
Naturally occurring radioactive substances in freshwater ecosystems pose potential radiological risks to both aquatic organisms and humans through dietary exposure. This study quantified the activity concentrations of 228Ra, 210Pb, 226Ra, 210Po and 40K in ten commonly consumed fish species from the Red River, Vietnam. The results showed significant inter-isotope variability, with 40K dominating all samples. Mean activity concentrations were 1.22 ± 0.2 (LOD–7.61) for 228Ra, 9.1 ± 1.7 (LOD–37.1) for 210Pb, 7.69 ± 1.04 (0.70–24.7) for 226Ra, 11.8 ± 0.9 (5.16–18.7) for 210Po, and 101 ± 8.6 (65–169) Bq.kg−1 for 40K. These variations reflect the different behaviors of the isotopes in aquatic environments. Negative correlation was observed between 226Ra and body weight, suggesting morphological influences on radioactivity absorption in fish. Radiation risk assessments revealed average annual effective doses of 0.105 mSv.y−1 (0.030–0.247 mSv.y−1), significantly lower than the safety threshold, with lifetime cancer risks ranging from 1.4 × 10−7 to 7.7 × 10−7, within acceptable limits. Ecological risk assessment using the ERICA Tool revealed total dose rates ranging from 0.2 to 10.4 µGy.h−1 (mean: 3.0 µGy.h−1), with the Zig-zag eel exceeding the screening threshold of 10 µGy·h−1, indicating a potential radiological risk for this species, while risks remained negligible for the remaining species. These findings enhance understanding of radionuclide dynamics in freshwater food webs and inform public health risk assessments for fish consumption in the Red River basin. The study establishes essential baseline data for regional radiation monitoring programs while contributing to biodiversity protection and food safety.
The presence of microplastics (MPs) in freshwater environments represents a growing environmental concern due to their persistence and ability to interact with contaminants such as pesticides. This study evaluated the influence of photodegradation on the toxicity of polyethylene microplastics (PE-MPs), alone and combined with the pesticide Imidacloprid (IMI), on the cladoceran Daphnia similis. Microparticles of polyethylene were aged under UV-C radiation for 94 days and subsequently characterized by Scanning Electron Microscopy (SEM) and Fourier-Transform Infrared Spectroscopy (FTIR). Surface analysis indicated polymer degradation through cracks and cavities, and increased surface roughness, while FTIR analyses showed polymer oxidation. Ecotoxicological results revealed that pristine MPs significantly reduced survival by 90
The present study investigates how host–parasite interactions influence mercury (Hg) distribution in the common toad (Bufo bufo) under contrasting environmental conditions. Using road-killed specimens as an ethical sampling source, Hg concentrations were quantified in multiple host tissues (muscle, liver, kidneys, lungs, and skin) and associated parasitic nematodes from two sites in eastern Slovakia with different contamination levels. In both populations, Hg accumulation followed a tissue-specific pattern, with the highest concentrations occurring in detoxification organs, particularly the kidneys and liver. Bayesian regression models revealed that habitat was the primary driver of Hg burdens, whereas biological factors, including sex and body condition factor, had no significant effect on host Hg concentrations. Parasitological analyses showed markedly higher infection levels at the contaminated site, with nematode prevalence reaching 100
Pesticide contamination in tropical agricultural soils represents a growing concern due to its impacts on environmental integrity and human health, particularly in transition zones where intensive agricultural converges with coastal wetlands. This study aimed to identify and quantify pesticide residues in soils from northwestern Mexico. Forty-eight composite soil samples were collected within a ≈ 30,000 ha polygon surrounding the study area in the municipality of Santiago Ixcuintla, one of the most agriculturally productive regions of the country. Sampling sites were preselected using remote sensing and geographic information systems. Pesticide extraction was performed using a modified QuEChERS method, and compounds were analyzed by UPLC-MS/MS and GC-TQ-MS/IT-MS. Of the 73 validated pesticides, 26 active ingredients (31.5
Wild boar (Sus scrofa) have been used as sentinel species for monitoring environmental metal contamination; however, developmental variation in internal metal distribution remains poorly understood in free-ranging wildlife. Here, we investigated organ-specific distribution patterns of copper (Cu), zinc (Zn), cadmium (Cd), and lead (Pb) by explicitly comparing fetal and non-fetal wild boar collected in western Japan. A total of 13 individuals, including eight fetuses from two pregnant females, the two pregnant females, one adult male, and two juveniles, were analyzed. Metal concentrations were determined in muscle, heart, liver, kidney, and femur using ICP-MS, and relative organ distributions were evaluated using non-metric multidimensional scaling and permutational multivariate analysis of variance. Developmental differences were apparent for Cu, Cd, and Pb, whereas Zn exhibited stable distribution patterns across stages, suggesting maintained physiological regulation. Fetal individuals showed pronounced hepatic enrichment of Cu, reflecting active placental transport and developmental hepatic storage. In contrast, Cd displayed renal dominance exclusively in non-fetal individuals, indicating postnatal accumulation, while Pb showed relatively greater femoral contributions in non-fetal individuals despite low absolute concentrations. Together, these results suggest that multi-organ distribution patterns provide useful insight into developmental metal partitioning beyond absolute concentration data and highlight the importance of incorporating developmental context into ecotoxicological assessments of free-ranging wildlife under natural exposure conditions.
The presence of potential toxic elements (PTEs) in water, soil, and plant resources poses a significant risk to human health. A field investigation on the content of PTEs and macronutrients in soils and 11 dominant plants (roots and shoots) from Ahangaran, a Pb–Zn mine in western Iran was carried out. The bioconcentration factors (BCF) for roots and shoots (BCF root, BCF shoot) based on both total and available PTEs and macronutrients in soils and translocation factors (TF) of 11 plant species were calculated. The average contents of total soil Cd, Cu, Ni, Pb, and Zn were 198, 124, 86, 38, and 5 times higher than their corresponding background values, respectively. The high and variable contents of P, K, Ca, and Mg, with lower Na, were observed across the plant species. Potential toxic elements like Pb, and Zn have high mobility indices in the soils, meaning these PTEs are more likely to be taken up by plants, potentially causing toxicity. Simultaneous presence of elevated levels of Pb, Fe, and Cd characterizes the contamination profile of the plants at the study locations. When various BCF values based on available PTEs in soils were taken into account, Stachys inflata and Senecio gallicus consistently function as effective phytostabilizers, exhibiting higher root accumulation than shoot accumulation and TF below 1 for all PTEs except Stachys inflata for Zn, making them the most reliable candidates for PTEs retention in roots. Conversely, Phlomis olivieri, Euphorbia szovitsii, and particularly Achillea millefolium consistently perform as phytoextractors across all six PTEs, displaying TF above 1 and high shoot bioconcentration factors, with Achillea millefolium showing exceptional TF (up to 3.03 for Fe). The native plant species identified in this study offer practical applicability for large-scale phytoremediation in similar semi-arid mining environments, enabling evidence-based selection of stabilizers for long-term PTEs containment and accumulators for active PTEs removal.
Global pollution by MPs has become an emerging concern, and recent studies have focused on analyzing their presence in wastewater treatment systems. One of the main challenges in this field is the absence of standardized methods, reference materials, and comparative data. This study aimed to develop, optimize, and validate a methodological framework for the extraction and identification of MPs in wastewater matrices. Reference MPs were produced from the polymers such as polyethylene terephthalate (PET), high-density polyethylene (HDPE), low-density polyethylene (LDPE), polyvinyl chloride (PVC), polypropylene (PP), and polystyrene (PS). To evaluate the integrity of these MPs after digestion, five Fenton reagent protocols were tested, and the carbonyl index was applied. For density-based separation, sodium chloride (NaCl) and zinc chloride (ZnCl2) solutions at different concentrations were used in a density separation device. The MPs obtained presented irregular fragments ranging from 0.1 to 3.0 mm. FTIR analysis performed before and after Fenton digestion showed no significant spectral deviations. The optimal Fenton conditions involved temperatures between 40 and 60 °C and 2 h of reaction time. ZnCl2 provided the best performance for particle recovery, ensuring high separation efficiency. The optimized methodology was subsequently validated using raw wastewater and dewatered sewage sludge collected from a full-scale WWTP. The validation demonstrated effective organic matter removal, successful recovery of spiked MPs, and reliable identification of native MPs present in the environmental samples. Overall, the proposed methodology proved to be a robust and reliable approach for the extraction and characterization of MPs in complex wastewater matrices, providing methodological support for future monitoring programs and contributing to the development of standardized protocols for MPs analysis.
Pesticide poisoning poses a continuing threat to wildlife, particularly avian species such as the Indian Peafowl Pavo cristatus, which holds ecological and cultural significance in India. Despite increasing reports of mass mortalities of the National bird across the country, confirmed and reported, cases of pesticide poisoning remain limited. In this study, eight mass mortality incidents involving death of 121 Indian Peafowls were investigated in Tamil Nadu between 2019 and 2021. Five separate tissue samples from each of 41 individuals were analysed for the presence of 45 pesticides using Liquid Chromatography–Tandem Mass Spectrometry (LC–MS/MS). In all cases, monocrotophos a highly toxic organophosphate pesticide was consistently detected in both tissue and bait samples at concentrations far exceeding known avian toxicity thresholds, indicating it as the cause of mortality. Concentration pattern was in the order; crop content (339.0 mg/Kg) > gut content (83.5 mg/Kg) > liver (1.59 mg/Kg) > kidney (1.00 mg/Kg) > muscle (0.45 mg/Kg). High concentration of monocrotophos were highest in crop and gut contents, suggesting oral ingestion through baited food grains recently. Additionally, flubendiamide, profenophos and carbendazim were also detected in one incident each, representing the first report of these pesticide residues in wild birds globally. These findings provide definitive toxicological evidence of pesticide-induced mortality in Indian Peafowls, reinforcing earlier suspicions from news and field reports. The study highlights the widespread possible misuse of the regulated pesticide and the urgent need for coordinated monitoring, stricter regulation, and community-level awareness to mitigate poisoning risks to non-target wildlife species in the country.
In this study we provide a comprehensive assessment of metal contamination in the Amazon River and its main tributaries, with a focus on urban contamination hotspots. A monitoring campaign was conducted across 40 sampling sites, analysing water concentrations of As, Cd, Cu, Cr, Fe, Mn, Pb, Ni, and Zn. Metal concentrations were evaluated against national and international water quality standards and compared between urbanized and less impacted regions to delineate pollution patterns. Additionally, an ecological risk assessment was conducted for individual metals and for metal mixtures using Species Sensitivity Distributions (SSDs). The results of this study show that Fe levels in the Amazon River were exceptionally high, with potential implications for aquatic species distribution across the basin. Exceedances of water quality standards were identified for four metals (Fe, Pb, Mn, Cu), with metal enrichment observed particularly in Manaus, Macapá, and Belém. Acute multi-substance potentially affected fraction (msPAF) of species in the Amazon River and its tributaries indicated insignificant risks, while chronic values ranged from 3
Small streams are important ecosystems in many areas globally, playing a crucial role in conserving biodiversity and providing ecosystem functions. However, owing to their high connectivity with surrounding landscapes, small streams are particularly susceptible to drivers of biodiversity decline. We report here the results of a monitoring program conducted between 2018 and 2021 at 18 sites in two stream catchments in southwest Germany, characterized by near-natural to highly anthropogenic land use. The monitoring program included 26 environmental, biological, and anthropogenic stressor variables describing the landscape context, aquatic ecosystem compartments, and riparian areas. We used ordination analysis and factor analysis of mixed data (FAMD) to analyse the relationship between variables and the effects of anthropogenic stressors on ecosystem structure and functioning. Results showed that near-natural upstream sites are substantially less impacted by anthropogenic stressors (e.g., pesticide pollution, morphological alterations) compared with agricultural and urban downstream sites. Biotic parameters, such as abundances and taxa richness of aquatic macroinvertebrates, fish, and carabid beetles, as well as leaf litter decomposition, exhibited overall high variations between sites. FAMD assigned the stream sites into four groups (i.e., forest, grassland, agriculture, viticulture), primarily based on land use and anthropogenic stressor influences and found anthropogenic stressors to often co-occur. Lower aquatic macroinvertebrate abundances and diversity, as well as reduced leaf litter decomposition, were observed at sites with higher anthropogenic disturbances. Overall, our findings suggest that small streams are subjected to multiple anthropogenic stressors, resulting in negative effects on both aquatic and riparian ecosystem health.
Estrone (E1) is one of the major natural estrogens influencing the reproductive system of vertebrates. However, it contaminates aquatic environments due to runoff from livestock waste and the discharge of domestic sewage. The aim of this study was to investigate the impact of exposure to environmentally relevant concentrations of E1 on oogenesis and spawning in zebrafish (Danio rerio). Fish were exposed, in duplicate, to 20, 200, and 2000 ng/L of E1 for 49 days for oogenesis analysis evaluating histology, sex steroids, morphometry, cell proliferation, apoptosis, immunohistochemistry for insulin-like growth factor (IGF) and inducible nitric oxide synthetase (iNOS). 17β-estradiol (E2) and 11-ketotestosterone (11-KT) were assessed by ELISA assay. Nitric oxide production was evaluated indirectly by quantifying nitrite. Results revealed that E1 exposure altered the proportions of ovarian follicles and somatic components, with significant increase of oogonia, follicular atresia and inflammatory infiltrate, and decrease of follicular cells. Levels of E2, and immunoreaction for IGF1 increased in E120 and 200 ng/L groups, but 11-KT, IGF2 and IGF1R were not altered. Nitrite concentrations in ovaries were significantly elevated in E1 200 and 2000 ng/L groups, but iNOS immunoreaction was not altered. These changes led to reduced egg production in all groups exposed to E1 and significatively decreased fertilization rates at 200 and 2000 ng/L. Overall, the findings demonstrate that E1, even at concentrations commonly found in surface waters, has detrimental effects on ovarian development, gametogenesis and reproductive success in zebrafish. Monitoring environmental estrogens levels in aquatic environments is therefore essential for fish conservation.
The use of artificial sediment in toxicity testing is required in studies assessing the impact of chemicals on benthic communities and testing chemicals for registration, as it offers increased repeatability and enhances consistency across bioassays and laboratories. While peat is conventionally incorporated into artificial sediment formulations for toxicological testing as the source of organic matter (OM), it is not a sustainable resource due to the long time it takes to form and the environmental impact of its extraction (carbon loss equivalent to 2.1 megatons of carbon dioxide emissions annually and potential release of other elements into the environment). In a preliminary effort, the usability of coconut husk as an alternative to peat in formulating artificial sediments for toxicity testing was assessed. Post-larval American lobsters (Homarus americanus) were exposed for 15 days to test sediment prepared with either peat or coconut husk as a source of OM. Three OM levels were tested: 0.5, 2.3, and 5
The Indian Bay of Bengal coast represents an important marine fishing area and serves as a significant source of dietary protein for consumers. These highlights growing concerns regarding the bioaccumulation of potentially toxic elements (PTEs) in fish and the associated environmental and human health risks linked to their consumption. Thus, this study aims to assess the accumulation of PTEs (As (arsenic), Sr (strontium), Ti (titanium), Be (beryllium), Cd (cadmium), Sb (antimony), Pb (lead), Hg (mercury), and Li (lithium)) in the muscles of three groupers. The highest value (29.88 ± 0.17 µg/g dry wt.) was observed in Epinephelus areolatus for As, and the lowest in E. bleekeri for Be (0.59 ± 0.20 µg/kg dry wt.). Multivariate analyses (PCA and HCA) confirmed a very close relationship among the As, Cd, Sb, Ti, and Be, while Pb, Hg, Th, and Li had their own specific habitats of bioaccumulation, implying distinct inputs of PTE. Health risk evaluations reveal that As was the most hazardous PTE, with the incidence of toxic effects and cancer risk, especially for habitual consumers. However, Ti is the most hazardous PTE, posing the highest non-carcinogenic risk of the studied species. Metal–protein docking further showed the strong binding of Pb2+ and Sr2+ to Nrf2, p53, and DNMT1, indicating potential disruption of redox regulation, genomic stability, and epigenetic control. The findings indicate that the intake of the specified grouper species poses a significant arsenic-related health risk to people, underscoring the necessity for ongoing monitoring and management of PTE contaminants in fisheries.
With increasing petroleum transport through coastal waters of the northeastern Pacific, understanding the ecological hazards of these mixtures is essential, yet the sublethal impacts on benthic filter feeders remain poorly characterized. The sub-chronic (7-d exposure) effects of crude oil (CO), marine diesel (MD), and diluted bitumen (DB) water-accommodated fractions (WAFs) on the scope for growth (oxygen consumption, food absorption efficiency, and clearance rate), as well as gonadal and digestive gland histopathology in Pacific oysters (Crassostrea gigas) were examined. Initial total polycyclic aromatic compound (TPAC) concentrations in WAFs were ranked CO > MD > DB, which also accumulated within oyster tissues in the same order. Sub-chronic exposures to different WAF dilutions of three petroleum products did not alter any measured endpoints in Pacific oysters. PACs accumulated in oyster tissues had a rapid depuration after the exposure period due to the potential biotransformation of parent hydrocarbons, which could elucidate the lack of significant adverse effects. These results indicate that adult C. gigas maintained physiological and tissue-level integrity under transient, declining petroleum exposure conditions, providing context for interpreting potential impacts of short-term nearshore spill scenarios.
Although evidence of adverse effects on human health is still scarce, exposure to pesticides has been shown to have a negative impact outside agroecosystems. We compared exposure to current use pesticides in one area surrounded by crop fields (Area 1) with exposure in another area located in the center of a city (Area 2). Both areas are located within one of the world’s most agriculturally productive regions. We analyzed the presence of glyphosate and its metabolite aminomethylphosphonic acid (AMPA) in urine samples from 86 female volunteers who had been environmentally exposed. Potential genotoxicity was tested using the micronucleus assay on buccal mucosa samples taken from the same donors. We also evaluated the residues of current use pesticides in drinking water and soil samples taken from public places in the two areas. All the analytical determinations were performed using ultra-high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS/MS). When glyphosate-positive samples were considered, significantly higher concentrations of glyphosate were found in Area 1 than in Area 2 (p = 0.02). An increase in the frequency of binucleated cells (p = 0.039) and linked nucleus (p = 0.035) was observed in women from Area 1 compared to those from Area 2. All the drinking water samples tested positive for at least two pesticide residues while 85
In this study, we assessed the genotoxicity induced by the exposure to polylactic acid microplastics (PLA-MPs) and/or antibiotic sulfamethoxazole (SMX) in the marine bivalve species, Pacific oyster Crassostrea gigas. Oysters were exposed for 14 days to three experimental conditions: PLA-MPs alone, SMX alone and mixing PLA-MPs + SMX at concentrations of five mg/L for PLA-MPs and 64 ng/L for SMX. To achieve this goal, the mediated genotoxicity was assessed by analyzing biomarker such as micronucleus frequency, the degree of DNA damage and changes in gene expressions in gill tissues of exposed oysters, while PLA-MPs accumulation were analyzed in gill tissues. Results show that PLA-MPs were the primary driver of microplastic accumulation in oyster gill tissues, whereas co-exposure PLA-MPs + SMX resulted in significant but reduced accumulation compared with PLA-MPs. Exposure to PLA-MPs or in combination with antibiotic SMX induced additional transmission pathways which lead to DNA damage and micronucleus formation. However, oysters co-exposed to PLA-MPs + SMX exhibited increased susceptibility to micronucleus in gill tissues, whereas the extent of DNA damage in oysters exposed to PLA-MPs alone was comparable to that observed under combined PLA-MPs + SMX exposure. Notably, combination of PLA-MPs and SMX amplified oxidative stress and detoxification responses in oysters through SMX modulating PLA-induced antioxidant signaling. This combination exacerbated suppression of key detoxification pathways, thereby potentially increasing susceptibility to genotoxic effects in contaminated aquatic environments. The research findings of this study provide relevant insights for toxicology related to bio-based microplastics.
In this study, for the first time, the distribution and fractionation of persistent organic pollutants (POPs) along both latitudinal and longitudinal transects are investigated simultaneously in surface soil. Distributions and fractionations of two isomers of hexachlorocyclohexane (HCH), α- and β-HCH, were analyzed in Chinese rural surface soil using a set of soil samples collected in 2005, 2012, and 2019 to study the lifecycle of the global/latitudinal fractionation of POPs. The primary longitudinal and secondary latitudinal distributions for each of α- and β-HCH and the primary longitudinal and secondary latitudinal fractionations formed together by these two isomers were observed concurrently in these three years. Based on these observations, we identified three stages for the lifecycle of global/latitudinal fractionation for α- and β-HCH. In Stage I, the primary fractionation of the two isomers mainly due to the distance from the source regions begins and intensifies with the fractionation pattern of the enrichment of the compounds having lower KOA (octanol-air partition ratio) values and the depletion of the compounds with higher KOA values. In Stage II, the secondary fractionation caused mainly by the air temperature gradient along latitudinal directions begins and intensifies, keeping the fractionation patterns appearing in the Stage I unchanging. In Stage III, the secondary fractionation is weakened primarily caused by local factors, degradation in particular, until its disappearance. The fractionation patterns appearing in the Stages I and II may change. Although the present study on the three stages for the development of latitudinal fractionation is for α- and β-HCH in Chinese surface soil. We believe that the findings presented in this study can provide instructive implications for the lifecycle of all polar cold-trapped persistent organic pollutants (PCT‑POPs) on a global scale.
The kairomone released by larvae of phantom midges in the genus Chaoborus induces a defensive morphology in Daphnia pulex. This morphology effectively reduces predation pressure and prevents a sharp decline in prey population density. Copper, an anthropogenic pollutant, inhibits the induction of defense mechanisms. However, the consequences of this inhibition at the population level are unclear. In the present study, we confirmed the inhibitory effect of copper on morphological defense using an individual-level experiment (Exp. 1) and further evaluated the effects in a bi-trophic population-level experiment (Exp. 2). In Exp. 1, the induction of neckteeth was inhibited by copper at a concentration of 53.9 nM (Cu2+ activity of 0.26 nM). In Exp. 2, no inhibition was detected at a total concentration of 100 nM (on day 12); however, inhibitory effects were observed as Cu2+ activity increased. These results indicate that the inhibition of morphological changes cannot be explained by the total copper concentration but by the concentration of free copper ions. The copper-induced suppression of morphological defense resulted in a decrease in the abundance of juvenile Daphnia due to increased predation pressure from Chaoborus larvae. In conclusion, these findings emphasize the potential for environmental copper pollution to alter population dynamics and prey–predator relationships in aquatic taxa.
The Fildes Peninsula (maritime Antarctica) is greatly affected by global warming and local human impacts since it is in one of the Antarctic regions with the highest intensity of human activity. To establish the effect of human activities on Fildes Peninsula lakes, we compared trends in diatom assemblages, bacterial communities and metal concentrations in sediment cores from two lakes close to human infrastructure with those in a more remote lake. In the two lakes close to stations and the airport, we found heavy metal enrichments and diatom teratologies, as well as notable changes in diatom assemblages in one of these lakes, roughly coincident with the time when the first two stations were built ( 1970). Due to the known association between diatom teratologies and metal enrichment, metal stress is a convincing explanation for these changes. Certain bacterial taxa determined to be indicators of pollution were also found to be more abundant in the impacted lakes in recent sediments (i.e., Hungateiclostridiaceae, OPB41, Anaerovorax and Leptolinea). Metal, diatom and bacteria changes observed in the lake more distant to infrastructure were more subtle and are likely related to climate change alone. Given the proximity of the affected lakes to the airport and roads, our data suggests that transportation infrastructure and activity on Fildes Peninsula is likely a key cause of contamination in the region’s ecosystems. This study provides important insights into how human activities and climate change have affected Fildes Peninsula aquatic ecosystems and how they may respond to future stressors.
The accumulation of metals in estuarine environments represents potential threats to both aquatic ecosystems and human health, highlighting the need for reliable in situ biomarkers of contamination. In the burrowing crab Neohelice granulata (Brachyura, Varunidae), previous studies have shown that biochemical markers may be influenced by seasonal fluctuations, limiting their usefulness for assessing spatial patterns of metal exposure. Recently, morphological variation among organisms has been proposed as a biomarker of environmental stress. Building on this framework, the present study aimed to evaluate whether metal contamination is associated with morphological changes in N. granulata populations from different sites of the Bahía Blanca estuary, Argentina. Concentrations of Cd, Cr, Cu, Fe, Pb, and Zn were measured in sediments and crab hepatopancreas, and their relationship with carapace and chelipeds morphology were analyzed by traditional and geometric morphometric approaches. Results revealed similar spatial patterns of metal loads in sediments and crab hepatopancreas, reflecting differences in anthropogenic impacts among sites. Moreover, significant morphological differences in carapaces were detected among crab populations, consistent with the observed gradient of metal contamination. These findings suggest that metal exposure may be associated with the morphological variation in N. granulata and highlight its potential use as a biomarker of environmental stress in estuarine crabs. Finally, geometric morphometrics of the carapace proved more effective than traditional chelipeds morphometrics in distinguishing crabs’ populations exposed to different levels of metal contamination.