Field-based molecular detection of Batrachochytrium dendrobatidis in critically endangered Atelopus toads and aquatic habitats in Ecuador.

Batrachochytrium dendrobatidis (Bd) is a lethal fungal species that parasitizes vertebrates and is associated with the worldwide decline of amphibian populations. The development of sensitive, rapid detection methods, particularly DNA-based techniques, is critical for effective management strategies. This study evaluates the efficacy of DNA extraction and a portable PCR device in a mountable field laboratory setup for detecting Bd near the habitats of three critically endangered Atelopus toad species in Ecuador. We collected skin swabs from Atelopus balios, A. nanay, and A. bomolochos, and environmental DNA (eDNA) samples from streams in Andean and coastal regions of Ecuador. For eDNA, a comparison was made with duplicates of the samples that were processed in the field and in a standard university laboratory. Our findings revealed Bd detection in eDNA and swabs from 6 of 12 water samples and 10 of 12 amphibian swab samples. The eDNA results obtained in the field laboratory were concordant with those obtained under campus laboratory conditions. These findings highlight the potential of field DNA-based monitoring techniques for detecting Bd in amphibian populations and their aquatic habitats, particularly in remote areas. Furthermore, this research aligns with the National Action Plan for the Conservation of Ecuadorian Amphibians and contributes to the global effort to control this invasive and deadly fungus.

A systematic review on metal contamination due to mining activities in the Amazon basin and associated environmental hazards.

Metal contamination associated with mining activities has been considered one of the main environmental pollution problems in the Amazon region. Understanding the levels of metal contamination from mining activities requires a good understanding of background metal concentrations, which may vary notably according to the geology/lithology characteristics of the region, soil type, and predominant biogeochemical processes. This review assessed 50 papers and reports published between 1989 and 2020 describing environmental concentrations of different metals and metalloids (As, Hg, Mn, Fe, Cd, Cu, Cr, Pb, Ni, and Zn) in water and sediments of mining and non-mining areas in five geographic regions of the Amazon basin. Metal enrichment caused by mining activities was calculated and exposure concentrations were compared with sediment and water quality standards set for the protection of aquatic life. Significant enrichments of Cd, Cu, Cr, Fe, Hg, Mn, Ni and Zn were observed in mining areas in both sediment and water. Regarding background levels in the different geographic regions, the highest prevalence of metal enrichment (i.e., concentrations 10 to 100-fold higher than mean background values) in sediment samples was found for Fe (100% of samples), Ni (90%), and Mn (69%). For water, high prevalence of metal enrichment occurred for Zn, Mn, and Fe (100% of samples), and for Hg (86%). Hg, Fe, Pb, Cu, Cd, Ni and Zn exceeded water and/or sediment quality standards in a significant number of samples in the proximity of mining areas. This study indicates that mining activities significantly contribute to water and sediment contamination across the Amazon basin, posing hazards for freshwater ecosystems and potentially having human health implications.

Effects of intensive agriculture and urbanization on water quality and pesticide risks in freshwater ecosystems of the Ecuadorian Amazon.

The Ecuadorian Amazon has experienced a significant land use change due to the demographic increase and the expansion of the agricultural frontier. Such changes in land use have been associated to water pollution problems, including the emission of untreated urban wastewater and pesticides. Here we provide the first report on the influence of urbanization and intensive agriculture expansion on water quality parameters, pesticide contamination and the ecological status of Amazonian freshwater ecosystems of Ecuador. We monitored 19 water quality parameters, 27 pesticides, and the macroinvertebrate community in 40 sampling locations of the Napo River basin (northern Ecuador), including a nature conservation reserve and sites in areas influenced by African palm oil production, corn production and urbanization. The ecological risks of pesticides were assessed using a probabilistic approach based on species sensitivity distributions. The results of our study show that urban areas and areas dominated by African palm oil production have a significant influence on water quality parameters, affecting macroinvertebrate communities and biomonitoring indices. Pesticide residues were detected in all sampling sites, with carbendazim, azoxystrobin, diazinon, propiconazole and imidacloprid showing the largest prevalence (>80% of the samples). We found a significant effect of land use on water pesticide contamination, with residues of organophosphate insecticides correlating with African palm oil production and some fungicides with urban areas. The pesticide risk assessment indicated organophosphate insecticides (ethion, chlorpyrifos, azinphos-methyl, profenofos and prothiophos) and imidacloprid as the compounds posing the largest ecotoxicological hazard, with pesticide mixtures potentially affecting up to 26-29% of aquatic species. Ecological risks of organophosphate insecticides were more likely to occur in rivers surrounded by African palm oil plantations, while imidacloprid risks were identified in corn crop areas as well as in natural areas. Future investigations are needed to clarify the sources of imidacloprid contamination and to assess its effects for Amazonian freshwater ecosystems.

Metal bioaccumulation and genotoxicity in Oreochromis niloticus reared in farming pools influenced by mining activities in Napo, in the Ecuadorian Amazonia.

Mining areas may suffer long-term metal contamination and represent harmful remnants of former mining activities. In the northern Amazon of Ecuador, former mining waste pits are used in Oreochromis niloticus (Nile tilapia) fish farming. Given the high consumption of this species by the local population, we aimed to estimate human consumption risks by determining Cd, Cu, Cr, Pb, and Zn tissue bioaccumulation (liver, gills, and muscle) and genotoxicity (micronucleus essay) in tilapia cultivated in one former mining waste pit (S3) and compare the findings to tilapias reared in two non-mining areas (S1 and S2); 15 fish total. Tissue metal content was not significantly higher in S3 than in non-mining areas. Cu and Cd were higher in the gills of tilapias from S1 compared to the other study sites. Higher Cd and Zn were detected in the liver of tilapias from S1 compared to the other sampling sites. Cu was higher in the liver of fish from S1 and S2, and Cr, in the gills of fish from S1. The highest frequency of nuclear abnormalities was observed in fish from S3, indicating chronic exposure to metals at this sampling site. The consumption of fish reared at the three sampling sites results in a 200-fold higher Pb and Cd ingestion than their maximum tolerable intake thresholds. Calculated estimated weekly intakes (EWI), hazard quotients (THQ), and Carcinogenic Slope Factors (CSFing) denote potential human health risks, indicating the need for continuous monitoring in this area to ensure food safety not only in areas affected by mining, but in general farms in the region.

Multi-level responses of oysters Crassostrea virginica for assessing organochlorine pesticides in a Ramsar coastal lagoon in southern Mexico.

Organochlorine pesticides (OCPs) have been intensively used without proper regulation and control in Latin America due to the prevalence of diseases and pests, thus posing potential risks to nontarget organisms. Initiatives for ecosystem preservation, such as to designate protected areas, may not be enough to avoid contamination by OCPs, considering that protected areas tend to be permeable to diffuse sources. Here, we investigate multi-level responses of the oyster Crassostrea virginica to OCPs in Laguna de Términos, a RAMSAR coastal lagoon in the southern Gulf of Mexico. For this aim, OCPs occurrence and concentrations in the water, sediment, and in oysters from 3 settlement banks were assessed. Enzymatic and non-enzymatic biochemical biomarkers were quantified in the oysters' mantle and digestive gland, and the human health risk due to oyster consumption was also evaluated. OCPs in water were below detection limits. Fourteen OCPs were detected in sediments (∑OCPs mean of 49 ngg-1) and 7 in oyster tissues (∑OCPs mean of 121 ngg-1). The occurrence of OCPs was related to the land uses along the watersheds of the rivers that drain into the lagoon. Biochemical responses were correlated with OCPs (∑HCH, ∑DDT, heptachlor and endosulfan) in sediment, and oyster tissues. OCPs in oyster tissues showed a strong association with pro-oxidant forces and oxidative stress responses (Superoxide dismutase, Catalase, Glutathione Peroxidase, and lipid peroxidation), and neurotoxicity (Acetylcholinesterase), suggesting that the current OCPs contamination exerts significant stress. Our study also shows that the consumption of oysters from the lagoon increases the potential human health risk. Considering that Laguna de Términos is a protected Ramsar site, we suggest that environmental protection measures should be increased and that a monitoring program for OCPs exposure is necessary to assess the effects on this ecosystem.

Occurrence of emerging contaminants in surface water bodies of a coastal province in Ecuador and possible influence of tourism decline caused by COVID-19 lockdown.

Emerging contaminants in water bodies is an issue of concern due to their impact on the ecosystem and human health. The COVID-19 pandemic has forced the implementation of protective measures such as social distancing, lockdowns, and remote work, which have affected the tourism influx. This study aimed to evaluate the occurrence of emerging pollutants in bodies of water in Esmeraldas, a coastal province of Ecuador, before and during the COVID-19 pandemic in a highly touristic region. For this purpose, surface waters from 14 beaches and ten river mouths were sampled at two-time points in November 2019 and November 2020. Compounds widely consumed in Ecuador: acetaminophen, caffeine, sodium diclofenac, trimethoprim, and sulfamethoxazole were extracted from water samples by solid phase extraction SPE and detected with a UPLC-QTOF-MS system. We found a decrease in the occurrence of caffeine from 100 % to 4.2 % of caffeine and 25 % to 0 % of diclofenac, likely related to the decline in tourist afflux due to the lockdown measures. Most of the compounds diminished in terms of frequency and/or concentration; however, as COVID-19 treatments make use of different pharmaceutical compounds such as antivirals, antibiotics, antiparasitics, or glucocorticoids, future studies should include these to assess their environmental impact.

Human health risk assessment of metals and metalloids in mining areas of the Northeast Andean foothills of the Ecuadorian Amazon.

Gold mining (GM) is a major source of metals and metalloids in rivers, causing severe environmental pollution and increasing the exposure risks to the residents of surrounding areas. Mining in Ecuadorian Amazonia has dramatically increased in recent years, but its impacts on Indigenous local populations that make use of rivers are still unknown. The aim of this study was to assess the risks to adults and children caused by the exposure to metals and metalloids in freshwater ecosystems contaminated with tailings released by GM activities in 11 sites of the upper Napo River basin, Ecuador. We selected a carcinogenic and a noncarcinogenic risk assessment method to estimate the hazard index (HI) and total cancer risk (TCR). The concentration of Ag, Al, As, Cd, Cu, Fe, Mn, Pb, Zn, B, and V in water and sediment samples was considered to assess the risks to human health. The calculated HI was 23-352 times greater than the acceptable limits in all sites for both children and adults. Mn and Fe were the main contributors (75% in water and 99% in sediment) to the total calculated risk based on the HI. The calculated TCR for children and adults exceeded approximately one to three times the permissible threshold in all sites. As and Pb contributed up to 93% of the total calculated risk based on TCR for both children and adults. This study demonstrates that the emission and mobilization of metals and metalloids caused by mining activities increase the risk to human health, to which we recommend further monitoring of freshwater contamination in the area and the implementation of preventive health management measures. Integr Environ Assess Manag 2023;19:706-716. © 2022 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

The synergistic effect of microplastic and malathion exposure on fiddler crab Minuca ecuadoriensis microplastic bioaccumulation and survival.

We assessed the combined effects of polyethylene microplastic (MP) and malathion (MLT) on the survival of the fiddler crab Minuca ecuadoriensis, and MP tissue bioaccumulation in four treatments following 120 h exposure: T1) Control; T2) MLT 50 mg L-1; T3) MP 200 mg L-1; and T4) MLT (50 mg L-1) + MP (200 mg L-1). The highest mortality (80%) was in T4, followed by T2 (28%) and no mortality was in T3. Higher MP bioaccumulation was observed in T4 (572 items g tissue-1) followed by T3 (70 items g tissue-1). Our findings indicate that the synergistic effect of MLT and MP increased M. ecuadoriensis bioaccumulative capacity and decreases survival. Thus, as MP contamination in aquatic environments is ubiquitous, our study raises a warning on the synergistic effects of MP with other environmental contaminants and serves as a baseline for further studies.

Microplastics in a tropical Andean Glacier: A transportation process across the Amazon basin?

Microplastic (MPs) contamination is ubiquitous in most terrestrial and aquatic ecosystems. Recently MPs have been reported at high altitudes which indicates that air masses can transport and deposit MPs in the surface snow of high mountain ecosystems, however, whether MPs typification and abundance can be influenced by direction and origin of air masses still remains an open question. Here we present the first report of MPs above 5000 m a.s.l from surface snow of a glacier in the tropical Andes. We collected surface snow along an elevational gradient, from 5000 to 5400 m a.s.l., in the Antisana Glacier, in the northern Andes cordillera of Ecuador to analyze MPs abundance and polymeric identification with the Fourier Transform Infrared (FTIR) and also to hypothesized the possible MPs sources in this remote area by comparing the oxygen and hydrogen stable isotopic ratio composition of the snow samples and by analyzing the wind direction. We observed an average of 131 ± 24 MPs L-1 in our samples. Fibers corresponded to 70% of all MP shapes; FTIR results showed that MPs composition mainly included polyurethane, polyethylene, polyamide, polyester, and high-density polyethylene in surface snow. There were no statistically significant differences of MPs abundance among sampled elevations, and the isotopic ratio composition did not differ among locations. Our results suggest that MP that accumulated in the glacier may be transported from the east, across the Amazonia, by the prevalent eastward air flow. The absence of industrial cities at least 2000 km further east from Antisana, indicates that the remote Andean glaciers could constitute important depositional zones for long-distance transported contaminants.

Microplastics in rivers and coastal waters of the province of Esmeraldas, Ecuador.

This study represents the first assessment of microplastic (MP) contamination in the coastal area of the Esmeraldas Province, Ecuador. MPs were quantified in 14 coastal waters in beaches with different urbanization level and in 10 rivers. The most abundant MP types were transparent fibres, brown fragments, grey fragments, transparent fragments, and black fragments, which together represented 84% of the total count. Coastal waters presented significantly higher quantities of MP than rivers. No difference in microplastic abundance was detected between beaches with higher and lower urban occupation, nor between beaches facing North or West. Our results indicate that MP contamination is widespread, and most likely transported from multiple sources. Our results can serve as a baseline for future MP monitoring in the area.

Integrating multiple lines of evidence to assess freshwater ecosystem health in a tropical river basin.

Degradation of freshwater ecosystems by uncontrolled human activities is a growing concern in the tropics. In this regard, we aimed at testing an integrative framework based on the IFEQ index to assess freshwater ecosystem health of river basins impacted by intense livestock and agricultural activities, using the Muchacho River Basin (MRB) as a case study. The IFEQ combine multiple lines of evidence such as riverine hydromorphological analysis (LOE 1), physicochemical characterization using ions and pesticides (LOE 2), aquatic macroinvertebrate monitoring (LOE 3), and phytotoxicological essays with L. sativa (LOE 4). Overall, results showed an important reduction in streamflow and an elevated increase in ion concentrations along the MRB caused by deforestation and erosion linked to agricultural and livestock activities. Impacts of the high ion concentrations were evidenced in macroinvertebrate communities as pollution-tolerant families, associated with high conductivity levels, represented 92 % of the total abundance. Pollution produced by organophosphate pesticides (OPPs) was critical in the whole MRB, showing levels that exceeded 270-fold maximum threshold for malathion and 30-fold for parathion, the latter banned in Ecuador. OPPs concentrations were related to low germination percentages of Lactuca sativa in sediment phytotoxicity tests. The IEFQ index ranged from 44.4 to 25.6, indicating that freshwater ecosystem conditions were "bad" at the headwaters of the MRB and "critical" along the lowest reaches. Our results show strong evidence that intense agricultural and livestock activities generated significant impacts on the aquatic ecosystem of the MRB. This integrative approach better explains the cumulative effects of human impacts, and should be replicated in other basins with similar conditions to help decision-makers and concerned inhabitants generate adequate policies and strategies to mitigate the degradation of freshwater ecosystems.

An Integrative Approach to Assess the Environmental Impacts of Gold Mining Contamination in the Amazon.

As the number of legal and illegal gold mining sites increases in the Andes-Amazonia region, integrative methods to evaluate the effects of mining pollution on freshwater ecosystems are of paramount importance. Here, we sampled water and sediments in 11 sites potentially affected by gold mining activities in the Napo province (Ecuador). The environmental impacts were evaluated using the following lines of evidence (LOEs): water physicochemical parameters, metal exposure concentrations, macroinvertebrate community response (AAMBI), and toxicity by conducting bioassays with Lactuca sativa and Daphnia magna. Dissolved oxygen and total suspended solids were under (<80%) and above (>130 mg/Ls) quality standards 65% of the sites. Ag, Al, As, Cd, Cu, Fe, Mn, Pb, and Zn in water and V, B, and Cr in sediments were detected above quality standards at sampled sites. Nine out of eleven sites were classified as having bad environmental quality based on the AAMBI. L. sativa seed germination in both water (37% to 70%) and sediment (0% to 65%) indicate significant toxicity. In five sites, neonates of D. magna showed a 25% reduction in survival compared to the control. Our integrated LOEs index ranked sites regarding their environmental degradation. We recommend environmental impact monitoring of the mining expansion at the Andes-Amazonia region using multiple LOEs.

Assessment of Microplastic and Organophosphate Pesticides Contamination in Fiddler Crabs from a Ramsar Site in the Estuary of Guayas River, Ecuador.

We assessed the concentration of organophosphate pesticides (OPs) and microplastics (MPs) in water and sediments from the burrows, and tissues of the fiddler crabs Leptuca festae and Minuca ecuadoriensis, from Isla Santay, a Ramsar site in the estuary of the Guayas River, Ecuador. MPs concentrations in the burrows were 660 ± 174.36 items kg-1 (w.w.) and 26 ± 1 items L-1 in collected sediments and water, respectively. Regarding OPs, water and sediment concentrations were up to 26 times above the USEPA thresholds for chronic exposure, indicating environmental risk. MPs were found in tissues collected from both species. The highest abundance was in the gills followed by the digestive tract and hepatopancreas. OPs concentrations in tissues were below the detection limits. Because fiddler crabs are chronically exposed to environmental contamination, they are suitable bioindicators to monitor Isla Santay and to comprehend human impacts in coastal environments of Ecuador.

Preliminary Assessment of Plastic Litter and Microplastic Contamination in Freshwater Depositional Areas: The Case Study of Puerto Misahualli, Ecuadorian Amazonia.

We quantify plastic litter (PL, > 2 cm) and microplastics (MP, < 5 mm) from the sediments of a beach formed at a riverine depositional area, at the upper Amazon River basin, Ecuador. In the collection area (4400 m2), the PL density was 0.045 items m-2, where low-density polyethylene bags were the prevalent PL. The beach was classified as "very clean" (Clean Coast Index (CCI) of 1.3 items m-2). Regarding MP, in 55 sampling stations, average MP concentrations ranged from 0 to 2200 items kg-1 of dry sediment (0.5-2 mm), and 0-4200 items kg-1 of dry sediment (2-5 mm). Blue fibers were the prevalent MP. Our results represent the first report to show the ubiquitous presence of PL and MP for the area. The monitoring and management of plastic disposal in freshwater beaches are necessary, as here we report a small part of an undocumented issue.

Assessing the Quality of Amazon Aquatic Ecosystems with Multiple Lines of Evidence: The Case of the Northeast Andean Foothills of Ecuador.

We assessed the quality of Andes-Amazonia streams in Ecuador impacted by gold mining (GM), discharges from inefficient sewage network in urban areas (UA), wastes from fish farming (FF) and from non-functional landfill (LF) and other few threats (FT). We selected three lines of evidence (LOE) that were used separately and integrated into a index: water quality (WQI) and macroinvertebrate community (AAMBI) indices and phytotoxicity tests. Streams affected by UA and LF had the lowest scores to WQI and phytotoxicity, and by GM had the lowest scores to AAMBI. Macroinvertebrate absence in GM should be considered as a warning signal of long-term mining impacts in the area. The integrated LOE index showed that sites with identified threats had 30%-53% stream quality decline compared to FT sites. The use of the selected LOE seems to be a useful tools for long-term monitoring and evaluation of this sensitive aquatic ecosystem.

Aerogels made of chitosan and chondroitin sulfate at high degree of neutralization: Biological properties toward wound healing.

In this study, highly neutralized, highly porous, and ultralight polymeric aerogels prepared from aqueous colloidal suspensions of chitosan (CS) and chondroitin sulfate (ChS) nanocomplexes, formulated as quasi-equimolar amounts of both, are described. These aerogels were designed as healing agents under the inspiration of minimizing the amount of matter applied to wounds, reducing the electrostatic potential of the material and avoiding covalent cross-linkers in order to decrease metabolic stress over wounds. Aerogels synthesized under these criteria are biocompatible and provide specific properties for the induction of wound healing. They do not affect neither the metabolic activity of cultured 3T3 fibroblasts nor the biochemical parameters of experimental animals, open wounds close significantly faster and, unlike control wounds, complete reepithelialization and scarring can be attained 14 days after surgery. Because of its hydration abilities, rapid adaptation to the wound bed and the early accelerator effect of wound closure, the CS/ChS aerogels appear to be functional inducers of the healing. Previous information show that CS/ChS aerogels improve wound bed quality, increase granulation tissue and have pain suppressive effect. CS/ChS aerogels are useful as safe, inexpensive and easy to handle materials for topical applications, such as skin chronic wounds. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2464-2471, 2018.

Antioxidants and the integrity of ocular tissues.

Oxygen-derived free radicals are normally generated in many pathways. These radicals can interact with various cellular components and induce cell injury. When free radicals exceed the antioxidant capacity, cell injury causes diverse pathologic changes in the organs. The imbalance between the generation of free radicals and antioxidant defence is known as oxidative stress. The eye can suffer the effect of oxidative damage due to the etiopathogenesis of some pathological changes related to oxidative stress. This paper reviews the role of oxidative stress in the onset and progression of damage in different eye structures, the involvement of the antioxidant network in protecting and maintaining the homeostasis of this organ, and the potential assessment methodologies used in research and in some cases in clinical practice.