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In Portugal, there are over 7800 children with life-limiting conditions. The context of pediatric palliative care represents a complex and distressing experience for families. Compared to parental caregivers and healthy siblings, grandparents are underexplored in the literature and clinical practice. The aim of the present study is to propose a psychological experience assessment tool of grandparents in this context. It consists of a sociodemographic and clinical data sheet and a semi-structured interview based on sharing a testimony with other grandparents. On the basis of the latter, 10 dimensions were explored through the grandparents' own perspective: representation of the illness; representation of the sick grandchild; changes in routine and life; family impact; grandparents' contributions to the family system; social support and coping strategies; emotional impact; triple concern; needs identification; and post-traumatic growth. The tool can be used in person or remotely and may be combined with other instruments. Its application enables a personalized identification of needs and challenges for each family, promoting the adjustment of the clinical intervention to their wellbeing and resilience from an eco-systemic perspective. The clinical tool is presented in detail and its importance in the context of research and systemic intervention is discussed.

Portugal has been identified as the European country with the most rapid evolution of Pediatric Palliative Care provision, where approximately 7800 children have life-limiting conditions. This is a highly complex experience not only for the children and their parental caregivers, but also for their healthy siblings and grandparents. The present descriptive-exploratory study seeks to contribute to the understanding of the psychological experience of life-limiting conditions in grandparents. A total of 19 families, consisting of 15 grandmothers and 4 grandfathers, completed a sociodemographic and clinical data sheet and a semi-structured interview was conducted in which they shared their testimony. The results of the thematic analysis highlighted an integrated view on 10 important dimensions in the grandparental experience and promoted creative responses by means of their own perspective. However, it has some limitations, such as the small sample size and the data collection procedure via telephone. The results contribute to the design of specific intervention methodologies in an ecosystemic approach and suggest further research to explore more protective factors and communication with health professionals. For psychological intervention, it is suggested considering the identification of individual and family resources that contribute to the activation of key processes in resilience and posttraumatic growth.

In Europe, Portugal has been identified as the country with the most rapid evolution of paediatric palliative care provision, which is a highly complex experience for families. The present descriptive-exploratory study seeks to contribute to the understanding of the psychological experience of life-limiting conditions in parent caregivers. A total of 14 families completed a sociodemographic and clinical data sheet and answered a structured online interview based on an incomplete narrative resulting from the Unwanted Guest Metaphor. A thematic analysis of the various narratives was performed through an inductive-deductive process. The results provide a holistic view of 10 essential dimensions in the parental psychological experience and contribute to the design of intervention methodologies in an eco-systemic approach. The importance of clear communication with health professionals, an awareness of the unpredictability of the disease, the desire for more self-care, the difficulty in understanding their children's needs and the threat implicit in everyday life are some of the main findings. This research emphasizes the importance of having opportunities of emotional expression and psychoeducation about anxiety' management, enhancing the perception of positive characteristics in children with palliative needs and creating time for the couple. The study has some limitations, such as the small sample size, and suggests that further research should explore the father's experience.

Background: Paediatric Palliative Care (PPC) has undergone rapid growth in Portugal, where there are over 7800 children with life-limiting conditions. This is a complex experience for families due to the ongoing threat and vulnerability caused by the emergence of an illness, and therefore several studies have tended to focus on the adaptation of parent caregivers. The aim of the present study is to present a psychological experience assessment protocol of parents in PPC. Methods: It consists of a socio-demographic and clinical questionnaire and a semi-structured interview based on an incomplete narrative deriving from the Unwanted Guest Metaphor . Results: On the basis of the latter, 10 dimensions of the experience in the parental subsystem were explored through parents' own perspective, namely: confrontation with the diagnosis; representation of the illness; emotional impact; day-to-day challenges; family impact; resources and social support; coping strategies; posttraumatic growth; representation of the sick child; and future perspectives. Discussion: The protocol can be used in person or remotely and its application enables the identification of specific needs and the establishment of psychotherapeutic goals and strategies for each family, thus enhancing their well-being and resilience, from an eco-systemic perspective. Conclusions: The protocol is presented in detail and its importance in the context of research and systemic intervention in PPC is discussed.

Background: Undergraduate students are a target for blood donation, as they constitute a young healthy and well-informed group., Aim: To understand motivations and attitudes underpinning the act of blood donation among undergraduate health science students., Materials and Methods: Cross-sectional study enrolling undergraduate students of a College of Health Sciences in the Northeast of Portugal (January/February, 2017). Data collection tool was a self-administered questionnaire covering questions about motivations (n = 8) and attitudes (n = 5) towards the act of blood donation. Participants were classified by donation status into donors (one previous donation) and non-donors (never donated). Multiple logistic regression models were used to assess the association between each attitude and motivation on donation status. Odds-ratio (OR) and respective 95% confidence intervals (95%CI) adjusted for potential confounders, were obtained., Results: Out of 362 participants, 12.7% (n = 46) had ever donated blood and 56.5% (n = 26) of them were regular donors. Out of 316 non-donors, 88.0% (n = 278) will donate blood under request. There were no differences between donors and non-donors regarding the attitudes towards blood donation. From all motivations only "be a civic duty" had a significant impact on donor status, such that participants reporting this motivation are more likely to be blood donors (OR = 2.58; IC95%:1.34-4.99) than their counterparts., Conclusion: This study revealed that 80.0% of undergraduate health science students are non-donors, but they are available to donate blood under request. Campaigns and advertising methods focused on the emergent needs for blood donation could play an important role in the recruitment of new donors among undergraduate students., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Metformin (MET) is among the most consumed pharmaceuticals worldwide. This compound has been frequently detected in fresh surface water. However, ecotoxicological information for MET is still too limited, particularly regarding chronic and behavioral data. This study aimed to help filling these knowledge gaps, by carrying out both acute and chronic studies with four different test organisms from three different trophic levels. We assessed different endpoints, including the swimming behavior of Danio rerio larvae. We also derived both short-term and long-term environmental quality standards (EQS) for the protection of freshwater pelagic biota towards MET adverse effects. A risk quotient (RQ) was calculated for MET in fresh surface water, considering a worst-case scenario. Daphnia similis was by far the most sensitive species evaluated. An EC 10 of 4.4 mg L -1 was obtained from the reproduction test with D. similis. A long-term EQS of 88 μg L -1 was derived and a RQ of 0.38 was obtained. An ecological risk is not expected for the chronic exposure of pelagic freshwater species to MET, considering the endpoints and the standard bioassays usually recommended in standard protocols. However, endocrine disruptive effects and potential interactive effects of MET with other co-occurring contaminants cannot be ruled out. To the best of our knowledge, this study presents the first data related with MET effects on population endpoints of D. similis and Hydra attenuata, as well as on the locomotor activity of D. rerio., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The detection and quantification of nanoparticles is a complex issue due to the need to combine "classical" identification and quantification of the constituent material, with the accurate determination of the size of submicrometer objects, usually well below the optical diffraction limit. In this work, the authors show that one of the most used analytical methods for silver nanoparticles, asymmetric flow field-flow fractionation, can be strongly influenced by the presence of dissolved organic matter (such as alginate) and lead to potentially misleading results. The authors explain the anomalies in the separation process and show a very general way forward based on the combination of size separation and size measurement techniques. This combination of techniques results in more robust AF4-based methods for the sizing of silver nanoparticles in environmental conditions and could be generally applied to the sizing of nanoparticles in complex matrices.

Using the bivalve macrofouler Corbicula fluminea, the suitability of in vitro testing as a stepping stone towards the improvement of control methods based on chemical mixtures was addressed in this study. In vitro cholinesterase (ChE) activity inhibition following single exposure of C. fluminea tissue to four model chemicals (the organophosphates dimethoate and dichlorvos, copper and sodium dodecyl phosphate [SDS]) was first assessed. Consequently, mixtures of dimethoate with copper and dichlorvos with SDS were tested and modelled; mixtures with ChE revealed synergistic interactions for both chemical pairs. These synergic combinations were subsequently validated in vivo and the increased control potential of these selected combinations was verified, with gains of up to 50% in C. fluminea mortality relative to corresponding single chemical treatments. Such consistency supports the suitability of using time- and cost-effective surrogate testing platforms to assist the development of biofouling control strategies incorporating mixtures.

Concentrations of persistent organic pollutants (POPs), including polybrominated diphenyl ethers (PBDEs), hexabromocyclododecanes (HBCDDs), polychlorinated biphenyls (PCBs), hexachlorocyclohexane isomers (HCHs), hexachlorobenzene (HCB), chlordane compounds (CHLs) and dichlorodiphenyltrichloroethane and its metabolites (DDTs), were measured in duplicate diet samples from 21 volunteers at a Portuguese academic community (University of Aveiro). Overall, the levels of the target compounds were low, with detection frequencies varying widely depending on the compounds and with brominated flame retardants (BFRs) registering the lowest detection frequencies. Among PCB congeners, nondioxin-like PCBs were predominant and detected in the majority of the samples. Organochlorine pesticides were also detected in the majority of the samples, with 100% detection for DDTs and HCHs. Estimated daily intakes (EDIs) were calculated using lower and upper bound estimations, and in both cases values were far below the currently established tolerable daily intakes for PCBs and OCs and the reference doses for PBDEs and HBCDDs., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Water quality criteria to protect aquatic life are not available for most disperse dyes which are often used as commercial mixtures in textile coloration. In this study, the acute and chronic toxicity of the commercial dye Disperse Red 1 (DR1) to eight aquatic organisms from four trophic levels was evaluated. A safety threshold, i.e. Predicted No-Effect Concentration (PNEC), was derived based on the toxicity information of the commercial product and the purified dye. This approach was possible because the toxicity of DR1 was accounting for most of the toxicity of the commercial mixture. A long-term PNEC of 60 ng L(-1) was proposed, based on the most sensitive chronic endpoint for Daphnia similis. A short-term PNEC of 1800 ng L(-1) was proposed based on the most sensitive acute endpoint also for Daphnia similis. Both key studies have been evaluated with the new "Criteria for Reporting and Evaluating ecotoxicity Data" (CRED) methodology, applying more objective criteria to assess the quality of toxicity tests, resulting in two reliable and relevant endpoints with only minor restrictions. HPLC-MS/MS was used to quantify the occurrence of DR1 in river waters of three sites, influenced by textile industry discharges, resulting in a concentration range of 50-500 ng L(-1). The risk quotients for DR1 obtained in this work suggest that this dye can pose a potential risk to freshwater biota. To reduce uncertainty of the derived PNEC, a fish partial or full lifecycle study should be performed., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The widespread use of pesticides in agriculture can lead to water contamination and cause adverse effects on non-target organisms. Brazil has been the world's top pesticide market consumer since 2008, with 381 approved pesticides for crop use. This study provides a comprehensive literature review on the occurrence of pesticide residues in Brazilian freshwaters. We searched for information in official agency records and peer-reviewed scientific literature. Risk quotients were calculated to assess the potential risk posed to aquatic life by the individual pesticides based on their levels of water contamination. Studies about the occurrence of pesticides in freshwaters in Brazil are scarce and concentrated in few sampling sites in 5 of the 27 states. Herbicides (21) accounted for the majority of the substances investigated, followed by fungicides (11), insecticides (10) and plant growth regulators (1). Insecticides are the class of major concern. Brazil would benefit from the implementation of a nationwide pesticide freshwater monitoring program to support preventive, remediation and enforcement actions.

Freshwater ecosystems are essential for humans; however, input of several types of contamination has led to the degradation of these ecosystems. Thus, it is urgent to assess their health to allow actions for prevention and remediation. The level of trace metals can be enhanced by natural or anthropogenic sources. Essential metals, such as copper and zinc, become toxic when present in the environment above threshold concentrations. To evaluate the physiological effects of these 2 essential metals for 2 freshwater detritivores, the shrimp Atyaephyra desmarestii and the amphipod Echinogammarus meridionalis, acute tests were performed. Forty-eight hour median lethal concentration (LC50) values were estimated for these species using static bioassays with copper and zinc. Sublethal assays for both metals with several phases were also done to evaluate the effects on feeding behavior. The LC50 values of copper for the shrimp A. desmarestii and amphipod E. meridionalis were 0.128 mg/L and 0.050 mg/L and those of zinc were 7.951 mg/L and 11.860 mg/L, respectively. The results indicated that copper is more toxic to both species. Only E. meridionalis showed deleterious effects of copper on feeding rate. Zinc showed some tendency for feeding inhibition in both species. Environ Toxicol Chem 2016;35:1442-1448. © 2015 SETAC., (© 2015 SETAC.)

The use of nanotechnology-based products is constantly increasing and there are concerns about the fate and effect on the aquatic environment of antimicrobial products such as silver nanoparticles. By combining different characterization techniques (asymmetric flow field-flow fractionation, single particle ICP-MS, UV-Vis) we show that it is possible to assess in detail the agglomeration process of silver nanoparticles in artificial seawater. In particular we show that the presence of alginate or humic acid differentially affects the kinetic of the agglomeration process. This study provides an experimental methodology for the in-depth analysis of the fate and behaviour of silver nanoparticles in the aquatic environment., (Copyright © 2015. Published by Elsevier Ltd.)

The equilibrium of the structure and functioning of freshwater ecosystems is dependent of detritivores that link all the other functional groups. The preference for feeding leaves with different diameters (particle size) and leaves with metal contamination (several concentrations of the essential metals copper and zinc) were determined for two detritivores, the decapod Atyaephyra desmarestii and the amphipod Echinogammarus meridionalis. Several no-choice and multi-choice assays were done to determinate which leaf diameter the amphipod and the decapod species would eat when they had or not had alternatives available and include a set of dual-choice assays with contaminated and uncontaminated foods. No significant preference was shown by either species relative to the diameter of leaves, either on no-choice or multi-choice assays. The presence of essential metals on food did not had any influence on the feeding choice of these organisms over the concentration range studied. Both showed no preference on ingesting food spiked with these essential metals, except E. meridionalis which preferred ingesting leaves with 2.19 μg.l(-1) of copper instead of uncontaminated leaves. For further works, despite no preference for leaves with a certain diameter, the leaves with 0.70 cm (0.385 cm(2)of area) and with 0.50 cm (1.767 cm(2) of area) should be used for A. desmarestii and E. meridionalis, respectively. Furthermore, to maintain E. meridionalis, the diet should include some percentage of copper in order to accomplish metabolic needs.

The use, environmental fate and ecological risks of antibiotics applied in tilapia cage farming were investigated in the Tha Chin and Mun rivers in Thailand. Information on antibiotic use was collected through interviewing 29 farmers, and the concentrations of the most commonly used antibiotics, oxytetracycline (OTC) and enrofloxacin (ENR), were monitored in river water and sediment samples. Moreover, we assessed the toxicity of OTC and ENR on tropical freshwater invertebrates and performed a risk assessment for aquatic ecosystems. All interviewed tilapia farmers reported to routinely use antibiotics. Peak water concentrations for OTC and ENR were 49 and 1.6 μg/L, respectively. Antibiotics were most frequently detected in sediments with concentrations up to 6908 μg/kg d.w. for OTC, and 2339 μg/kg d.w. for ENR. The results of this study indicate insignificant short-term risks for primary producers and invertebrates, but suggest that the studied aquaculture farms constitute an important source of antibiotic pollution., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

In vivo studies with Daphnia magna remain inconclusive as to whether engineered nanoparticles (NPs) are internalized into tissues after ingestion. Here we used a three-pronged approach to study the in vivo retention and efflux kinetics of 20 nm citrate stabilized Au NPs ingested by this key aquatic species. Daphnids were exposed to suspended particles (600 μg L(-1)) for 5 h after which they were depurated for 24 h in clean water containing algae. Light microscopy was used to follow the passage of Au NPs through the gastrointestinal tract, Au body burdens were determined by ICP-MS (inductively coupled plasma mass spectrometry), and transmission electron microscopy (TEM) was used to examine the presence and distribution of Au NPs in tissues. Results revealed that the elimination of Au NPs was bi-phasic. The fast elimination phase lasted<1h and the rate constant at which Au (of Au NPs) was eliminated was 1.12 ± 0.34 h(-1) (±SE) which accounted for ∼75% of the ingested Au. The remaining ∼25% of the ingested Au NPs was eliminated at a 100-fold slower rate. TEM analysis revealed that Au NPs in the midgut were in close proximity to the peritrophic membrane after 1 and 24h of depuration. There were no observations of Au NP uptake at the microvilli. Thus, although Au NPs were retained in the gut lumen, there was no observable internalization into the gut epithelial cells. Similar to carbon nanotubes and CuO NPs, our findings indicate that in daphnids the in vivo retention of Au NPs does not necessarily result in their internalization., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Metals are released into freshwater ecosystems from natural and anthropogenic sources, compromising their structural and functional equilibrium. As early warning tools, cholinesterases (ChEs) are usually used to assess the effects of organophosphate and carbamate pesticides, but are also known to be inhibited by metals. The objectives of this work were to characterise the activity of ChE present in the amphipod Echinogammarus meridionalis and the shrimp Atyaephyra desmarestii and to evaluate the in vivo effects of the metals copper and zinc in their ChE activity. To achieve this, firstly the activity of ChE forms were characterised using different in vitro assays with substrates and selective inhibitors. Then, the in vivo effects of 48 h exposures to increasing concentrations of copper and zinc on ChE activity were determined. The ChE form present in both species was acetylcholinesterase (AChE) since both revealed preference for the acetylthiocholine iodide substrate, total inhibition with eserine, the inhibitor of ChEs, and with 1,5-bis(4-allyldimethylammoniumphenyl)-pentan-3-one dibromide, the specific inhibitor of AChE, and presented insensitivity to iso-OMPA, a specific inhibitor of butyrylcholinesterase. The activity of ChEs was inhibited by zinc exposures in the amphipod species, but was not affected by copper. Exposure to copper and zinc did not affect ChEs activity in the shrimp at the concentrations tested. This work is a relevant contribution as foundation for the use of AChE in freshwater crustaceans in further studies including biomonitoring campaigns in different contamination scenarios.

The biocide glutaraldehyde (GA) is widely used as a disinfectant and sterilizing agent against bacteria and virus in hospital and veterinary facilities. GA or its metabolites may reach aquatic ecosystems due to incomplete or inadequate treatment of wastewaters. Data about GA effects at lethal and a sub lethal level to non-target organisms is needed so that a risk assessment to aquatic ecosystems can be done. Thus, in this work a battery of toxicity tests with primary producers, primary consumers and secondary consumers were performed and a species sensitive distribution (SSD) for GA was built. Moreover, effects on biomarkers (catalase, lactate dehydrogenase, glutathione-S-transferase, and cholinesterase) were measured in Danio rerio embryos and adults. Primary consumers (Thamnocephalus platyurus 24h - EC50=3.6 mg/l; Daphnia magna 48 h - EC50=6.6 mg/l) and D. rerio adults (96 h - LC50=5.5mg/l) were slightly more sensitive to GA than D. rerio embryos (96 h - LC50=22.2mg/l) and primary producers (Lemna minor 168 h - EC50=73.8 mg/l; Pseudokirchneriella subcapitata 72h - EC50=12.3mg/l; Chlamydomonas reinhardtii 72 h - EC50=14.6 mg/l; Chlorella vulgaris 72 h - EC50=31.3mg/l). However, no significant differences between the trophic levels were found and general HC5 and HC50 values of 0.6 and 11.4 mg/l were respectively estimated. Despite the low GA toxicity to D. rerio embryos, hatching delay and malformations were found (96 h - EC50=11.9 mg/l). For biomarkers, an inhibition of lactate dehydrogenase activity was observed in embryos whereas an inhibition in catalase, lactate dehydrogenase and glutathione-S-transferase activities was observed in adults. Thus, GA is moderately toxic (doses>1mg/l) to aquatic organisms, independently of the trophic level. However, considering the varied range of effects depending on the life stage and organism tested and relatively low HC5 value of 0.6 mg/l, mesocosm and chronic toxicity tests seem to be the next step in direction of more realistic scenarios of GA risk assessment in aquatic ecosystems., (© 2013.)

House dust is a repository and concentrator of many chemical and biological agents including fungi. Considering that dust acts as a long-term reservoir of airborne fungi and that cumulative exposure is more relevant to potential health problems than single-day or short-term exposure, characterization of fungal communities in dust samples is of paramount importance. In the present study, the fungal composition of Portuguese house dust samples was determined. A total of 28 samples were obtained from vacuum cleaner deposits from households located in central Portugal. DNA was extracted from dust samples and fungal communities were analyzed using a culture-independent polymerase chain reaction (PCR)- denaturing gradient gel electrophoresis (DGGE) approach. Cultural analyses were also performed in order to identify the viable fungi species present in selected samples. Fungal diversity, reported as the number of operational taxonomic units (OTU), varied between 9 and 56 OTU. This analysis of viable fungi showed that Aspergillus was the most abundant genus, followed by Penicillium, Mucor, and Rhizomucor. Trichoderma, Chrysosporium, Fusarium, Rhizopus, and Stachybotrys were found in a limited number of houses. Our results demonstrated that dust is, in fact, home for a diverse and heterogeneous fungal community and that some of the species found are known allergic agents with severe negative impacts on human health.

Chlorhexidine is among the most used biocides in Europe, however its toxicity to aquatic organisms is scarcely known. The main objective of this study was to assess the lethal and sub lethal effects of chlorhexidine digluconate (ChD) on four aquatic model organisms: the bacteria Vibrio fischeri, the algae Pseudokirchneriella subcapitata, the crustacean Daphnia magna and the embryos of the fish Danio rerio. ChD was very toxic to algae and crustaceans, with a 72 h-EC50 of 62.2 μg/l and a 48 h-EC50 of 45.0 μg/l, respectively. Toxicity to fish embryos and the bacteria was lower, with a 96 h-EC50 of 804.0 μg/l and a 15 min-EC50 of 1,694.0 μg/l, respectively. Concerning sub lethal effects on D. magna (feeding inhibition) a 6 h-EC50 of 503.7 μg/l was obtained. In fish, ChD caused developmental abnormalities, namely alterations in the amniotic fluid (48 h-EC20 of 753.6 μg/l) and early hatching. Moreover, enzymatic biomarkers on fish embryos showed an induction of cholinesterase activity in all ChD tested concentrations (80-900 μg/l). The catalase activity was also induced at the highest concentration tested (900 μg/l) whereas no changes were observed for glutathione-S-transferase and lactate dehydrogenase activities. The toxicity of ChD to the algae and crustacean raises concerns about its potential effects in aquatic food webs, since these organisms are in the base of trophic chains, and highlights the need for further studies on ChD toxicity to aquatic organisms.

Antibiotics have been widely used in human and veterinary medicine to treat or prevent diseases. Residues of antibiotics have been found in aquatic environments, but their effects on fish have been not properly investigated. This work aimed to assess the sub-lethal effects of oxytetracycline and amoxicillin on zebrafish development and biomarkers. Embryos and adults were exposed during 96 h to amoxicillin and oxytetracycline following OECD guidelines. Tissues of adults and pools of embryos were used for catalase, glutathione-S-transferases and lactate dehydrogenase determinations. Amoxicillin caused premature hatching (48 h-EC50=132.4 mg/l) whereas oxytetracycline cause delayed hatching of embryos (72 h-EC50=127.6 mg/l). Moreover, both antibiotics inhibited catalase and induced glutathione-S-transferases in zebrafish adults. However, only oxytetracycline induced lactate dehydrogenase. Short-term effects of antibiotics were observed at high doses (mg/l) indicating that physiological impairment in fish populations is unlike to occur. However, effects of chronic exposures to low doses of ABs must be investigated., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Pet cats and dogs have been successfully used as indicators of environmental pollution by a great variety of chemicals, including metals. However, information on mercury (a well know priority environmental pollutant) concentrations in household pets tissues and/or organs is scarce. Thus, in the present work we quantified total mercury (Hg(Total)) in blood and hair samples from twenty-six household dogs. The obtained results disclose relatively low levels of total mercury in the surveyed dogs, with values ranging from 0.16 to 12.38 ng g(-1) in blood; and from 24.16 to 826.30 ng g(-1) in hair. Mercury concentrations were independent of gender, age and diet type. A highly significant positive correlation was established between total mercury in blood and hair, validating the latter as a surrogate, non-invasive matrix for mercury exposure evaluation. Additionally, the obtained blood to hair ratio (200) is similar to the one described for humans reinforcing the suitability of dogs as sentinels. Overall, the determination of total mercury levels in dogs' hair samples proved to be a good screening method for the estimation of mercury burden in this species. We propose the quantification of Hg(Total) in hair as a screening method for sentinels like household pets to be performed in routine veterinary visits.

Risk assessment of metals in the environment is performed mainly with toxicity evaluations on single metals, which is largely inadequate since these substances occur in mixtures. The development of models predicting combined toxic effects on the basis of the concentration-response relationships of individual compounds has emerged as an answer. In the present study, metal effects on post-exposure anorexia (the concept of FdC(50)--concentration causing 50% of feeding inhibition--is implemented) in Echinogammarus marinus, a widely distributed gammarid amphipod, were assessed and compared with modelled ones obtained through the application of the concentration addition (CA) model, which represents a reasonable worst-case scenario for the risk assessment of metal mixtures. Data were validated using in situ experiments performed along a latitudinal gradient (Iceland, Scotland and Portugal) aiming at establishing a geographic profile of autochthonous population susceptibilities to metals. For all of the metals studied concentrations in the water column at exposure sites were in good agreement with feeding inhibition levels. Models gave low to relatively high percentage agreement between predictions and experimental data. Boreal populations demonstrated higher susceptibility to single metals, but not to mixture exposures. Meridional populations denoted lower susceptibilities with higher FdC(50).

Nile tilapia Oreochromis niloticus at 9 days post-hatch were exposed in semi-static experiments to the carbamate insecticide carbofuran, which is applied in agricultural systems in Brazil. Although the molecular mechanism of carbofuran toxicity is well known, a detailed understanding of the ecological mechanisms through which carbofuran effects can propagate towards higher levels of biological organization in fish is incomplete. Mortality rates were quantified for larvae exposed for 96 h to 8.3, 40.6, 69.9, 140, 297 and 397 μg/L carbofuran, and the LC(50) 96 h was 214.7 μg/L. In addition, the biochemical biomarker cholinesterase inhibition and behavioral biomarkers related to vision, swimming, prey capture and predator avoidance were quantified in individual larvae, as well as their growth in weight. The behavioral parameters were quantified by analysis of digitally recorded videos of individual larvae within appropriate experimental setups. The activity of the enzyme cholinesterase decreased after exposure to carbofuran with a lowest observed effects concentration (LOEC) of 69.9 μg/L. Visual acuity deficits were detected after carbofuran exposure with a LOEC of 40.6 μg/L. Swimming speed decreased with carbofuran exposure, with a LOEC of 397.6 μg/L. The number of attacks to prey (Daphnia magna nauplii) decreased in larvae exposed to carbofuran, with a LOEC of 397.6 μg/L. Growth in weight was significantly reduced in a dose dependent manner, and all carbofuran groups exhibited a statistically significant decrease in growth when compared to controls (p<0.05). The number of predator attacks necessary to capture larvae decreased after exposure to carbofuran, and the LOEC was 69.9 μg/L. These results show that exposure of sensitive early life stages of tilapia O. niloticus to sublethal concentrations of carbofuran can affect fundamental aspects of fish larval ecology that are relevant to recruitment of fish populations, and that can be better understood by the application of behavioral biomarkers., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Trichlorfon (TCF) is one of the most used veterinary pharmaceuticals not only to fight infestations but also as a preventive measure worldwide. The high concentrations used generate concerns about environmental and human health. In this work we assessed the acute toxicity of this compound to non-target organisms belonging to different trophic levels: Danio rerio (early life stages and adults), Daphnia magna and algae (Pseudokirchneriella subcapitata and Chlorella vulgaris), and studied the potential of the biomarkers cholinesterase (ChE), glutathione-S-transferase (GST), lactate dehydrogenase (LDH) and catalase (CAT) to assess sub-lethal effects of trichlorfon in zebrafish and daphnids. The fish embryo test followed the OECD draft guideline FET and was based on the exposure of newly fertilized eggs to 0, 2.5, 5.0, 10, 20, 40, 80 and 160 mg/L of TCF for 5 days; the fish acute test followed the OECD guideline 203 and was based on the exposure of adult fish to 0, 2.5, 5, 10, 20, 40, 60 and 80 mg/L of TCF for 4 days; Daphnia sp. immobilization assay followed the OECD guideline 202 and was based on the exposure of juvenile daphnids to 0, 0.1, 0.3, 0.5, 0.7, 0.9, 1 and 2 μg/L of TCF for 2 days and the algae growth inhibition assay followed the OECD guideline 201 and was based on the exposure of the two species to 0, 1, 3.2, 10, 32, 100 and 300 mg/L of TCF for 4 days. Biomarker levels were measured after 96 h exposure to TCF in zebrafish early life stages and adults and after 48 h exposure in D. magna. Tested organisms seem to have dissimilar sensitivities towards TCF exposure. D. magna (48 h-LC(50)=0.29 μg/L) was the most sensitive organism, followed by early life stages and adults of zebrafish (96 h-LC(50)=25.4 and 28.8 mg/L, respectively) and finally by the algae P. subcapitata (96 h-LC(50)=274.5 mg/L) and C. vulgaris (no effect observed). As daphnids are a source of food for organisms of higher trophic levels, the impairment on its population is prone to have consequences in the entire ecosystem. The biomarker activities measured in daphnids and fish seemed to be useful tools in the assessment of trichlorfon effects, especially ChE activity which was the most sensitive biomarker tested for all organisms. Trichlorfon was teratogenic for zebrafish embryos leading to anomalies in the absorption of the yolk sac, spine bending and pericardial oedemas. The present research suggests that further work is urgently needed in order to monitor environmental concentrations of trichlorfon and to test the long term effects of environmentally realistic concentrations of this compound., (Copyright © 2011 Elsevier B.V. All rights reserved.)

In twin pregnancy studies, molecular genetic techniques have rarely been used to determine zygosity, despite their known precision and accuracy. The present work aimed to assess the power of discrimination in zygosity assessment, using a set of microsatellite markers that were routinely used for aneuploidy screening by multiplex-PCR in a prenatal context. Rapid aneuploidy screening using a group of 20 microsatellite markers (STRs) located on chromosomes 13, 18, 21 and X has been performed in our lab for over 10 years, with a total of approximately 1,500 samples studied to date. A retrospective analysis of the 257 prenatal samples from multiple pregnancies was carried out. A subset of 14 cases presenting theoretical monozygosity were re-evaluated by the use of biostatistics tools accessed via the ZygProb website. Further monozygosity determination relative to dizygosity was calculated, given an estimated overall error value of 0.093%. The results show that monozygosity had been correctly determined in all our previously studied twins. This work demonstrates that accurate zygosity assessment can be achieved with the same STRs applied in aneuploidy screening with a high power of discrimination and a matching probability of over 99.999999%.

We determined the seroprevalence of Toxoplasma gondii and associated risk factors among 963 pregnant women attending an obstetric hospital in Fortaleza, Brazil. Seroprevalences of IgG and IgM against T. gondii were 68.6% (95% confidence interval [CI] = 65.6-71.6%) and 0.5% (95% CI = 0.06-1.0%), respectively. Seroprevalence of IgG was high in women less than 25 years of age (91.7%) and in low-income women (odds Ratio [OR] = 1.40, 95% CI = 1.02-1.90). Multivariate regression analysis showed that consumption of homemade water ice (adjusted OR = 1.49, 95% CI = 1.09-2.04), vegetables washed with untreated water (adjusted OR = 1.43, 95% CI = 1.05-1.94), consumption of chicken (adjusted OR = 1.49, 95% CI = 1.12-2.0), and dog ownership (adjusted OR= 1.46, 95% CI = 1.07-1.98) were factors associated with IgG seropositivity. Young women in northeastern Brazil living under poor socioeconomic conditions are at highest risk for acquiring infection with T. gondii. Oocyst contamination of water and soil must be addressed in future prevention strategies.

Direct and indirect effects of the fungicide carbendazim on ecosystem structure and functioning were studied < or =8 weeks after application (nominal concentrations: 0, 3.3, 33, 100, and 1000 microg/L) to outdoor microcosms in Thailand. Direct effects on macroinvertebrates are discussed in detail in a separate article. The present article presents the effects on other end points and discusses the hypothesized ecologic effect chain. Negative treatment effects on the zooplankton community were only recorded for the highest carbendazim treatment (NOEC(community) = 100 microg/L). The rotifer Keratella tropica, cladocerans (Moina micrura, Ceriodaphnia cornuta, and Diaphanosoma sp.), and cyclopoid copepods were decreased or even eliminated at this treatment level. The decrease in zooplankton and macroinvertebrate abundances was accompanied by an increase in numbers of several tolerant invertebrates, presumably caused by a release from competition and predation. The death of sensitive invertebrates probably also led to an overall decreased grazing pressure because increased levels of chlorophyll-a and bloom of the floating macrophyte Wolffia sp. were noted. The increase in primary producers is discussed to be the probable cause of changes in physicochemical water conditions, eventually resulting in an anoxic water layer during the last 3 weeks of the experiment. This is likely to have resulted in decreased invertebrate abundances noted in that period. Furthermore, the decreased decomposition of Musa (banana) leaves observed 8 weeks after application is considered to be the indirect effect of a decreased microbial activity resulting from these anoxic water conditions, rather than a direct toxic effect of carbendazim.

Studies investigating the use of biomarkers in pesticide risk assessment have greatly increased in recent years; however, issues concerning the ecological meaning of enzymatic responses have proved controversial. Ideally a good biomarker response should be modulated by the environmental contaminants alone and demonstrate a predictable behavior towards certain types of toxins. As these premises are rarely observed, the present study aims to outline research that has contributed to an understanding of the behavior of two widely used biomarkers, cholinesterase and glutathione-S-transferase, describing environmental and biotic factors that affect their response in freshwater invertebrates. Studies were performed in the main classes of aquatic invertebrates with these biomarkers and conclusions were reached concerning their behavior towards the main classes of pesticides. Links between biomarker responses and conventional endpoints were evaluated so that ecological relevance could be attributed to enzymatic responses. Toxicity of mixtures was investigated, and cases of synergism and antagonism were pointed out as factors changing the expected toxicity of aquatic systems and leading to misinterpretations of biomarker responses. Finally, the use of biomarkers as a tool for biomonitoring and in situ assays was investigated, with discussion of advantages and disadvantages of their use.

In November 2002, the tanker 'Prestige' released about 19,000 tonnes of a heavy fuel oil (no. 6) before sinking with about 58,000 tonnes of its cargo, 135 miles from Cabo Finisterra (Spain). A considerable part of the released fuel oil reached the Galician coast, causing a heavy black tide and an ecological disaster. Although the black tide did not reach the NW coast of Portugal, it is possible that some of the fuel oil or its components also arrived to this area directly through the sea water and/or indirectly through the food chain. Therefore, the aim of this study was to investigate possible changes in two widely used biomarkers, the activity of the enzymes cholinesterases (ChE) and glutathione S-transferases (GST), of three molluscs (Mytilus galloprovincialis, Nucella lapillus and Monodonta lineata) from wild populations of the NW Portuguese coast in relation to the 'Prestige' oil spill. Molluscs were collected seasonally before (autumn 2002) and after (winter 2002/2003), spring and summer 2003) the oil spill at several sites along the Portuguese NW coast. Enzymatic activities determined before the accident were compared with those determined at different times after the oil spill taking into consideration abiotic factors. Information from different parameters was integrated by Redundancy Analysis and Principal Response Curves (PRC). Results show that GST and ChE activities were influenced by abiotic factors. Despite this influence, the results of PRC analysis also suggest that some of the fuel oil reached the NW Portuguese coast changing the patterns of ChE and GST activities of local populations of rocky shore species. Furthermore, the present study highlights the need of long-term monitoring with wild populations to assess both historical and punctual effects of pollution in the marine environment.

The present paper discusses the fate of the fungicide carbendazim (nominal concentrations: 0, 3.3, 33, 100 and 1000 microg L(-1)) and its effects on the macroinvertebrate community in outdoor microcosms set up in Thailand. Fate and threshold values were subsequently compared with those noted in temperate model ecosystem studies in order to evaluate to which extent toxicity data generated in temperate regions for compounds like carbendazim may be extrapolated to tropical areas. Carbendazim disappeared faster from the water column than reported in temperate model ecosystem studies, which is explained by a possibly greater microbial breakdown of the fungicide under the tropical conditions tested. The No Observed Effect Concentration (NOEC) calculated in the present study for the macroinvertebrate community was the same as recorded in a study carried out in The Netherlands (3.3 microg L(-1)). However, most sensitive representatives in the latter study were "worm-like" taxa, whereas water boatmen (Corixidae) were the most sensitive macroinvertebrates in the present study. The ecological realism of a tropical risk assessment based on temperate organisms that are less sensitive than local macroinvertebrates may be questionable. Recommendations for future research include testing the possibility to rear sensitive representatives of indigenous Thai Corixidae in the laboratory and conducting model ecosystem studies evaluating a wider range of pesticides on a larger tropical geographical scale.

In aquatic environments, many factors determine the overall impact of a toxic event, constituting "exposure scenarios" that must be understood so that risk assessment strategies can be successfully applied. In this work, an experimental design was planned to investigate the influence of the organism age, duration of exposure and subsequent recovery period (after cessation of toxic exposure) on chlorpyrifos or carbendazim toxicity to Kiefferulus calligaster. The endpoints studied were cholinesterase (ChE) activity and glutathione S-transferase (GST) activity. The influence of age and exposure time and the existence of recovery depend on the chemical and the biomarkers tested, suggesting that no general rule can be applied when trying to assess overall toxicity of pesticides and that detailed knowledge has to be acquired on a case-to-case basis.

Effects of a single application of the photosynthesis-inhibiting herbicide linuron (0, 15, 50, 150, and 500 microg/L) on the ecology of outdoor plankton-dominated microcosms were studied in Thailand. As a result of the decreased photosynthesis, DO and pH decreased while EC, alkalinity and nutrient concentrations increased. Chlorophytes belonging to the genera Scenedesmus, Coelastrum and Pediastrum were the most sensitive taxa, whereas several other chlorophytes, diatoms, and cryptophytes increased in abundance. Tolerant taxa appeared to be less digestable for several zooplankton taxa, which subsequently decreased in abundances. Chamaesiphon sp. (Cyanobacteria) was the most susceptible periphyton species. As a consequence of functional redundancy, effects of the herbicide on the chlorophyll-a content of periphyton and especially phytoplankton did not always reflect the effects noted on community level.

This paper compares the fate and effects of linuron in an outdoor plankton-dominated microcosm study carried out in Thailand with those reported in temperate model ecosystem studies evaluating linuron and other photosynthesis-inhibiting herbicides. Lower linuron concentrations disappeared slightly faster from the water compartment compared to temperate conditions, which appears to be related with the experimental design rather than differences in climatic conditions. Sensitivity of primary producers and zooplankton were similar for the climatic regions, whereas effects on ecosystem functioning were less pronounced in tropical microcosms. Recovery potential of affected endpoints appears higher for tropical ecosystems compared to their temperate counterparts. These findings support the use of toxicity data generated in temperate countries in the tropics. Recommendations for the methodology of tropical model ecosystem experiments are discussed.

The fate and effects of the insecticide chlorpyrifos were studied in plankton-dominated, freshwater microcosms in Thailand. Disappearance rates of chlorpyrifos from the water column in the present study were similar to those in temperate regions. Insecticide accumulation in the sediment was relatively small, with the major part in the top layer (depth, 1.5 cm). Application of chlorpyrifos led to significant changes in freshwater biological communities. Clam shrimps (Conchostraca) and the cladoceran Moina micrura were the most susceptible species (no-observed-effect concentration [NOEC], 0.1 microg/L) and macroinvertebrates the most sensitive community (NOEC, 0.1 microg/L). These results are in agreement with those from semifield experiments with chlorpyrifos in temperate regions. The results of an in situ bioassay were used to calculate a NOEC of 0.1 microg/L and a 48-h median lethal concentration of 0.6 microg/L for M. micrura, which are similar to toxicity values reported for Daphnia magna in studies in temperate regions. Overall, these findings support the use of toxicity data from temperate regions for the risk assessment of low-persistent insecticides like chlorpyrifos for aquatic communities in tropical regions.

The drainage of abandoned mines leads to several ecological problems, particularly the acidification of surface freshwater systems and heavy metal contamination. In order to study the possibility of using the behavioural early warning responses of Portuguese indigenous benthic invertebrates to detect an acute short-term pulse of Acid Mine Drainage (AMD), experiments with the Multispecies Freshwater Biomonitor MFB were performed and locomotion and ventilation were measured as endpoints. AMD was collected from the "São Domingos" mine (Southeast Portugal) and the following species were selected: Echinogammarus meridionalis (Pinkster, 1973), Hydropsyche pellucidula (Curtis, 1834) and Choroterpes picteti (Eaton, 1870). For simulating the pulsed exposure, AMD was added to river water where invertebrates were collected and pH was lowered until reaching 3.5. The effects of H+ and heavy metals were discriminated using HCl positive controls. In addition to behaviour, mortality was registered. E. meridionalis was the most sensitive species in terms of mortality and behavioural endpoints, followed by C. picteti and H. pellucidula. E. meridionalis early warning responses consisted of increased locomotion with subsequent increase in ventilation, whereas for C. picteti only an increase in locomotion was observed. H. pellucidula showed no early warning responses. This work demonstrates the suitableness of using benthic invertebrates' behavioural early warning responses for detecting spikes of pollutants like AMD.

Steroid compounds have been globally detected in surface waters. The ecological impacts of these biologically active chemicals are largely unknown. Toxicity of testosterone and 4-hydroxyandrostenedione was assessed for the freshwater cladoceran Daphnia magna. Acute toxicity tests showed that 6.20 mg L(-1) of testosterone, the highest concentration tested, did not have effect on the daphnids, whereas 4-hydroxyandrostenedione had an EC(50) of 4.26 mg L(-1). Chronic toxicity tests were carried out using survival, body length, fecundity, and fertility as endpoints. Long-term testosterone exposure reduced D. magna fecundity and fertility at concentrations ranging from 0.31 to 2.48 mg L(-1). The significant decrease in fecundity was associated with an increase in aborted eggs. Long-term 4-hydroxyandrostenedione exposure at 0.84 mg L(-1) increased the mortality of the neonates. The chronic toxicity effects were observed at concentrations higher than the measured environmental concentrations of these compounds. Nevertheless, the reproductive impairment of the daphnids is likely to occur at environmental levels as an ultimate response to long-term exposure.

This paper describes the effects of a single and a repeated application of the organophosphorus insecticide chlorpyrifos on zooplankton and phytoplankton communities in outdoor microcosms in Thailand. Treatment levels of 1 microg L(-1) were applied once or twice with a 2-week interval. Both treatments led to a significant decrease in cladocerans followed by an increase in rotifers, although the extent by which species were affected was different. Ceriodaphnia cornuta was the most responding cladoceran after the first treatment, while Moina micrura responded most to the second. This is explained by differences in the growth phase of M. micrura at the time of application and an increase in Microcystis abundance over the course of the experiment. Several phytoplankton taxa either increased or decreased as a result of the chlorpyrifos-induced changes in zooplankton communities. Even though chlorpyrifos disappeared fast from the water column, effects on plankton communities persisted till the end of the experiment (42 days) when the insecticide concentrations had dropped below the detection limit. This was presumably due to the increasing population trend of Microcystis, favouring rotifers over cladocerans.

The aims of this study were to verify the suitability of in situ tests using the tropical midge Kiefferulus calligaster and to evaluate the most sensitive endpoint for the assessment of aquatic pesticide contamination. In situ tests were carried out in freshwater drainage channels (farm channels) that supply vegetable crops and receive considerable pesticide spray drift, and at channels outside farms (main channels). Moreover a pesticide-free farm was used as reference site. The endpoints analysed were: survival of the larvae, body length increment, capsule width increment, cholinesterase activity and glutathione S-transferase activity. Seasonal change was investigated as rainy season and dry season. Deleterious effects were observed at some farms especially during the rainy season when farmers apply heavier doses of pesticides. However, high mortality rates observed in main channels suggest that these water bodies are also affected by other impacts besides pesticide use. This work shows the potential of the in situ assay with K. calligaster as a tool for the environmental quality assessment of tropical aquatic ecosystems.

Initial surgical reconstruction for hypoplastic left heart syndrome (HLHS) is associated with satisfactory outcomes only in a few referral centers. Moreover, there is a persistent high-risk period for sudden death while the patient waits for the next surgical procedure. The development of a less invasive approach, so-called "hybrid," postponing a major surgery outside the neonatal period, might reduce the immediate and late surgical burden on these patients. This is a retrospective study of a contemporary series of patients with HLHS seen in two separate institutions. Patients with HLHS or its variants who underwent a "hybrid" management were included in the study. Data are described as the mean and standard deviation or absolute numbers and percentage, as appropriate. From January 2004 to June 2006, 15 patients (10 male; 5 +/- 3.8 days old and 2.9 +/- 0.5 kg) were included in the study. Ten had both mitral and aortic atresia; the ascending aorta and atrial septal defect measured 2.5 +/- 1.4 and 4.9 +/- 1.2 mm, respectively. There were six hospital survivors after stage I (mortality rate 60%). During the interstage period, all but one patient needed additional procedures. One patient died of bacterial meningitis 4 months after stage I. Four patients were submitted to stage II operation at 6.6 +/- 0.5 months of age and one is waiting for the operation. All four required early reinterventions for pulmonary artery stenosis. Only one was discharged home and was not yet submitted to the third stage. The hybrid approach for HLHS was associated with poor results in this early experience from two independent institutions in a developing country. This might have been related to infrastructure and technical problems, as well as our own learning curve. Institutions working under the same conditions might face similar problems during their initial experience.

Background: Few data are available on quality of life after surgical repair of tetralogy of Fallot (ToF), and on its relationship to right ventricle function., Methods: Patients with at least 1 year of follow-up evaluation after surgery were studied. Right ventricle function was evaluated by the Doppler-derived myocardial performance index (MPI), and health-related quality of life (HRQL) was assessed by the Child Health Questionnaire, Parent Form-50. Findings were compared with those for an age- and sex-matched group of healthy children., Results: The study enrolled 35 successfully repaired ToF patients with 4.9 years (range, 3-7 years) of follow-up evaluation after surgery and 36 healthy children. The MPI demonstrated right ventricle dysfunction in patients compared with controls (0.34; range, 0.26-0.49 vs 0.2; range, 0.13-0.27; p < 0.01), although HRQL for the psychosocial domain was similar between patients and healthy children (summary score, 47.9; range, 45-52 vs 47.5; range, 44.5-50; p = 0.17). A trend for poorer physical area results was observed in patients (summary score, 44; range, 31-52 vs 48.5; range, 45.7-51.7; p = 0.06). Neither physical nor psychosocial summary scores for HRQL had any correlation with right ventricle MPI., Conclusions: Right ventricle dysfunction is present in postoperative ToF patients. The psychosocial domain of HRQL is preserved after surgery. A trend for worse results was observed in the physical domain. The right ventricle function is not related to quality of life after surgical repair of ToF.

In this study, a tree (Populus nigra L.) has been presented as data logger of mercury release in aquatic environments using tree rings chemistry to provide chronological historical monitoring of mercury discharge from a chlor-alkali industrial effluent to a coastal lagoon. Tree rings (Populus nigra L.) as mercury data logger is suggested by mercury accumulation trends in the tree rings reflecting the industrial plant capacity increments in the early stages of mercury discharges and enhancing industrial plant controls to minimize mercury discharges in the last two decades after imposed global regulations on mercury emissions.

In this study we evaluate the ability of an in situ bioassay with Chironomus riparius larvae, using larval development and growth as endpoints, to biomonitor water quality and to assess the biological recovery of metal contaminated freshwater ecosystems of mine areas that are subject of restoration measures. The bioassay was carried out in streams located near an abandoned goldmine in North Portugal, throughout an environmental rehabilitation of the mine (2002-2004). During this period, a decrease in the inhibition of larval growth in the metal contaminated stream was observed. The bioassay was also performed in streams located near an active tungsten mine in Central Portugal. Larval growth and development were highly inhibited in the stream that receives acid drainage from the tungsten mine and treated water from the AMD treatment station. The results indicate that the bioassay can be used to evaluate the efficiency of environmental restoration measures in mining areas.

Two prevailing theoretical models: concentration addition (CA) and independent action (IA), predict mixture toxicity on the basis of known toxicities of the mixture components. To date, both models have been in most occasions evaluated using unicellular in vivo responses or biochemical in vitro responses. However, when considering more complex models such as the whole organism physiology or life-history traits, the dominant ecotoxicological mode of action, based on the exposure concentrations at which various toxicological effects become operative at the level of whole organism, should be considered. Offspring production in Daphnia magna is driven by the resources acquired from food, and the number of live offspring produced by an organism is the result of two independent factors: the number of eggs produced and the percentage of eggs that survive egg development. In this study joint toxicity effects on offspring production in D. magna were tested using binary mixtures of toxic contaminants known to specifically impair food acquisition (lambda-cyhalothrin and cadmium) or to cause egg mortality during development (3,4 dichloroaniline). Tests were performed using a simplified 10-day reproduction assay initiated with gravid females. The results obtained indicate that irrespective of their primary pharmacological mode of action, the joint toxicity of cadmium and lambda-cyhalothrin was predicted by the CA model, whereas the joint toxicity of cadmium and 3,4 dichloroaniline was predicted by the IA model. Thus, the results obtained indicate that for life-history traits such as reproduction responses, the dominant ecotoxicological rather than the pharmacological mode of action should be used for predicting joint mixture effects.

We used bioassays employing head capsule width and body length increase of Chironomus riparius larvae as end points to evaluate metal contamination in streams. Bioassays were performed in situ near an abandoned Portuguese goldmine in the spring of 2003 and 2004. Bioassays also were performed under laboratory conditions with water and sediment collected from each stream to verify if laboratory bioassays could detect in situ toxicity and to evaluate the relative contribution of sediment and water to overall toxicity. We used field sediments with control water and control sediments with field water to discriminate between metal contamination in water and sediment. Field water with dry and sieved, organic matter-free, and nontreated sediments was used to determine the toxicity of heavy metals that enter the organism through ingested material. In both in situ and laboratory bioassays, body length increase was significantly inhibited by metal contamination, whereas head capsule width was not affected. Body length increase was more affected by contaminated sediment compared to contaminated water. The lowest-effect level of heavy metals was observed in the dry and sieved sediment that prevented ingestion of sediment particles by larvae. These results suggest that body length increase of C. riparius larvae can be used to indicate the impact of metal contamination in rivers. Chironomus riparius larvae are more affected by heavy metals that enter the organism through ingested sediment than by heavy metals dissolved in the water column. Nevertheless, several factors, such as the particle size and organic matter of sediment, must be taken into account.

This study was conducted to investigate the potential use of biomarkers in bioassays with chironomids to assess contamination by pesticides in temperate and tropical climates. Two species of midge were studied, the widespread Chironomus riparius and the tropical Kiefferulus calligaster (Kieffer, 1911). Preliminary studies included investigations of the effects of temperature on larval development and the influence of larval age on normal variability of cholinesterase (ChE) and glutathione S-transferase (GST) activities and protein content. In the second phase, the influence of two abiotic factors particularly important in tropical conditions (temperature and oxygen concentration) and of the organophosphorous (OP) insecticide dimethoate on biochemical and conventional endpoints was investigated. Results showed that K. calligaster is morphologically and physiologically similar to C. riparius and for both, the time of larval development decreases with the increase of temperature. Moreover, 3rd and 4th instars appeared to be the most suitable for biomarkers determinations. ChE activity seems to be valuable biomarker regarding temperature and dissolved oxygen (DO) variations, while some caution should be taken when using GST as an environmental biomarker, since it shows some dependence of these parameters. C. riparius was more sensitive to dimethoate than K. calligaster suggesting that the use of bioassays with the former species in tropical conditions may overestimate the toxicity of OP pesticides to autochthonous species. When testing sub-lethal effects of dimethoate to C. riparius, ChE activity showed to be a very sensitive parameter detecting significant effects at the lowest concentration that caused emergence delay of larvae, suggesting that it is an ecologically relevant parameter.

Lethal and sublethal effects of Cadmium and Zinc on two freshwater crustaceans were investigated. The 96-h LC50 for Cadmium and Zinc were 42.20 microg l(-1) and 5.43 mg l(-1) for Atyaephyra desmarestii, and 36.17 microg l(-1) and 4.61 mg l(-1) for Echinogammarus meridionalis, respectively. An increase in metal concentrations at sublethal levels resulted in significant reductions of the feeding rate of both species. The LOECs in the chronic feeding assays for Zinc were 1.29 mg l(-1) for A. desmarestii and 0.4 mg l(-1) for E. meridionalis. For Cadmium the LOEC was 6.53 microgl(-1) for both species. The results suggest the potential use of these two crustacean species as test organisms in ecotoxicological tests. Feeding assays appear to be a rapid, cheap and effective tool to be used in biomonitoring studies of Portuguese freshwaters.

A bioassay with Chironomus riparius larvae, using larval development and growth as endpoints, was carried out inside a rice field and in the adjacent wetland channel in Portugal, during pesticide treatments (molinate, endosulfan and propanil) to determine impact caused by pesticide contamination in freshwater ecosystems. The bioassay was also performed under laboratory conditions, to assess whether in situ and laboratory bioassays demonstrated comparable results. Growth was inhibited by concentrations of endosulfan (2.3 and 1.9 microgL(-1) averages) in water from rice field in both the field and laboratory, and by concentrations of endosulfan (0.55 and 0.76 microgL(-1) averages) in water from the wetland channel in the laboratory bioassay, while development was not affected. C. riparius larvae were not affected by molinate and propanil concentrations. The results indicate that endosulfan treatments in rice fields may cause an ecological impairment in adjacent freshwater ecosystems. The results also indicate that laboratory testing can be used to assess in situ toxicity caused by pesticide contamination.

As a result of mining activities the exposure of metal sulphides to oxidation takes place with consequent release of acid mine drainage (AMD). Biomonitoring instruments have been proven to have the best deterrent effect upon polluters. A new approach in online biomonitoring, with the Multispecies Freshwater Biomonitortrade mark (MFB), was developed combining behavioural and feeding responses of the Portuguese indigenous benthic shredder, Echinogammarus meridionalis (Pinkster, 1973) (Crustacea, Amphipoda). These endpoints, along with mortality, were measured and analyzed for a gradient of sublethal doses of AMD. Original river water was used as the control and three doses of treatments were attained by adding increasing volumes of AMD to the control. The increase in AMD concentration and concomitant decrease in pH and increase in the concentration of most metals was followed by an overall increase of the mortality, decrease of locomotion/feeding activity and inhibition of the feeding rate. Mortality was observed in the two highest concentrations of AMD. Significant decrease in average locomotion/feeding activity took place in the second treatment. Although an inhibition of feeding was observed along the gradient of AMD concentration only in the highest concentration the feeding rate was significantly reduced.