Gabriela Rodríguez-Fuentes

Biomarkers and pollutants in the Nile Tilapia, Oreochromis niloticus, in four lakes from San Miguel, Chiapas, Mexico

The effects of the oil industry and environmental pollutants present in four lakes from Reforma, Chiapas, Mexico, were assessed via ethoxyresorufin O-deethylase activity (EROD), PAH metabolites, and butyrylcholinesterase activity (BChE) in Nile Tilapia, Oreochromis niloticus. Fish were collected during two seasons (the rainy season in September 2000 and the dry season in May 2002) from the lakes Caracol, Rio, Enmedio, and Limon. Fish were sacrificed and EROD activities, hydrocarbons, organochlorine pesticide and heavy metal concentrations were determined in the liver, while PAH metabolites and cholinesterase activity were measured in bile and muscle, respectively. Hydrocarbon concentrations in fish liver were moderately high, as compared with results reported for the other species in different coastal ecosystems in Mexico. The highest concentrations of total hydrocarbons were found in fish captured in Lake Limon, and the lowest in Lake Caracol. The highest EROD and PAH metabolites were found in fish collected from Limon and Enmedio lakes. A Spearman test showed significant negative correlations between EROD and BChE activity with fish weight. EROD correlated with total hydrocarbons, the metabolites of pyrene and benzo[a]pyrene, and negatively with polychlorinated biphenyls (PCBs), endosulphan, hexachlorobenzene (HCB), and pentachloroanisol. BChE activity in the four lakes was lower than the activity measured in laboratory cultured tilapias, indicating the presence of anti-cholinergic pollutants in the area; Caracol was the lake with the lowest enzyme activity. This study demonstrated that high concentrations of contaminants can cause effects on fish metabolism, and that tilapia can be used as a test organism in tropical ecosystems.

Evaluation of the estrogenic and oxidative stress effects of the UV filter 3-benzophenone in zebrafish (Danio rerio) eleuthero-embryos

Personal care products have been detected in superficial waters, representing an environmental risk to the biota. Some studies indicated that 3-benzophenone (3BP) alters hormones, inducing vitellogenesis and having adverse effects on fish reproduction. Other studies have reported generation of free radicals and changes in antioxidant enzymes. Therefore, the aim of the present study was to test acute exposure to 3BP at concentrations within and beyond that found environmentally to provide important toxicological information regarding this chemical. We evaluated the effect of 3BP on vitellogenin 1 (VTG1) gene expression and the transcription of the enzymes catalase (CAT), superoxide dismutase (SOD) or glutathione peroxidase (GPx), which are involved in cellular redox balance. Zebrafish eluthero-embryos (168hpf) were exposed to 1,10, 100, 1000µg/L 3BP, in addition to a negative control and a 0.1% ethanol control for 48h. The results of our study indicated a positive significant correlation between exposure concentrations and VTG1 expression (r=0.986, p=0.0028) but only 1000µg/L 3BP produced a significant increase from control. Acute exposure showed no significant differences in transcription levels of CAT, SOD or GPx at the tested conditions. Nevertheless, a trend toward increase in GPx expression was observed as a positive significant correlation (r=0.928, p=0.017) was noted.

Impacts of oxidative stress on acetylcholinesterase transcription, and activity in embryos of zebrafish (Danio rerio) following Chlorpyrifos exposure

Organophosphate pesticides cause irreversible inhibition of AChE which leads to neuronal overstimulation and death. Thus, dogma indicates that the target of OP pesticides is AChE, but many authors postulate that these compounds also disturb cellular redox processes, and change the activities of antioxidant enzymes. Interestingly, it has also been reported that oxidative stress plays also a role in the regulation and activity of AChE. The aims of this study were to determine the effects of the antioxidant, vitamin C (VC), the oxidant, t-butyl hydroperoxide (tBOOH) and the organophosphate Chlorpyrifos (CPF), on AChE gene transcription and activity in zebrafish embryos after 72h exposure. In addition, oxidative stress was evaluated by measuring antioxidant enzymes activities and transcription, and quantification of total glutathione. Apical effects on the development of zebrafish embryos were also measured. With the exception of AChE inhibition and enhanced gene expression, limited effects of CPF on oxidative stress and apical endpoints were found at this developmental stage. Addition of VC had little effect on oxidative stress or AChE, but increased pericardial area and heartbeat rate through an unknown mechanism. TBOOH diminished AChE gene expression and activity, and caused oxidative stress when administered alone. However, in combination with CPF, only reductions in AChE activity were observed with no significant changes in oxidative stress suggesting the adverse apical endpoints in the embryos may have been due to AChE inhibition by CPF rather than oxidative stress. These results give additional evidence to support the role of prooxidants in AChE activity and expression.

Biological effects of environmental pollutants in American Oyster, Crassostrea virginica: a field study in Laguna de Terminos, Mexico

The response of Crassostrea virginica to a complex mixture of toxic contaminants was studied at four sites in Laguna de Terminos, Mexico. Contaminants assessed were heavy metals, organochlorine compounds, and hydrocarbons. Biomarkers (cholinesterase activity, neutral red retention, and metallothionein), histopathology and prevalence of Perkinsus marinus were used to evaluate the effects of contaminants. The resultant contaminant body burden was moderately as high as a whole, and biomarker levels were also moderate. Oedema in the gills and systemic haemocytosis were the only histopathological lesions observed. P. marinus was never evidenced in the oyster tissues examined. Statistical results from principal components analysis show that metallothioneins are positively correlated with copper and chromium, neutral red and oedema with chlordanes, pentachloroanisol, drins, polychlorobiphenyls and total pesticides, total hydrocarbons and polycyclic aromatic hydrocarbons, whereas cholinesterase activity is negatively correlated to Cu and Cr. The biomarkers used were sensitive indicators responding to moderate levels of pollutants.

Characterization of cholinesterases in the damselfish Sergeant major (Abudefduf saxatilis)

Cholinesterase (ChE) activity has been used for many years as a biomarker of exposure to organophosphate and carbamate pesticides. Recent studies have demonstrated that there could be biological factors that determine ChE type and levels; thus, juvenile Sergeant major (Abudefduf saxatilis) ChE enzymes were biochemically characterized. ChE enzymes found in the head and trunk were evaluated for their substrate preference and sensitivity to selective inhibitors. The use of the head and trunk was chosen as a strategy to reduce dissection time and to ensure sample uniformity between stations. The results indicated that there are two types of ChE enzymes in the head: acetylcholinesterase (AChE) and atypical butyrylcholinesterase (BChE) that exhibits intermediate characteristics of human AChE and BChE activities. Atypical BChE is predominantly found in the trunk. The results also indicated that the ChE activity found in A. saxatilis may be used as a biomarker in studies monitoring the Mexican Caribbean.

Chronic effect of nitrite on the rearing of the white shrimp Litopenaeus vannamei in two salinities

Abstract The aim of this study was to determine the effects of nitrite on the growth and survival of the white shrimp L. vannamei in two different salinities. Nitrite concentrations tested in salinity 8 g/L were 0 (control), 2.5, 5.0, 10.0, and 20.0 mg NO2−-N/L, and in salinity 24 g/L were 0 (control), 5.0, 10.0, 20.0, and 40.0 mg NO2−-N/L. For these experiments, 30 experimental units with 30 L of useful volume were stocked with 20 juvenile L. vannamei (8.0 ± 0.50 g), corresponding to a stocking density of 100 shrimp/m2, and cultivated for an experimental period of 30 days. A significant difference was found between the control and treatment groups with respect to growth and survival. The 2.5 mg NO2−-N/L treatment showed the best performance indexes in salinity 8 g/L, while the best growth performance indexes were found in the control and 5.0 mg NO2−-N/L treatments in salinity 24 g/L. Total mortality was observed in the 10 and 20 mg NO2−-N/L treatment groups from salinity 8 g/L and in the 40 mg NO2−-N/L treatment group in salinity 24 g/L. This study determined that concentrations of nitrite of up to 2.5 and 10 mg/L are acceptable for the rearing of L. vannamei in salinities of 8 and 24 g/L, respectively.

Cholinesterases in Gambusia yucatana: Biochemical Characterization and its Relationship with Sex and Total Length

Since several reports have indicated that cholinesterases (ChE) type and distribution is species specific and that in some species there is a relationship among gender, size and ChE activities, characterization has been suggested. The aim of the present study was to characterize the ChE present in head and muscle of Gambusia yucatana (using selective substrates and inhibitors) and to find its relationship with total length or gender. Results indicated that the ChE present in G. yucatana is an acetylcholinesterase (AChE) with high sensitivity to BW284C51 and an atypical smaller Km with butyrylthiocholine. Scatterplots indicated that there is no linearity between total length and AChE in male or female wild mosquitofish. There were no sex differences in AChE activities. Results indicated significant differences between a single collection site in the Yucatan peninsula and depurated organisms. This study emphasized the importance of characterizing ChE before usage in biomonitoring.

Gene expression in caged fish as indicators of contaminants exposure in tropical karstic water bodies.

Karstic areas in Yucatan are very permeable, which allows contaminants to move rapidly into the aquifer. In the present study, we evaluated gene expression of vitellogenin (VTG) and cytochrome P-450 1A (CYP1A) in caged juvenile zebrafish deployed for 15 days in 13 different water bodies, cenotes and aguadas, throughout karstic region of the Yucatan peninsula. Gene expression was evaluated using qRT-PCR. Results indicated induction of VTG in 7 water bodies with respect to reference cage. The highest relative VTG expression, about 3000 times higher than reference cage, was found in an aguada close to a cattle farm. CYP1A induction with respect to reference cage was observed in 3 water bodies, all of them located near villages or used for tourist activities. Pollutants and biomarkers of effect should be monitored in these water bodies in order to have a better understanding of the actual levels of pollutants that are present at Yucatans aquifer and the potential risk to human and environmental health.

Sexual maturation and embryonic development in octopus: use of energy and antioxidant defense mechanisms using Octopus mimus as a model.

Sexual maturation and reproduction influence the status of a number of physiological processes and consequently the ecology and behaviour of cephalopods. Using Octopus mimus as study model, the present study was focused in the changes in biochemical composition that take place during gonadal maturation of octopus females and its consequences in embryo and hatchlings characteristics, putting special attention to energetic metabolites, digestive enzymes and antioxidant defence mechanisms. To do that, a total of 32 adult females of Octopus mimus were sampled during ovarian maturation and the biochemical composition (metabolites and digestive enzymes) of digestive gland (DG) and ovaries (only metabolites) were followed during physiological and functional maturation. Levels of protein (Prot), triacyl glycerol (TG), cholesterol (Chol), glucose (Glu) and glycogen (Gly) were evaluated. In DG also the activity of alkaline and acidic enzymes was measured. Simultaneously, groups of eggs coming form mature females were also sampled along development, and metabolites (Prot, TG, Glu, Gly, TG, Chol), digestive enzymes activity (Lipases, alkaline and acidic), antioxidant defence mechanisms and radical oxygen species (ROS) were evaluated. Results obtained showed that ovarium is a site for reserve of some nutrients for reproduction. Presumably, TG where stored at the beginning of the maturation processes followed by Chol, both at the same time were energetically supported by Glu, derived from Gly following gluconeogenic pathways. Also, was observed that embryos during organogenesis nutrients and enzymes (metabolic, digestive and REDOX system) where maintained without significant changes and in a low activity. Results obtained in the present study shows that that activity could be not energetically costly. In contrast, was observed that during the embryo growth there were mobilization of nutrients and activation of the metabolic and digestive enzymes, joint with increments in consumption of yolk and glycogen, and reduction in molecules associated with oxidative stress, allowing paralarvae hatch with the antioxidant defence mechanisms ready to support the ROS production.