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.

Microplastics (MPs) in the marine environment are ubiquitous. The ingestion of these pollutants by marine organisms has drawn global attention. This work studies the distribution pattern and characteristics of MPs found in the body of the clam Donax cuneatus and its environment in order to understand the possible relationship between the MP concentration in the environment (water and sediment) and that in the clam’s body. Samples of D. cuneatus were collected from the coast between Vembar and Periyathazhai in Tuticorin district along GoM. MP concentrations range from 0.6 to 1.3 items/g (wet weight) in clams, 10–30 items/l in water, and 24–235 items/kg in sediment. Small-sized clams contain the highest concentration of MPs. Hence it is hypothesised that allometric relationship exists between body size and MP concentration, depending on the surface-area to volume ratio. MP abundance in clam body has a clear, positive, significant correlation with MP abundance in sediment but not with abundance of MP in water. Microplastics of fiber type with size 100–250 μm have a predominant presence in clams. The study identified ten types of polymers, of which polyethylene is the most common polymer in all sample types. FTIR-ATR spectra and surface morphology indicate that most of the microplastics have been strongly weathered. Energy dispersive X-ray spectroscopy analysis detected heavy metals associated with MPs like Cd, Pb, Cu, Zn, Ni and Fe. Filter-feeding clams like Donax sp. can provide valuable information on the spatial patterns of MP distribution, and so can act as bio-indicators in monitoring MP pollution in coastal areas.

Kriging interpolation and land use regression (LUR) have characterized the spatial variability of long-term nitrogen dioxide (NO₂), but there has been little research on combining these two methods to capture small-scale spatial variation. Furthermore, studies predicting NO₂ exposure are almost exclusively based on traffic-related variables, which may not be transferable to Taiwan, a typical Asian country with diverse local emission sources, where densely distributed temples and restaurants may be important for NO₂ levels. To advance the exposure estimates in Taiwan, a hybrid kriging/LUR model incorporates culture-specific sources as potential predictors. Based on 14-year NO₂ observations from 73 monitoring stations across Taiwan, a set of interpolated NO₂ values were generated through a leave-one-out ordinary kriging algorithm, and this was included as an explanatory variable in the stepwise LUR procedures. Kriging interpolated NO₂ and culture-specific predictors were entered in the final models, which captured 90% and 87% of NO₂ variation in annual and monthly resolution, respectively. Results from 10-fold cross-validation and external data verification demonstrate robust performance of the developed models. This study demonstrates the value of incorporating the kriging-interpolated estimates and culture-specific emission sources into the traditional LUR model structure for predicting NO₂, which can be particularly useful for Asian countries.

Much attention is currently paid to microplastic (MP) pollution, particularly in marine systems. There is increasing concern regarding the potential toxicity of MPs to organisms at the physiological and morphological levels. However, little is known about the impact of MPs on aquatic life, despite their ubiquitous presence in freshwater ecosystems. In this study, the aquatic plant Utricularia vulgaris was exposed to 1, 2 and 5 μm polystyrene fluorescent MP particles at concentrations of 15, 70 and 140 mg/L for 7 days. The toxic effects of MPs on the growth rate and morphological and physiological characteristics of U. vulgaris were assessed. The results showed that the relative growth rates and the functional traits of leaves (morphological and photosynthetic) were significantly inhibited at a high concentration of MP particles (140 mg/L) when compared to the control group. The impacts on growth performance were likely due to bioaccumulation of MPs in the bladders, as shown by confocal microscopy. Furthermore, the antioxidative enzyme activities showed that high concentrations of MPs induce high ecotoxicity and oxidative damage to U. vulgaris. Thus, U. vulgaris has the potential to be an excellent bioindicator of MP pollution in freshwater ecosystems and should further be applied in ecological risk assessments of the effects of MPs on higher aquatic plants.

The large accumulation of heavy metals in the soil surrounding steel factories has become a severe environmental problem. However, few studies have focused on how the earthworm gut microbiota responds to heavy metals in the soil. This study used research sites at a steel factory in Nanjing, China, to investigate how the soil bacterial community and earthworm gut microbiota respond differently to heavy metal contamination using Illumina high-throughput sequencing targeting 16S rRNA genes. The bacterial community of earthworm guts showed a distinct structure compared with that of the soil, featuring a higher relative abundance of Proteobacteria (45.7%) and Bacteroidetes (18.8%). The bacterial community in the earthworm gut appeared more susceptible to heavy metal contamination compared with the soil community. For example, we identified 38 OTUs (Operational taxonomic units) significantly influenced by contamination among 186 abundant OTUs in the soil, whereas 63 out of the 127 abundant OTUs in the earthworm gut were altered significantly under contamination. This susceptibility may be partly explained by the lower alpha diversity and distinct microbial interactions in the gut. In addition, the accumulation of heavy metals also stimulated the growth of potential plant growth promoting bacteria (PGPB) in the earthworm gut, especially those related to indole-3-acetic acid (IAA) and 1-aminocyclopropane-1-carboxylic acid deaminase (ACCD) production, which may potentially benefit the phyto-remediation of heavy metals. These results contribute to our understanding of the soil biota and its interactions under heavy metal contamination and may provide further insights into the phyto-remediation of metal-contaminated soil.

Large benthic foraminifers (LBFs) are significant contributors to coral island formation in the Pacific Ocean. In recent years, the population of LBFs has decreased because of the increase in anthropogenic influences, such as wastewater (WW) discharge. To implement efficient mitigation measures, pollution tolerance in LBFs should be understood. However, the effects of WW on LBFs and their symbionts have not yet been demonstrated. This study examined the changes in the photosynthetic efficiency (Y[II]) of Calcarina gaudichaudii and Baculogypsina sphaerulata in response to WW by using a pulse-amplitude-modulation fluorometer. These LBFs were exposed to WW with different dilution levels for 22 days. The Y(II) values of the LBFs were found to deteriorate within 1–2 days. However, the Y(II) values both deteriorated and were enhanced in the experiments, thus indicating that WW contains both harmful and beneficial components. Baculogypsina sphaerulata showed an earlier response and greater sensitivity to WW and a higher epibiont infestation than C. gaudichaudii. This result can be attributed to the differences in the physiological and morphological responses of distinct LBFs. A sequencing analysis of 18S rDNA confirmed that the dominant eukaryotic symbionts in the two LBFs studied were Ochrophyta and Labyrinthulomycetes. These eukaryotic symbionts were released and attached as epibionts onto LBFs that were exposed to WW, thus leading to an increase in inactive LBFs. The Shannon–Weaver and Simpson diversity indices revealed that eukaryotic symbiont communities decreased in biodiversity after exposure to WW because of the abundance of algal symbionts. On the basis of these results, we conclude that WW, even with 10,000 × dilution, causes a decrease in active LBF populations owing to the release of eukaryotic symbionts, the decrease in biodiversity, and the infestation of epibionts even though Y(II) is temporarily enhanced. These responses are more significant in B. sphaerulata than in C. gaudichaudii.

Recently, sulfate radical-based advanced oxidation processes (SR-AOPs) have been studied extensively for the removal of pollutants, however, few researches focused on the activation of bisulfite by nanoscale zerovalent iron (nZVI), especially, surface reaction mechanism and sulfate radical-mediated degradation pathway have not been elucidated in detail. In this study, influencing factors, the kinetics, transformation pathway and mechanism of triphenyl phosphate (TPHP) degradation in the nZVI/bisulfite system were systematically discussed. Compared with Fe²⁺, nZVI was found to be a more efficient and long-lasting activator of bisulfite via gradual generation of iron ions. The optimal degradation efficiency of TPHP (98.2%) and pseudo-first-order kinetics rate constant (kₒbₛ = 0.2784 min⁻¹) were obtained by using 0.5 mM nZVI and 2.0 mM bisulfite at the initial pH 3.0. Both Cl⁻ and NO₃⁻ inhibited the degradation of TPHP and the inhibitory effect of Cl⁻ was stronger than that of NO₃⁻ due to the higher reaction rate of Cl⁻ with •SO₄⁻. Furthermore, SEM, XRD and XPS characterization revealed that a thin passivation layer (Fe₂O₃, Fe₃O₄, FeOOH) deposited on the surface of fresh nZVI and a few iron corrosion products generated and assembled on the surface of reacted nZVI. Radical quenching tests identified that •SO₄⁻ was the dominant reactive oxidative species (ROS) for TPHP removal. Based on HRMS analysis, six degradation products were determined and a sulfate radical-mediated degradation pathway was proposed. In a word, this study revealed that the nZVI/bisulfite system had a great potential for the TPHP elimination in waterbody.

Pharmaceuticals and personal care products (PPCPs) are a class of emerging contaminants commonly detected in environmental waters worldwide. Although reports about their detection in aquatic environments are increasing, limited studies show their effects on holometabolous insects. In this study, acute and chronic exposure to naproxen (0.02, 41, 82, 164, 382, 656, and 1312 mg L⁻¹) and propylparaben (0.02, 25, 50, 100, 250, 500, and 1000 mg L⁻¹) were evaluated in Aedes aegypti L. Acute exposure to naproxen (≥0.02 mg L⁻¹) and propylparaben (≥0.02 mg L⁻¹) reduced egg eclosion. Propylparaben (≥250 mg L⁻¹) caused significant larval mortality but naproxen did not even at the highest experimental concentration used. LC₅₀ for naproxen and propylparaben in larvae were 1100 mg L⁻¹ and 182.6 mg L⁻¹, respectively. Naproxen (≥0.02 mg L⁻¹) and propylparaben (≥0.02 mg L⁻¹) reduced pupation. Emergence was also reduced by naproxen (≥164 mg L⁻¹) and propylparaben (≥0.02 mg L⁻¹). The fecundity of females was significantly reduced due to chronic exposure to naproxen (≥0.02 mg L⁻¹). There was also a reduction in the fecundity of females due to chronic propylparaben exposure but it was statistically insignificant in the concentrations used. In the F1 generation eggs, only 100 mg L⁻¹ propylparaben reduced eclosion. Eclosion and larval survival were sensitive to acute exposure, particularly to propylparaben. The reduced pupation and emergence indicated a delay in the progression of the life cycle. Chronic exposure also indicated a reduction in fecundity. F1 eggs exhibited tolerance to the negative effect of subsequent exposure. Our findings suggest that propylparaben can affect Ae. aegypti more negatively than naproxen.

Human interferences have caused groundwater contamination in alluvial aquifers which subsequently affects the health of exposed population. In the present study, 74 groundwater samples from the semi-arid region of Panipat district, falling under Yamuna sub-basin, India was evaluated to know the potential non-carcinogenic human health risk in local adult and child population. The major objective of the present study was to know the non-carcinogenic human health risk due to intake of fluoride and nitrate contaminated water, using two different approaches: deterministic and probabilistic (Monte Carlo simulation). The values of hazard quotient (HQ) determined by deterministic as well as probabilistic approach were nearly identical. The hazard index (HI) value of 40.8% samples was above the unity in case of adults while 69.7% samples indicated HI value greater than unity for children thus indicating children are more prone to non-carcinogenic health risk than the adult population. Sensitivity analysis was performed to identify the influence of the non-carcinogenic human health risk predictor variables for the prediction of risk and concentration factor (CF) was the most influential variable. Multivariate statistical techniques were employed to know the positive and negative relationship of fluoride and nitrate with other parameters. Results of principal component analysis/factor analysis (PCA/FA) indicated that the concentration of fluoride is controlled by the presence of calcium due to their negative correlation in groundwater samples. The hierarchical agglomerative cluster analysis (HCA) also supported the outcome of PCA/FA and both indicated anthropogenic sources of fluoride and nitrate in groundwater.

Butylated hydroxytoluene (BHT) is one of the most frequently used synthetic phenolic antioxidants added to food and consumer products such as plastics as a preservative. Due to its high production volume, BHT has been detected in aquatic environments, raising concerns about sub-lethal toxicity. However, there are limited toxicological data for BHT, especially in fish. In this study, zebrafish embryos were exposed to BHT at concentrations ranging 0.01–100 μM for up to 6 days post fertilization (dpf). Acute toxicity was assessed, and experiments revealed that BHT had a 96 h LC50 value of 57.61 μM. At sub-lethal doses (0.1–60 μM), BHT markedly decreased heart rates of zebrafish embryos at 48 h and 72 h by ∼25–30%. Basal and maximal respiration of zebrafish embryos at 24 hpf were decreased by 59.3% and 41.4% respectively following exposure to 100 μM BHT. Behavior in zebrafish was measured at 6 dpf following exposures to 0.01–10 μM BHT. Locomotor behaviors (e.g. total distance moved and velocity) were significantly increased in larvae at doses higher than 0.1 μM BHT. In addition, dark-avoidance behavior was decreased following exposure to 0.01 μM BHT, while conversely, it was increased in zebrafish exposed to 0.1 μM BHT. To investigate potential underlying mechanisms that could explain behavioral changes, transcripts involved in dopamine signaling were measured. Relative expression of dat mRNA was increased in larval fish from the 0.01 μM BHT treatment, while there were no effects on dat mRNA levels at higher concentrations. The mRNA levels of drd3 were decreased in zebrafish from the 1 μM BHT treatment. Taken together, BHT can affect the expression of the dopamine system, which is hypothesized to be related to the abnormal anxiety-associated behavior of larval zebrafish.