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High temperatures alter the physiological condition of Octopus maya embryos, juveniles, and adults, and the time of exposure could have a key role in their thermal tolerance. The present study evaluates the effects of temperature and exposure time on octopus juveniles obtained from a thermally stressed female and a control female when exposed to optimal (25 °C) and high temperatures (30 °C) for 20 and 30 days, respectively. The results showed a transgenerational temperature effect that was expressed with low survival, depressed routine resting and high metabolic rates. Moreover, a collapse of antioxidant defense enzymes and high levels of oxidative damage products were detected in juveniles from thermally stressed females. Stress was lethal for animals acclimated at 30 °C, while the performance of juveniles acclimated at optimal temperature (25 °C) was conditioned by high oxidative stress levels and a reduction of the high metabolic rate (HMR) even after 30 days of experiment. In contrast, juveniles from the non-thermally stressed female had an optimal performance when acclimated at 25 °C but at 30 °C, they had a comparatively higher HMR during the first 8 days. These results suggest energy surplus in those animals to escape from warming scenarios before experiencing oxidative damage accumulation. Further studies should confirm if epigenetic alterations could be involved.

The major nesting sites for the endangered green turtle (Chelonia mydas) are in Campeche, Mexico: Carmen, Isla Aguada, and Sabancuy. Although they are in a natural reserve, these nesting sites are threatened by agricultural activities and oil extraction. This study aimed to determine the presence and concentration of organochlorine pesticides (OCPs) and polycyclic aromatic hydrocarbons (PAHs) in plasma and eggs and to assess the antioxidant response in plasma of nesting C. mydas from the southern Gulf of Mexico. Using censored statistics allowed us to do a realistic calculation incorporating the presence of non-detects in the analysis. A few contaminants (α+β+γ-HCH, trans-chlordane, 4,4′-DDE, 4,4′-DDT, methoxychlor, naphthalene, acenaphthylene, 2-Bromo naphthalene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, and dibenz[a,h]anthracene) were detected in plasma and their corresponding eggs, but correlations were not-significant. Spatial differences in concentrations of persistent organic pollutants (POPs) among nesting sites may reflect differences in foraging areas. Approximately 30% of the POPs in plasma and 60% of the POPs in eggs correlated with the biomarkers of oxidative stress. Detection of POPs in C. mydas indicated that plasma and eggs are suitable matrices to assess POPs concentrations.

Anthropogenic global warming generates profound metabolic alterations in marine ectotherm invertebrates capable of leading a wide range of these species to extinction. To evaluate the cross-generational effect of thermal stress on the cephalopod Octopus maya, groups of females were exposed at 24 and 30 °C until spawn. After, embryos of each female group were incubated at 24 and 30 °C allowing for evaluating the transgenerational effects on embryos exposed to high and low temperatures. We analyzed the morphology, oxygen consumption, antioxidant mechanisms, and oxidative stress indicators of the embryos. The results demonstrate that thermally stressed females produced smaller eggs with lower yolk content as observed in nonthermally stressed females. Also was observed that embryos from females acclimated at 30 °C had lower body weight and higher respiratory rates when compared with nonthermal stressed females. Embryos from females acclimated at 30 °C showed a collapse of the antioxidant defense system measured as lower both catalase activity and total glutathione concentrations. Additionally, glutathione-s transferase activity increased in embryos incubated at 30 °C and in females maintained at high temperatures in a clear deleterious and cross-generational effect of thermal stress on this octopus species. No changes were observed in the activity of B-esterases in octopus embryos linked with the thermal stress of females. Embryos from thermally stressed females had smaller sizes, less yolk, and higher metabolic rates. Additionally, a collapse in the antioxidant defense system was observed indicating they were unable to control the high load of ROS and oxidative damage, which was partially acquired by maternal inheritance.

No ecotoxicological information exists on phenanthrene (Phe) exposure in cephalopods, animals of commercial and ecological importance. This study investigated the effect of Phe on two B-esterases, Acetylcholinesterase (AChE) and Carboxylesterases (CbE), in Octopus maya embryos. Octopus embryos were exposed to different treatments: control (seawater), solvent control (seawater and DMSO 0.01%), 10 and 100 µg/L of Phe. AChE and CbE activities were measured at different developmental stages (blastula, organogenesis, and growth). B-esterase activities increased in control and solvent control as the embryos developed, showing no statistically significant differences between them. On the other hand, the embryos exposed to Phe had significant differences from controls, and between the high and low concentrations. Our results indicate that B-esterases are sensitive biomarkers of exposure to Phe in O. maya. Still, complementary studies are needed to unravel the toxicodynamics of Phe and the implications of the found inhibitory effect in hatched organisms.

The effect of biofloc (BFT) and clear water (CW) at low (ls) and high-water salinity (hs) in wild juveniles of Penaeus setiferus were evaluated. Four treatments were implemented: hsBFT, lsBFT, hsCW, and lsCW. After 45 days, final weight (FW), weight gain (WG), hepatosomatic index (HSI), and survival rate (SR) were evaluated. Antioxidant activity in muscle and hepatopancreas were measured: superoxide dismutase (SOD), catalase (CAT), glutathione-S transferase (GST), lipid peroxidation (LPO), protein oxidation (PO). The FW and SR was low for lsCW (P < 0.05), whereas the WG and HSI was high in hsBFT and lsBFT (P < 0.05). The integrative biomarker response index (IBR) showed differences between muscle (M) and hepatopancreas (HP). Results showed an increase of SOD and CAT measured from HP for lsBFT treatment. For M, CAT and GST were high only in BFT (regardless of the water salinity). LPO determined from M and HP recorded the high values at hs and ls, respectively. The PO determined in HP showed substantial damage in BFT (regardless of the salinity), whereas the same determination from M was only important for lsCW treatment. When the analyses of variance (ANOVA) were performed, SOD, LPO, and PO were significant in response to salinity (P < 0.05). In contrast, for CAT, the higher activity was due to the rearing system and salinity interaction (P < 0.05). Results suggest that BFT could diminish the stress by stimulating the antioxidant system to maintain the redox balance through a higher enzyme activity and decrease damage in the proteins and lipids of the cells.

Benzophenone-3 (BP-3) is one of the most used UV filters. The present study aimed to evaluate the toxic effects of BP-3 during embryo stages of zebrafish four hours post-fertilization (4hpf). Embryos were exposed to 0, 1, and 10 µg L−1 of BP-3 for 72 h. We investigated biochemical and molecular biomarkers of neurotoxicity (AChE) and the antioxidant system (gene expression of catalase, CAT, superoxide dismutase, SOD, glutathione peroxidase, GPX, the concentration of total glutathione, GSH, and lipid hydroperoxides, LPO). Results indicated that the acute exposure to BP-3 in zebrafish embryos did not show significant differences in survival, hatching rate, or antioxidant system biomarkers. In contrast, there were significant differences associated with AChE gene expression and activity.

Current anthropogenic global warming generates profound metabolic alterations in marine ectotherm invertebrates capable of leading a wide range of these species to extinction. The most worrying and devastating consequence may be that the effect of thermal stress overpasses the individual generations. To evaluate the transgenerational effect of thermal stress on the cephalopodOctopus maya, this study experimentally tests morphology, respiratory metabolism, antioxidant mechanisms, and oxidative stress indicators of the embryos incubated at two temperatures (24 and 30°C) produced by females acclimated at 24 and 30°C. The results demonstrate that, regardless of their incubation temperature, embryos from females acclimated at 30°C are smaller, show more accelerated development, and higher respiratory rates than those from females acclimated at 24°C. These embryos confirmed a greater oxidative stress degree, as well as an increased amount of soluble carbonylated proteins and catalase activity as the main enzyme during the activation development stage (even the highest in the embryos incubated at 30°C). Finally, a collapse of the antioxidant defense system was observed, measured as lower both CAT activity and GSH concentrations. Additionally, soluble carbonylated proteins reduced and GST activity increased in embryos incubated at 30°C from females maintained at high temperatures in a clear deleterious and transgenerational effect of thermal stress on this octopus species.

Introducción: Los eventos adversos (EA) son un acontecimiento imprevisto no intencional que causa daño a los pacientes. En neonatología, existe mayor prevalencia de EA debido a la fragilidad de los pacientes. Objetivo: Analizar la literatura científica publicada entre los años 2011-2021 sobre el manejo de los EA en servicios de Neonatología en América. Metodología: Revisión bibliográfica. Bases de datos: PUBMED, EBSCO, Google Scholar y Scielo. Descriptores DeCS: “Administración de la Seguridad”, “Neonatología”, “Prevención y Control”, “Errores Médicos”, “Seguridad del Paciente”, “Unidad de Cuidados Intensivos Neonatales” en español, inglés y portugués. Operadores booleanos: “AND” y “OR”. Lectura crítica considerando criterios de inclusión: artículos sobre tipos, control y manejo de EA en neonatología; países de América. Resultados: Se seleccionaron 18 artículos: 14 de EE. UU y 4 de Brasil. Los EA más frecuentes son: mal almacenamiento, mala técnica de administración, dilución errónea y error en la prescripción de medicamentos. Las causas más comunes de EA son: carga laboral y estrés. Estrategias de prevención: buena higiene y asepsia, uso correcto de EPP, adecuada dotación de personal, tecnología de códigos de barra para control de medicamentos, liderazgo, trabajo en equipo, comunicación, capacitación y educación al personal de salud. Una correcta notificación permite detectar y monitorear los EA, lo cual perfecciona su manejo. Discusión/Conclusión: Con sistemas de notificación y protocolos adecuados el manejo de los EA es más eficaz, disminuyendo la probabilidad de daño a los recién nacidos.

Anticipating, identifying, and prioritizing strategic needs represent essential activities by research organizations. Decided benefits emerge when these pursuits engage globally important environment and health goals, including the United Nations Sustainable Development Goals. To this end, horizon scanning efforts can facilitate identification of specific research needs to address grand challenges. We report and discuss 40 priority research questions following engagement of scientists and engineers in North America. These timely questions identify the importance of stimulating innovation and developing new methods, tools, and concepts in environmental chemistry and toxicology to improve assessment and management of chemical contaminants and other diverse environmental stressors. Grand challenges to achieving sustainable management of the environment are becoming increasingly complex and structured by global megatrends, which collectively challenge existing sustainable environmental quality efforts. Transdisciplinary, systems‐based approaches will be required to define and avoid adverse biological effects across temporal and spatial gradients. Similarly, coordinated research activities among organizations within and among countries are necessary to address the priority research needs reported here. Acquiring answers to these 40 research questions will not be trivial, but doing so promises to advance sustainable environmental quality in the 21st century. Environ Toxicol Chem 2019;38:1606–1624. © 2019 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

This study aimed to evaluate possible modification of some immune‐related genes (hemocyanin, alpha2‐macroglobulin, prophenoloxidase and penaeidin3a) at the transcriptional level and signs of disease and mortality in Litopenaeus vannamei previously reared in biofloc and challenged with different Vibrio harveyi densities (10³–10⁸ CFU per ml). Gene expression was up‐ or downregulated based on bacterial density, indicating that defence mechanisms were activated to remove the pathogen. The genes analysed were not affected in the control group (without bacteria) or in the 10³ CFU per ml group. The prophenoloxidase gene was expressed in the 10⁴ CFU per ml group, whereas transcripts of the alpha2‐macroglobulin gene were detected in the 10⁵ and 10⁶ CFU per ml groups. However, the penaeidin3a and hemocyanin genes were upregulated in the 10⁷ and 10⁸ CFU per ml groups, in which signs of disease and mortality in shrimp were evident. These results suggest that the genes involved in the shrimp immune response fluctuate according to pathogen density, and when this density increases (e.g. 10⁷ and 10⁸ CFU per ml), the activated disease tolerance and defence mechanisms are inefficient. For this reason, disease and mortality were observed in the shrimp regardless of the previous condition of the biofloc.

The inhibition of cholinesterase (ChE) activity has been used as a biomarker of exposure to organophosphate and carbamate insecticides. ChE of nesting female green turtles (Chelonia mydas) were biochemically characterized using two substrates, acetylthiocholine iodide and butyrylthiocholine iodide, and three ChE inhibitors (eserine sulfate, BW284C51 and iso-OMPA). The results indicated that BChE is the predominant plasma ChE in female C. mydas, but with atypical properties that differ from those found in human BChE. Eggs from green turtles nesting at two sites in Laguna de Terminos contained µg g−1 (wet weight) quantities of organochlorine (OC) pesticides. Drins (aldrin, dieldrin, endrin, endrin ketone, endrin aldehyde) were found at the highest concentrations with no significant differences in the concentrations in eggs collected at the two sampling sites. A negative relationship was found between levels of OC pesticides in eggs and BChE activity in the plasma of female turtles laying the eggs. Since OC pesticides are not cholinesterase inhibitors, we hypothesized that this inverse relationship may be related to an antagonistic effect between OCs and organophosphate pesticides and mobilization of OCs from the fatty tissues of the female turtles into their eggs. However, further study is required to verify the hypothesis. It is also possible that other contaminants, such as petroleum hydrocarbons are responsible for the modulation of cholinesterase activity in female turtles.

Tropical populations of marine species are predicted to be the most impacted by global warming because they are likely adapted to a narrow range of temperatures in their local environment. In the present study, we investigated the thermal range at which activity metabolic rate (AMR) is maximal, pejus, and critical in the spiny lobster Panulirus argus using different ecophysiological biomarkers. Lobsters were acclimated at constant temperatures (18, 22, 26 and 30 °C; N = 150) at laboratory conditions for 35 days; after that time thermal biology was evaluated. Results obtained demonstrated that, in P. argus the temperature where AMR was maximal does not coincide with temperature that promotes the maximum growth rate (26 °C). In fact, the preferred temperature, determined in a horizontal gradient was around the mean value of 27.5 °C, 5.5 °C above the temperature where animals showed their maximum metabolic scope, growth and thermal preference. Lactate and redox imbalance showed its maximum values in animals acclimated at 30 °C indicating that lobsters where in pejus thermal ranges. Although lobsters are adapted to tolerate temperatures beyond 28 °C, results indicated that temperature and exposure time play important roles in the way organisms respond to thermal challenges. It is likely that the difference in the responses of different ectothermic organisms and the thermal response is linked to the way these effects are measured. These results can help generate predictive models to anticipate changes in the P. argus fishery derived from likely changes in distribution and abundance of this species under warming scenarios of tropical oceans.

Most of our understanding on embryonic development in cephalopods comes from studies using constant temperature. Even though spawning generally occurs at relatively stable environments, eggs and embryos can be exposed over short periods of time to high temperature anomalies caused by oceanic warming. Therefore, in this study we examined the effects of increasing temperatures on growth, respiratory metabolism, and antioxidant defense mechanisms to define the thermal limits of Octopus maya embryos in a changing environment. Ramp temperatures consisted in increments of 1 °C every 5 days from 24 °C to 30 °C. The thermal limit of embryos at ramp temperatures was detected at 27 °C with metabolic rate increments between 63% and 69%, compared with the metabolic rate observed at control temperature (24 °C). Based on the calculated integrated biomarker response (IBR) was possible to detect that at the thermal limit, an increment on oxygen consumption was observed joint with reactive oxygen species (ROS). The IBR analysis indicated that above 27 °C embryos experienced changes in the oxidative system and were not able to recover. This study suggests that in a scenario of rapidly increasing temperatures after a short winter, O. maya embryos may suffer irreversible effects that are likely to negatively affect recruitment and the population dynamic. The thermal sensitivity of O. maya embryos, revealed in the present work, indicates that this species has an important potential for biomonitoring of the effects of environmental warming. Changes in the population dynamics of this species should be further studied since they can be used as a proxy for warming of the southern Gulf of Mexico ecosystems.

In May and June 2015, moderate and severe lesions were observed in Litopenaeus vannamei reared in clear seawater while, at the same time, lesions in shrimp reared in biofloc were considerably fewer. The signs of disease included anorexia, lethargy, melanization, expanded chromatophores, luminescence and necrotic areas in the uropods, suggesting a possible vibriosis. However, lesions observed in shrimp reared in biofloc disappeared after a certain time and without mortality in tanks, whereas mortality and severe signs continued to be observed in shrimp reared in clear seawater. To treat the possible vibriosis, oxytetracycline was administered only in clear seawater tanks, but the results were not successful. Bacterial cultures from hepatopancreas tissues of shrimp from both rearing systems confirmed a vibriosis outbreak only in the clear seawater system. Subsequently, Vibrio harveyi, Vibrio rotiferianus, Photobacterium sp. and Photobacterium damselae were identified from bacterial culture previously isolated for both rearing systems by molecular methods. Shewanella sp. was isolated and identified only in biofloc. To understand the possible pathogenicity and resistance mechanisms of the Vibiro strains for both rearing systems, pathogenicity (toxR) and oxytetracycline resistance-related genes (tet(B), tet(D), tet(G)) were determined. Although these genes were expressed for both rearing systems, biofloc proved to have the ability to control the development of the disease, in comparison to clear water, where the vibriosis was evident regardless of the administration of oxytetracycline as a treatment.

Ground characteristics in the Yucatan Peninsula make recovery and treatment of wastewater very expensive. This situation has contributed to an increase of pollutants in the aquifer. Unfortunately, studies related to the effects of those pollutants in native organisms are scarce. The aim of this work was to obtain partial sequences of widely known genes used as biomarkers of pollutant effect in Gambusia yucatana and Gambusia sexradiata. The studied genes were: cytochrome P450 1A (CYP1A); vitellogenin (VTG); metallothionein (MT), and two housekeeping genes, 18S and β-actin. From reported sequences of Gambusia affinis, primers were designed and amplification was done in the local Gambusia species exposed for 48 h to gasoline (100 µL/L, stirred for 24 h pre-exposure). Preliminary results revealed partial sequences of all genes with an approximate average length of 200 bp. BLAST analysis of found sequences indicated a minimum of 97% identity with reported sequences for G. affinis or Gambusia holbrooki showing great similarity.

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 tr...

The abundance of the alien, Indo-Pacific damselfish Neopomacentrus cyanomos on an oil-loading platform in the southwest Gulf of Mexico indicates that widely distributed platforms could facilitate the expansion of its geographic range across the western and northern fringes of the Gulf. From there it likely will spread to other areas of the Greater Caribbean. The lionfish example demonstrates that it eventually happens, and can do so rapidly. Reduced temperature effects on the physiology of this species were examined to better predict its survivability in the northern Gulf during winter, when sea surface temperatures fall as low as 15 °C along the coast. Overall, our results show that when the degree of experimental temperature decline was large and rapid, no compensation occurred and the stress response observed mostly reflected cellular processes that minimized damage. Integrated biomarker response values were significantly different between fish rapidly exposed to colder vs. warmer temperatures (declines of −4 °C each day, from 26 to 14 °C), reflected in higher values of blood metabolites and routine metabolic rates observed in fish exposed to 14 and 18 °C respectively, and lower activity of all enzymes, lower protein carbonylation, and higher oxidative damage to lipids in fish exposed to 14 °C. While the physiological proxies responded to minimize damage during the rapid-decrease experiment, the same proxies reflected the consequences of compensation when fish were thermally challenged after a 45 days acclimation at 18 °C. In this case, lower values of blood metabolites and high antioxidant levels and indicators of damages underpinned its pejus lower range. Based on the results of the present work, it seems clear that low winter SSTs in the northern Gulf will slow down the colonization of the inshore area of N. cyanomos . We suggest that the use of physiological cellular stress markers on specimens acquired at the beginning of an invasion should be implemented in new standardized experimental protocols, including both rapid increases/decreases of temperature and post-acclimation temperature challenges, to assess the invasiveness potential of aquatic species such as this.

Sexual maturation and reproduction influence the status of a number of physiological processes and consequently the ecology and behavior of cephalopods. Using Octopus mimus as study model, the present study was focused on 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 defense 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.

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.

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.

Sexual maturation and reproduction influence the status of a number of physiological processes and consequently the ecology and behaviour of cephalopods. UsingOctopus mimusas a study model, the present work was focused in the changes in biochemical compound and activity that take place during gonadal maturation of females and its consequences in embryo and hatchlings characteristics. To do that, a total of 31 adult females ofO. mimuswere sampled to follow metabolites (ovaries and digestive gland) and digestive enzyme activities (alkaline and acidic proteases) during physiological and functional maturation. Levels of protein (Prot), triacylglyceride (TG), cholesterol (Chol), glucose (Glu), and glycogen (Gly) were evaluated. Groups of eggs coming from mature females were also sampled along development and after hatching (paralarvae of 1 and 3 days old) to track metabolites (Prot, TG, Glu, Gly, TG, Chol), digestive enzymes activity (Lipase, alkaline proteases, and acidic proteases), and antioxidant/detoxification defence indicators with embryos development. Based on the data obtained, we hypothesized that immature females store Chol in their ovaries, probably from the food they ingested, but switch to TG reserves at the beginning of the maturation processes. At the same time, results suggest that these processes were energetically supported by Glu, obtained probably from Gly breakdown by gluconeogenic pathways. Also, was observed that embryos metabolites and enzyme activities (digestive and antioxidant/detoxification enzymes) where maintained without significant changes and in a low activity during the whole organogenesis, meaning that organogenesis is relatively not energetically costly. In contrast, after organogenesis, a mobilization of nutrients and activation of the metabolic and digestive enzymes was observed, together with increments in consumption of yolk and Gly, and reduction in lipid peroxidation. Derived from our results, we also have the hypothesis that reactive oxygen species (ROS) were produced during the metabolic processes that occurs in ovarian maturation. Those ROS may be in part transferred to the egg provoking a ROS charge to the embryos. The elimination of ROS in embryos started when the activity of the heart and the absorption of the yolk around stages XIV and XV were evident. Altogether, these processes allowed the paralarvae to hatch with buffered levels of ROS and with the antioxidant defence mechanisms ready to support further ROS production derived from paralarvae higher life stage requirements (feeding and metabolic demands).

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.

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.

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 Yucatan’s aquifer and the potential risk to human and environmental health.

Earlier reports have demonstrated that recombinant flavin-containing monooxygenase 1 (FMO1) catalyzes the oxidation of the organophosphate pesticide fenthion to (+)-fenthion sulfoxide in a stereoselective fashion. In order to elucidate the absolute configuration of the sulfoxide metabolite produced, we established an efficient synthesis of both enantiomers of fenthion sulfoxide, which were transformed into chiral fenoxon sulfoxides using a two-step protocol. The use of chiral oxidants, namely, N-(phenylsulfonyl)(3,3-dichlorocamphoryl) oxaziridines, afforded enantioenriched fenthion sulfoxides with high ee (>82%) from the parent sulfide. Single recrystallizations afforded chiral fenthion sulfoxides with >99% ee, measured by chiral HPLC analysis. The absolute configuration of the (+)-sulfoxide generated from fenthion metabolism by FMO1 was determined to be (R)-(+)-fenthion sulfoxide, confirmed by X-ray crystallographic analysis of the (S)-(-)-antipode. Inhibition of human recombinant (hrAChE) and electric eel (eeAChE) acetylcholinesterase were assayed with fenthion, fenoxon, and the racemates and enantiomers of fenthion sulfoxide and fenoxon sulfoxide. Results revealed stereoselective inhibition with (R)-(+)-fenoxon sulfoxide when compared with that of (S)-(-)-fenoxon sulfoxide (IC50 of 6.9 and 6.5 microM vs 230 and 111 microM in hrAChE and eeAChE, respectively). Fenthion sulfoxide (R or S enantiomers) did not present anti-AChE properties. Although the stereoselective sulfoxidation of fenthion to (R)-(+)-fenthion sulfoxide by FMO represents a detoxification pathway, the results of this study support the notion that subsequent oxidative desulfuration of (R)-(+)-fenthion sulfoxide (in vivo) may represent a critical bioactivation pathway, resulting in the production of (R)-(+)-fenoxon sulfoxide, a potent AChE inhibitor.

Flavin-containing monooxigenases (FMOs) are a polymorphic family of drug and pesticide metabolizing enzymes, found in the smooth endoplasmatic reticulum that catalyze the oxidation of soft nucleophilic heteroatom substances to their respective oxides. Previous studies in euryhaline fishes have indicated induction of FMO expression and activity in vivo under hyperosmotic conditions. In this study we evaluated the effect of hypersaline conditions in rat kidney. Male Sprague–Dawley rats were injected intraperitoneal with 3.5 M NaCl at a doses ranging from 0.3 cm3/100 g to 0.6 cm3/100 g in two separate treatments. Three hours after injection, FMO activities and FMO1 protein was examined in the first experiment, and the expression of FMO1 mRNA was measured in the second experiment from kidneys after treatment with NaCl. A positive significant correlation was found between FMO1 protein expression and plasma osmolarity (p < 0.05, r = 0.6193). Methyl-p-tolyl sulfide oxidase showed a statistically significant increase in FMO activity, and a positive correlation was observed between plasma osmolarity and production of FMO1-derived (R)-methyl-p-tolyl sulfoxide (p < 0.05, r = 0.6736). Expression of FMO1 mRNA was also positively correlated with plasma osmolality (p < 0.05, r = 0.8428). Similar to studies in fish, these results suggest that expression and activities of FMOs may be influenced by hyperosmotic conditions in the kidney of rats.

A cDNA encoding a hepatic isoform of flavin-containing monooxygenase (hFMO) (EF063736) containing an open reading frame of 1792 base pairs (bp) and encoding 554 amino acids was cloned and sequenced from liver mRNA of rainbow trout (Oncorhynchus mykiss). The genomic sequence of hFMO was also characterized and was 4.379 kilobases, possessing 10 exons and 9 introns (EU519462). Structural analysis of the promoter region showed several cis-acting elements including putative glucocorticoid and osmoregulatory response elements, which have been reported to be functionally related to induction of flavin-containing monooxygenase (FMO) proteins in vertebrates. The amino acid sequence showed 74% identity to a putative FMO gene from fugu (Takifugu rubripes; Q6ZZY9), 52 to 55% to zebrafish (Brachydanio rerio; Q5RGM6, Q5RGM3, Q6TLD2, Q7T1D7) FMO5, and 54 and 50% to human FMO1 (Q01740), FMO3 (P49326), and FMO5 (P49326). Southern blot analysis using a 180-bp fragment of the hFMO cDNA indicated at least seven potential genes. Treatment of primary trout hepatocytes with cortisol and sodium chloride for 24 h enhanced hFMO expression. Expression of hFMO was not detected in untreated or solute-treated primary cultures of gill epithelial cells, suggesting tissue-specific expression of hFMO. Induction of hFMO is consistent with the occurrence of cis-osmoregulatory and glucocorticoid response elements identified in the 5′-upstream sequence, indicating regulation of hFMO in response to hypersaline conditions and the osmoregulatory hormone cortisol.

A large number of organophosphorous insecticides (OPs) are chiral compounds, and yet enantioselectivity in their environmental fate and effects is rarely addressed. In the present study, we isolated individual enantiomers of three OPs, profenofos, fonofos, and crotoxyphos, and evaluated enantioselectivity in their inhibition of acetylcholinesterase (AChE). Acetylcholinesterase inhibition by the enantiomers and racemates was determined in vivo in the aquatic invertebrate Daphnia magna and in Japanese medaka (Oryzias latipes) as well as in vitro with electric eel (Electrophorus electricus) and human recombinant AChEs. The overall results showed variable sensitivity between AChE enzymes from different species as well as variable magnitude of enantioselectivity in enzyme inhibition. The (-)-enantiomer of profenofos was 4.3- to 8.5-fold more inhibitory to AChE in vivo, whereas (-)-fonofos was 2.3- to 29-fold more potent than the corresponding (+)-enantiomer. The (+)-enantiomer of crotoxyphos was 1.1- to 11-fold more inhibitory to AChE than the (-)-enantiomer. In contrast, the in vitro results showed (+)-profenofos to be 2.6- to 71.8-fold more inhibitory than the (-)-enantiomer and (-)-crotoxyphos to be 1.6- to 1.9-fold more active than the (+)-enantiomer. The reversed direction of enantioselectivity observed between the in vivo and in vitro assays suggests enantioselectivity within toxicodynamic processes such as uptake, biotransformation, or elimination. Findings from the present study provide evidence of enantioselectivity in the AChE inhibition of chiral OPs in nontarget organisms and indicate the need to consider enantiomers individually when assessing environmental risk of these chiral pesticides.

Two cholinesterases (ChE) present in vertebrates, acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are distributed in tissues in species-specific patterns. English sole (Pleuronectes vetulus, ES) and Hornyhead turbot (Pleuronichthys verticalis, HT) muscle ChE activities were measured in fish from a location near a municipal wastewater outfall and a far-field reference site off the coast of Southern California. ES muscle ChE correlated with total fish length (r=-0.3271) in fish from two sites. No significant differences were observed between gender or sampling sites. Substrate preference assays showed that both ChEs were found in muscle with BChE representing about 30% of the total ChE activity. Data resulting from the use of selective inhibitors suggested the presence of an atypical BChE. Higher sensitivity to pesticide inhibition was observed in ChEs from smaller fish. In HT muscle, ChE activities presented significant differences between males and females, but no correlation was found with total length. No significant differences were observed between sampling sites. Both ChEs were also present with BChE activity constituting 90% of the total ChE activity. Selective inhibitor results indicated the presence of an atypical ChE. Male ChE was more sensitive to pesticide inhibition. These data indicate that species, size, and gender dependent differences in ChE may contribute to differences in susceptibility to ChE-inhibiting toxicants encountered in the marine environment.