Responses of bacterial and three sub-microeukaryote communities in the water of white shrimp Penaeus vannamei aquaculture ponds in two polyculture models.

Polyculture operations in freshwater aquaculture ponds can disrupt microbial communities. High-throughput sequencing was used to assess the impact of polyculture operations on bacterial and three sub-microeukaryote communities (fungi, zooplankton, and eukaryotic phytoplankton) in Penaeus vannamei aquaculture ponds containing oriental river prawns and giant freshwater prawns, respectively. The results showed that the bacterial community was less sensitive than the microeukaryote communities to both the polyculture activity and environmental variations. The polyculture of giant freshwater prawns rather than oriental river prawns was the primary factor affecting the beta diversity of the three sub-microeukaryote communities. This may be due to the larger biomass of the polyculture varieties of giant freshwater prawns compared with oriental river prawns. The polyculture activity of giant freshwater prawns with a higher density and that of oriental river prawns with a lower density increased the stochasticity of the community assembly of the three sub-microeukaryote communities. It also affected the topological properties of the microbial communities, including greater correlations between ecosystem elements, and reducing the correlations among zooplanktons. The eukaryotic phytoplankton was the only microbial community that could also be explained by nutrient variation (mainly the total nitrogen). This highlights the potential role of the eukaryotic phytoplankton as a suitable indicator of the effects of nutrient input into ecosystems.

Bacterial Community Structure and Diversity in the Aqueous Environment of Shihou Lake and its Relationship with Environmental Factors.

Microorganisms are sensitive to changes in the external environment and are often used as indicators to monitor and reflect water quality. Using Illumina MiSeq sequencing, the characteristics of the microbial community in Shihou Lake water at different time points were analyzed and the key environmental factors affecting the bacterial community were identified. The microbial community diversity in Shihou Lake water was rich and showed significant differences over time. The main bacterial phyla were the Cyanobacteria, Proteobacteria, Actinobacteria, Verrucomicrobia, Bacteroidetes, Chloroflexi, Planctomycetes, Firmicutes, Chlorobi, WS6 and Saccharibacteria. The relative abundance of these major phyla in the sample accounted for 97.83%-99.07% of the total abundance; Cyanobacteria had the highest relative abundance, accounting for 13.07%-44.61% of the total, and the abundance of each dominant phylum was significantly different at different time points. The Shannon and Simpson indexes showed that the diversity of each month was as follows: August > October > July > September > November. The Chao1 and Ace indexes indicated that the order of richness was: November > October > July > August > September. Beta diversity analysis found significant differences in the samples from month to month. Environmental factors such as temperature, total nitrogen, chlorophyll-a, permanganate index, nitrite, pH and ammonia nitrogen had significant effects on microbial community structure.

Hepatic oxidative stress, DNA damage and apoptosis in adult zebrafish following sub-chronic exposure to BDE-47 and BDE-153.

Polybrominated diphenyl ethers (PBDEs) are ubiquitous and prolific contaminant in both the abiotic and biotic environment because of the wide industrial applications of these chemicals. In the present study, the effects of 2,2',4,4'-tetrabrominateddiphenyl ether (BDE-47) and 2,2',4,4',5,5'-hexabromodiphenyl ether (BDE-153) exposure on the induction of hepatic oxidative stress, DNA damage, and the expression of apoptosis-related genes in adult zebrafish were investigated. The activities of antioxidant enzymes, such as catalase and superoxide dimutase, significantly increased when adult zebrafish was exposed to various concentrations of BDE-47 and BDE-153 for 7 and 15 days. BDE-47 and BDE-153 elicited significant alterations in zebrafish 7-Ethoxyresorufin-O-deethylase activity at 3, 7, or 15 days of exposure. In addition, the significant increase in comet assay parameters of zebrafish hepatocytes in a concentration-dependent manner indicated BDE-47 and BDE-153 induced DNA damage, probably due to observed oxidative stress. Furthermore, a monotonically upregulation of p53 and Caspase3, which are apoptotic-regulated genes, and decreased expression ratio of the anti-apoptotic B-cell lymphoma/leukaemia-2 and Bcl2-associated X protein genes for all BDE-47 and BDE-153 treatments at 7 and 15 days indicated apoptosis induction in zebrafish liver. Our findings help elucidate the mechanisms of BDE-47- and BDE-153-induced toxicity in zebrafish hepatocytes.

Gut microbiota analysis of juvenile genetically improved farmed tilapia (Oreochromis niloticus) by dietary supplementation of different resveratrol concentrations.

The genetically improved farmed tilapia (GIFT, Oreochromis niloticus) is cultured widely for production of freshwater fish in China, while streptococcosis, likely related to pathogenic infections, occurs frequently in juvenile, mother, and operated GIFT. The gut microbiota plays an important role in nutrient digestibility in animals, and resveratrol (RES) has been used in feed for different freshwater fish species. Therefore, understanding changes in the tilapia gut microbiota across different concentrations of dietary RES supplementation is extremely important. The gut microbiota population in tilapia at 45 d after supplementation with different concentrations (0, 0.025, 0.05, 0.1 g/kg) of dietary RES was assessed by 16S rDNA gene sequencing. A total of 5445 operational taxonomic units were identified from all samples, and 14 phyla and 81 families were identified from all fecal samples. The bacteria of the phylum Firmicutes were significantly enriched in the 0.025 g/kg RES group when compared with the controls. Proteobacteria, Firmicutes and Cyanobacteria were the most dominant three phyla in all samples. With the increasing concentrations, the proportion of beneficial microbial taxa (Acetobacteraceae and Methylobacteriaceae) increased, whereas the proportion of harmful microbial taxa decreased, eg. Streptococcaceae except for 0.1 g/kg RES groups. RES did not affect the richness and diversity in tilapia gut microbiota. These findings provide information on the diversity and differences in GIFT gut microbiota database, and may contribute to developing strategies for management of diseases and long-term sustainability of O. niloticus culture.

Methanogenic community compositions in surface sediment of freshwater aquaculture ponds and the influencing factors.

Aquaculture ponds represent ecologically relevant environments to study the community composition and diversity of methanogenic assemblages, as well as their interactions with cultivated species and chemical indicators. In this study, aquaculture ponds with crab (Eriocheir sinensis), oriental river prawn (Macrobrachium nipponense), perch (Micropterus salmonides) and Wuchang fish (Parabramis pekinensis) were sampled, and Illumina high-throughput sequencing was used to investigate the methanogenic communities. The results revealed that the abundant methanogenic orders in surface sediment were Methanomicrobiales, Methanosarcinales and Methanocellales. The relative abundance of Methanocellales was higher in crab and prawn ponds as compared to other ponds. Methanogenic 16S rRNA gene abundance and beta diversity of the community was affected by the cultivated species. Methanogenic communities in aquaculture ponds with higher contents of total nitrogen and organic matter had decreased species richness, while those with higher contents of ammonia and nitrite had an overall decreased abundance of methanogens and their respective diversities. Overall, in addition to the differences in cultivated species, the consequent differences in farming practices including the types and amounts of feeds used, the contents of total nitrogen, organic matter, ammonia and nitrite could all influence the methanogenic community in surface sediment of aquaculture ponds.

Hepatic transcriptome analysis of juvenile GIFT tilapia (Oreochromis niloticus), fed diets supplemented with different concentrations of resveratrol.

The GIFT (Genetically Improved Farmed Tilapia) tilapia, Oreochromis niloticus, is cultured widely for the production of freshwater fish in China. Streptococcosis, which is related to pathogenic infections, occurs frequently in juvenile and adult female GIFT individuals. Resveratrol (RES) has been used in feed to control these infections in freshwater tilapia. To address the effects of RES on tilapia, we used high-throughput RNA sequencing technology (RNA-Seq, HiSeq. 2500) to explore the global transcriptomic response and specific involvement of hepatic mRNA of juvenile O. niloticus fed with diets containing different concentrations of (0, 0.025, 0.05, and 0.1g/kg) RES. A total of > 24,513,018 clean reads were generated and then assembled into 23,244 unigenes. The unigenes were annotated by comparing them against non-redundant protein sequence (Nr), non-redundant nucleotide (Nt), Swiss-Prot, Pfam, Gene Ontology database (GO), Clusters of Orthologous Groups (COG) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases, and 12,578 unigenes were annotated to the GO database. A total of 1444 (0.025g/kg RES), 1526 (0.05g/kg RES), and 3135 (0.1g/kg RES) genes were detected as significant differentially expressed genes (DEGs), when compared with the controls. A total of 6 (0.025 vs 0.05g/kg RES), 19 (0.025 vs 0.1g/kg RES), and 124 (0.05 vs 0.1g/kg RES) genes were detected as significant DEGs. Six genes, including dnah7x1, sox4, fam46a, hsp90a, ddit4, and nmrk2, were associated with an immune response. These findings provide information on the innate immunity of GIFT and might contribute to the development of strategies for the effective management of diseases and long-term sustainability of O. niloticus culture.

Archaeal community compositions in tilapia pond systems and their influencing factors.

Archaea, like the bacterial communities are gradually being realized as key players in the biogeochemical progress of water ecosystems. In this study, tilapia aquaculture ponds were used for an in-depth understanding of archaeal community compositions in water and surface sediment. Some of the main functions, as well as the communities' response patterns, to time variations, pond differences and some physio-chemical parameters were investigated. The results revealed the dominant phylum in both the water and surface sediment, as Euryarchaeota, while, the most abundant classes were: Halobacteria and Methanomicrobia respectively. Significant differences in the archaeal community compositions in the water and surface sediment, were observed in the early stages of cultivation, which became minimal at the later stage of the GIFT tilapia cultivation. Additionally to the differences in the most abundant classes, more OTUs were observed in water samples than in surface sediment samples. The methane generation could be attributed to the large proportion of methanogens found in both pond water and in the surface sediment. Furthermore, the archaeal community compositions in water and the surface sediment were shaped mainly by temporal variations and pond differences respectively. In the pond water, the archaeal community compositions were highly co-related to the concentration changes of ammonia, sulfate and total nitrogen; while in the surface sediment, the correlation to the content changes was significant in total phosphorus. The archaeal community compositions in surface sediment should be considered as an indicator for future environmental capacity studies in aquaculture.

Dietary supplementation with rutin has pro-/anti-inflammatory effects in the liver of juvenile GIFT tilapia, Oreochromis niloticus.

Dietary supplementation with rutin may have some pharmacological qualities including anti-inflammatory effects. Kupffer cell activation resulted in increased transcription of pro- and anti-inflammatory cytokines. The main purpose of this study was to investigate the pro- and anti-inflammatory activities in juvenile freshwater tilapia, Oreochromis niloticus, in response to 0.1 or 0.3 g/kg dietary supplementation of rutin. Results showed that hepatic IgM, anti-inflammatory-cytokines, and pro-inflammatory cytokines were significantly decreased in groups treated with high doses of rutin. Hepatic IgM and anti-inflammatory cytokines (IL-10 and IFN-γ) transcripts were significantly decreased, whereas the transcripts of the pro-inflammatory cytokines, TNFα and IL-1ß were significantly decreased, whereas IL-8 was significantly increased. The number of Kupffer cells in rutin-treated groups was significantly decreased, and scanning electron micrographs showed that rutin enriched the number of gut microvilli and secretion pits. With the phenomena of cell apoptosis occurred in the rutin groups, the present study demonstrated that optimum levels of rutin may be beneficial but excessive level may cause liver impairment, which may be absorbed by the gut and then transported to the liver.

Dietary risk ranking for residual antibiotics in cultured aquatic products around Tai Lake, China.

Antibiotics are widely used in aquaculture and therefore may be present as a dietary risk in cultured aquatic products. Using the Tai Lake Basin as a study area, we assessed the presence of 15 antibiotics in 5 widely cultured aquatic species using a newly developed dietary risk ranking approach. By assigning scores to each factor involved in the ranking matrices, the scores of dietary risks per antibiotic and per aquatic species were calculated. The results indicated that fluoroquinolone antibiotics posed the highest dietary risk in all aquatic species. Then, the total scores per aquatic species were summed by all 15 antibiotic scores of antibiotics, it was found that Crab (Eriocheir sinensis) had the highest dietary risks. Finally, the most concerned antibiotic category and aquatic species were selected. This study highlighted the importance of dietary risk ranking in the production and consumption of cultured aquatic products around Tai Lake.

Effect of methomyl on sex steroid hormone and vitellogenin levels in serum of male tilapia (Oreochromis niloticus) and recovery pattern.

Tilapia were exposed to sub-lethal concentrations of 0, 0.2, 2, 20 or 200 µg/L for 30 days, then transferred to methomyl-free water for 18 days. E2 , T, 11-KTand VTG in serum were examined. There were no significant changes in all the parameters in serum of tilapia exposed to 0.2 µg/L and 2 µg/L methomyl compared to the control. However, 20 µg/L and 200 µg/L have the potential to disrupt the endocrine system of male tilapia, as shown by its ability to increase VTG and E2 and decrease T and 11-KT in serum. Thus it would appear the no observed adverse effect level for sexual steroid hormones of methomyl is lower than 2 µg/L. Recovery data showed that the effects produced by 20µg/L were reversible but not at 200µg/L. Furthermore, the sensitivity of above parameters to methomyl followed the order of VTG>E2 >11-KT>T>GSI, suggesting VTG being the better biomarkers.

Characterizing bacterial communities in tilapia pond surface sediment and their responses to pond differences and temporal variations.

Bacterial community compositions in the surface sediment of tilapia ponds and their responses to pond characteristics or seasonal variations were investigated. For that, three ponds with different stocking densities were selected to collect the samples. And the method of Illumina high-throughput sequencing was used to amplify the bacterial 16S rRNA genes. A total of 662, 876 valid reads and 5649 operational taxonomic units were obtained. Further analysis showed that the dominant phyla in all three ponds were Proteobacteria, Bacteroidetes, Chloroflexi, and Acidobacteria. The phyla Planctomycetes, Firmicutes, Chlorobi, and Spirochaetae were also relatively abundant. Among the eight phyla, the abundances of only Proteobacteria, Bacteroidetes, Firmicutes, and Spirochaetae were affected by seasonal variations, while seven of these (with the exception of Acidobacteria) were affected by pond differences. A comprehensive analysis of the richness and diversity of the bacterial 16S rRNA gene, and of the similarity in bacterial community composition in sediment also showed that the communities in tilapia pond sediment were shaped more by pond differences than by seasonal variations. Linear discriminant analysis further indicated that the influences of pond characteristics on sediment bacterial communities might be related to feed coefficients and stocking densities of genetically improved farmed tilapia (GIFT).

Effects of Microcystis on Hypothalamic-Pituitary-Gonadal-Liver Axis in Nile Tilapia (Oreochromis niloticus).

In the present study, Nile tilapia (Oreochromis niloticus) were used to assess the endocrine disruption potential of Microcytis aeruginosa. Male Nile tilapia were exposed to lyophilized M. aeruginosa or purified microcystin-LR (8.3 µg/L) for 28 days. The levels of serum hormones (17ß-estradiol and testosterone) and transcripts of selected genes in the hypothalamus-pituitary-gonadal-liver axis were analyzed. The results showed that serum hormones were significantly up-regulated, and transcripts of 13 genes (GHRH, PACAP, GH, GHR1, GHR2, IGF1, IGF2, CYP19a, CYP19b, 3ß-HSD1, 20ß-HSD, 17ß-HSD1 and 17ß-HSD8) were significantly altered after Microcytis exposure. These results indicate that fish reproduction can be altered in a Microcystis bloom-contaminated aquatic environment.

Responses of glutathione-related antioxidant defense system in serum of Nile tilapia (Oreochromis niloticus) exposed to sublethal concentration of methomyl and recovery pattern.

Tilapia were exposed to sublethal concentrations of 0, 0.2, 2, 20, or 200 µg/L for 30 days, and then transferred to methomyl-free water for 18 days. GST, GPx, GR, GSH, and GSSG in tilapia serum were examined at 0, 6, 12, 18, 24, and 30 days after methomyl exposure and at 18 days after transferring to methomyl-free water. There were no significant changes in antioxidants activities and contents in serum of tilapia exposed to 0.2 µg/L. Significant increases in GST, GR, GPx, and GSSG accompanied by a decrease in GSH were observed following methomyl exposure to 2, 20, or 200 µg/L, suggesting the presence of oxidative stress. Thus, it would appear the 0.2 µg/L methomyl might be considered the no observed adverse effect level. Recovery data showed that the effects produced by lower concentration of 20 µg/L were reversible but not at the higher 200 µg/L concentration.

Intra-species differences shape differences of enrofloxacin residues and its degradation products in tilapia: A precise risk assessment.

The increasing use and abuse of antibiotics in agriculture and aquaculture necessitates a more thorough risk assessment. We first advocate a precise assessment that subdivides the assessment scope from interspecies to intraspecific levels. Differences in ENR residues and degradation within the intraspecific category were simultaneously explored. This study chose red and GIFT tilapia, both belonging to the intra-specific category of tilapia, for an enrofloxacin (ENR) exposure experiment. Red tilapia had a lower area under the curve (AUC) representing drug accumulation, indicating a notably shorter withdrawal period (7 days) compared to GIFT tilapia (31.4 days) in the edible parts. While four potential transformation pathways were proposed for ENR in tilapia, red tilapia had fewer detected degradation products (6 items) than GIFT tilapia (10 items), indicating a simpler transformation pathway in red tilapia. Predictive assessments using the Toxtree model revealed that of the four extra degradation products in GIFT tilapia, two may possess carcinogenic and mutagenic properties. Overall, differences were observed in ENR residues and degradation within the intraspecific category, with red tilapia presenting lower risks than GIFT tilapia. This work suggests a new strategy to perfect the methodology for antibiotic risk assessment and facilitate systematic antibiotic administration management in the future.

Responses of Digestive, Antioxidant, Immunological and Metabolic Enzymes in the Intestines and Liver of Largemouth Bass (Micropterus salmoides) under the Biofloc Model.

To investigate the activities of intestinal digestive enzymes, liver antioxidant enzymes, immunological enzymes, and glucometabolic enzymes in largemouth bass (Micropterus salmoides) under the biofloc model, an experiment was conducted in 300-liter glass tanks. The experiment comprised a control group, which was fed a basal diet, and a biofloc group, where glucose was added to maintain a C/N ratio of 15. Each group had three parallel setups, with a stocking density of 20 fish per tank. The experiment ran for 60 days, employing a zero-water exchange aquaculture model. The results showed that at the end of the culture period, there were no significant differences between the initial weight, final weight, WGR, SGR, and SR of the biofloc group and the control group of largemouth bass (p > 0.05), whereas the lower FCR and the higher PER in the biofloc group were significant (p < 0.05); intestinal α-amylase, trypsin, and lipase activities of largemouth bass in the biofloc group were significantly increased by 37.20%, 64.11%, and 51.69%, respectively, compared with the control group (p < 0.05); liver superoxide dismutase and catalase activities, and total antioxidant capacity of largemouth bass in the biofloc group were significantly increased by 49.26%, 46.87%, and 98.94% (p < 0.05), while the malondialdehyde content was significantly reduced by 19.91% (p < 0.05); liver lysozyme, alkaline phosphatase, and acid phosphatase activities of largemouth bass in the biofloc group were significantly increased by 62.66%, 41.22%, and 29.66%, respectively (p < 0.05); liver glucokinase, pyruvate kinase, glucose-6-phosphate kinase, pyruvate kinase, glucose-6-phosphatase, and glycogen synthase activities were significantly increased by 46.29%, 99.33%, 32.54%, and 26.89%, respectively (p < 0.05). The study showed that the biofloc model of culturing largemouth bass can not only enhance digestive enzyme activities, antioxidant capacity, and immune response but can also promote the process of glucose metabolism and reduce feeding costs. This study provides data support for healthy culturing of largemouth bass in future production, provides a theoretical reference for optimizing the biofloc technology culture model, and is crucial for promoting the healthy and green development of aquaculture.

Environment consistently impact on aquaculture: The predominant source of residual pollutants in cultured Chinese mitten crab (Eriocheir sinensis) across China.

Advancements in monitoring and operation of aquaculture environments has minimized the concentrations of some residual pollutants in cultured aquatic products. However, currently most aquatic products are "farmed", and relationships among residual pollutants in tissues of crabs were still unclear. In this study, 64 typical pollutants, including 25 antibiotics, 15 metal, 23 organochlorine pesticides, and one dioxin-like compound inducing hydrocarbon-receptor (AHR) activity were measured in Chinese mitten crab (Eriocheir Sinensis) risks of consumption assessed and ranked. The superposition of properties including severity and relative potency of effects and parameters describing persistence and exposure along with rates of usage and identification of groups most likely to be exposed were assessed in combination to rank likelihood of dietary exposure and probabilities of adverse effects for each contaminant. The results indicated that the total scores per pollutants found that Cadmium (Cd), Heptachlor epoxide (HEPE), dioxin TEQ exhibited the greatest scores and explained the severity of dietary risk, while source analysis found that the three main pollutants resulted from the ambient environment and are not due to specific aquaculture processes. In summary, environment is still the predominant source of residual pollutants in cultured Chinese mitten crab across China.

Multidimensional occurrence and diet risk of emerging contaminants in freshwater with urban agglomerations.

Freshwater systems near highly urbanized areas are extremely susceptible to emerging contaminants (ECs), yet their stereoscopic persistence in aquatic ecosystems and related risks remain largely unknown. Herein, we characterized the multi-mediums distribution of 63 ECs in Baiyangdian Lake, the biggest urban lake in the North of China. We identified variations in the seasonal patterns of aquatic EC levels, which decreased in water and increased in sediment from wet to dry seasons. Surprisingly, higher concentrations and a greater variety of ECs were detected in reeds than in aquatic animals, indicating that plants may contribute to the transferring of ECs. Source analysis indicated that human activity considerably affected the distribution and risk of ECs. The dietary risk of ECs is most pronounced among children following the intake of aquatic products, especially with a relatively higher risk associated with fish consumption. Besides, a comprehensive scoring ranking method was proposed, and 9 ECs, including BPS and macrolide antibiotics, are identified as prioritized control pollutants. These findings highlight the risks associated with aquatic ECs and can facilitate the development of effective management strategies.

Aquaculture activities influencing the generation of geosmin and 2-methylisoborneol: a case study in the aquaculture regions of Hongze Lake, China.

Contamination by odor substances such as geosmin (GSM) and 2-methylisoborneol (2-MIB) was examined in the cultured water from aquaculture farming in the region of the Hongze Lake in 2022, and some factors influencing residual levels of them in the water were analyzed. Geographically, high concentrations of GSM were located mainly in the north and northeast culture areas of the lake, while those of 2-MIB were found in the northeast and southwest. Analysis of the water in the enclosure culture revealed significant differences in the concentrations of GSM and 2-MIB among the cultured species. The mean concentrations of GSM in culture water were ranked in the order: crab > the four major Chinese carps > silver and bighead carp, and silver and bighead carp > crab > the four major Chinese carps for 2-MIB. The concentration of GSM was significantly higher at 38.99 ± 18.93 ng/L in crab culture water compared to other fish culture water. Significant differences were observed in GSM concentrations between crab enclosure culture and pond culture, while 2-MIB levels were comparable. These findings suggest that cultural management practices significantly affect the generation of odor substances. The taste and odor (T&O) assessment revealed that the residual levels of GSM and 2-MIB in most samples were below the odor threshold concentrations (OTCs), although high levels of GSM and 2-MIB in all water bodies were at 30.9% and 27.5%, respectively. Compared with the corresponding data from other places and the regulation guidelines of Japan, USA, and China, the region in the Hongze Lake is generally classified as a slightly T&O area, capable of supporting the aquaculture production scale.

Citizen science in action: Time-resolved immunofluorescence-based field detection of antibiotics with portable analytical kit.

Citizen scientist-based environmental monitoring and public education are becoming increasingly popular. However, current technologies for antibiotic-based novel contaminant identification are still restricted to laboratory sample collection and analysis due to detection methodologies and apparatus limitations. This study developed a time-resolved immunofluorescence-based simultaneous field-based assay for ciprofloxacin (CIP) and enrofloxacin (ENR) that matches test results to geographic locations. The assay helps the public understand the potential levels of antibiotic exposures in their environments and helps them take appropriate action to reduce risk. The assay was developed using smartphones and social software in addition to rapid testing. The method uses a portable, low-cost analytical kit with a smartphone app to build a field-based detection platform for the detection and analysis of ENR and CIP in water and aquatic products. The methodological evaluation was good, with detection limits of 0.4 ng/mL and 0.5 ng/g for ENR in water and fish, and quantification limits of 1.2 ng/mL and 1.4 ng/g, with recoveries of 89.0 %-101.0 % and 78.0 %-97.0 %. For CIP in water and fish, the limits of detection were 0.3 ng/mL and 0.4 ng/g, the limits of quantification were 0.9 ng/mL and 1.2 ng/g, and the recoveries were 75.0 %-91.0 % and 72.0 %-89.0 %, both with coefficients of variation <15 %. These limits were sufficient to prevent the two antibiotics from crossing over during simultaneous detection. The assay was validated using real samples to assess the effectiveness of the assay platform in field deployments, and the results were consistent with those obtained through liquid chromatography-tandem mass spectrometry (LC-MS) and enzyme-linked immunoassay (ELISA) techniques. In addition, the TRFIA assay process requires less time, uses more portable instruments, and is less complex than traditional methods. This study provides a new scientific, accurate, and rapid detection method for antibiotic detection by citizen scientists, helping scientists to obtain a wider range of data and providing more opportunities to solve scientific problems.

Significant decline of water pollution associated with inland fishery across China.

Water pollution seriously threatens the sustainable development of fisheries in China. To inform effective pollution control policies, a comprehensive understanding of the fishery environment status is needed. However, nationwide data on the temporal changes of major pollutants in the fishery waters of China are scarce. This study collected data on the major water pollutants, including total nitrogen, total phosphorus, heavy metals, and total petroleum hydrocarbons (TPHs), from 2003 to 2017 to evaluate dynamic changes in the inland fishery water environment across China. We discovered that the levels of four heavy metals (Cu, Zn, Pb, and Cd) and TPH decreased during the 15-year period, corresponding to the reduced national discharge of pollution sources from 2003 to 2015. However, nitrogen and phosphorus levels in the inland fishery waters showed no significant changes during this period. A comparative analysis of water quality in different periods indicated that these improvements were highly associated with effective measures for water pollution control in China. In addition, the decline in pollution was consistent among the three regions of China (north, west, and southeast) from 2003 to 2017, while southeast China exhibited the weakest pollution mitigation among the three regions. These findings suggest that the inland fishery water quality improved during 2003-2017, but still faced eutrophication risk.