This study investigates the occurrence and distribution of microplastics in water, sediment, and crayfish samples within pond and rice-crayfish co-culture breeding modes in Jianli prefecture, China. Microplastics in environmental and biological samples were systematically extracted by CaCl₂ solution, digested by H₂O₂ and KOH, and identified by μ-FTIR. A cleansing treatment for crayfish was performed in pure water before dissection and microplastic accumulation in different tissues (gill, stomach, gut, and flesh) of non-cleansed and cleansed crayfish were compared. The average microplastic abundances were 1.3 ± 0.1–2.5 ± 0.1 particles/L, 0.03 ± 0.01–0.04 ± 0.02 particles/g, and 0.17 ± 0.07–0.92 ± 0.19 particles/individual in water, sediment, and crayfish samples, respectively. Microplastics were detected in all studied crayfish tissues, except the flesh. There were no significant differences in microplastic abundances in water (P = 0.82), sediment (P = 0.90), and crayfish (P = 0.47 for non-cleansed samples; P = 0.30 for cleansed samples) between two breeding modes despite the detection of relatively higher microplastic abundances in the samples from the pond breeding mode. Microplastic accumulation in non-cleansed crayfish stomachs and guts (0.71 ± 0.18 particles/individual) was higher (P < 0.01) than that recorded in their gills (0.13 ± 0.06 particles/individual). Moreover, microplastics present in the stomachs and guts of cleansed crayfish were significantly less abundant (P < 0.01) than in non-cleansed crayfish, although this was not observed in the gills (P = 0.99). The majority of microplastic particles in this study had fiber-like shapes, blue and transparent colors, a size smaller than 1 mm, and polymer types of PP:PE and PE. The results demonstrate that microplastics in the environment can accumulate in the internal tissues of crayfish, which may pose a potential risk to humans through food consumption without the removal of the gills, stomach, and guts. This study provides valuable information for understanding microplastic accumulation in the different tissues of crayfish and the potential risk of human exposure to microplastics from crayfish as a food supplement.

Antibiotic resistance genes (ARGs) are emerging contaminants that pose a potential risk to human health worldwide. In this study, a metagenomic analysis was performed to investigate the distribution of ARGs in paddy field ecosystems, crayfish monoculture pond ecosystems, and rice-crayfish cultivation field ecosystems. The results showed that MacB and BcrA are two dominant ARGs, and macrolide is the dominant antibiotic not only in the water, but also in the sediment and gut of crayfish, in both the crayfish monoculture and the rice-crayfish cultivation ecosystems. Meanwhile, some lower-abundance ARGs in the water and sediment of crayfish cultivation ecosystems were significantly different in their abundance than those in rice paddy fields, and the ARGs in crayfish culture system and rice paddy fields showed even higher dissimilarities of diversity. Comprehensive analysis with redundancy analysis（RDA）and the distribution of dominant ARGs showed that the dissimilarity was related to the higher concentrations of total nitrogen (TN), total phosphorus (TP), chlorophyll a (Chla), permanganate index (CODMₙ₎, and nitrate in the water of rice paddy fields, and was related to the higher contents of N, P, K, and organic matter (OM) in the sediment of rice paddy fields. The source trackers of ARGs within the crayfish cultivation ponds and the rice-crayfish cultivation fields showed that the sediment in crayfish cultivation ponds mainly played the role of the ‘sink’ for ARGs, and the water and gut of crayfish mutually contributed to the high rates of ARGs. The ARG contribution rates of crayfish gut and sediment decreased and increased, respectively, in rice-crayfish cultivation fields and in crayfish monoculture ponds, which might be related to the lower crayfish biomass and the lower water depth in rice-crayfish cultivation fields.

As the two most commonly used organophosphorus herbicides, glyphosate (Gly) and glufosinate-ammonium (Glu) have unique properties for weed control and algae removal in aquaculture. However, the occurrences and health risks of Gly and Glu in aquaculture ponds are rare known. This study aimed to investigate the occurrences of Gly, AMPA (primary metabolity of Gly) and Glu in surface water, sediment and aquatic products from the grass carp (ctenopharyngodon idella), crayfish (procambarus clarkii) and crab (eriocheir sinensis) ponds around Lake Honghu, the largest freshwater lake in Hubei province, China where aquaculture has become the local pillar industry. Three age groups (children, young adults, middle-aged and elderly) exposure to these compounds through edible aquatic products (muscle) consumption were also assessed by target hazard quotient (THQ) method. The results indicated that Gly, AMPA and Glu were widely occurred in surface water, sediment and organisms in the fish, crayfish and crab ponds. AMPA was more likely to accumulate in the intestine of aquatic products than Gly and Glu. According to the total THQ value (1.04＞1), muscle consumption of grass carp may pose potential risk to children.

The effect of selenium (Se) on the reproductive system has been investigated in both humans and vertebrates, but few studies of female fertility and reproduction in invertebrate have been reported. This study is aimed to investigate the effect of SeMet on growth performance and reproductive system after crayfish were fed with graded levels of dietary SeMet (0, 1.49, 3.29, 10.02, 30.27 or 59.8 μg Se/g dry weight) for 60 days. Crayfish treated with the high levels of SeMet (10.02, 30.27 and 59.76 μg Se/g) exhibited decreasing FW and CL in both male and female. Interestingly, Se accumulation was higher in ovary than in other tissues, suggesting that ovary may serve as a target organ for Se accumulation. We found that dietary Se concentration of 10.02 μg Se/g significantly improved the spawning rate, promoted the synchronized spawning, and up-regulated the expressions of mRNA of cdc2 and vitellogenin, with significantly increased E2 and VTG concentrations in hemolymph of female crayfish. However, a marked decrease of the E2 contents and spawning rate was observed in the groups treated with 30.27 and 59.76 μg Se/g diets. In conclusion, the results of this study indicated that the Se had maximum accumulation in ovary, affecting the reproductive capacity by intervening the expression of cdc2 and vitellogenin in the reproductive system. The LOAEL to induce FW was observed in crayfish fed with 10.02 μg Se/g diet, and its value can cause toxicity within the range of natural concentration, so the addition of Se in the feed should be within 10.02 μg Se/g.

Metal pollution is one of the main environmental threats in freshwater ecosystems. Aquatic animals can accumulate these substances and transfer them across the food web, posing risks for both predators and humans. Accumulation patterns strongly vary depending on the location, species, and size (which in fish and crayfish is related to age) of individuals. Moreover, high metal concentrations can negatively affect animals’ health. To assess the intraspecific relationship between metal accumulation and size and health (proxied by the body condition) of individuals, the concentration of 14 metals (Al, As, Cd, Co, Cr, Cu, Fe, Hg, Mg, Mn, Ni, Pb, Se, Zn) was analyzed in six alien species from the highly anthropogenically altered Arno River (Central Italy): five fish (Alburnus alburnus, Pseudorasbora parva, Lepomis gibbosus, Ictalurus punctatus, and Silurus glanis) and one crayfish (Procambarus clarkii). We found that in P. clarkii, Cu was negatively related to size, as well as Al in L. gibbosus and Mg for adult I. punctatus. Positive size-dependent relationships were found for Hg in L. gibbosus, Fe in S. glanis, and Cr in juvenile I. punctatus. Only Co and Mg in S. glanis were found to negatively correlate with individual health. Since metal concentrations in animal tissue depend on trade-offs between uptake and excretion, the few significant results suggest different types of trade-offs across different species and age classes. However, only predatory fish species (L. gibbosus, I. punctatus, and S. glanis) presented significant relationships, suggesting that feeding habits are one of the primary drivers of metal accumulation.

Environmental pollution is one of the most serious causes of degradation of Mediterranean wetlands. Mercury (Hg) and cadmium (Cd) are of particular concern due to biomagnification. Here, we used red swamp crayfish (Procambarus clarkii) to monitor the spatial and temporal patterns of these two metals in a Portuguese rice field system. We sampled the crayfish in three different sites and three different time periods in the Sado River Basin (Portugal). We measured temperature, pH, total dissolved solids and conductivity in the water. Hg and Cd were measured in the crayfish abdominal muscle tissue and exoskeleton. In muscle, a spatial pattern was found for the accumulation of Cd while for Hg, only a temporal pattern emerged. The spatial pattern for Cd seemed to reflect the mining history of the sites, whereas the temporal pattern for Hg seemed related to the flooding of rice fields. We suggest that this flooding process increases Hg bioavailability.

Freshwater crayfish are bioindicators of environmental pollution, often used for the assessment of heavy metal (HM) presence in the tissues, a time-consuming and expensive task. In this study, we propose the use of the vibrational spectroscopy to detect in a fast, non-destructive and sensitive way the presence of HM in the cephalothorax exoskeleton of the freshwater crayfish. Incorporation of HM into the cephalothorax exoskeleton was investigated under controlled laboratory conditions. In particular, the cephalothorax exoskeleton of five crayfish species (Astacus leptodactylus, Procambarus clarkii, Austropotamobius pallipes, Faxonius limosus, and Pacifastacus leniusculus) was analyzed by attenuated total reflection–Fourier transformed infrared (ATR–FTIR) spectroscopy in the presence or absence of cadmium (Cd), chromium (Cr), lead (Pb), nickel (Ni), and zinc (Zn) up to 4 weeks at various concentrations (0.01, 0.1, 1, 10, ppm). The ATR–FTIR profile of the crayfish cephalothorax exoskeleton was compatible with the presence of amorphous calcium carbonate, chitin, and proteins. The incubation with the HM revealed two main modifications: the shift of the peak from 859 to 872 cm⁻¹ and the appearance of a peak at 712 cm⁻¹. Both are ascribable to the HM interaction with calcium carbonate. The absorbance of both peaks increased along with the time of incubation, and the HM concentration. We conclude that ATR–FTIR analysis can be a useful, quick, and cost-sensitive tool to detect HM presence in the crayfish cephalothorax exoskeleton. However, it has to be regarded as a non-specific analytical technique for assessing HM contamination, since it is unable to discriminate between different HM.

The influence from the manufacturing of compact fluorescent lamps (CFL) on mercury (Hg) speciation and distribution in river catchments nearby a typical CFL manufacturing area in China was investigated. Water, sediment, river snail (Procambarus clarkii), and macrophyte (Paspalum distichum L.) samples were collected. Total Hg (THg) and methylmercury (MeHg) concentrations in water ranged from 1.06 to 268 ng · L⁻¹ and N.D. −2.14 ng · L⁻¹, respectively. MeHg was significantly positively correlated with THg in water. THg and MeHg in sediment ranged from 15.0 to 2480 and 0.06 to 1.85 ng · g⁻¹, respectively. River snail samples exhibited high concentrations of THg (206–1437 ng · g⁻¹) and MeHg (31.4–404 ng · g⁻¹). THg and MeHg concentrations in root of P. distichum L. were significantly higher than those in shoot, indicating that THg and MeHg in the plant were mainly attributed to root assimilation. A very high bioaccumulation factor (20.9 ± 22.1) for MeHg in P. distichum L was noted, suggesting that P. distichum L. might have a potential role in phytoremediating MeHg contaminated soil due to its abnormal uptake capacity to MeHg.

There is a need to develop bioassessment tools that can diagnose the effects of individual stressors that can have multiple ecological effects. Using nuclear magnetic resonance (NMR)-based metabolomics, our experiments aimed to identify the sensitivity of metabolites to changes in food availability and dissolved oxygen (DO) concentrations, and compare these results to identify metabolites that may differentiate between the effects of these two stressors. Forty-eight, laboratory-raised, red swamp crayfish (Procambarus clarkii) were randomly assigned and exposed to one of three food availability or DO treatment levels (high, normal, low). Starved crayfish had lower amounts of amino acids than fed crayfish, suggesting catabolic effects of starvation on tail muscle tissue for energy requirements. In contrast, crayfish exposed to hypoxic conditions experienced changes in abundance of metabolites primarily associated with energy metabolism. Tail muscle was the only tissue sensitive to food and DO stress, suggesting the need to select tissues for monitoring appropriately. Our evaluation of environmental metabolomics as a tool for bioassessment indicates that several identified metabolites in crayfish tail muscle may be able to diagnose food and oxygen stress. Further study is required to determine if these metabolic effects are linked with changes of individual fitness and higher levels of biological organization, such as population size.

Bioindicators can reflect the effects of pollutants on their metabolism, being widely used to assess environmental stress. In this sense, the crab Procambarus clarkii has been previously proposed to monitor the contamination in Doñana National Park (southwest Spain) using conventional biomarkers. In this work, a metabolomic approach based on direct infusion mass spectrometry, which allows an easy and quick study of a large number of metabolites in a single run, was used for pollution assessment of this area, considering the biological response of this organism to contamination. In addition, metal accumulation in crab tissues was determined. Thus, the integrated analysis of metabolomic and metallomic data enabled the study of metabolic response of the organism against pollution. Several metabolites were discovered as potential biomarkers of pollution, such as decreased levels of carnosine, alanine, niacinamide, acetoacetate, pantothenic acid, ascorbate, glucose-6-phosphate, arginine, glucose, lactate, phospholipids, and tryglicerides, as well as elevated levels of acetyl carnitine, phosphocholine, choline, and uric acid. In this way, metal-induced toxicity could be related to metabolic impairments, principally oxidative stress, metabolic dysfunction, and dyslipidemia.