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A closer look on the variety and abundance of the faecal resistome of wild boar
2022
Dias, Diana | Fonseca, Carlos | Mendo, Sónia | Caetano, Tânia
Antimicrobial resistance (AMR) is a serious problem for public and animal health, and also for the environment. Monitoring and reporting the occurrence of AMR determinants and bacteria with the potential to disseminate is a priority for health surveillance programs around the world and critical to the One Health concept. Wildlife is a reservoir of AMR, and human activities can strongly influence their resistome.The main goal of this work was to study the resistome of wild boar faecal microbiome, one of the most important game species in Europe using metagenomic and culturing approaches. The most abundant genes identified by the high-throughput qPCR array encode mobile genetic elements, including integrons, which can promote the dissemination of AMR determinants. A diverse set of genes (n = 62) conferring resistance to several classes of antibiotics (ARGs), some of them included in the WHO list of critically important antimicrobials were also detected. The most abundant ARGs confer resistance to tetracyclines and aminoglycosides. The phenotypic resistance of E. coli and Enterococcus spp. were also investigated, and together supported the metagenomic results.As the wild boar is an omnivorous animal, it can be a disseminator of AMR bacteria and ARGs to livestock, humans, and the environment. This study supports that wild boar can be a key sentinel species in ecosystems surveillance and should be included in National Action Plans to fight AMR, adopting a One Health approach.
显示更多 [+] 显示较少 [-]Arsenic resistance and horizontal gene transfer are associated with carbon and nitrogen enrichment in bacteria
2022
Neethu, C.S. | Saravanakumar, C. | Purvaja, R. | Robin, R.S. | Ramesh, R.
Coastal waters are confluences receiving large amounts of point and non-point sources of pollution. An attempt was made to explore microbial community interactions in response to carbon, nitrogen and metal pollution. Additionally, experiments were designed to analyze the influence of these factors on horizontal gene transfer (HGT). Shift in bacterial diversity dynamics by arsenic stress and nutrient addition in coastal waters was explored by metagenomics of microcosm setups. Phylogenetic analysis revealed equal distribution of Gammaproteobacteria (29%) and Betaproteobacteria (28%) in control microcosm. This proportional diversity from control switched to unique distribution of Gammaproteobacteria (44.5%)> Flavobacteria (17.7%)> Bacteriodia (11.92%)> Betaproteobacteria (11.52%) in microcosm supplemented with carbon, nitrogen and metal (C + N + M). Among metal-stressed systems, alpha diversity analysis indicated highest diversity of genera in C + N + M followed by N + M > C+M> metal alone. Arsenic and ampicillin sensitive E. coli XL1 blue and environmental strains (Vibrio tubiashii W85 and E. coli W101) were tested for efficiency of uptake of plasmid (P) pUCminusMCS (arsBᴿampᴿ) under varying stress conditions. Transformation experiments revealed that combined effect of carbon, nitrogen and metal on horizontal gene transfer (HGT) was significantly higher (p < 0.01) than individual factors. The effect of carbon on HGT was proved to be superior to nitrogen under metal stressed conditions. Presence of arsenic in experimental setups (P + M, P + N + M and P + C + M) enhanced the HGT compared to non-metal counterparts supplemented with carbon or nitrogen. Arsenic resistant bacterial isolates (n = 200) were tested for the ability to utilize various carbon and nitrogen substrates and distinct positive correlation (p < 0.001) was found between arsenic resistance and utilization of urea and nitrate. However, evident positive correlation was not found between carbon sources and arsenic resistance. Our findings suggest that carbon and nitrogen pollution in aquatic habitats under arsenic stress determine the microbial community dynamics and critically influence uptake of genetic material from the surrounding environment.
显示更多 [+] 显示较少 [-]Deposition and mobilization of viruses in unsaturated porous media: Roles of different interfaces and straining
2021
Zhang, Wenjing | Wu, Shengyu | Qin, Yunqi | Li, Shuo | Lei, Liancheng | Sun, Simiao | Yang, Yuesuo
The vadose zone is the first natural layer preventing groundwater pollution. Understanding virus transport and retention in the vadose zone is necessary. The effects of different interfaces and mechanisms on virus transport and retention were investigated by studying Escherichia coli phage migration in laboratory-scale columns under unsaturated conditions. The E. coli phage was used as a model virus. Colloid filtration theory, extended Derjagin–Landau–Verwey–Overbeek theory and two−site kinetic deposition model were used to calculate fitted parameters and interaction energies to assess virus retention at different interfaces. The collector diameters and the size of E. coli phages in the influent and effluent were compared to assess the effect of straining. The results indicated that the roles of solid–water interfaces (SWIs) and air–water interfaces (AWIs) in retaining E. coli phages are strongly controlled by the moisture content and hydrochemical conditions. Decreasing the moisture content and increasing the ionic strength (IS) of the suspension increased E. coli phage retention. At suspension ISs of 0.01 or 0.03 M and various moisture contents, E. coli phages were mainly retained at the SWIs rather than AWIs. When the IS was increased to 0.06 M, the viruses were strongly retained by becoming attached to both SWIs and AWIs. The role of straining in virus retention could not be ignored. Viruses were retained more at the SWIs and less straining occurred under acidic conditions than under neutral or alkaline conditions. This was mainly because of the effects of the pH and IS on surface charges and the model virus particle size. This study has important implications for modeling and predicting virus transport in soil affected by rainfall, snowmelt, and human activities (e.g., irrigation and artificial groundwater recharging).
显示更多 [+] 显示较少 [-]Identification of three metallothioneins in the black soldier fly and their functions in Cd accumulation and detoxification
2021
Zhang, Jie | Shi, Zhihui | Gao, Zhenghui | Wen, Yiting | Wang, Wanqiang | Liu, Wen | Wang, Xiaoping | Zhu, Fen
The black soldier fly (BSF) Hermetia illucens has a strong tolerance to cadmium stress. This helps to use BSF in entomoremediation of heavy metal pollution. Rich metallothionein (MT) proteins were thought to be important for some insects to endure the toxicity of heavy metal. We identified and characterized three MTs genes in BSF (BSFMTs), including BSFMT1, BSFMT2A, and BSFMT2B. Molecular modeling was used to predict metal binding sites. Phylogenetic analysis was used to identify gene families. Overexpression of the recombinant black soldier fly metallothioneins was found to confer Cd tolerance in Escherichia coli. Finally, functions of BSFMTs in BSF were explored through RNA interference (RNAi). RNAi results of BSFMT2B showed that the larval fresh weight decreased significantly, and the larvae mortality increased significantly. This study suggests that BSFMTs have important properties in Cd detoxification and tolerance in BSF. Further characterization analyses of physiological function about metallothioneins are necessary in BSF and other insects.
显示更多 [+] 显示较少 [-]An assessment of contamination fingerprinting techniques for determining the impact of domestic wastewater treatment systems on private well supplies
2021
Fennell, Christopher | Misstear, Bruce | O’Connell, David | Dubber, Donata | Behan, Patrice | Danaher, Martin | Moloney, Mary | Gill, Laurence
Private wells in Ireland and elsewhere have been shown to be prone to microbial contamination with the main suspected sources being practices associated with agriculture and domestic wastewater treatment systems (DWWTS). While the microbial quality of private well water is commonly assessed using faecal indicator bacteria, such as Escherichia coli, such organisms are not usually source-specific, and hence cannot definitively conclude the exact origin of the contamination. This research assessed a range of different chemical contamination fingerprinting techniques (ionic ratios, artificial sweeteners, caffeine, fluorescent whitening compounds, faecal sterol profiles and pharmaceuticals) as to their use to apportion contamination of private wells between human wastewater and animal husbandry wastes in rural areas of Ireland. A one-off sampling and analysis campaign of 212 private wells found that 15% were contaminated with E. coli. More extensive monitoring of 24 selected wells found 58% to be contaminated with E. coli on at least one occasion over a 14-month period. The application of fingerprinting techniques to these monitored wells found that the use of chloride/bromide and potassium/sodium ratios is a useful low-cost fingerprinting technique capable of identifying impacts from human wastewater and organic agricultural contamination, respectively. The artificial sweetener acesulfame was detected on several occasions in a number of monitored wells, indicating its conservative nature and potential use as a fingerprinting technique for human wastewater. However, neither fluorescent whitening compounds nor caffeine were detected in any wells, and faecal sterol profiles proved inconclusive, suggesting limited suitability for the conditions investigated.
显示更多 [+] 显示较少 [-]Electrocatalytic inactivation of antibiotic resistant bacteria and control of antibiotic resistance dissemination risk
2021
Liu, Haiyang | Hua, Xiuyi | Zhang, Ya-nan | Zhang, Tingting | Qu, Jiao | Nolte, Tom M. | Chen, Guangchao | Dong, Deming
Antibiotic resistance in environmental matrices becomes urgently significant for public health and has been considered as an emerging environmental contaminant. In this work, the ampicillin-resistant Escherichia coli (AR E. coli) and corresponding resistance genes (blaTEM₋₁) were effectively eliminated by the electrocatalytic process, and the dissemination risk of antibiotic resistance was also investigated. All the AR E. coli (∼8 log) was inactivated and 8.17 log blaTEM₋₁ was degraded by the carbon nanotubes/agarose/titanium (CNTs/AG/Ti) electrode within 30 min. AR E. coli was inactivated mainly attributing to the damage of cell membrane, which was attacked by reactive oxygen species and subsequent leakage of intracellular cytoplasm. The blaTEM₋₁ was degraded owing to the strand breaking in the process of electrocatalytic degradation. Furthermore, the dissemination risk of antibiotic resistance was effectively controlled after being electrocatalytic treatment. This study provided an effective electrocatalytic technology for the inactivation of antibiotic resistant bacteria and control of antibiotic resistance dissemination risk in the aqueous environment.
显示更多 [+] 显示较少 [-]Visible light driven exotic p (CuO) - n (TiO2) heterojunction for the photodegradation of 4-chlorophenol and antibacterial activity
2021
Gnanasekaran, Lalitha | Pachaiappan, Rekha | Kumar, P Senthil | Hoang, Tuan K.A. | Rajendran, Saravanan | Durgalakshmi, D. | Soto-Moscoso, Matias | Cornejo-Ponce, Lorena | Gracia, F.
The treatment of industrial waste and harmful bacteria is an important topic due to the release of toxins from the industrial pollutants that damage the water resources. These harmful sources frighten the life of every organism which was later developed as the carcinogenic and mutagenic agents. Therefore, the current study focuses on the breakdown or degradation of 4-chlorophenol and the antibacterial activity against Escherichia coli (E. coli). As a well-known catalyst, pure titanium-di-oxide (TiO₂) had not shown the photocatalytic activity in the visible light region. Hence, band position of TiO₂ need to be shifted to bring out the absorption in the visible light region. For this purpose, the n-type TiO₂ nanocrystalline material's band gap got varied by adding different ratios of p-type CuO. The result had appeared in the formation of p (CuO) – n (TiO₂) junction synthesized from sol-gel followed by chemical precipitation methods. The optical band gap value was determined by Kubelka-Munk (K-M) plot through UV–Vis diffusive reflectance spectroscopy (DRS). Further, the comprehensive mechanism and the results of photocatalytic and antibacterial activities were discussed in detail. These investigations are made for tuning the TiO₂ catalyst towards improving or eliminating the existing various environmental damages.
显示更多 [+] 显示较少 [-]Predicting mixture toxicity and antibiotic resistance of fluoroquinolones and their photodegradation products in Escherichia coli
2020
Wang, Dali | Ning, Qing | Dong, Jiayu | Brooks, Bryan W. | Yau, Ching
Antibiotics in the environment usually co-exist with their transformation products with retained toxicity, raising concerns about environmental risks of their combined exposure. Herein, we reported a novel predictive approach for evaluating the individual and combined toxicity for photodegradation products of fluoroquinolone antibiotics (FQs). Quantitative structure-activity relationship (QSAR) models with promising predictive performance were constructed and validated using experimental data obtained with 13 FQs and 78 mixtures towards E. coli. A structural descriptor reflecting the interaction among FQ molecules and the target protein was employed in the QSAR models, which was obtained through molecular docking and thus provided a rational mechanistic explanation for these models. The predicted results indicated that the degradation products displayed varying degrees of changes compared to the parent FQs, while the combined toxicity of FQs and their degradation products was mostly additive. Furthermore, following UV irradiation the degradation products displayed elevated capacity of inducing resistance mutations in E. coli, though their overall toxicity was reduced. This result highlights the implications of antibiotic degradation products on resistance development in bacteria and stresses the importance of considering such impacts during environmental risk assessments of antibiotics.
显示更多 [+] 显示较少 [-]Photoassisted degradation of 2,2′,4,4′-tetrabrominated diphenyl ether in simulated soil washing system containing Triton X series surfactants
2020
Huang, Kaibo | Liu, He | He, Jinglei | Li, Yan | Wang, Rui | Tang, Ting | Tao, Xueqin | Yin, Hua | Dang, Zhi | Lu, Guining
This study aims to use ultraviolet (UV) irradiation to decompose polybrominated diphenyl ethers (PBDEs) in the elutes and then reuse the surfactants. The results indicate that UV can remove 2,2′,4,4′-tetrabrominated diphenyl ether (BDE-47) from surfactant eluents and Triton X series surfactants also can remove BDE-47 from the soil. Triton X-100 (TX-100) is the most promising surfactant during the washing and photodegradation processes. Quench experiments suggest that both ¹O₂ and OH• were involved in the TX-100 decomposition but only ¹O₂ is responsible for the degradation of BDE-47. In analysis of the photoproducts of BDE-47 by Gas Chromatography Mass Spectrum (GC-MS) and Liquid Chromatography High Resolution Mass Spectrometry (LC-HRMS), BDE-47 was mainly debrominated to the lower-brominated BDEs and then oxidized to ring-opening products. The little loss of TX-100 can mainly be attributed to the breakage of polyethylene oxide (PEO) chain. Nevertheless, the washing wastes treated by UV light can exhibit higher solubility for BDE-47 than before, indicating they can be reused for BDE-47 removal from soil. The toxicity assessment experiments were performed using Escherichia coli (E.coli) as an indicator. The results indicate that the removal of BDE-47 by UV irradiation can reduce the toxicity of eluent.
显示更多 [+] 显示较少 [-]Consecutive ultrafiltration and silica adsorption for recovery of extracellular antibiotic resistance genes from an urban river
2020
Liu, Miaomiao | Hata, Akihiko | Katayama, Hiroyuki | Kasuga, Ikuro
The dissemination of antibiotic resistance (AR) has attracted global attention because of the increasing antibiotic treatment failure it has caused. Through natural transformation, a live bacterium takes up extracellular DNA (exDNA), which facilitates AR dissemination. However, recovery of exDNA from water samples is challenging. In this study, we validated a consecutive ultrafiltration-based protocol to simultaneously recover intracellular DNA (inDNA), dissolved exDNA (Dis_exDNA, dissolved in the bulk water), and adsorbed exDNA (Ads_exDNA, adsorbed to the surfaces of suspended particles). Using hollow fiber ultrafiltration (HFUF), all DNA fractions were concentrated from environmental water samples, after which Dis_exDNA (supernatant) was separated from inDNA and Ads_exDNA (pellets) using centrifugation. Ads_exDNA was washed off from the pellets with proteinase K and sodium phosphate buffer. Dis_exDNA and Ads_exDNA were further concentrated using centrifugal ultrafiltration, from which silica binding was performed. inDNA was extracted from washed pellets with a commercial kit. For inDNA, HFUF showed recovery efficiencies of 96.5 ± 18.5% and 88.0 ± 2.0% for total cells and cultured Escherichia coli, respectively (n = 3). To represent all possible DNA fragments in water environment, exDNA with different lengths (10.0, 4.0, 1.0, and 0.5 kbp) were spiked to test the recovery efficiencies for Dis_exDNA. The whole process achieved 62.2%–62.9% recovery for 10 and 4 kbp exDNA, and 38.8%–44.5% recovery for 1.0 and 0.5 kbp exDNA. Proteinase K treatment enhanced the recovery of Ads_exDNA by 4.0–10.7 times. The protocol was applied to water samples from an urban river in Tokyo, Japan. The abundance of AR genes (ARGs) in inDNA, Dis_exDNA, and Ads_exDNA increased downstream of wastewater treatment plants. ARGs in Ads_exDNA and Dis_exDNA accounted for 1.8%–26.7% and 0.03%–20.9%, respectively, of the total DNA, implying that Ads_exDNA and Dis_exDNA are nonnegligible potential pools for the horizontal transfer of ARGs.
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