Application of ecotoxicological tools to evaluate the quality status of mangroves under restoration in the Yucatán Peninsula, Mexico.

Ecotoxicological tools, namely biomarkers and bioassays, may provide insights on the ecological quality status of mangroves under restoration. We investigated how 1) physicochemical parameters and water bioassays using Artemia franciscana; and 2) quantification of sublethal (osmoregulatory capacity, biochemical, and oxidative stress) and individual biomarkers (density, length-weight relationship [LWR], parasitic prevalence) in the sentinel fiddler crab Minuca rapax, can improve restoration indicators in mangroves from the Yucatán Peninsula, Southern Gulf of Mexico. We showed that water quality was improved with restoration, but still presented toxicity. Regarding sublethal biomarkers, M rapax from restored areas lower osmotic regulatory capacity, higher oxidative stress, and showed lipid peroxidation. As to the individual biomarkers, the density, LWR, and the prevalence of parasites in M. rapax was higher in restored areas. The use of bioassays/biomarkers were useful as early warning indicators to better assess the health of mangroves under restoration.

Integrated evaluation of the biological response of the earthworm Eisenia fetida using two glyphosate exposure strategies: soil enriched and soils collected from crops in Southeastern Mexico.

Under laboratory conditions, the toxicological effects of pesticides tend to be less variable and realistic than under field conditions, limiting their usefulness in environmental risk assessment. In the current study, the earthworm Eisenia fetida was selected as a bioindicator for assessing glyphosate toxic effects in two different trials to solve this dilemma. In Trial 1, the worms were exposed for 7 and 14 days to concentrations of a commercial glyphosate formulation (1 to 500 mg a.i. kg-1) currently used in the field. In Trial 2, the worms were kept in nine soils collected from different plots with crops for 14 days of exposure. In both experiments, glutathione S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and acetylcholinesterase (AChE) activities and contents of lipid peroxidation (LPO) were evaluated. In T1, the glyphosate formulation produced a 40% inhibition of AChE activity and a significant increase in GST, SOD, CAT, and GPx activities and LPO contents in E. fetida on day 7. In T2, higher concentrations of glyphosate were detected in the soils of soybean, papaya, and corn (0.92, 0.87, and 0.85 mg kg-1), which induced a positive correlation between the levels of glyphosate residues with GST, SOD, CAT, GPx, and LPO and a negative correlation with AChE. These findings indicate that crop soils polluted with glyphosate elicited higher oxidative stress than under laboratory conditions, confirmed by IBRv2, PCA, and AHC analyses.

Biochemical Response of the Endogeic Earthworm (Balanteodrilus extremus) Exposed to Tropical Soils.

This work evaluated the biochemical responses of the endogeic earthworm Balanteodrilus extremus exposed for 14 and 48 days (d) to soils collected from two tropical agricultural systems: maize-sorghum (MS) and soybean-sorghum (SS). A soil without agricultural management (WAM) and the use of pesticides was selected as a reference. The presence of organochlorine (OC) and organophosphate (OP) pesticide residues was quantified in MS and SS soils. Biomarkers of detoxification [glutathione S transferase (GST)], neurotoxicity [acetylcholinesterase (AChE)] and oxidative stress [superoxide dismutase (SOD), catalase (CAT) and lipoperoxidation (LPO)] were evaluated in B. extremus. The concentration of OP pesticide residues was higher in SS than in MS. Activity of AChE in B. extremus exposed to SS soil for 14 d was significantly more inhibited (78%) than in MS soil (68%). B. extremus has been shown to be a good bioindicator of contaminated soils in tropical regions.

Synergistic effects of microplastic and lead trigger physiological and biochemical impairment in a mangrove crab.

Microplastics (MP) are vectors for other environmental contaminants, such as metals, being a considerable problem, especially in the aquatic ecosystem. To investigate the combined effects of MP (high density polyethylene) with lead (Pb), we exposed the mangrove fiddler crab Minuca vocator to Pb (50 mg L-1), and MP (25 mg L-1) alone and in mixture, for 5 days. We aimed to determine Pb and MP bioaccumulation, as well as physiological (oxygen consumption and hemolymph osmolality) and biochemical (superoxide dismutase, catalase, glutathione peroxidase, and lipid peroxidation) traits effects. Co-exposure of MP and Pb significantly increased the bioaccumulation of Pb, but reduced MP tissue accumulation. Regarding the physiological traits, increasing osmolality and oxygen consumption rates compared to the control were observed, particularly in the combined Pb and MP exposure. As to biochemical traits, the combination of Pb and MP induced the most significant responses in the enzymatic profile antioxidant enzyme activity. The catalase (CAT), glutathione peroxidase (GPx), and dismutase superoxide (SOD) decreased compared to individual exposure effects; the combination of MP and Pb had a synergistic effect on promoting lipid peroxidation (LPO). The co-exposure of MP and Pb acted synergistically when compared to the effects of the isolated compounds. Due to the increasing MP contamination in mangroves, more severe physiological and biochemical effects can be expected on mangrove crabs exposed to metal contamination.

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.

Distribution of heavy metals in crop soils from an agricultural region of the Yucatan Peninsula and biochemical changes in earthworm Eisenia foetida exposed experimentally.

Heavy metals (HM) are natural components of agricultural soils. However, excessive use of agrochemicals (fertilizers and pesticides) can increase the concentration of these elements, making them harmful to crops and soil biota. Therefore, this study aimed to evaluate the concentrations of 6 HM (Cr, Mn, Cu, Zn, Al, and Fe) in soils collected from a typical agricultural region of the Yucatan Peninsula, Mexico, and relate their effects on weight, biotransformation (EROD, GST), and oxidative stress (SOD, CAT, LPO, and MT) in the earthworm Eisenia foetida. The results showed different concentrations of HM accumulation in soils, in the following order, Fe (598.74 ± 223.81 µg g-1), Al (145.30 ± 33.54 µg g-1), Cr (88.38 ± 36.23 µg g-1), Mn (69.53 ± 22.48 µg g-1), Zn (54.10 ± 12.04 µg g-1), and Cu (24.91 ± 12.40 µg g-1), the soils with maize, soybean, and chihua squash crops being the ones with the highest distribution and availability of these elements. These concentrations of metals did not produce mortality. However, Cr, Fe, and Al induced a significant biomass loss of E. foetida. The integrated biomarker response version 2 revealed that soil with maize cultivation was affected the most, with the earthworms exposed to this soil showing greater variations in the dominant biological responses including EROD, TBARS, SOD, and MT (IBRv2 = 9.73). PCA analysis indicated a strong positive relationship between these oxidative stress biomarkers and Cu and Zn concentrations. These findings provide a reference guide that should be part of the monitoring and management programs for different agricultural types in the region under study.

Banded tetra (Astyanax aeneus) as bioindicator of trace metals in aquatic ecosystems of the Yucatan Peninsula, Mexico: Experimental biomarkers validation and wild populations biomonitoring.

Bioindicator organisms are important tools in environmental monitoring studies. Understanding this, the overall goal of the present study was to evaluate the sensitivity and viability of the native fish species Banded tetra, (Astyanax aeneus; Günther, 1860), widely spread in the aquatic ecosystems of the Yucatan Peninsula in Mexico, as a bioindicator organism. In order to do this, we performed a bioassay at sublethal concentrations using copper (CuSO4) to experimentally evaluate and validate the relationship between the trace metals and oxidative stress biomarkers response [(catalase (CAT), lipoperoxidation content (LPO)], detoxification [(glutathione S-transferase (GST), metallothionein content (MT)] and neurotoxicity (AChE) in muscle of A. aeneus. Results showed changes in biomarkers after 96 h: Catalase activity (CAT) was significantly higher above 1.5 and 2 mg/L (154.35 and 172.50% increase, respectively); lipid peroxidation contents (TBARS), GST activity, and MT content were very similar to CAT activity at 1.5 and 2 mg/L of Cu. In terms of neurotoxicity, AChE activity was significantly inhibited at 0.1 mg/L (64%; p < 0.001) and 0.5 mg/L (44%; p < 0.001) of Cu. Based on the bioassay results, we performed a trace metal monitoring campaign in muscle of A. aeneus caught in 15 sites with different anthropogenic activities, during the summer of 2017, to establish a baseline of trace metals pollution in the state of Campeche. A. aeneus showed the highest trace metal accumulation in the following order: Al > Fe > Mn > Zn > Cu > Hg > Cr > Pb > Cd > V > As, while sites were arrange as follows: Xnoha lagoon > Palizada River > Candelaria River > Ululmal > Maravillas > López Mateos. PCA showed a cluster between biomarkers (GST, CAT, TBARS, and MT) and concentration of metals (Cd, Cu, Fe, Zn, Hg and Cr). Conversely, AChE inhibition was not related to a specific metal, but highest inhibitions (>50%) were present in those sites with intensive agricultural practices. These results determined that, based on its physiological response and trace metal bioaccumulation, Astyanax aeneus can be considered a good bioindicator for evaluating the presence of trace metals in tropical aquatic systems of the Yucatan Peninsula.

Stomach flushing technique applied to quantify microplastics in Crocodilians.

The impact of microplastics on wildlife is a recent problem for which methods to evaluate exposure still need development. Being able to identify and quantify microplastics (particles < 5 mm) in the gastric contents of live crocodiles allows us to evaluate exposure, at both individual and population level, and also its contribution as transporter of other contaminants. The method was validated to determine and quantify microplastics in crocodile stomach contents recovered during an experiment where a known amount of this contaminant was given to crocodiles via oral administration. Through stomach flushing we were able to recover more than 80 % of the total volume of microplastic administrated to each crocodile. In summary, the method used during the experiment consists of 1) immobilization of the crocodile; 2) extraction of microplastics from stomach contents obtained through stomach flushing; 3) separation, identification and quantification of recovered microplastic fragments using microscopy and FTIR. •Low cost method that uses a small number of materials, does not take long to produce results and can easily be performed in the field or the laboratory.•Effective in extracting stomach contents (95 %).•High (>80 %) and good (>60 %) recovery efficiencies within two and four days after ingestion of microplastics by crocodiles.

Occurrence and seasonal distribution of microplastics and phthalates in sediments from the urban channel of the Ria and coast of Campeche, Mexico.

Microplastics (MPs) are part of a global problem affecting all countries including those with smaller populations. Coastal sediments are a reservoir of pollutants generated in basins. Phthalate esters (PAEs) are used as additives to make plastics stronger and more flexible. Urban ecosystems are the most susceptible to receive and transport pollutants. In this study, we determined the presence of MPs and PAEs in coastal sediments and in an urban and natural drainage system, known as the "Ria" in San Francisco de Campeche city. Plastics were detected in all samples taken from both regions, with plastics ranging in size from 0.3 to 5 mm in diameter. This indicates that they are particles of plastics, paints, and fibers, which may come from boats and nylon nets or synthetic products. The greatest abundance of MPs was detected on the coast with quantities of 1392 items/m2 (dry weight), which is double than that seen in the urban channel. As for PAEs, the highest concentrations were determined in two sites: M7A with 18.292 µg g-1 dw and PE with 21.702 µg g-1 dw, with the Ría channel being the area with the highest concentration of PAEs totaling 49.315 µg g-1 dw. The amounts of MPs detected in the sediments of the coast and the Ria vary significantly among the sites (p = 0.001) and seasons (p = 0.001), with the rainy season being the season where the greatest amount of MPs were detected. Likewise, the concentrations of ∑PAEs varied significantly among seasons (p = 0.0002) and sampling sites (p = 0.0001), with the west sub-basin being the site where the greatest concentration of PAEs was detected. A strong correlation was found between the ∑PAEs and MPs, which indicates that the conditions of the system favor the release and extended presence of PAEs on the coast.

Glyphosate Residues in Groundwater, Drinking Water and Urine of Subsistence Farmers from Intensive Agriculture Localities: A Survey in Hopelchén, Campeche, Mexico.

The use of pesticides in Mexican agriculture creates an interest in learning about the presence of these substances in different environmental matrices. Glyphosate (GLY) is an herbicide widely used in the state of Campeche, located in the Mayan zone in the western Yucatan peninsula. Despite the fact that GLY is considered a non-toxic pesticide to humans, its presence in water bodies through spillage, runoff, and leaching are a risk to human health or biota that inhabit these ecosystems. In the present study, glyphosate residues were determined in groundwater, bottled drinking water, and the urine of subsistence farmers from various localities of the Hopelchén municipality in Campeche. Determination of GLY was carried out using Enzyme-Linked Immunosorbent Assay (ELISA). The highest concentrations of GLY were observed in the groundwater (1.42 µg/L) of Ich-Ek and urine (0.47 µg/L) samples of subsistence farmers from the Francisco J. Mújica communities. The glyphosate concentrations in groundwater and bottled drinking water indicate an exposure and excessive use of glyphosate in these agricultural communities. This is one of the first studies that reports glyphosate concentration levels in human urine and bottled drinking water in México and in the groundwater in the Yucatan Peninsula as part of a prospective pilot study, to which a follow-up will be performed to monitor this trend over time.

Oxidative stress response in the skin mucus layer of Goodea gracilis (Hubbs and Turner, 1939) exposed to crude oil: A non-invasive approach.

The skin of the fish is the foremost target of oxidative stress due to the generation of Reactive Oxygen Species (ROS) originated in the environment and in the skin itself. In this study, a non-destructive assay was developed to evaluate the effects of crude oil (0.0001-0.1mg/L, 96h) on oxidative stress response in the Skin Mucus Layer (SML) of the dusky splitfin goodeid (Goodea gracilis). The response in the SML was compared with recognized target organs through the Integrated Biomarker Response (IBRv2) and a slight addition to the method was proposed. Crude oil was extremely toxic and elicited a clear induction of ROS in the SML, as in the brain, liver and muscle. By the exposure to crude, a significant change in the activities of Superoxide Dismutase (SOD), Catalase (CAT), Glutathione Peroxidase (GPx) as well as on lipid peroxidation (TBARS) and carbonyl protein (RCO) levels was detected. Also, increases in the activity of EROD were found. The general IBRv2 proposed in this study (gIBRv2) showed that oil causes the higher oxidative response in the SML (60.049) under different concentrations of petroleum, which was greater in the brain (56.749), muscle (56.561) and liver (55.775). The results of the study revealed an organ-specific antioxidant defense response that was dependent on the load of petroleum. These results contributed to the understanding of the complexity of oxidative stress response in fish exposed to crude oil using the Skin Mucus Layer as a target for environmental monitoring studies.

Usefulness of oxidative stress biomarkers evaluated in the snout scraping, serum and Peripheral Blood Cells of Crocodylus moreletii from Southeast Campeche for assessment of the toxic impact of PAHs, metals and total phenols.

In this study, we assessed the effects of inorganic and organic pollutants [As, Cu, Fe, Mn, Pb, Zn, PAHs (11 compounds) and total phenols] from a panel of biomarkers [O2, H2O2, thiobarbituric acid reactive substances (TBARS), carbonyl proteins (RCO), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and total cytochrome P450 activities] evaluated in the Snout Scraping (SS), Serum (S) and Peripheral Blood Cells (PBC) of the Morelet's crocodile (Crocodylus moreletii) inhabiting the reference locality (Lake Mocu) and polluted locality (Champoton River) using Principal Component Analysis (PCA). In male crocodiles from the reference site, only H2O2 in PBC was related to levels of fluoranthene on the Keel of Caudal Scales (KCS), but, in females, no association was detected. In contrast, a sex-linked response was detected in specimens from the polluted locality. Levels of benzo[a]pyrene, benzo[a]anthracene, chrysene, pyrene, phenanthrene, acenaphthene, Zn, Cu, and Pb in KCS of the female crocodil were related to the oxidative stress biomarkers on PBC, incluing the total CYP450 activity and levels of O2, H2O2 in serum. However, in male crocodiles, the oxidative stress in SS and in the serum (TBARS, RCO, CAT, GPx), and SOD in PBC was related to As, Pb, Cu, Fe, and benzo[a]pyrene water concentrations and to the burdens of As, Fe, Mn, indeno[1,2,3cd]pyrene in KCS. These results confirm the usefulness of minimal or non-invasive methods of evaluating the oxidative stress response for the environmental monitoring program on the wild Morelet's crocodile that is subject to special protection in Mexican guidelines.

Fat-associated lymphoid cluster in Cyprinus carpio: Characterisation and its relation with peritoneal haemangiosarcoma.

FALC cells are natural helper cells producing Th2-type cytokines, which express c-kit, Sca-1, IL7R and CD45 in mouse and human. These cells are involved in allergic responses and contribute to the inflammatory reactions of adipose tissue; however, a lack of information prevails about the presence of these cells in other species. The aim of the study was to identify and characterise FALC cells in the common carp (Cyprinus carpio) using immunohistochemistry and molecular biology techniques as well as to explore their relationships with their microenvironment. Histological description of the FALC was performed using H&E and polyclonal antibodies were used against cell-surface markers such as c-kit, Sca-1 and CD45. Furthermore, gene expression of c-kit, Sca-1 and IL7R was assessed. C. carpio FALC cells express the same surface markers reported in FALC of the mouse at both the pre- and post-transcriptional level. By exposure to the soluble fraction of helminths, FALC cells produce abundant Th2 cytokines (IL-5, IL-6 and IL-13) but do not synthesise IL-1α. Additionally, FALC cells probably participate in vascular remodelling of the intestine vessels, inducing tumours because a malignant haemangiosarcoma in the peritoneal cavity was found. In this tumour, abundant FALC with their characteristic cell-surface markers were detected. The findings of this study suggest the involvement of some proto-oncogenes such as c-kit and Sca-1, and the deregulation of Src kinases modulated by CD45 present in C. carpio FALC with the ontogeny of peritoneal haemangiosarcoma in this fish species.

Relationship between biomarkers and endocrine-disrupting compounds in wild Girardnichthys viviparus from two lakes with different degrees of pollution.

Despite great efforts worldwide to evaluate the effects of endocrine-disrupting compounds (EDCs) in fish, there is little information available about the interactions of EDCs with the disruption of the sexual endocrine axis in fish species with matrotrophic viviparity and intraluminal gestation. To understand these interactions, six sampling campaigns were performed within a period of 1 year in two lakes with different degrees of pollution. A battery of biomarkers of the oestrogenic response was assessed in the liver [vitellogenin, CYP 1A1, epoxide hydrolase activity, and metallothioneins (MT)] and MT in the head of Girardinichthys viviparus. Linear correlation analysis and canonical correspondence analysis were performed to explore the relationship between the oestrogenic response with EDCs and with metals. The biomarker responses were assessed using the water content of EDCs (oestrone, 17-ß-oestradiol, oestriol, 17-α-ethinyl oestradiol, total phenols, bisphenol A, nonyl phenol, octyl phenol), as well as the PAHs indene[1,2,3-c,d]pyrene, naphthalene, pyrene, benzo[a]anthracene, benzo[k]fluoranthene and benzo[a]pyrene) and metals (Cu, Fe, Mn, Pb and Zn). Greater disruption of the sexual endocrine axis occurred in fish of both sexes inhabiting the polluted lake whose effects were apparently influenced by CYP 1A1 activity and by 17-α-ethinyl oestradiol. In addition, non-estrogenic mechanisms in the hypothalamus and pituitary glands in male fish were observed, elicited by endogenous levels and the water concentration of Pb. In contrast, in females from the less polluted lake, VTG induction was related to exogenous oestrogens. The disruption of the hypothalamic-pituitary-gonadal axis is a complex process influenced by both endogenous and exogenous factors and contributes to male feminisation by exposure to EDCs.

Biomarkers involved in energy metabolism and oxidative stress response in the liver of Goodea gracilis Hubbs and Turner, 1939 exposed to the microcystin-producing Microcystis aeruginosa LB85 strain.

Goodea gracilis is an endemic fish that only habitats in some water bodies of Central Mexico that are contaminated with cyanobacteria-producing microcystins (MC); however, a lack of information on this topic prevails. With the aim to generate the first approximation about the physiological changes elicited by cyanobacterium that produce MC congeners in this fish species, specimens born in the laboratory was exposed for 96 h to cell densities of 572.5, 1145, 2290, 4580, and 9160 × 10(6) cells of Microcystis aeruginosa strain LB85/L, and a set of novel endpoint related to hepatic gluconeogenesis (ADH/LDH) and pro-oxidant forces O2., H2 O2 ) in addition to biomarkers of oxidative damage and antioxidant response was evaluated in the liver. Results suggest that high inhibition of protein serine/threonine phosphatase (PP) may trigger many metabolic processes, such as those related to hepatic gluconeogenesis (ADH/LDH) and pro-oxidant O2⋅, H2 O2 , TBARS, ROOH, RC=O) as well as antioxidant (SOD, CAT, GPx) response to oxidative stress. Particularly, we observed that inhibition of LDH and PP, and H2 O2 increase and TBARS production were the key damages induced by high densities of M. aeruginosa. However, changes between aerobic and anaerobic metabolism related with ROS metabolism and ADH/LDH balance are apparently an acclimation of this fish species to exposure to cyanobacteria or their MCs. Fish species living in environments potentially contaminated with cyanobacteria or their MCs possess mechanisms of acclimation that allow them to offset the damage induced, even in the case of fish that have never been exposed to MCs.

The relationship between the bioactivation and detoxification of diazinon and chlorpyrifos, and the inhibition of acetylcholinesterase activity in Chirostoma jordani from three lakes with low to high organophosphate pesticides contamination.

In fish, a number of studies have linked acetylcholinesterase (AChE) inhibition with exposure to organophosphate pesticides (OPs); however, evidence suggests the need to study aspects related to the bioactivation and detoxification of OPs, since their neurotoxicity is dependent on these processes. Thus, the study aim was to examine the relations between chlorpyrifos (CPF) and diazinon (DZN) bioactivation by hepatic CYP450 izoenzymes (CYP 2B6, CYP 2C19, CYP 3A4) and detoxification by aryl esterases and oxonases with brain and muscle AChE activity in Chirostoma jordani from three lakes with low to high OPs contamination in water and sediments. We found two patterns of bioactivation in vitro: (i) in fish from a lake with high CPF pollution, the main isoenzymes involved in this process were CYP 2C19>CYP 2B6>CYP 3A4, and (ii) in fish captured in a lake with a high concentration of DZN, the isoenzymes were CYP 3A4>CYP 2C19>CYP 2B6. Bioactivation is shown in this study to be fundamental in brain and muscle AChE inhibition in vivo. The rate of bioactivation of CPF was lower than for DZN. CPF bioactivation was accompanied by reduced detoxification and higher neurotoxicity, which was inversely dependent on the environmental contamination of CPF. Detoxification was also inversely correlated with environmental contamination by CPF, and was higher with diazoxon than chlorpyrifos-oxon. Oxonases were the most relevant enzymes involved in detoxification. The current findings suggest a series of strategies between the bioactivation and detoxification of OPs that allowed the survival of C. jordani despite of OPs pollution levels.

Pro-oxidant and antioxidant response elicited by CH2Cl2, CHCl3 and BrCHCl2 in Goodea gracilis using non-invasive methods.

The development of non-invasive methods aimed to evaluate the effects of many toxicants is required. Although there are some studies conducted in successful ways, a lack of information prevails especially for those substances that could be formed autochthonously in the water bodies, such as halomethanes (HMs). In this study, induction of pro-oxidant forces (CH2O, O2, H2O2), oxidative stress (TBARS, RCO) and antioxidant defenses (superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) in the skin mucus layer regarding to the liver of Goodea gracilis exposed to CH2Cl2, CHCl3 and BrCHCl2 were evaluated, in addition to the hepatic cytochrome P450 (CYP 2E1) and glutathione S-transferase theta (GSTT) activities. Regardless of the implicit toxicity involved in the bioactivation of the HMs, carried out by the CYP 2E1 and GST, it was noticeable that this process induces oxidative stress. The usefulness of the mucus layer for the evaluation of the oxidative stress response was demonstrated, despite some peculiar characteristics concerning induction of oxidative stress in liver and skin mucous layer. However, for the understanding of the induction of reactive oxygen species in both targets it is essential to evaluate the activity of antioxidant defenses; otherwise the interpretation of toxic effects elicited by HMs would be erroneous. In the skin mucus layer, lower activities of the enzymes involved in antioxidant defense than in liver were observed. The evaluation of the biomarkers in the skin mucus layer involved in the oxidative stress is useful due the consistent response regarding to concentration of the HMs.

Relations of oxidative stress in freshwater phytoplankton with heavy metals and polycyclic aromatic hydrocarbons.

The phytoplankton is the basis of food webs and also, may bioconcentrate different toxic substances. This phenomenon is well documented, but there are few studies on the toxic effects on the phytoplankton community. In the current study the relation of oxidative stress (TBARS, ROOH, RC=O) and antioxidant defenses (activities of SOD, CAT, GPx and GST) of two phytoplankton communities with BCF of heavy metals (Cu, Fe, Mn, Pb, Zn) and of PAHs (naphthalene, pyrene, indenol, benzo[a]pyrene, benzo[a]anthracene, benzo[b]fluoranthene) was tested. Three sampling surveys were conducted bimonthly on the surface and bottom at different sampling points in the lakes Menor and Mayor of the 2nd section of Chapultepec Park. Also negative and positive controls obtained in the laboratory were included. Toxicity relationships were analyzed using the integrated biomarker response (IBR). Both green algae and cyanobacteria dominated. The contents of ROOH and protein oxidation assessed as RC=O were higher in the Lago Menor, a water body that contains ancient sediments. Through the IBR it was demonstrated that these damages were influenced by Pb, indenol and benzo[b]fluoranthene. In contrast, TBARS content was higher in Lago Mayor, which has sediment in formation. Through IBR it was estimated that Cu, Fe, Mn, Pb, indenol and benzo[b]fluoranthene were related to this damage. However, oxidative stress was accompanied by an induction of CAT and SOD, in contrast, GPx and GST had low or null activity. The field data were similar to the positive controls. We demonstrated for the first time that, although the phytoplankton suffers oxidative stress elicited by metals and PAHs, this community is able to counter this damage through antioxidant defenses. The effects of organic or inorganic toxics in phytoplankton depend on their bioavailability that is modulated by the sediment and also by its physicochemical properties and the characteristics of the abiotic medium.

Implications of cytochrome 450 isoenzymes, aryl-esterase and oxonase activity in the inhibition of the acetylcholinesterase of Chirostoma jordani treated with phosphorothionate pesticides.

Organophosphate pesticides must be metabolized by cytochrome-P450 isoenzymes such CYP 2C19 as CYP 3A4 to induce neurotoxicity, but damage apparently depends on the activity of aryl esterases of the oxonase type that are involved in detoxication of these compounds. However, information on this subject is not available in fish. Chirostoma jordani has sustained significant population reductions, probably due to changes in land-use as well as pesticide impact; nevertheless, no specific studies demonstrating this are available. This study shows for the first time that the activity of cytochrome-P450 isoenzymes (CYP 2B6, CYP 2C19, CYP 3A4) in C. jordani is involved in diazinon and chlorpyrifos bioactivation. However, higher toxicity of chlorpyrifos cannot be explained solely because its bioactivation. Differences in toxicity between both pesticides are due to the activity of aryl esterases and oxonases that are responsible for oxon detoxication. Both hepatic enzymes metabolize diazoxon more efficiently than chlorpyrifos oxon. At lethal concentrations, detoxication is particularly important since mortality was lower with diazinon (LC50=1.5 µg/L) than with chlorpyrifos (LC50=0.17 µg/L). At sublethal levels, maximum acetylcholinesterase inhibition took place at 4h in both brain and muscle and was of lower magnitude in diazinon-treated fish. This is due to the higher affinity of both aryl esterases for diazoxon, which allows higher detoxication rates and therefore greater recovery of acetylcholinesterase activity.