Insights into the effects of anthropogenic activities on oil reservoir microbiome and metabolic potential.

Microbial communities have long been observed in oil reservoirs, where the subsurface conditions are major drivers shaping their structure and functions. Furthermore, anthropogenic activities such as water flooding during oil production can affect microbial activities and community compositions in oil reservoirs through the injection of recycled produced water, often associated with biocides. However, it is still unclear to what extent the introduced chemicals and microbes influence the metabolic potential of the subsurface microbiome. Here we investigated an onshore oilfield in Germany (Field A) that undergoes secondary oil production along with biocide treatment to prevent souring and microbially induced corrosion (MIC). With the integrated approach of 16 S rRNA gene amplicon and shotgun metagenomic sequencing of water-oil samples from 4 production wells and 1 injection well, we found differences in microbial community structure and metabolic functions. In the injection water samples, amplicon sequence variants (ASVs) belonging to families such as Halanaerobiaceae, Ectothiorhodospiraceae, Hydrogenophilaceae, Halobacteroidaceae, Desulfohalobiaceae, and Methanosarcinaceae were dominant, while in the production water samples, ASVs of families such as Thermotogaceae, Nitrospiraceae, Petrotogaceae, Syntrophaceae, Methanobacteriaceae, and Thermoprotei were also dominant. The metagenomic analysis of the injection water sample revealed the presence of C1-metabolism, namely, genes involved in formaldehyde oxidation. Our analysis revealed that the microbial community structure of the production water samples diverged slightly from that of injection water samples. Additionally, a metabolic potential for oxidizing the applied biocide clearly occurred in the injection water samples indicating an adaptation and buildup of degradation capacity or resistance against the added biocide.

Non-regulated haloaromatic water disinfection byproducts act as endocrine and lipid disrupters in human placental cells.

The disinfection of drinking water generates hundreds of disinfection byproducts (DBPs), including haloaromatic DBPs. These haloaromatic DBPs are suspected to be more toxic than haloaliphatic ones, and they are currently not regulated. This work investigates their toxicity and ability to interfere with estrogen synthesis in human placental JEG-3 cells, and their genotoxic potential in human alveolar A549 cells. Among the haloaromatic DBPs studied, halobenzoquinones (2,6-dichloro-1,4-benzoquinone (DCBQ) and 2,6-dibromo-1,4-benzoquinone (DBBQ)) showed the highest cytotoxicity (EC50: 18-26 µg/mL). They induced the generation of very high levels of reactive oxygen species (ROS) and up-regulated the expression of genes involved in estrogen synthesis (cyp19a1, hsd17b1). Increased ROS was linked to significant depletion of polyunsaturated lipid species from inner cell membranes. The other DBPs tested showed low or no significant cytotoxicity (EC50 ≥ 100 µg/mL), while 2,4,6-trichloro-phenol (TCP), 2,4,6-tribromo-phenol (TBP) and 3,5-dibromo-4-hydroxybenzaldehyde (DCHB) induced the formation of micronuclei at concentrations much higher than those typically found in water (100 µg/mL). This study reveals the different modes of action of haloaromatic DBPs, and highlights the toxic potential of halobenzoquinones, which had a significant impact on the expression of placenta steroid metabolism related genes and induce oxidative stress, implying potential adverse health effects.

Response mechanisms of pipe wall biofilms in water supply networks under different disinfection strategy pressures and the effect of mediating halogenated acetonitrile formation.

Residual chlorine and biofilm coexistence is inevitable in drinking water transmission and distribution networks. Understanding the microbial response and its mediated effects on disinfection byproducts under different categories of residual chlorine stress is essential to ensure water safety. The aim of our study was to determine the response of pipe wall biofilms to residual chlorine pressure in chlorine and chloramine systems and to understand the microbially mediated effects on the formation and migration of haloacetonitriles (HANs), typical nitrogenous disinfection byproducts. According to the experimental results, the biofilm response changes under pressure, with significant differences noted in morphological characteristics, the extracellular polymeric substances (EPS) spatial structure, bacterial diversity, and functional abundance potential. Upon incubation with residual chlorine (1.0 ± 0.2 mg/L), the biofilm biomass per unit area, EPS, community abundance, and diversity increased in the chloramine group, and the percentage of viable bacteria increased, potentially indicating that the chloramine group provides a richer variety of organic matter precursors. Compared with the chloramine group, the chlorination group exhibited increased haloacetonitrile formation potential (HANFP), with Rhodococcus (43.2%) dominating the system, whereas the prediction abundance of metabolic functions was advantageous, especially with regard to amino acid metabolism, carbohydrate metabolism, and the biodegradation and metabolism of foreign chemicals. Under chlorine stress, pipe wall biofilms play a stronger role in mediating HAN production. It is inferred that chlorine may stimulates microbial interactions, and more metabolites (e.g., EPS) consume chlorine to protect microbial survival. EPS dominates in biofilms, in which proteins exhibit greater HANFP than polysaccharides.

Efficacy of microbicidal actives and formulations for inactivation of Lassa virus in suspension.

The World Health Organization's R&D Blueprint list of priority diseases for 2022 includes Lassa fever, signifying the need for research and development in emergency contexts. This disease is caused by the arenavirus Lassa virus (LASV). Being an enveloped virus, LASV should be susceptible to a variety of microbicidal actives, although empirical data to support this expectation are needed. We evaluated the virucidal efficacy of sodium hypochlorite, ethanol, a formulated dual quaternary ammonium compound, an accelerated hydrogen peroxide formulation, and a p-chloro-m-xylenol formulation, per ASTM E1052-20, against LASV engineered to express green fluorescent protein (GFP). A 10-µL volume of virus in tripartite soil (bovine serum albumin, tryptone, and mucin) was combined with 50 µL of disinfectant in suspension for 0.5, 1, 5, or 10 min at 20-25 °C. Neutralized test mixtures were quantified by GFP expression to determine log10 reduction. Remaining material was passaged on Vero cells to confirm absence of residual infectious virus. Input virus titers of 6.6-8.0 log10 per assay were completely inactivated by each disinfectant within 1-5 min contact time. The rapid and substantial inactivation of LASV suggests the utility of these microbicides for mitigating spread of infectious virus during Lassa fever outbreaks.

Isolation and characterization of Rhodococcus sp. GG1 for metabolic degradation of chloroxylenol.

The coronavirus disease 2019 (COVID-19) pandemic has significantly increased the demand of disinfectant use. Chloroxylenol (para-chloro-meta-xylenol, PCMX) as the major antimicrobial ingredient of disinfectant has been widely detected in water environments, with identified toxicity and potential risk. The assessment of PCMX in domestic wastewater of Macau Special Administrative Region (SAR) showed a positive correlation between PCMX concentration and population density. An indigenous PCMX degrader, identified as Rhodococcus sp. GG1, was isolated and found capable of completely degrading PCMX (50 mg L-1) within 36 h. The growth kinetics followed Haldane's inhibition model, with maximum specific growth rate, half-saturation constant, and inhibition constant of 0.38 h-1, 7.64 mg L-1, and 68.08 mg L-1, respectively. The degradation performance was enhanced by optimizing culture conditions, while the presence of additional carbon source stimulated strain GG1 to alleviate inhibition from high concentrations of PCMX. In addition, strain GG1 showed good environmental adaptability, degrading PCMX efficiently in different environmental aqueous matrices. A potential degradation pathway was identified, with 2,6-dimethylhydroquinone as a major intermediate metabolite. Cytochrome P450 (CYP450) was found to play a key role in dechlorinating PCMX via hydroxylation and also catalyzed the hydroxylated dechlorination of other halo-phenolic contaminants through co-metabolism. This study characterizes an aerobic bacterial pure culture capable of degrading PCMX metabolically, which could be promising in effective bioremediation of PCMX-contaminated sites and in treatment of PCMX-containing waste streams.

Evolution of Chlorhexidine Susceptibility and of the EfrEF Operon among Enterococcus faecalis from Diverse Environments, Clones, and Time Spans.

Chlorhexidine (CHX) is widely used to control the spread of pathogens (e.g., human/animal clinical settings, ambulatory care, food industry). Enterococcus faecalis, a major nosocomial pathogen, is broadly distributed in diverse hosts and environments facilitating its exposure to CHX over the years. Nevertheless, CHX activity against E. faecalis is understudied. Our goal was to assess CHX activity and the variability of ChlR-EfrEF proteins (associated with CHX tolerance) among 673 field isolates and 1,784 E. faecalis genomes from the PATRIC database from different sources, time spans, clonal lineages, and antibiotic-resistance profiles. The CHX MIC (MICCHX) and minimum bactericidal concentration (MBCCHX) against E. faecalis presented normal distributions (0.5 to 64 mg/L). However, more CHX-tolerant isolates were detected in the food chain and recent human infections, suggesting an adaptability of E. faecalis populations in settings where CHX is heavily used. Heterogeneity in ChlR-EfrEF sequences was identified, with isolates harboring incomplete ChlR-EfrEF proteins, particularly the EfrE identified in the ST40 clonal lineage, showing low MICCHX (≤1mg/L). Distinct ST40-E. faecalis subpopulations carrying truncated and nontruncated EfrE were detected, with the former being predominant in human isolates. This study provides a new insight about CHX susceptibility and ChlR-EfrEF variability within diverse E. faecalis populations. The MICCHX/MBCCHX of more tolerant E. faecalis (MICCHX = 8 mg/L; MBCCHX = 64 mg/L) remain lower than in-use concentrations of CHX (≥500 mg/L). However, increased CHX use, combined with concentration gradients occurring in diverse environments, potentially selecting multidrug-resistant strains with different CHX susceptibilities, signals the importance of monitoring the trends of E. faecalis CHX tolerance within a One Health approach. IMPORTANCE Chlorhexidine (CHX) is a disinfectant and antiseptic used since the 1950s and included in the World Health Organization's list of essential medicines. It has been widely applied in hospitals, the community, the food industry, animal husbandry and pets. CHX tolerance in Enterococcus faecalis, a ubiquitous bacterium and one of the leading causes of human hospital-acquired infections, remains underexplored. Our study provides novel and comprehensive insights about CHX susceptibility within the E. faecalis population structure context, revealing more CHX-tolerant subpopulations from the food chain and recent human infections. We further show a detailed analysis of the genetic diversity of the efrEF operon (previously associated with E. faecalis CHX tolerance) and its correlation with CHX phenotypes. The recent strains with a higher tolerance to CHX and the multiple sources where bacteria are exposed to this biocide alert us to the need for the continuous monitoring of E. faecalis adaptation toward CHX tolerance within a One Health approach.

Transcriptomic Responses of Salmonella enterica Serovars Enteritidis in Sodium Hypochlorite.

Salmonella enterica serovars Enteritidis (S. Enteritidis) can survive extreme food processing environments including bactericidal sodium hypochlorite (NaClO) treatments generally recognized as safe. In order to reveal the molecular regulatory mechanisms underlying the phenotypes, the overall regulation of genes at the transcription level in S. Enteritidis after NaClO stimulation were investigated by RNA-sequencing. We identified 1399 differentially expressed genes (DEG) of S. Enteritidis strain CVCC 1806 following treatment in liquid culture with 100 mg/L NaClO for 20 min (915 upregulated and 484 downregulated). NaClO stress affects the transcription of genes related to a range of important biomolecular processes such as membrane damage, membrane transport function, energy metabolism, oxidative stress, DNA repair, and other important processes in Salmonella enterica. First, NaClO affects the structural stability of cell membranes, which induces the expression of a range of outer and inner membrane proteins. This may lead to changes in cell membrane permeability, accelerating the frequency of DNA conversion and contributing to the production of drug-resistant bacteria. In addition, the expression of exocytosis pump genes (emrB, yceE, ydhE, and ydhC) was able to expel NaClO from the cell, thereby increasing bacterial tolerance to NaClO. Secondly, downregulation of genes related to the Kdp-ATPase transporter system (kdpABC) and the amino acid transporter system (aroP, brnQ and livF) may to some extent reduce active transport by bacterial cells, thereby reducing their own metabolism and the entry of disinfectants. Downregulation of genes related to the tricarboxylic acid (TCA) cycle may drive bacterial cells into a viable but non-culturable (VBNC) state, resisting NaClO attack by reducing energy metabolism. In addition, significant upregulation of genes related to oxidative stress could mitigate damage caused by disinfectants by eliminating alkyl hydroperoxides, while upregulation of genes related to DNA repair could repair damage to bacterial cells caused by oxidative stress. Therefore, this study indicated that S. Enteritidis has genomic mechanisms to adapt to NaClO stress.

Antiviral Activity of Olanexidine-Containing Hand Rub against Human Noroviruses.

Human norovirus (HuNoV) is the leading cause of epidemic and sporadic acute gastroenteritis worldwide. HuNoV transmission occurs predominantly by direct person-to-person contact, and its health burden is associated with poor hand hygiene and a lack of effective antiseptics and disinfectants. Specific therapies and methods to prevent and control HuNoV spread previously were difficult to evaluate because of the lack of a cell culture system to propagate infectious virus. This barrier has been overcome with the successful cultivation of HuNoV in nontransformed human intestinal enteroids (HIEs). Here, we report using the HIE cultivation system to evaluate the virucidal efficacy of an olanexidine gluconate-based hand rub (OLG-HR) and 70% ethanol (EtOH70%) against HuNoVs. OLG-HR exhibited fast-acting virucidal activity against a spectrum of HuNoVs including GII.4 Sydney[P31], GII.4 Den Haag[P4], GII.4 New Orleans[P4], GII.3[P21], GII.17[P13], and GI.1[P1] strains. Exposure of HuNoV to OLG-HR for 30 to 60 s resulted in complete loss of the ability of virus to bind to the cells and reduced in vitro binding to glycans in porcine gastric mucin. By contrast, the virucidal efficiency of EtOH70% on virus infectivity was strain specific. Dynamic light scattering (DLS) and electron microscopy of virus-like particles (VLPs) show that OLG-HR treatment causes partial disassembly and possibly conformational changes in VP1, interfering with histo-blood group antigen (HBGA) binding and infectivity, whereas EtOH70% treatment causes particle disassembly and clumping of the disassembled products, leading to loss of infectivity while retaining HBGA binding. The highly effective inactivation of HuNoV infectivity by OLG-HR suggests that this compound could reduce HuNoV transmission. IMPORTANCE Human noroviruses (HuNoVs) are highly contagious and cause nonbacterial acute gastroenteritis in all age groups worldwide. Since the introduction of rotavirus vaccines, HuNoVs have become the leading cause of diarrheal illness in children. These viruses are very stable in the environment and resistant to common disinfectants. This study evaluated the virucidal efficacy of a new disinfectant, olanexidine-based hand rub (OLG-HR), against HuNoV strains in an ex vivo human intestinal stem cell-derived enteroid (HIE) cultivation system. Exposure of multiple HuNoV strains to OLG-HR for 30 to 60 s resulted in complete loss of infectivity and binding to HBGAs, possibly due to partial disassembly and conformational changes in the major virus capsid (VP1). By comparison, the virucidal efficiency of EtOH70% was strain specific, leading to loss of infectivity while retaining HBGA binding. These findings show the utility of the ex vivo HIE cultivation system to test the effectiveness of disinfectants and report a highly effective product.

Effect of Biocides on the Activity of Extracellular Enzymes Produced by Degrader Fungi from Technical Objects Used in Tropical Climates (Vietnam).

Activity of extracellular enzymes was assessed in 20 strains of microscopic fungi involved in biodegradation of technical objects exploited under tropical climate conditions (Vietnam). It was found that 19 strains possessed catalase activity, 18 strains had phenol oxidase activity, and eight strains had protease activity. The effect of industrial biocides on the activity of these enzymes was also assessed. The biocides Bior-1, Bioneutral A 10, and Bioneutral A 101 were shown to inhibit the enzymatic activity to various extent. All biocides inhibited extracellular catalase activity in most fungal strains studied. The inhibition of protease and phenol oxidase activity of same test strains was less pronounced. The response to biocides varied at the strain level; its characteristics could differ significantly even between strains of the same species. In several cases, it was observed that exposure to biocides resulted in an increase in enzyme activity.

Toxicological effects induced by the biocide triclosan on Pseudokirchneriella subcapitata.

Triclosan, a widely used biocide broadly found in aquatic environments, is cause of concern due to its unknown effects on non-targets organisms. In this study, a multi biomarker approach was used in order to evaluate the 72 h-effect of triclosan on the freshwater alga Pseudokirchneriella subcapitata (Raphidocelis subcapitata). Triclosan, at environmental relevant concentrations (27 and 37 µg L-1), caused a decrease of proliferative capacity, which was accompanied by an increase of cell size and a profound alteration of algae shape. It was found that triclosan promoted the intracellular accumulation of reactive oxygen species, the depletion of non-enzymatic antioxidant defenses (reduced glutathione and carotenoids) and a decrease of cell metabolic activity. A reduction of photosynthetic pigments (chlorophyll a and b) was also observed. For the highest concentration tested (37 µg L-1), a decrease of photosynthetic efficiency was detected along with a diminution of the relative transport rate of electrons on the photosynthetic chain. In conclusion, triclosan presents a deep impact on the microalga P. subcapitata morphology and physiology translated by multiple target sites instead of a specific point (cellular membrane) observed in the target organism (bacteria). Additionally, this study contributes to clarify the toxicity mechanisms of triclosan, in green algae, showing the existence of distinct modes of action of the biocide depending on the microalga.

Trimester-Specific Blood Trihalomethane and Urinary Haloacetic Acid Concentrations and Adverse Birth Outcomes: Identifying Windows of Vulnerability during Pregnancy.

BACKGROUND: Some disinfection by-products (DBPs) are reproductive and developmental toxicants in laboratory animals. However, studies of trimester-specific DBP exposure on adverse birth outcomes in humans are inconsistent. OBJECTIVE: We examined whether trimester-specific blood and urinary biomarkers of DBP were associated with small for gestational age (SGA), low birth weight (LBW), and preterm birth. METHODS: A total of 4,086 blood and 3,951 urine samples were collected across pregnancy trimesters among 1,660 mothers from Xiaogan City, China. Blood samples were quantified for biomarkers of trihalomethanes (THMs): chloroform (TCM), bromodichloromethane, dibromochloromethane, and bromoform. Urine samples were quantified for biomarkers of haloacetic acids (HAA): dichloroacetic acid and trichloroacetic acid. Birth outcomes were abstracted at delivery from medical records. We used Poisson regression models with log link functions to estimate risk ratios (RRs) and 95% confidence intervals (CIs) for SGA, LBW, and preterm birth across tertiles (or categories) of DBP biomarker concentrations measured across pregnancy trimesters. We also examined the relative exposure differences across gestation comparing adverse outcomes with normal births using mixed-effects models. RESULTS: Blood TCM concentrations in the second trimester were associated with an elevated risk of SGA comparing middle vs. lowest (RR, 2.34; 95% CI: 1.02, 5.35) and highest vs. lowest (RR, 2.47; 95% CI: 1.09, 5.58) exposure groups. Third-trimester blood TCM concentrations were also associated with an increased risk of SGA comparing the second tertile with the first (RR, 2.61; 95% CI: 1.15, 5.92). We found that maternal blood TCM concentrations were significantly higher for SGA compared with non-SGA births across the period from 23 to 34 wk gestation. Other blood and urinary DBP biomarkers examined were unrelated to SGA, LBW, or preterm birth. CONCLUSION: Blood TCM concentrations in mid to late pregnancy were associated with an increased risk of SGA, whereas other biomarkers of DBPs examined across pregnancy were not associated with birth outcomes. https://doi.org/10.1289/EHP7195.

Comparing in vivo data and in silico predictions for acute effects assessment of biocidal active substances and metabolites for aquatic organisms.

The purpose of this study was to determine the acute toxicity in aquatic organisms of one biocidal active substance and six metabolites derived from biocidal active substances and to assess the suitability of available QSAR models to predict the obtained values. We have reported the acute toxicity in sewage treatment plant (STP) microorganisms, in the freshwater microalgae Pseudokirchneriella subcapitata and in Daphnia magna following OECD test methods. We have also identified in silico models for acute toxicity of these trophic levels currently available in widely recognized platforms such as VEGA and the OECD QSAR ToolBox. A total of six, four and two models have been selected for Daphnia, algae and microorganisms, respectively. Finally, we have compared the in silico and in vivo data for the seven compounds plus two previously assayed biocidal substances. None of the compounds tested were toxic for Daphnia and STP microorganisms. For microalgae, CGA71019 (1,2,4 triazole) presented an ErC50 value of 38.3 mg/L. The selected in silico models have provided lower EC50 values and are therefore more conservative. Models from the OECD QSAR ToolBox predicted values for 7 out of 9 and for 4 out of 9 chemicals for Daphnia and P. subcapitata, respectively. No predictive models were identified in such platform for STP microorganism's acute effects. In terms of models's specificity, biocide-specific models, developed from curated datasets integrated by biocidal active substances and implemented in VEGA, perform better in the case of microalgae but for Daphnia an alternative, non biocide-specific has revealed a better performance. For STP microorganisms only biocide-specific models have been identified.

The methylisothiazolinone and methylchloroisothiazolinone metabolite N-methylmalonamic acid (NMMA) in urine of children and adolescents in Germany - Human biomonitoring results of the German Environmental Survey 2014-2017 (GerES V).

Mixtures of methylisothiazolinone and methylchloroisothiazolinone are used as biocides in cosmetics, cleaning agents, and water-based paint. A biomonitoring method to evaluate exposure to these compounds was developed using N-methylmalonamic acid (NMMA), the main metabolite of both, methylisothiazolinone and methylchloroisothiazolinone, as the exposure biomarker. First-morning void urine samples (N = 2078) of 3- to 17-year-old children and adolescents living in Germany were analysed for concentrations of NMMA in the population representative German Environmental Survey for Children and Adolescents GerES V (2014-2017). NMMA was quantified in almost all samples, with a geometric mean concentration of 6.245 µg/L (5.303 µg/gcrea) and a 95th percentile of 15.0 µg/L (12.6 µg/gcrea). Urinary concentrations could not be related to self-reported application of specific cleaning agents or personal care products, leaving potential, specific sources of exposure unrevealed as most products relevant for isothiazolinone exposure are used ubiquitously. For the first time, reference values can be derived for urinary NMMA for children and adolescents in Germany, facilitating a more substantiated exposure assessment.

Metabolism of Benzalkonium Chlorides by Human Hepatic Cytochromes P450.

Benzalkonium chlorides (BACs) are widely used as disinfectants in cleaning products, medical products, and the food processing industry. Despite a wide range of reported toxicities, limited studies have been conducted on the metabolism of these compounds in animal models and none in human-derived cells or tissues. In this work, we report on the metabolism of BACs in human liver microsomes (HLM) and by recombinant human hepatic cytochrome P450 (CYP) enzymes. BAC metabolism in HLM was NADPH-dependent and displayed apparent half-lives that increased with BAC alkyl chain length (C10 < C12 < C14 < C16), suggesting enhanced metabolic stability of the more lipophilic, longer chain BACs. Metabolites of d7-benzyl labeled BAC substrates retained all deuteriums and there was no evidence of N-dealkylation. Tandem mass spectrometry fragmentation of BAC metabolites confirmed that oxidation occurs on the alkyl chain region. Major metabolites of C10-BAC were identified as ω-hydroxy-, (ω-1)-hydroxy-, (ω, ω-1)-diol-, (ω-1)-ketone-, and ω-carboxylic acid-C10-BAC by liquid chromatography-mass spectrometry comparison with synthetic standards. In a screen of hepatic CYP isoforms, recombinant CYP2D6, CYP4F2, and CYP4F12 consumed substantial quantities of BAC substrates and produced the major microsomal metabolites. The use of potent pan-CYP4 inhibitor HET0016, the specific CYP2D6 inhibitor quinidine, or both confirmed major contributions of CYP4- and CYP2D6-mediated metabolism in the microsomal disappearance of BACs. Kinetic characterization of C10-BAC metabolite formation in HLM demonstrated robust Michaelis-Menten kinetic parameters for ω-hydroxylation (Vmax = 380 pmol/min/mg, Km = 0.69 µM) and (ω-1)-hydroxylation (Vmax = 126 pmol/min/mg, Km = 0.13 µM) reactions. This work illustrates important roles for CYP4-mediated ω-hydroxylation and CYP2D6/CYP4-mediated (ω-1)-hydroxylation during the hepatic elimination of BACs, an environmental contaminant of emerging concern. Furthermore, we demonstrate that CYP-mediated oxidation of C10-BAC mitigates the potent inhibition of cholesterol biosynthesis exhibited by this short-chain BAC.

The effects of biodegradation on the characteristics and disinfection by-products formation of soluble microbial products chemical fractions.

Soluble microbial products (SMPs) discharged into rivers from sewage treatment plants may increase the health risk for downstream drinking water by acting as a precursor of DBPs. Biotransformation or biodegradation could alter the characteristics of SMPs and affect the subsequent formation of DBPs. This study observed the relative contribution of chemical fractions in SMPs and explored the biodegradation of each fraction and their effect on disinfection by-products (DBPs) formation in surface water. The hydrophilic acid (HPIA) and hydrophobic acid (HPOA) constituted the major portion of the SMPs, which were dominated by fulvic acid and humic acids. The transphilic acid (TPIA) and hydrophobic bases (HPOB) were relatively minor but it contained a relative substantial portion of protein-like materials in SMPs. TPIA and HPOB produced insignificant amounts of DBP corresponding to 13% and 14% in the original samples, but they were collectively responsible for 50% of the DBPs yield. Much larger amounts of hydrophobic fractions were utilized than hydrophilic fractions after biodegradation. The increase in SUVA values indicating aromatic structures, except for HPOA fraction, was observed after biodegradation. The protein-like materials in both the HPOA and HPIA fractions and polycarboxylate-type humic acid in the HPIA fraction decreased but the enrichment of HPOA (MW > 100 kDa) and TPIA (MW < 1 kDa) was observed after biodegradation. The production of = C-H in HPIA fraction and the appearance of double peak at 1100 cm-1 in TPIA and HPOB fractions occurred after biodegradation. In overall level, microorganisms effectively utilized DBP precursors from HPIA, HPOA and HPOB fractions but increased the DBPs precursors from the TPIA fraction. TPIA and HPOB fractions had higher DBP yield with chlorine but the DBPs yield of HPIA and HPOA changed little after biodegradation.

Natural biocide disrupts nestmate recognition in honeybees.

Honeybee colonies are under the threat of many stressors, biotic and abiotic factors that strongly affect their survival. Recently, great attention has been directed at chemical pesticides, including their effects at sub-lethal doses on bee behaviour and colony success; whereas the potential side effects of natural biocides largely used in agriculture, such as entomopathogenic fungi, have received only marginal attention. Here, we report the impact of the fungus Beauveria bassiana on honeybee nestmate recognition ability, a crucial feature at the basis of colony integrity. We performed both behavioural assays by recording bee guards' response towards foragers (nestmate or non-nestmate) either exposed to B. bassiana or unexposed presented at the hive entrance, and GC-MS analyses of the cuticular hydrocarbons (CHCs) of fungus-exposed versus unexposed bees. Our results demonstrated that exposed bees have altered cuticular hydrocarbons and are more easily accepted into foreign colonies than controls. Since CHCs are the main recognition cues in social insects, changes in their composition appear to affect nestmate recognition ability at the colony level. The acceptance of chemically unrecognizable fungus-exposed foragers could therefore favour forager drift and disease spread across colonies.

Removal of Chloroxylenol Disinfectant by an Activated Sludge Microbial Community.

Chloroxylenol (CHL) is an antimicrobial ingredient that is frequently used in antiseptics/disinfectants for skin (e.g. hand soap) and non-living surfaces. CHL is an alternative to triclosan and triclocarban, the use of which has recently been banned in some countries. Accordingly, the more widespread use of CHL may significantly increase its occurrence and level in aquatic environments in the near future, eventually resulting in potential ecological risks. Wastewater treatment plants (WWTPs) may be a point source of CHL in natural environments due to extensive discharge through urban waste stream disposal. While the satisfactory removal of CHL in WWTPs is critical for maintaining healthy aquatic ecosystems, the extent of CHL removal and whether CHL causes system upset/failure in WWTPs currently remain unknown. In the present study, we conducted bioreactor operation and batch experiments to investigate the fate and effects of CHL and elucidate the mechanisms underlying degradation at various levels from environmentally relevant to high levels (0.5-5 mg L-1). Bioreactors partially removed CHL (44-87%) via a largely biological route. Microbial association networks constructed using 16S rRNA gene sequencing data revealed selective enrichment and a correlation between Sphingobium and CHL, implying its involvement in the biological breakdown of CHL through dehalogenation and ring hydroxylation pathways. The present results provide insights into the behavior and effects of CHL in activated sludge communities and important information for the sustainable management of CHL that may be an emerging issue in the urban water cycle.

Bioaccumulation and risks of 24 personal care products in plasma of wild fish from the Yangtze River, China.

We used a hybrid precipitation method to simultaneously extract and analyze 24 personal care products (PCPs), including 16 biocides, 4 synthetic musks, and 4 benzotriazoles, in the plasma of fish. The method's performance was validated for plasma samples with and without ß-glucuronidase/aryl-sulfatase hydrolysis. The recoveries were in the range of 70-120% for most of the PCPs, except N,N-diethyl-3-methylbenzamide (DEET), clotrimazole (CTZ), miconazole and itraconazole at spiking concentration of 20 and 5 ng/mL. The quantification limits ranged between 0.89 and 17.9 ng/mL (hydrolyzed plasma) and 0.85-18.5 ng/mL (non-hydrolyzed plasma), except CTZ at 77.5 ng/mL and 76.3 ng/mL. Totally, 13 PCPs were detected in plasma samples of fish collected from the Yangtze River, with a maximum concentration of 58.4 ng/mL (galaxolide). Compounds with the phenol hydroxyl groups of parabens or triclosan in hydrolyzed plasma showed higher concentrations than those in unhydrolyzed plasma with the ratio of conjugation (glucuronides + sulfates) forms up to 86%. The median values for the logarithm of bioaccumulation factors were between 1.39 and 4.15, which were 2-3 orders of magnitude higher than the theoretical logarithm of bioconcentration factors. Using the fish plasma model, the effect ratios (effect concentration/measured plasma concentration ratios) of tonalide, galaxolide, benzotriazole, triclosan, and DEET reached 0.35, 4.15, 3.78, 7.52, and 9.24, respectively. These are recognized as priority chemicals for further risk assessment.

Disinfection Byproducts Bind Human Estrogen Receptor-α.

Disinfection byproducts are formed during most drinking water treatment and presently number >800, some of which are implicated in human health outcomes including bladder cancer and infertility, with unknown mechanisms of action. In particular, it is not yet understood whether these compounds can disrupt the estrogen-signaling pathway through binding to the human estrogen receptor (ER). In the present study, 21 disinfection byproducts, selected for their predicted involvement in endocrine-related diseases and their structural diversity, were individually evaluated for their binding affinity to the human ER and in silico, and then a subset of these chemicals was studied in binary mixtures with the known weak estrogen, 4-n-nonylphenol. Individually, 9 of the 21 disinfection byproducts were able to weakly bind to the ER, with affinities ranging from log median inhibitory concentration values of -3.83 to -2.19 M. In binary mixtures, the chemicals followed concentration addition, with their weak binding affinities having little contribution to the overall mixture affinity. These results demonstrate the variety of small-molecule disinfection byproduct structures that are capable of binding to the ER, and that their weak binding can still be of importance when overall human exposure to mixtures of disinfection byproducts in disinfected drinking water is considered. Environ Toxicol Chem 2019;9999:1-9. © 2019 SETAC.

Bioavailability of soluble microbial products as the autochthonous precursors of disinfection by-products in aerobic and anoxic surface water.

Soluble microbial products (SMPs), as a major part of the effluent organic matter discharged into surface water, may affect the formation of disinfection by-products (DBP) in downstream drinking water treatment plants. In this study, excitation emission matrix fluorescence with parallel factor analysis (EEM-PARAFAC), infrared spectroscopy (IR), high performance size-exclusion chromatography (HPSEC) and 16SrRNA high-throughput sequencing were used to investigate the aerobic and anoxic bioavailability of SMPs in surface water and evaluate their influences on DBP formation upon chlorination in a subsequent drinking water plant. In this study, SMPs were utilized by enriched microbial communities such as Bacteroidetes and Proteobacteria, but the accumulation of SUVA was pronounced during the two oxygen conditions. Biodegraded SMPs had higher humic substructures and lower protein-like components. Due to the presence of SMPs, microbial community compositions were influenced during biodegradation. Moreover, DO was the main factor in biodegradation of SMPs, thus affecting a series of processes, such as microbial compositions, properties of SMPs, DBP formation and reactivity. DBP formation potential decreased after anoxic and aerobic incubations. However, SMPs after aerobic degradation had higher DBP reactivity meanwhile the opposite was found for anoxic incubation. Based on the analysis of IR and HPSEC, it was found that some new substrates or intermediates with MW (220 KDa, <1 KDa) during microbial incubation may contribute to the formation of trihalomethane (THMs), chloral hydrate (CH), dichloroacetonitrile (DCAN) and trichloronitromethane (TCNM) in each DBP sampling episode.