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The Science of the Total Environment Jul 2024The spread of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in humans, animals and environment is a growing threat to public health....
Characterisation of microbial communities and quantification of antibiotic resistance genes in Italian wastewater treatment plants using 16S rRNA sequencing and digital PCR.
The spread of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in humans, animals and environment is a growing threat to public health. Wastewater treatment plants (WWTPs) are crucial in mitigating the risk of environmental contamination by effectively removing contaminants before discharge. However, the persistence of ARB and ARGs even after treatment is a challenge for the management of water system. To comprehensively assess antimicrobial resistance dynamics, we conducted a one-year monitoring study in three WWTPs in central Italy, both influents and effluents. We used seasonal sampling to analyze microbial communities by 16S rRNA, as well as to determine the prevalence and behaviour of major ARGs (sul1, tetA, bla, bla, bla, bla) and the class 1 Integron (int1). Predominant genera included in order: Arcobacter, Acinetobacter, Flavobacterium, Pseudarcobacter, Bacteroides, Aeromonas, Trichococcus, Cloacibacterium, Pseudomonas and Streptococcus. A higher diversity of bacterial communities was observed in the effluents compared to the influents. Within these communities, we also identified bacteria that may be associated with antibiotic resistance and pose a significant threat to human health. The mean concentrations (in gene copies per liter, gc/L) of ARGs and int1 in untreated wastewater (absolute abundance) were as follows: sul1 (4.1 × 10), tetA (5.2 × 10), bla (1.1 × 10), bla (2.1 × 10), bla (1.1 × 10), bla (9.4 × 10), and int1 (5.5 × 10). The mean values in treated effluents showed reductions ranging from one to three log. However, after normalizing to the 16S rRNA gene (relative abundance), it was observed that in 37.5 % (42/112) of measurements, the relative abundance of ARGs increased in effluents compared to influents. Furthermore, correlations were identified between ARGs and bacterial genera including priority pathogens. This study improves our understanding of the dynamics of ARGs and provides insights to develop more effective strategies to reduce their spread, protecting public health and preserving the future efficacy of antibiotics.
Topics: Wastewater; Italy; RNA, Ribosomal, 16S; Waste Disposal, Fluid; Bacteria; Drug Resistance, Microbial; Microbiota; Genes, Bacterial; Environmental Monitoring; Polymerase Chain Reaction; Drug Resistance, Bacterial; Anti-Bacterial Agents; Water Microbiology
PubMed: 38750766
DOI: 10.1016/j.scitotenv.2024.173217 -
Water Research Jun 2024Waterborne pathogens are threatening public health globally, but profiling multiple human pathogenic bacteria (HPBs) in various polluted environments is still a...
Waterborne pathogens are threatening public health globally, but profiling multiple human pathogenic bacteria (HPBs) in various polluted environments is still a challenge due to the absence of rapid, high-throughput and accurate quantification tools. This work developed a novel chip, termed the HPB-Chip, based on high-throughput quantitative polymerase chain reactions (HT-qPCR). The HPB-Chip with 33-nL reaction volume could simultaneously complete 10,752 amplification reactions, quantifying 27 HPBs in up to 192 samples with two technical replicates (including those for generating standard curves). Specific positive bands of target genes across different species and single peak melting curves demonstrated high specificity of the HPB-Chip. The mixed plasmid serial dilution test validated its high sensitivity with the limit of quantification (LoD) of averaged 82 copies per reaction for 25 target genes. PCR amplification efficiencies and R coefficients of standard curves of the HPB-Chip averaged 101 % and 0.996, respectively. Moreover, a strong positive correlation (Pearson' r: 0.961-0.994, P < 0.001) of HPB concentrations (log copies/L) between HPB-Chip and conventional qPCR demonstrated high accuracy of the HPB-Chip. Subsequently, the HPB-Chip has been successfully applied to absolutely quantify 27 HPBs in municipal and hospital wastewater treatment plants (WWTPs) after PMA treatment. A total of 17 HPBs were detected in the 6 full-scale WWTPs, with an additional 19 in the hospital WWTP. Remarkably, Acinetobacter baumannii, Legionella pneumophila, and Arcobacter butzler were present in the final effluent of each municipal WWTP. Overall, the HPB-Chip is an efficient and accurate high-throughput quantification tool to comprehensively and rapidly quantify 27 HPBs in the environment.
PubMed: 38941866
DOI: 10.1016/j.watres.2024.121927 -
Water Research Apr 2024Membrane biofouling is a challenge to be solved for the stable operation of the seawater reverse osmosis (SWRO) membrane. This study explored the regulation mechanism of...
Membrane biofouling is a challenge to be solved for the stable operation of the seawater reverse osmosis (SWRO) membrane. This study explored the regulation mechanism of quorum sensing (QS) inhibition on microbial community composition and population-level behaviors in seawater desalination membrane biofouling. A novel antibiofouling SWRO membrane (MA_m) by incorporating one of quorum sensing inhibitors (QSIs), methyl anthranilate (MA) was prepared. It exhibited enhanced anti-biofouling performance than the exogenous addition of QSIs, showing long-term stability and alleviating 22 % decrease in membrane flux compared with the virgin membrane. The results observed that dominant bacteria Epsilon- and Gamma-proteobacteria (Shewanella, Olleya, Colwellia, and Arcobacter), which are significantly related to (P ≤ 0.01) the metabolic products (i.e., polysaccharides, proteins and eDNA), are reduced by over 80 % on the MA_m membrane. Additionally, the introduction of MA has a more significant impact on the QS signal-sensing pathway through binding to the active site of the transmembrane sensor receptor. It effectively reduces the abundance of genes encoding QS and extracellular polymeric substance (EPS) (exopolysaccharides (i.e., galE and nagB) and amino acids (i.e., ilvE, metH, phhA, and serB)) by up to 50 % and 30 %, respectively, resulting in a reduction of EPS by more than 50 %, thereby limiting the biofilm formation on the QSI-modified membrane. This study provides novel insights into the potential of QSIs to control consortial biofilm formation in practical SWRO applications.
Topics: Biofouling; Quorum Sensing; Biofilms; Extracellular Polymeric Substance Matrix; Osmosis; Microbiota; Seawater; Membranes, Artificial; Water Purification
PubMed: 38402750
DOI: 10.1016/j.watres.2024.121358 -
The Science of the Total Environment Jul 2024Bioaerosols released from municipal wastewater treatment plants (MWWTPs) contain pathogenic microorganisms, if dispersed into the atmosphere, which pose potential health...
Bioaerosols released from municipal wastewater treatment plants (MWWTPs) contain pathogenic microorganisms, if dispersed into the atmosphere, which pose potential health risks to humans. In this study, the concentrations and size distribution of bioaerosol, factors on the bioaerosol emission, exposure risk, and microbial composition in different treatment units of a MWWTP were investigated. The results showed that bioaerosol was released to different degrees in each treatment unit, with the concentrations of bioaerosol varied widely, ranging from 978 to 3710 CFU/m. FG and PST were primary bioaerosol emission sources in MWWTP. COD concentration, wind speed (WS) and relative humidity (RH) significantly influenced bioaerosol concentrations. The proportion of inhalable particles (< 4.7 μm) ranged from 51.35 % to 83.33 %, and bioaerosol emitted from WWTP caused a non-carcinogenic risk to children by the exposure risk assessment (HI > 1), which need to be paid more attention. Bacterial, fungal and actinomycete aerosols were detected in each treatment unit of MWWTP. Among these bioaerosols, bacterial aerosol was dominant. Importantly, several pathogenic bacteria including Sphingobium, Brevundimonas, Romboutsia, Arcobacter, Acinetobacter, and Mycobacterium were identified within the airborne bacteria population, most of which originated from wastewater or sludge, particularly in the ambient air of AeT. Pathogenic bacteria from MWWTP should be studied further to determine their long-term behavior and possible health risks.
Topics: Aerosols; Air Microbiology; Risk Assessment; Wastewater; Environmental Monitoring; Waste Disposal, Fluid; Air Pollutants; Bacteria; Humans
PubMed: 38729365
DOI: 10.1016/j.scitotenv.2024.173096 -
Yakugaku Zasshi : Journal of the... 2024Neural activity generates essential responses, such as thinking, memory formation, and muscle contraction. It is controlled by the well-coordinated activity of various...
Neural activity generates essential responses, such as thinking, memory formation, and muscle contraction. It is controlled by the well-coordinated activity of various cation-selective channels of the cell membrane. The divalent cation block plays an essential role in various tetrameric ion channels. For example, N-methyl-D-aspartic acid receptors, which are tetrameric ion channels involved in memory formation, are inhibited by magnesium ions. Divalent cations are thought to bind in the ion pathway of the ion channel and as a consequence block the channel current, however, direct observation of such a block has not been reported yet. As a consequence, the behavior of these blocking divalent cations remains poorly understood. NavAb, a similar tetrameric sodium channel cloned from Arcobacter butzleri, is one of the most structurally analyzed tetrameric channels that is not inhibited by divalent cations. In this study, we elucidated the molecular mechanism of the divalent cation block by reproducing the divalent cation block in NavAb. The X-ray crystal structure of divalent-cation-block mutants show electron density in the ion transmission pathway of the divalent cation blocked mutants, indicating that the mutations increasing the hydrophilicity of the inner vestibule of the pore domain enable a divalent cation to stack into the ion pathway. In molecular dynamics simulations, the stacked calcium ion repels the sodium ions near the channel lumen's entrance at the selective filter's bottom. These results suggest the primary process of the divalent cation block mechanism in tetrameric cation channels and suggest a process of functional acquisition in ion channel evolution.
Topics: Arcobacter; Cations, Divalent; Crystallography, X-Ray; Magnesium; Molecular Dynamics Simulation; Mutation; Sodium Channels
PubMed: 38692927
DOI: 10.1248/yakushi.23-00178-3