Unveiling the hidden risks: Pesticide residues in aquaculture systems.

The present study systematically assessed the presence and ecological risks of 79 pesticides in various aquaculture systems, namely pond aquaculture (PA), greenhouse aquaculture (GA), and raceway aquaculture (RA) at different aquaculture stages, along with evaluating the pesticide removal of four tailwater treatment systems. Sixteen herbicides and two fungicides were identified, with the total concentrations ranging from 8.33 ng/L to 3248.45 ng/L. The PA system demonstrated significantly higher concentrations (p < 0.05) and a wider range of pesticide residues compared to the GA and RA systems. Prometryn, simetryn, atrazine, and thifluzamide were found to be the predominant pesticides across all three aquaculture modes, suggesting their significance as pollutants that warrant monitoring. Additionally, the findings indicated that the early aquaculture stage exhibits the highest levels of pesticide concentration, underscoring the importance of heightened monitoring and regulatory interventions during this phase. Furthermore, among the four tailwater treatment systems analyzed, the recirculating tailwater treatment system exhibited the highest efficacy in pesticide removal. A comprehensive risk assessment revealed minimal ecological risks in both the aquaculture and tailwater environments. However, the pesticide mixtures present high risks to algae and low to medium risks to aquatic invertebrates and fish, particularly during the early stages of aquaculture. Simetryn and prometryn were identified as high-risk pesticides. Based on the prioritization index, simetryn, prometryn, diuron, and ametryn are recommended for prioritization in risk assessment. This study offers valuable data for pesticide control and serves as a reference for the establishment of a standardized pesticide monitoring and management system at various stages of aquaculture.

Isolation of Capillaries from Small Amounts of Mouse Brain Tissue.

The integrity of the blood-brain barrier (BBB) is essential for the normal functioning of the central nervous system (CNS). Isolated brain capillaries are essential for analyzing changes in protein and gene expression at the BBB under physiological and pathological conditions. The standard methods for isolating brain capillaries require the use of at least one or more mouse brains in order to obtain sufficient quantity and purity of brain capillaries. Here, we describe an optimized protocol for isolating and purifying capillaries from tiny amounts of mouse cerebral cortex using manual homogenization, density gradient centrifugation, and filtration while preserving the structural integrity and functional activity of microvessel fragments. Western blotting showed that proteins expressed at the BBB were enriched in mouse brain capillaries isolated by the optimized method compared to cerebral cortex protein homogenates. This approach can be used for the analysis of a variety of rare mouse genetic models and can also help the investigators to understand regional differences in susceptibility to pathological phenomena such as ischemia and traumatic brain injury. This will allow the investigators to better understand the physiology and pathology of the BBB.

Comparative antioxidant and metabolomic analysis for the identification of differential response of mussel (Mytilus coruscus) to four succinate dehydrogenase inhibitor fungicides.

Succinate dehydrogenase inhibitor fungicides (SDHIs) are frequently detected in the marine environment. However, studies on the toxicity of SDHIs to marine organisms, Mytilus coruscus (M. coruscus), are poorly reported. Therefore, the antioxidant activities and metabolomic response of four SDHIs, namely, boscalid (BC), thifluzamide (TF), fluopyram (FO), and bixafen (BIX), to (M. coruscus), were comprehensively investigated. The antioxidant activity of BC and TF was significantly increased (p<0.05), whereas those of FO and BIX were significantly decreased. Furthermore, metabolite discriminations among M. coruscus to four SDHIs were illustrated by an untargeted metabolomics approach. A total of 52, 50, 93, and 129 differential metabolites were obtained for BC, TF, FO, and BIX. KEGG of the different metabolites show that the four SDHIs had differential effects on the metabolic pathways of M. coruscus. The current study demonstrated four SDHIs triggered glucose metabolism, lipid metabolism, tricarboxylic acid cycle, and oxidative phosphorylation processes and caused the disruption of nutrient and energy conversion processes in mussels. Finally, five biomarkers were screened by analyzing common differential metabolites that emerged from the four SDHI exposures, which could be used for risk assessment of marine ecosystem exposure to SDHIs. Our results demonstrated the use of metabolomics to understand the potential mechanisms of toxicity of four SDHIs to mussels and to identify potential targets for future targeted risk assessment.

Accumulation and elimination properties and comparative toxicity of fluxapyroxad in juvenile and adult large yellow croaker (Larimichthys crocea).

Fluxapyroxad (FX), a succinate dehydrogenase inhibitor fungicide, has been detected in global marine and aquatic organisms. However, as a new pollutant, its biotoxicity and ecological risks to marine aquatic organisms are unclear. The accumulation and elimination processes and toxic effects of FX on Larimichthys crocea (L. crocea) at environmental concentrations were assessed. FX (1.0 µg/L) was rapidly enriched and persisted prolonged in L. crocea muscle and FX is highly toxic to juvenile L. crocea with the 96 h LC50 of 245.0 µg/L. Furthermore, the toxic effects of FX on juvenile L. crocea and adults L. crocea were compared and analyzed. In contrast to those of adult L. crocea, juvenile L. crocea showed a stronger oxidative stress response and rescued liver damage in terms of antioxidant enzyme activity, energy supply, and liver damage to FX. Transcriptomic analysis also showed that drug metabolism was activated. In the adult L. crocea, the disturbance of the energy metabolism, oxidative respiration, TCA cycle, and lipid metabolism genes were firstly found. The results revealed the accumulation and elimination pattern and ecotoxicological hazards of FX to L. crocea, which provided important theoretical basis for the study of environmental risks caused by new pollutants to marine organisms.

High-throughput analytical methodology of monoalkyl phthalate esters and the composite risk assessment with their parent phthalate esters in aquatic organisms and seawater.

A sensitive, robust, and highly efficient analytical methodology involving solid phase extraction coupled to ultra-high performance liquid chromatography tandem mass spectrometry was successfully established to detect 13 monoalkyl phthalate esters (MPAEs) in aquatic organisms and seawater. After the organisms were preprocessed using enzymatic deconjugation with ß-glucuronidase, extraction, purification, and qualitative and quantitative optimization procedures were performed. Under optimal conditions, the limits of detection varied from 0.07 to 0.88 µg/kg (wet weight) and 0.04-1.96 ng/L in organisms and seawater, respectively. Collectively, MPAEs achieved acceptable recovery values (91.0-102.7%) with relative standard deviations less than 10.4% and matrix effects ranging from 0.93 to 1.07 in the above matrix. Furthermore, MPAEs and phthalate esters were detected by the developed methodology and gas chromatography-triple quadrupole tandem mass spectrometer in practical samples, respectively. Mono-n-butyl phthalate and mono-iso-butyl phthalate were the most predominant congeners, accounting for 24.8-35.2% in aquatic organisms and seawater. Comprehensive health and ecological risks were higher after the MPAEs were incorporated than when phthalate esters were considered separately, and greater than their risk threshold. Therefore, the risks caused by substances and their metabolites in multiple media, with analogous structure-activity relationships, should be considered to ensure the safety of aquatic organisms and consumers.

Horizontal and Vertical Transmission of a Mycovirus Closely Related to the Partitivirus RhsV717 That Confers Hypovirulence in Rhizoctonia solani.

Rhizoctonia solani virus717 (RhsV717) was isolated from the Rhizoctonia solani (R. solani) AG-2 strain Rhs717. This study isolated a virus designated as Rhizoctonia solani partitivirus BS-5 (RsPV-BS5) from the R. solani AG-3 strain BS-5, the causal agent of tobacco target spot disease. The virus was identified as a strain of RhsV717. Transmission electron microscopy (TEM) images showed that RsPV-BS5 had virus particles with a diameter of approximately 40 nm. Importantly, it can be horizontally transmitted through hyphal anastomosis and vertically transmitted via sexual basidiospores. Furthermore, this study demonstrated that RsPV-BS5 infection significantly impedes mycelial growth and induces hypovirulence in tobacco leaves. Thus, RsPV-BS5 presents a promising avenue for biocontrolling tobacco target spot disease. Transcriptome analysis unveiled differential expression of four genes related to cell wall-degrading enzymes between two isogenic strains, 06-2-15V and 06-2-15. These findings shed light on the molecular mechanism through which RsPV-BS5 reduces host pathogenicity.

Environmental profile, potential sources, and ecological risk of polycyclic aromatic hydrocarbons in a typical coastal bay and outer bay area.

As a class of persistent organic pollutants, polycyclic aromatic hydrocarbons (PAHs) are widely present and accumulate in multimedia environments. The pollution characteristics, spatiotemporal distribution, potential sources, influencing factors, and ecological risks of 16 PAHs were investigated in the water-sediment system of the Hangzhou Bay and outer bay area (HZB and OBA, respectively). The total concentrations of 16 PAHs (∑PAHs) were 220 ± 97.0 and 130 ± 36.0 ng/L in the seawater and 343 ± 179 and 505 ± 415 µg/kg (dry weight) in the sediments of the HZB and OBA, respectively. The pollution level of PAHs in the HZB seawater was higher than that in the OBA seawater, but the opposite result was found in the sediments. Moreover, ∑PAHs exhibited high temporal variability in the HZB seawater (rainy season > dry season), whereas ∑PAHs in the sediments showed no significant difference between seasons. The molecular diagnostic ratio method was used to identify pollution sources and showed that the PAHs in seawater came from different pollution sources (fuel combustion and petroleum), whereas the PAHs in the sediments originated from coincident sources (mixed combustion). Correlation analysis revealed that temperature was positively related to ∑PAHs, whereas salinity was negatively related to seawater ∑PAHs values. Ecological risk assessment demonstrated that the potential for adverse ecological effects was low to moderate in seawater but moderate to high in the sediments.

Simultaneous determination and dietary intake risk assessment of 60 herbicide residues in aquatic products.

A sensitive and rugged analytical method was first established to simultaneously determine 60 herbicides in aquatic products with gas chromatography-tandem mass spectroscopy (GC-MS/MS). After extraction with acetonitrile (MeCN), NaCl and anhydrous Na2SO4 were added, concentrated supernatants were directly passed through the Carb/NH2 solid phase extraction column. Then, the cartridge was rinsed with elution solution (MeCN/toluene, 3:1, v/v), followed by GC-MS/MS analysis with multiple reaction monitoring. An excellent linearity (1.0-100.0 µg/L) with R2 value of ≥0.9991 was obtained, and the limits of quantification were 0.018-3.852 µg/kg. Satisfactory recoveries (70.8 %-117.6 %) with RSDs below 11.0 % of herbicide residues were obtained at spiked levels of 0.010-0.050 mg/kg. Furthermore, herbicide residues in actual aquatic products were analyzed, and the acute/chronic risk assessment of dietary exposure was carried out. The wide use of herbicides for controlling weed and removing moss and harmful algae may obviously increase the risk of contamination of the aquaculture environment and fishery products. Therefore, considerable attention and more research are necessary to monitor residue levels for herbicides in aquatic products and ensure the quality of marine products and consumer safety.

Progress and opportunities for metal-organic framework composites in electrochemical sensors.

Metal-organic framework composites have the advantages of large surface area, high porosity, strong catalytic efficiency and good stability, which provide a great possibility of finding excellent electrode materials for electrochemical sensors. However, MOF composites still face various challenges and difficulties, which limit their development and application. This paper reviews the application of MOF composites in electrochemical sensors, including MOF/carbon composites, MOF/metal nanoparticle composites, MOF/metal oxide composites and MOF/enzyme composites. In addition, the application challenges of MOF composites in electrochemical sensors are summarized. Finally, the application prospect for MOF composites is considered to promote the synthesis of more MOF composites with excellent properties.

Complete genome sequence of a novel fusarivirus from Rhizoctonia solani AG-3 PT strain 3P-2-2.

Here, we describe a novel mycovirus, tentatively designated as "Rhizoctonia solani fusarivirus 6" (RsFV6), which was discovered in Rhizoctonia solani AG-3 PT strain 3P-2-2. The virus has a single-stranded positive-sense RNA (+ssRNA) genome of 6141 nucleotides containing two open reading frames (ORFs) and a poly(A) tail. ORF1 encodes a large polypeptide of 1,862 amino acids (aa) with conserved RNA-dependent RNA polymerase (RdRp) and helicase (Hel) domains. ORF2 encodes a putative 167-aa protein of unknown function. BLASTp searches revealed that the ORF1-encoded polypeptide showed the highest sequence similarity (70.67% identity) to that of Rhizoctonia solani fusarivirus 3 (RsFV3), which was isolated from Rhizoctonia solani AG-2-2LP. Multiple sequence alignments and phylogenetic analysis based on RdRp and Hel sequences indicated that RsFV6 could be a novel member of the genus Alphafusarivirus family Fusariviridae.

Transcriptome sequencing and metabolite analysis reveal the single and combined effects of microplastics and di-(2-ethylhexyl) phthalate on Peneaus vannamei.

Due to the rising usage of plastics, plastic debris are present throughout marine ecosystems and detrimentally affects marine biota. Additionally, plastics likely result in elusive toxicity effects due to addition of plasticizers. The aim of the present study was to reveal the potential effects and mechanism of microplastics (MPs), di-(2-ethylhexyl) phthalate (DEHP) and copollution of MPs and DEHP (MPs-DEHP) on Peneaus vannamei (P. vannamei) juveniles regarding oxidative stress, transcriptomics and metabolomics. MPs, DEHP and MPs-DEHP significantly induced the activities of superoxide dismutase (SOD) and catalase (CAT); MPs and DEHP have an antagonistic effect for malondialdehyde (MDA); suggesting that disorders of the antioxidant defence systems. 13, 133 and 58 differentially expressed genes and 21, 82 and 39 differentially expressed metabolites were responsible for the distinction of MPs, DEHP and MPs-DEHP groups, respectively. The combination of transcriptomic and metabolomic analyses showed that MPs, DEHP and MPs-DEHP exposure disturbed amino acid and lipid metabolism, and further induced inflammatory responses and dysfunction of purine metabolism. Furthermore, the presence of MPs might alleviate the biotoxicity of DEHP in P. vannamei. These findings provide new insights into the single and combined toxicological effects of MPs and additives for marine biota.

Functional differentiation of bmp2a and bmp2b genes in zebrafish.

Bone morphogenetic protein 2 plays an important role in the regulation of osteoblast proliferation and differentiation. Phylogenetic analysis showed that the bmp2 ortholog evolved from the same ancestral gene family in vertebrates and was duplicated in teleost, which were named bmp2a and bmp2b. The results of whole-mount in situ hybridization showed that the expression locations of bmp2a and bmp2b in zebrafish were different in different periods (24 hpf, 48 hpf, 72 hpf), which revealed potential functional differentiation between bmp2a and bmp2b. Phenotypic analysis showed that bmp2a mutations caused partial rib and vertebral deformities in zebrafish, while bmp2b-/- embryos died massively after 12 hpf due to abnormal somite formation. We further explored the expression pattern changes of genes (bmp2a, bmp2b, smad1, fgf4, runx2b, alp) related to skeletal development at different developmental stages (20 dpf, 60 dpf, 90 dpf) in wild-type and bmp2a-/- zebrafish. The results showed that the expression of runx2b in bmp2a-/- was significantly downregulated at three stages and the expression of other genes were significantly downregulated at 90 dpf compared with wild-type zebrafish. The study revealed functional differentiation of bmp2a and bmp2b in zebrafish embryonic and skeletal development.

Molecular characterization of a novel mitovirus from Rhizoctonia solani AG-4 HGIII strain XMC-IF.

The nucleotide sequence of a viral double-stranded RNA (dsRNA) from Rhizoctonia solani AG-4 HGIII strain XMC-IF (designated as "Rhizoctonia solani mitovirus 106", RsMV-106) was determined. The complete sequence was 2794 bp in length with a 57.50% A + U content and contained a large open reading frame (ORF) when the fungal mitochondrial genetic code was used. The ORF potentially encodes a 95.76-kDa protein containing a conserved domain of an RNA-dependent RNA polymerase (RdRp). BLASTp analysis revealed that the RdRp domain of RsMV-106 shared 47.52-73.24% sequence identity with those of viruses of the genus Duamitovirus and was most similar (73.24% identity) to that of Alternaria alternata mitovirus 1 (AaMV1). Phylogenetic analysis showed that RsMV-106 is a novel member of the genus Duamitovirus, family Mitoviridae. This is the first report of the full genome sequence of a mitovirus associated with R. solani AG-4 HGIII.

Development and application of tricolor ratiometric fluorescence sensor based on molecularly imprinted nanoparticles for visual detection of dibutyl phthalate in seawater and fish samples.

A tricolor ratiometric fluorescence sensor was fabricated by mixing blue- and red-emission molecularly imprinted quantum dots (MIP-QDs) with green-emission quantum dots at the optimal ratio. The MIP-QDs were synthesized by coating CdSe/ZnS QDs in polymer by inverse microemulsion method. Compared with single-emission or dual-emission sensors, the tricolor ratiometric fluorescence sensor provided a wider range of color variations for visual DBP detection. The ratio fluorescence value I530/(I450 + I630) of the tricolor ratiometric fluorescence sensor linearly changed within the concentration of 2.0-20.0 × 103 µg/L DBP. The correlation coefficient was 0.9910, and the limits of detection were 1.0 µg/kg and 0.65 µg/L in fish and seawater, respectively. Meanwhile, the fluorescence color gradually changed from purple to plum to pink to salmon to yellowish green and finally to green. The recoveries of DBP in fish and seawater were 84.3 %-91.4 % and 88.3 %-110.3 %, respectively. Moreover, no obvious differences were observed between the detection results of the tricolor ratiometric fluorescence sensor and gas chromatography-tandem mass spectrometry. The tricolor ratiometric fluorescence sensor constructed herein provides an ideal choice for rapid and intuitive DBP detection in environmental and aquatic products.

Organophosphate esters in the mariculture ecosystem: Environmental occurrence and risk assessments.

Most investigations on organophosphate esters (OPEs) are conducted predominantly in a separate biological or abiotic medium, and few joint analyses have been performed in the mariculture ecosystem based on yearly sampling. Herein, we investigated the occurrence, load estimation, phase distribution, source diagnostics, and risks of 20 OPEs in seawater, sediment, and aquaculture organisms from a typical mariculture area in China. The total of these OPEs (∑OPEs) ranged within 3.97-1068 ng/L, 0.39-65.5 ng/g (dw), and 4.09-16.3 ng/g (ww) in seawater, sediment and organisms, respectively. Chlorinated OPEs were the predominant congeners detected in seawater, whereas alkyl-OPEs were the leading contributors in sediment and biological samples. Seasonal variations of ∑OPEs in seawater were more distinct than those in sedimentary environments. Load estimation indicated that approximately 70% of the OPEs in the study area existed in the water bodies. Source identification performed using the U.S. EPA positive matrix factorization indicated that polyurethane foam/plastics and hydraulic oil made the greatest contributions in seawater, whereas chemical production was the predominant source in sediment. Indices of ecological and health risks of OPEs were lower than their risk threshold, indicating that the OPEs detected in this study posed a low risk to the aquatic environment and human health.

NHC-alcohol adduct-mediated photocatalytic deoxygenation for the synthesis of 6-phenanthridine derivatives.

NHC-alcohol adduct-mediated deoxygenation of alcohols under photocatalytic conditions is described. This process provides various alkyl radicals, which can react with 2-isocyanobiaryls to afford 6-substituted phenanthridine derivatives in moderate to good yields. This method offered the first example of directly using alcohols as radical sources for 6-phenanthridine synthesis.

A green and rapid analytical method for determination of kitasamycin in animal feedstuffs by ultra-high performance liquid chromatography tandem mass spectrometry.

A rapid, simple, highly efficiency analytical method for detecting kitasamycin A1, A4, A5, and A13 in different feedstuffs was successfully developed by combining enhanced matrix removal (EMR) lipid cartridge and ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). After extraction with acetonitrile, the sample supernatants were directly passed through the EMR lipid cartridge. Then, the cartridge was rinsed and eluted with acetonitrile and methanol, respectively, followed by UHPLC-MS/MS analysis with positive mode using multiple reaction monitoring. Optimized pretreatment procedure without solvent conversion, multiple nitrogen drying steps and activated cartridge before loading, and no significant interference were found during the analysis of different types of animal feedstuffs. Excellent sensitivity (Limit of quantification, LOQ) of kitasamycin A1, A4, A5, and A13 was 1.1-2.0 µg/kg. Satisfactory recoveries of kitasamycin A1, A4, A5, and A13 in different feedstuffs were from 74.0% to 98.8%, with the relative standard deviations (RSDs) below 10.4%, and good linear correlation coefficient (r)>0.9990 in the matrix matched standard curve range of 0.02-50.0 µg/L. Results demonstrated that the developed method exhibited excellent linearity, accuracy, precision, sensitivity, and the feasibility of using this method in kitasamycin determination of animal feedstuffs. The method was evaluated using the greenness analysis method through Eco-Scale assessment tool.

Highly selective molecularly imprinted-electrochemiluminescence sensor based on perovskite/Ru(bpy)32+ for simazine detection in aquatic products.

A novel molecularly imprinted electrochemiluminescence (MIECL) sensor based on the luminescence of molecularly imprinted polymer-perovskite (MIP-CsPbBr3) layer and Ru(bpy)32+ was fabricated for simazine detection. MIP-CsPbBr3 layers were immobilized onto the surface of glassy carbon electrode as the capture and signal amplification probe, and Ru(bpy)32+ and co-reactant tripropylamine exhibited stronger electrochemiluminescence (ECL) emission. Under optimal conditions, the ECL signal of the MIECL sensor was linearly quenched, with the logarithm of simazine concentration ranging from 0.1 µg/L to 500.0 µg/L, correlation coefficient of 0.9947, and limit of detection of 0.06 µg/L. The practicality of the developed MIECL sensor method for simazine determination in aquatic samples was validated. Excellent recoveries of 86.5 %-103.9 % with relative standard deviation below 1.6 % were obtained for fish and shrimp samples at three different spiked concentrations. The MIECL sensor exhibited excellent selectivity, sensitivity, reproducibility, accuracy, and precision for simazine determination in actual aquatic samples.

Occurrence, source, and ecological risk assessment of organochlorine pesticides and polychlorinated biphenyls in the water-sediment system of Hangzhou Bay and East China Sea.

The pollution characteristics, potential sources, and potential ecological risk of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) were investigated in the Hangzhou Bay (HZB) and East China Sea (ECS). Total OCPs concentration ranged from 2.62 to 102.07 ng/L and 4.41 to 75.79 µg/kg in the seawater and sediment samples, with PCBs concentration in the range of 0.40-51.75 ng/L and 0.80-45.54 µg/kg, respectively. The OCPs were positively correlated with nutrients, whereas PCBs presented a negative correlation. The newly imported dichlorodiphenyltrichloroethane (DDT) in HZB is mainly the mixing of technical DDT and dicofol sources. The PCB source composition is more likely related to the mixture of Kanechlor 300, 400, Aroclor 1016, 1242, and Aroclor 1248. Risk assessment results indicate that OCPs posed low risk in seawater. The potential risk of DDTs in the sediments is a cause of concern.

Contaminant occurrence, spatiotemporal variation, and ecological risk of organophosphorus flame retardants (OPFRs) in Hangzhou Bay and east China sea ecosystem.

Occurrence, spatiotemporal variation, sources, and ecological risks of 20 organophosphorus flame retardants (OPFRs) in water and sediments from Hangzhou Bay (HZB) and its adjacent East China Sea (ECS) were investigated in this study. The concentrations of OPFRs (∑OPFR) in water ranged from 0.51 ng/L to 885 ng/L, with chlorinated OPFRs having the highest value. For sediments, ∑OPFR ranged from 2.93 ng/g, dry weight (dw) to 37.8 ng/g, dw. The ∑OPFR in the water and sediments of HZB in summer was significantly (p < 0.05) higher than that in autumn. Additionally, the pollution of OPFRs in HZB was higher than that in ECS, and the high-concentration areas appeared in the north and south banks of HZB and near the coast of ECS. Principal component analysis-multiple linear regression showed that the OPFRs in this region were mainly from industrial products (e.g., polyurethane foam/paint/coating/textiles/product processing). In terms of aquatic environments, ecological risks were in a low (∑RQs<0.1) to moderate (0.1<∑RQs<1) level, with regard to median exposure levels, a moderate risk (0.1<∑RQs<1) was found in the sediments during autumn. This study can provide new insights into the OPFR pollution characteristic and ecological risk in a specific eco-environment.