Effescts of acute diclofenac exposure on intestinal histology, antioxidant defense, and microbiota in freshwater crayfish (Procambarus clarkii).

In the present study, we exposed Procambarus clarkii to different doses (0, 1, and 10 mg/L) of diclofenac (DCF). Meanwhile, we investigated the effects of exposure to DCF on intestinal histology, antioxidant defense, and microbial communities in P. clarkii. The results showed DCF caused histological changes in the intestines. Additionally, DCF induced significant changes in the expression of antioxidant genes including Mn-sod, cat, gst, and gpx. High-throughput sequencing of 16 S rRNA gene revealed DCF changed the diversity, richness, and composition of intestinal microbial communities. The relative abundances of the predominant phyla Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria showed significant changes at the phylum level after treatment with DCF. At the genus level, the most predominant genera with marked differences in abundance were Lucibacterium, Shewanella, Bacteroides, Anaerorhabdus, Aeromonas, Acinetobacter, Clostridium XlVb, Arcobacter, Bosea, and so on. To conclude, treatment with DCF could cause intestinal histological damage, induce significant changes of the expression of intestinal antioxidant genes, and impact the composition of intestinal microbiota in P. clarkii. This research will provide novel insights into the toxic effects of DCF on aquatic crustaceans.

Microcystin-LR-induced changes of hepatopancreatic transcriptome, intestinal microbiota, and histopathology of freshwater crayfish (Procambarus clarkii).

As a hepatotoxin, microcystin-LR (MC-LR) poses a great threat to aquatic organisms. In this research, the hepatopancreatic transcriptome, intestinal microbiota, and histopathology of Procambarus clarkii (P. clarkii) in response to acute MC-LR exposure were studied. RNA-seq analysis of hepatopancreas identified 372 and 781 differentially expressed genes (DEGs) after treatment with 10 and 40 µg/L MC-LR, respectively. Among the DEGs, 23 genes were immune-related and 21 genes were redox-related. GO functional enrichment analysis revealed that MC-LR could impact nuclear-transcribed mRNA catabolic process, cobalamin- and heme-related processes, and sirohydrochlorin cobaltochelatase activity of P. clarkii. In addition, the only significantly enriched KEGG pathway induced by MC-LR was galactose metabolism pathway. Meanwhile, sequencing of the bacterial 16S rRNA gene demonstrated that MC-LR decreased bacterial richness and diversity, and altered the intestinal microbiota composition. At the phylum level, after 96 h, the abundance of Verrucomicrobia decreased after treatment with 10 and 40 µg/L MC-LR, while Firmicutes increased in the 40 µg/L MC-LR-treated group. At the genus level, the abundances of 15 genera were significantly altered after exposure to MC-LR. Our research demonstrated that MC-LR exposure caused histological alterations such as structural damage of hepatopancreas and intestines. This research provides an insight into the mechanisms associated with MC-LR toxicity in aquatic crustaceans.

Effects of cadmium on intestinal histology and microbiota in freshwater crayfish (Procambarus clarkii).

In this study, Procambarus clarkii (P. clarkii) were exposed to different concentrations (0, 2, 5 and 10 mg/L) of cadmium (Cd). We studied the effects of Cd exposure on intestinal histology and microbiota in P. clarkii. The results demonstrated that exposure to Cd caused histological alterations in the intestines of P. clarkii. Meanwhile, high-throughput sequencing analysis revealed that Cd exposure could alter the richness, diversity, and composition of intestinal microbiota in P. clarkii. At the phylum level, the relative abundances of the prevalent phyla Firmicutes, Proteobacteria, Bacteroidetes, Fusobacteria, and Actinobacteria changed significantly after exposure to Cd. At the genus level, the most prevalent genera with significant difference in relative abundance were Bacteroides, Clostridium XlVb, Hafnia, Buttiauxella, Shewanella, Anaerorhabdus, Alistipes, Arcobacter, Azoarcus, Chryseobacterium, and so on. Furthermore, functional prediction analysis of intestinal microbial communities showed that Cd exposure could significantly alter the pathways related to metabolism, diseases, cellular processes, and so on. Taken together, exposure to Cd could induce intestinal histological damage and affect intestinal microbiota composition and functions of P. clarkii. Our study can be an important step toward a better understanding of the toxic effects of Cd on aquatic crustaceans.

Comprehensive assessments of ecological states of Songhua River using chemical analysis and bivalves as bioindicators.

The aim of this study was to compare environmental pollution and ecological states of two different areas of the Songhua River areas: near Harbin City and Tongjiang City, located at a distance of about 500 km downstream. The anthropogenic pollution concentrations of heavy metals (HM) were determined. The results showed that concentrations of eight metals (Cd, Cr, Cu, Fe, Mn, Ni, Pb, and Zn) in the water were in the range of 0.001-0.588 mg/L for Tongjiang and 0.001-0.508 mg/L for Harbin, while that in sediments were in the range of 0.67-1575.37 mg/kg for Tongjiang and 0.07-5617.13 mg/kg for Harbin, respectively. Bivalves from tested sites exposed to environmental pollution exhibited significantly different physiological states. The latter was assessed using the method of physiological loading, based on measuring the recovery time (Trec) of heart rate (HR) after removal of the load. Trec in mussels from Harbin was recorded in the range of 151 to 234 min, while that from Tongjiang was only 115 min. Cd, Cu, Pb, and Zn in mollusk soft tissues were also determined for Harbin and Tongjiang, respectively. The metal pollution index (MPI) and bioconcentration factor (BCF) in the mollusks were calculated for each metal. BCF in the mussels from the Tongjiang area was lower than that from the Harbin area. Physiological testing, as well as the concentration of HM in water, and sediment, and also the bioaccumulation of HM in tissue showed that the ecological state of the Tongjiang area was better than that of Harbin. Apparently, after more extensive studies, a methodological approach of assessing the ecological state of water areas, based on physiological state testing of aboriginal mollusks, could be used in the monitoring of pollution effects in water bodies and streams.

Cadmium-induced oxidative stress, histopathology, and transcriptome changes in the hepatopancreas of freshwater crayfish (Procambarus clarkii).

Cadmium (Cd) is a common contaminant in environment. Crayfish are considered suitable for indicating the impact of heavy metals on the environment. However, there is limited information on the mechanisms causing damage to the hepatopancreas of Procambarus clarkii exposed to Cd. We exposed adult male P. clarkii to 2.0, 5.0, and 10.0 mg/L Cd for 24, 48, and 72 h to explore Cd toxicity. Afterwards, we measured bioaccumulations in the hepatopancreas and determined malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST). Additionally, the hepatopancreas histopathology was analyzed and the transcriptome analysis of the P. clarkii hepatopancreas under Cd stress was conducted. The results revealed that hepatopancreas could accumulate Cd in a time- and dose-dependent manner. Cd induced significant changes in MDA content and antioxidant enzyme activity. Severe histological alterations were observed in crayfish hepatopancreas. After 72 h exposure to 2.0, 5.0, and 10.0 mg/L Cd, transcriptome analysis identified 1061, 747, and 1086 differentially expressed genes (DEGs), respectively. Exposure to 5.0 mg/L Cd inhibited heme binding, tetrapyrrole binding, iron ion binding and activity of oxidoreductase and sulfotransferase, while exposure to 10.0 mg/L Cd enhanced the export of matters from nucleus. In the hepatopancreas treated with 10.0 mg/L Cd, pathways related to diseases and immune system were significantly enriched. Meanwhile, 31, 31, 24, 7, and 12 identified DEGs were associated with the oxidation-reduction process, immune system, ion homeostasis, digestion and absorption, and ATPases, respectively. Our study provides comprehensive information for exploring the toxic mechanisms of Cd and candidate biomarkers for aquatic Cd risk evaluation.

Investigation of chloramine-T impact on crayfish Astacus leptodactylus (Esch., 1823) cardiac activity.

The crayfish play an essential role in the biomonitoring and may reflect ambient water quality through the biochemical, behavioural and physiological reactions. To assess whether narrow-clawed crayfish Astacus leptodactylus can respond by heart rate changes to presence in water of such biocide as chloramine-T, adult males were exposed to its low (2 and 5 mg L(-1)), moderate (10 mg L(-1), commonly used in industry and aquaculture) and exceeded (20 and 50 mg L(-1)) concentrations. In addition, a physical stress test evaluated energy expenditure following the chemical trials. Three key reactions (cardiac initial, first-hour and daily prolonged exposure) were discussed with particular focus on crayfish initial reaction as the most meaningful in on-line water quality biomonitoring. After short-term exposure to both chloramine-T concentrations, crayfish were found to respond rapidly, within 2-5 min. According to heart rate changes, the 1-h exposure did not adversely affect crayfish at either concentration, as well as during daily exposure to 10 mg L(-1). As assessed by the heart rate, the 24-h exposure to 50 mg L(-1) of chloramine-T was toxic for crayfish and led to substantial loss of energy that became apparent during subsequently conducted physical stress. The results supported a hypothesis that crayfish vital functions are connected with environment they inhabit closely enough to serve as biological monitors. Crayfish were tolerant to short-term chloramine-T exposure, while rapid crayfish reaction to an increased chemical level indicated their high sensitivity, an essential attribute of real-time environmental assessment.

Sediment contaminants and contaminant levels and biomarkers in caged mussels (Mytilus trossulus) in the southern Baltic Sea.

Sediment from four southern Baltic Sea locations and caged mussels were analyzed for PCBs, organochlorine pesticides and metals (Cu, Zn, Cd, Pb, Hg). In mussels, which were additionally analyzed for 16 individual PAHs, a set of biomarker responses was measured to derive an integrated biomarker response (IBR) index as a part of integrative assessment of pollution in this area. Concentrations of PCBs, ΣDDT, and metals in sediment and mussels, greatest within the Gulf of Gdansk area, showed decreasing gradients outwards from the Gulf. Sediment quality quotients (SQQs) and biological impact quotients (BIQs), reflecting on sediment- and mussel-accumulated contaminants' potential for biological effects, respectively, pointed out to the Gulf of Gdansk to be of greatest concern among the examined sites. The IBRs corresponded poorly with the SQQs and BIQs, nevertheless, provided a line of evidence indicative of biological effects of contaminants to support more complex processes of environmental status assessment.