the distribution and diversity of antibiotic resistance genes (ARGs) and metal resistance genes (MRGs) in the sediment of Daihai Lake using high-throughput sequencing and metagenomic analysis. The results indicated that all sampling sites had similar bacterial community structures, with Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes being the most abundant. A total of 16 ARG types containing 111 ARG subtypes were deposited in the sediment. Among the resistance genes to bacitracin, multidrug, macrolide-lincosamide-streptogramin (MLS), tetracycline, beta-lactam, and sulfonamide were the dominant ARG types, accounting for 89.9–94.3% of the total ARGs. Additionally, 15 MRG types consisting of 146 MRG subtypes were identified. In all samples, MRGs of the same type presented resistance to Pb, Ni, Hg, W, Zn, Ag, Cr, Fe, As, Cu, and multimetals. Overall, the distribution and diversity of antibiotic and metal resistance genes showed no significant differences in the samples. Plasmids (91.03–91.82%) were the most dominant mobile genetic elements in the sediments of Daihai Lake. Network analysis indicated that the target ARGs and MRGs were significantly positively correlated with the microorganisms. Potential hosts for various ARGs and MRGs include Proteobacteria, Euryarchaeota, Actinobacteria, Chloroflexi, and Bacteroidetes.
Show more [+] Less [-]Antibiotic resistome profile based on metagenomics in raw surface drinking water source and the influence of environmental factor: A case study in Huaihe River Basin, China
2019
Bai, Ying | Ruan, Xiaohong | Xie, Xianchuan | Yan, Zhongyue
The contamination with antibiotic resistance genes (ARGs) in raw drinking water source may pose a direct threat to human health. In this study, metagenomics sequencing and analysis were applied to investigate the ARG pattern in 12 drinking water sources in upper and middle reach of Huaihe River Basin, China. Based on the redundant analysis and multi-linear regression model, location, specific microbial taxa, number of livestock and health facilities significantly influenced the ARG profile in drinking water sources. Besides the cluster effect of ARG in samples from plain and bedrock mountain areas, the samples from fracture aquifer areas also showed a distinctive biogeographic pattern with that from porous aquifer areas. Putative ARGs host Opitutus and Flavobacterium were the enriched biomarkers in plain and fracture aquifer area respectively, which mainly carried bacitracin, multidrug, beta-lactam and tetracycline ARGs. This result illuminated that both natural background and anthropogenic activities in the watershed influenced the ARG profile in natural freshwater system significantly. The low MGEs abundance and absence of pathogen revealed a low ARG dissemination risk in sampled drinking water sources, while Polynucleobacter was an abundant ARGs host and was significantly related to the ARG profile, which indicated that specific bacteria was responsible for ARGs propagation and accumulation in surface freshwater system. Further researches are needed to assess human exposure to raw drinking water source and the potential risk, as well as the species interaction in microbial community and its impact on ARG propagation under oligotrophic condition.
Show more [+] Less [-]Metagenome sequencing to unveil the resistome in a deep subtropical lake on the Yunnan-Guizhou Plateau, China
2020
Pan, Xiong | Lin, Li | Zhang, Weihong | Dong, Lei | Yang, Yuyi
Erhai Lake in the Yunnan-Guizhou Plateau, a typical subtropical deep lake, experienced long-time input of nutrients and pollutants, which provides beneficial conditions for evolution and spread of resistance genes. In this study, the profile of bacterial community, antibiotic and metal resistance genes in sediments of Erhai Lake were analyzed via high-thought sequencing of 16S rRNA and metagenomic DNA. Proteobacteria, Firmicutes, Nitrospirae, and Bacteroidetes were found to be the main composition of the bacterial community. Multidrug-, bacitracin-, macrolide-lincosamide-streptogramin (MLS)- and tetracycline resistance genes were the primary antibiotic resistance gene (ARG) types with high relative abundance, whereas Multi-metal-, and arsenic- resistance genes were the primary metal resistance gene (MRG) types. The effects of nitrogen and phosphorus on the abundance of ARGs and MRGs depended on the different types. Some specific ARG (fosmidomycin resistance genes) and MRG types (multi-metal resistance genes) also showed significant geographic distribution. Bacterial community was the main driver shaping the resistome. Nutrients also played an essential role in structuring the bacterial community and resistome in the sediments of Erhai Lake. This study sheds light on the distribution and fate of resistome under a high load of nitrogen and phosphorus in a deep lake.
Show more [+] Less [-]Bioplastic accumulates antibiotic and metal resistance genes in coastal marine sediments
2021
Di Cesare, Andrea | Pinnell, Lee J. | Brambilla, Diego | Elli, Giulia | Sabatino, Raffaella | Sathicq, María B. | Corno, Gianluca | O'Donnell, Colin | Turner, Jeffrey W.
The oceans are increasingly polluted with plastic debris, and several studies have implicated plastic as a reservoir for antibiotic resistance genes and a potential vector for antibiotic-resistant bacteria. Bioplastic is widely regarded as an environmentally friendly replacement to conventional petroleum-based plastic, but the effects of bioplastic pollution on marine environments remain largely unknown. Here, we present the first evidence that bioplastic accumulates antibiotic resistance genes (ARGs) and metal resistance genes (MRGs) in marine sediments. Biofilms fouling ceramic, polyethylene terephthalate (PET), and polyhydroxyalkanoate (PHA) were investigated by shotgun metagenomic sequencing. Four ARG groups were more abundant in PHA: trimethoprim resistance (TMP), multidrug resistance (MDR), macrolide-lincosamide-streptogramin resistance (MLS), and polymyxin resistance (PMR). One MRG group was more abundant in PHA: multimetal resistance (MMR). The relative abundance of ARGs and MRGs were strongly correlated based on a Mantel test between the Bray-Curtis dissimilarity matrices (R = 0.97, p < 0.05) and a Pearson's analysis (R = 0.96, p < 0.05). ARGs were detected in more than 40% of the 57 metagenome-assembled genomes (MAGs) while MRGs were detected in more than 90% of the MAGs. Further investigation (e.g., culturing, genome sequencing, antibiotic susceptibility testing) revealed that PHA biofilms were colonized by hemolytic Bacillus cereus group bacteria that were resistant to beta-lactams, vancomycin, and bacitracin. Taken together, our findings indicate that bioplastic, like conventional petroleum-based plastic, is a reservoir for resistance genes and a potential vector for antibiotic-resistant bacteria in coastal marine sediments.
Show more [+] Less [-]Metagenomic exploration reveals a differential patterning of antibiotic resistance genes in urban and peri-urban stretches of a riverine system
2021
Rajput, Vinay | Yadav, Rakeshkumar | Dharne, Mahesh S.
Antimicrobial resistance in the riverine ecosystem of urban areas is an alarming concern worldwide, indicating the importance of molecular monitoring to understand their patterning in urban and peri-urban areas. In the present study, we evaluated the influence of urban rivers on the connected peri-urban rivers of a riverine system of India in the context of antibiotic resistance genes. The rivers traversing through urban (Mula, Mutha, Pawana, and Ramnadi) and peri-urban stretches (Bhima and Indrayani) form the riverine system of Pune district in Maharashtra, India. The MinION-based shotgun metagenomic analysis revealed the resistome against 26 classes of antibiotics, including the last line of antibiotics. In total, we observed 278 ARG subtypes conferring resistance against multiple drugs (40%), bacitracin (10%), aminoglycoside (7.5%), tetracycline (7%), and glycopeptide (5%). Further, the alpha diversity analysis suggested relatively higher ARG diversity in the urban stretches than peri-urban stretches of the riverine system. The NMDS (non-metric multidimensional scaling) analysis revealed significant differences with overlapping similarities (stress value = 0.14, p-value = 0.004, ANOSIM statistic R: 0.2328). These similarities were reasoned by assessing the influence of downstream sites (sites at the outskirts of Pune city; however, directly impacted), which revealed significant differences in the ARG contents of urban and peri-urban stretches (stress value = 0.14, p-value = 0.001, ANOSIM statistic R: 0.6137). Overall, we detected the dissemination of antibiotic resistance genes from the polluted urban rivers into the peri-urban rivers located downstream in the connected riverine system potentially driven by anthropogenic activities.
Show more [+] Less [-]A comprehensive dynamic kinetic model for the UVC/H2O2 process: application to zinc bacitracin degradation in wastewater as a case study
2021
Metolina, Patrícia | Teixeira, Antonio Carlos Silva Costa
A dynamic kinetic model is presented for the UVC/H₂O₂-driven process. The model comprises 103 reactions, including background species, such as HCO₃–/CO₃²–, NO₂–, NO₃–, SO₄²⁻, Cl⁻, and H₂PO₄⁻/HPO₄²/PO₄³– anions, and effluent organic matter (EfOM) was validated based on experimental data obtained for the photooxidation of the nonribosomal peptide antibiotic zinc bacitracin (Zn-Bc, 34 μmol L⁻¹). The set of ordinary differential equations for 38 species was combined with the molar balances describing the recirculating tubular photoreactor used. Predictions for the photolytic and UVC/H₂O₂ processes confirmed the good agreement with experimental data, enabling the estimation of fundamental kinetic parameters, such as the direct photolysis quantum yield (Ф₂₅₄ ₙₘ, Zₙ₋Bc = 0.0143 mol Einstein⁻¹) and the second-order rate constants for the reactions of Zn-Bc with HO•, HO₂•, and O₂•⁻ radicals (2.64 × 10⁹, 1.63 × 10³, and 1.49 × 10⁴ L mol⁻¹ s⁻¹, respectively). The predicted optimum process conditions correspond to [H₂O₂]₀ = 6.8 mmol L⁻¹ and a specific photon emission rate of 11.1 × 10⁻⁶ Einstein L⁻¹ s⁻¹. Zn-Bc photooxidation was significantly impacted by wastewater constituents, particularly EfOM and HCO₃–/CO₃²– (i.e., alkalinity), resulting in a degradation rate about 32% lower compared to that obtained in deionized water. In particular, EfOM acts as a strong radical scavenger and inner filter. In addition, simulations pointed out the continuous tubular photochemical reactor as the best configuration for treating Zn-Bc-containing wastewater. This study hence provides a comprehensive modeling approach, especially useful for predicting the effect of complex water matrices on the performance of the UVC/H₂O₂ treatment process.
Show more [+] Less [-]Characterization of multiple antibiotic resistance of culturable microorganisms and metagenomic analysis of total microbial diversity of marine fish sold in retail shops in Mumbai, India
2018
Naik, OnkarA. | Shashidhar, Ravindranath | Rath, Devashish | Bandekar, JayantR. | Rath, Archana
Marine fish species were analyzed for culturable and total metagenomic microbial diversity, antibiotic resistance (AR) pattern, and horizontal gene transfer in culturable microorganisms. We observed a high AR microbial load of 3 to 4 log CFU g⁻¹. Many fish pathogens like Providencia, Staphylococcus, Klebsiella pneumoniae, Enterobacter, Vagococcus, and Aeromonas veronii were isolated. Photobacterium and Vibrio were two major fish and human pathogens which were identified in the fish metagenome. Other pathogens that were identified were Shewanella, Acinetobacter, Psychrobacter, and Flavobacterium. Most of these pathogens were resistant to multiple antibiotics such as erythromycin, kanamycin, neomycin, streptomycin, penicillin, cefotaxime, bacitracin, rifampicin, trimethoprim, ciprofloxacin, and doxycycline with a high multiple antibiotic resistance index of 0.54–0.77. The fish microflora showed high prevalence of AR genes like bla TEM, Class I integron, tetA, aph(3′)-IIIa, ermB, aadA, and sul1. Nineteen of 26 AR isolates harbored Class I integrons showing high co-resistance to trimethoprim, kanamycin, doxycycline, and cefotaxime. Mobile R-plasmids from 6 of the 12 AR pathogens were transferred to recipient E. coli after conjugation. The transconjugants harbored the same R-plasmid carrying bla CTX₋M, dfr1, tetA, bla TEM, and cat genes. This study confirms that fish is a potential carrier of AR pathogens which can enter the human gut via food chain. To the best of our knowledge, this is the first study in the Indian subcontinent reporting a direct evidence of spread of AR pathogens to humans from specific marine fish consumption.
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