Biomedical subjects

Rui Ribeiro

Publications and source records attributed to Rui Ribeiro.

18 recordsLinked to original sources

PubMed2006

Effects of estuarine sediment contamination on feeding and on key physiological functions of the polychaete Hediste diversicolor: Laboratory and in situ assays.

This study aimed at integrating postexposure feeding and some biochemical parameters in the responses of the estuarine polychaeta, Hediste diversicolor, to controlled laboratory exposure conditions and to in situ exposures scenario of sediment contamination. Since H. diversicolor feeding may be considered as a major rate-limiting step in the processing of detritus in European estuaries, a reduction in feeding activity may have implications not only at the individual and population level of the species but also in detritus processing and in organic matter decomposition rates at the ecosystem level. The biochemical parameters were chosen as indicators of four key physiological functions: neurotransmission, metabolic condition, detoxification processes and antioxidant defences. The Mira and Sado estuaries, located in the Southwest coast of Portugal and classified as undisturbed and impacted, respectively, were selected as sites for this study. A significant depression in H. diversicolor postexposure feeding (from 30 to 70%) was consistently detected in all impacted sediments, supporting the sensitivity and responsiveness of feeding as a sublethal toxicity endpoint. Alongside with a reduced energy intake, an increased rate of organisms' anaerobic metabolism, as evidenced by an enhancement of lactate dehydrogenase activity (up to 1.5-fold), suggested a rapid need of additional energy to ameliorate chemical stress. Moreover, oxidative stress was shown to be an important mechanism of toxicity of the impacted sediments in H. diversicolor, as evidenced by a marked reduction in the glutathione redox status (up to 6.5-fold) and an increase in lipid peroxides levels (up to 2.3-fold) in organisms exposed to the most impacted sediments. Results of the in situ assay, conducted to assess the ecological relevance of sediment laboratory toxicity estimates and their application to make valid field extrapolations, revealed a lack of agreement in the response of catalase in organisms exposed to moderate impacted sediments. Our results support the utility of integrating responses at individual and sub-individual level to evaluate potential toxicant-induced changes in key physiological functions of H. diversicolor and to interpret their potential ecological consequences.

Animals↗

PubMed2006

Genetic adaptation to metal stress by natural populations of Daphnia longispina.

Loss of genetic diversity in natural populations as a result of chemical contamination has been reported in some studies. Here, four field populations of Daphnia longispina, two from sites historically impacted by acid mine drainage (AMD) and two from reference sites, were used to address four objectives: (1) identify differences in sensitivity between the stressed and reference populations; (2) distinguish between the components responsible for those differences (environmental influence vs genetic determination); (3) determine if genetically determined responses of reference and stressed populations converge from lethal to sublethal levels of contamination; and (4) evaluate losses of variability in genetically determined resistance by the stressed populations. Lethal and sublethal assays were carried out by exposing nonacclimated and acclimated neonates to AMD-contaminated waters and to copper dissolved in an artificial medium. Results indicate that both nonacclimated and acclimated individuals from the stressed populations are significantly less sensitive to AMD-contaminated waters than those from the reference populations, at both lethal and sublethal levels. The hypothesis of a convergence from lethal to sublethal responses was confirmed.

Adaptation, Physiological↗

PubMed2006

An in situ assay with the microalga Phaeodactylum tricornutum for sediment-overlying water toxicity evaluations in estuaries.

Given the urgent need for ecologically relevant toxicity tools specific to estuarine systems, the main goal of the present study was to develop and evaluate the potential of an in situ algal assay for estuarine sediment-overlying water toxicity evaluations using the microalga Phaeodactylum tricornutum immobilized in alginate beads. With its simple and cost-effective design, the assay apparatus, coupled with the use of microalgae immobilized in alginate beads, allowed rapid and easy deployment as well as retrieval of assay chambers. The effectiveness of the proposed in situ assay was investigated by deploying it at two reference and six contaminated Portuguese estuaries. By deploying open chambers both with and without an artificial nutrient source and closed chambers with an artificial seawater medium, it was possible to discriminate toxic effects from those caused by differences in temperature, light, and nutrient conditions across sites. A reduction in algal growth was found at contaminated sites relative to reference sites. The strontium alginate beads used for microalgae immobilization were found to be appropriate to perform in situ assays, showing no signs of disruption or dissolution after up to 8 d of exposure in estuarine sediment-overlying waters. The proposed in situ assay was shown to be a useful tool for estuarine sediment-overlying water toxicity testing to provide relevant data concerning primary production to estuarine ecological risk assessment studies.

Environmental Monitoring↗

PubMed2005

An in situ postexposure feeding assay with Carcinus maenas for estuarine sediment-overlying water toxicity evaluations.

This study developed and evaluated a short-term sublethal in situ toxicity assay for estuarine sediment-overlying waters, with the crab Carcinus maenas (L.) based on postexposure feeding. It consisted of a 48-h in situ exposure period followed by a short postexposure feeding period (30 min). A precise method for quantifying feeding, using the Polychaeta Hediste (Nereis) diversicolor Müller as food source, was first developed. The sensitivity of the postexposure feeding response was verified by comparing it to that of lethality, upon cadmium exposure. The influence of environmental conditions prevailing during exposure (salinity, temperature, substrate, light regime, and food availability) on postexposure feeding was also addressed. The potential of this in situ assay was then investigated by deploying organisms at ten sites, located in reference and contaminated Portuguese estuaries. Organism recovery ranged between 90% and 100% and a significant postexposure feeding depression (16.3-72.7%) was observed at all contaminated sites relatively to references.

Animals↗

PubMed2005

Optimization of a pressurization methodology for extracting pore-water.

Sediment toxicity can be assessed by conducting pore-water toxicity assays with standard water column organisms. Several methods have been developed for sampling pore-water. Centrifugation and pressurization methods are recommended when large volumes of pore-water are required to perform toxicity assays. Nevertheless, these methods involve sediment transportation and storage in laboratory, which can alter sediment toxicity. Therefore, an extraction method for large volumes that could be employed in the field site would be highly desirable. This study aimed to optimize and further evaluate an existing sediment pressurizing device with low construction costs, easy to carry and operate in the field, and presenting minimal chemical reactivity. The latter characteristic was achieved by lining the device interior with Teflon, by using large pore filters (50 microm), and by using an inert gas (nitrogen). Pore-water extraction efficiency and the toxicities of pore-water samples obtained by pressurization and by refrigerated centrifugation were compared. An artificial sediment (70% sand, 20% kaolin and 10% alpha-cellulose) spiked with an alcohol (phenol), a surfactant (SDS), a metal (copper), an organophosphate pesticide (parathion), and a natural sediment contaminated with acid mine drainage, were assayed for toxicity using Microtox assays. Sediment pressurization was found to be as efficient to extract pore-water as centrifugation, being more cost effective and adequate for field use.

Chemistry Techniques, Analytical↗

PubMed2005

Resistance to metal contamination by historically-stressed populations of Ceriodaphnia pulchella: environmental influence versus genetic determination.

Field populations of daphnids historically-stressed by metal contamination may show increased resistance to those contaminants. This study was undertaken aiming to confirm/infirm three main hypotheses: (1) field populations living in historically-impacted environments are more tolerant to metal stress than populations from reference sites; (2) resistance differences are genetically-determined, i.e., differences persist after controlling for environmental and maternal effects, by acclimating cloned lineages to similar conditions; and (3) resistance to stress in field populations living in historically-impacted environments is due to the disappearance of sensitive individuals rather than the appearance of highly resistant ones, i.e., the shift in the central tendency of resistance is linked to a decrease in the range of population resistance and not to an increased upper limit of the population resistance. Three populations of the cladoceran Ceriodaphnia pulchella Sars in Southern Portugal were sampled; one of which has been historically-stressed by acid mine drainage (AMD) from an abandoned cupric-pyrite mine and two from reference sites within the same watershed. To assess if resistance differences were genetically-determined, the three populations were acclimated for at least five generations under the same controlled conditions. Assays with AMD contaminated water samples were performed with both non-acclimated and acclimated individuals from all studied populations. Reproduction results in sub-lethal assays revealed significant differences between the reference and stressed populations. Significant differences in resistance to lethal levels of toxicity were observed for both non-acclimated and acclimated populations, individuals from population I being more resistant than those from reference populations. The existence of genetically-determined sensitivity differences was attested by the presence of significant differences in resistance to lethal levels of toxicity in acclimated individuals from reference and stressed populations. Results from cumulative mortality assays revealed that sensitive individuals were most probably present in the original population, but no conclusion could be draw about the presence of extreme resistant individuals in the historically-stressed population. Finally, it was shown that responses among populations converged from high to low levels of contamination.

Adaptation, Physiological↗

PubMed2005

Genetically determined resistance to lethal levels of copper by Daphnia longispina: association with sublethal response and multiple/coresistance.

The development of resistance to toxicants has been reported for a wide range of organisms. Two questions were addressed in the present study: Are genetically determined resistance responses at lethal levels (min to h) of copper associated with responses at extreme lethal (h to d) and sublethal levels and does genetically determined resistance to lethal levels of copper confer resistance to lethal levels of other chemicals? Twelve cloned lineages of Daphnia longispina, differing in their resistance to copper, were exposed to sublethal concentrations of copper and to lethal concentrations of copper, zinc, cadmium, hydrogen ions, and a pyrethroid insecticide (deltamethrin). Three kinds of toxicity assays were carried out: Survival time (death occurs in min to h; only for metals and hydrogen ions), cumulative mortality (death occurs in h to d; for all tested chemicals), and feeding depression assays (only for copper). A correlation between different levels of toxicity was observed only for extreme and moderate lethal responses to copper, and no correlation was found between lethal and sublethal levels of copper. Multiple resistance to lethal levels of toxicants was observed only for the pair copper/zinc.

Animals↗

PubMed2005

Short-term sublethal (sediment and aquatic roots of floating macrophytes) assays with a tropical chironomid based on postexposure feeding and biomarkers.

This study proposes assays with a freshwater chironomid, Chironomus xanthus, distributed over South America, based on subindividual (acetylcholinesterase activity) and individual (survival and postexposure feeding) level endpoints. Sediment and aquatic-rooted floating macrophyte assays were developed, due to the importance of both substrates for toxicant exposure in subtropical/tropical environments. Assays were evaluated under realistic exposure scenarios by simulating a runoff over an agricultural field dosed with deltamethrin. In situ assays were performed within microcosms to discriminate the effects of deltamethrin from those of additional potential stress factors (organism handling and caging, microcosms, use of aquatic roots, and runoff per se). A laboratory sediment assay based on feeding was conducted with samples from the microcosms. In all assays, both sublethal endpoints were responsive to deltamethrin and more sensitive than survival. Survival and feeding were more sensitive in situ than in the laboratory. In the in situ sediment assays, both sublethal endpoints were within a similar range of sensitivity; they were significantly inhibited as of the lowest Decis dose, from approximately 20 to 70%. In situ feeding was more sensitive in the sediment than in the macrophyte assay, where it was inhibited significantly only at the two highest Decis doses (up to approximately 60%). Larval performance was not influenced significantly by any of the other potential stress factors.

Acetylcholinesterase↗

PubMed2005

A short-term sublethal in situ toxicity assay with Hediste diversicolor (Polychaeta) for estuarine sediments based on postexposure feeding.

This study evaluated a short-term sublethal endpoint for in situ toxicity assays for estuarine sediments, based on postexposure feeding of the polychaete Hediste (Nereis) diversicolor Müller. A method for precisely quantifying postexposure feeding rates of H. diversicolor was established under laboratory conditions using Artemia franciscana Kellog nauplii. The sensitivity of the postexposure feeding response to copper was investigated by comparing postexposure feeding rates to growth and lethality. The 48-h and 96-h median lethal concentration (LC50) of copper were 241 and 125 microg/L, respectively, whereas the 48-h median inhibitory concentration (IC50) for postexposure feeding and the 20-d IC50 for growth were 52 and 25 microg/L of copper, respectively. The influence of different exposure conditions (substrate, temperature, salinity, food availability, and light) on H. diversicolor postexposure feeding was assessed; temperature and salinity were found to influence significantly postexposure feeding. The effectiveness of the proposed in situ assay was investigated by deploying it at two reference and six contaminated Portuguese estuaries. A 48-h exposure period was followed by a 1-h postexposure feeding period. High organism recoveries (89-100%) were obtained. Postexposure feeding was depressed significantly (17-90%) at all contaminated sites relatively to reference sites. The proposed in situ assay with H. diversicolor was shown to be a potential useful tool for estuarine sediment toxicity testing.

Animals↗

PubMed2005

Functional and structural impact of linuron on a freshwater community of primary producers: the use of immobilized algae.

An approach in determining ecosystem integrity and stress on ecosystem level is to assess processes within ecosystems. The aim of the present study was to evaluate the potential use of an in situ assay with immobilized Chlorella vulgaris as an indicator of effects on ecosystem functioning with regard to primary production. The herbicide linuron, applied in concentrations of 20, 60, and 180 microg linuron/L, was used to induce direct effects on primary producers. Direct and indirect changes in structure and function within outdoor model ecosystems of 3 m3 were monitored. The intermediate and highest concentration of linuron caused a decline in growth of the macrophyte Elodea sp., resulting in a significant increase of concentrations of nutrients. The increase in concentrations of nutrients caused an indirectly stimulated growth of immobilized C. vulgaris at the intermediate concentration, whereas similar concentrations of nutrients, at the highest treatment, did not stimulate Chlorella growth. It appeared that the direct effect of linuron on C. vulgaris growth was masked by nutrient availability at the intermediate but not at the highest linuron concentration. The observed immobilized algal growth was an accumulated effect of toxic and trophic pressures within the ecosystem, resulting in an integrative endpoint to detect actual impairment of ecosystem function.

Biological Availability↗

PubMed2004

[Herpes gestationis].

Herpes Gestationis is a serious dermatological disease, albeit rare, associated to pregnancy or to the trophoblast diseases. Contrary to what the name suggests, it is not a viral disease but an auto-immune disease. We present the clinical case of a 38 year-old woman to whom a case of Herpes Gestationis was diagnosed when she was 15 weeks pregnant and whom has been treated with corticosteroids and antihistamine's showing positive results and without major complications for the mother or the embryo. The authors are undertaking a review of the existing literature, based on this clinic case.

Chlorpheniramine↗

PubMed2004

An in situ bioassay integrating individual and biochemical responses using small fish species.

The interest in the ecological relevance of risk assessments and, thus, in in situ bioassays has been increasing in the last years. The present study developed a time- and cost-effective in situ bioassay, aiming at obtaining, in a short period of time and with a minimum of resources, a set of ecologically relevant toxicological information in a site-specific approach. Poecilia reticulata and Gambusia holbrooki were chosen as test species. Post-exposure feeding inhibition and the biomarkers acetylcholinesterase, lactate dehydrogenase and glutathione S-transferases were the endpoints tested. The battery of biomarkers as a whole was sensitive to the in situ exposure in an acid mine drainage impacted effluent, although responses varied between test species. Post-exposure feeding inhibition was the most sensitive endpoint, and its association with biomarker responses was discussed. The linkage between individual responses, such as feeding, and biomarkers suggested that, at least in this case, biomarkers can be relevant at higher levels of biological organization. Altogether, the proposed short-term in situ bioassay seems to be a promising tool, since it represents a reasonable compromise between sensitivity, time/cost-effectiveness and ecological relevance.

Acetylcholinesterase↗

PubMed2004

An in situ bioassay for freshwater environments with the microalga Pseudokirchneriella subcapitata.

This study was designed to evaluate the suitability of an in situ microalgal bioassay with Pseudokirchneriella subcapitata to be used in freshwater environments. The assay potential was investigated by deploying it in a system impacted by acid mine drainage. Water samples were collected to perform a laboratory assay also. P. subcapitata was viewed to be a good option for the in situ assay because it grew well and according to control acceptability criteria when immobilized in calcium alginate beads. A reduction in algal growth was apparent at both impacted sites demonstrating assay sensitivity: the site closest to the effluent discharge was clearly impacted and the one further downstream appeared to be moderately impacted. Results from the laboratory assay, designed to distinguish effects of nutrient differences across sites from those due to the effluent, confirmed the in situ responses. Results are discussed in light of the significance of combining information from different assessment tools, namely in situ and laboratory assays, as well as water-quality parameters, particularly at sites that are moderately impacted.

Analysis of Variance↗

PubMed2004

Avoidance of copper contamination by field populations of Daphnia longispina.

Almost all terrestrial and aquatic assays that are accepted as standardized, or that have been proposed, involve the exposure of confined organisms to toxicants. If avoidance (sensu evasion, displacement) of contaminants occurs in real situations, then bioassays involving forced exposure severely underestimate pernicious effects of contamination. Two main objectives were achieved in this study: To verify the occurrence of avoidance of copper by cladocerans; to determine the association between avoidance and other toxicity endpoints (lethality and feeding depression), and therefore, to ascertain if fully acclimated individuals from a reference field population exhibited a genetically determined avoidance markedly different from those belonging to a historically metal-stressed population. Twelve cloned lineages of the cladoceran Daphnia longispina, collected from two field populations, were selected according to their lethal sensitivity to copper and acclimated to controlled conditions for more than 30 generations. A 1.1-m test chamber with five compartments was built, allowing the establishment of a dissolved toxicant gradient and the free movement of individuals. In the absence of any toxicant, juveniles from each cloned lineage distributed themselves randomly along the test chamber and furthermore, no significant differences were observed between the two replicates, attesting the repeatability of this novel assay. All lineages showed significant avoidance to copper when exposed to a gradient from 3 to 87 microg/L. The most sensitive lineages to lethal levels of copper began avoiding this metal earlier than resistant ones. An intense association was observed between other endpoints and avoidance; furthermore, avoidance was determined to be much more sensitive than lethality. Therefore, avoidance assays should be recommended as a complementary tool in ecological risk assessments and effluent biomonitoring because such assays can provide cost-effective and ecologically relevant information.

Animals↗

PubMed2004

A phytoplankton growth assay for routine in situ environmental assessments.

This study proposes an ecologically relevant and cost-effective phytoplankton growth assay for routine in situ toxicity assessments. Assay procedures were developed applying, to the extent possible, the rationale behind the design of standard algal assays. Chlorella vulgaris was selected as test species because it grows well immobilized in alginate beads and has a wide geographic distribution. The performance of the assay in a freshwater system impacted by acid mine drainage demonstrated the suitability of assay chambers and procedures. The test system, made of inexpensive materials, allowed the rapid and easy deployment of the assay. The deployment of extra chambers at reference sites provided the ability to periodically check whether algal growth had already reached recommended growth criteria (time at which the assay should end). By deploying chambers filled with control medium at all sites, temperature was identified to explain 95% of the variation in growth. By using an artificial nutrient source shown capable of promoting algal growth according to recommended standards, toxicity from the mine effluent was distinguish from in situ nutrient limitation effects. The very good agreement (r2 = 90%) between mean in situ growth rates estimated by microscopy and by spectrophotometry and their similar coefficient of variation showed the latter to be a suitable straightforward methodology for assay endpoint estimation.

Biological Assay↗

PubMed2004

Avoidance tests with Collembola and earthworms as early screening tools for site-specific assessment of polluted soils.

Avoidance tests with earthworms and collembolans were conducted to demonstrate their feasibility as early screening tools for assessing the toxic potential of metal-polluted soils. Four different soils, with different degrees of metal contamination, were obtained from an abandoned mining area. All possible paired combinations were assessed for an avoidance response by the organisms. Data revealed that both species were able to avoid the most contaminated soils at the center of the ore extraction and treatment areas compared to those collected further away from the mine. However, earthworms and springtails differed in sensitivity to metals, especially when testing the two most polluted soils that had different contamination profiles. Earthworms exhibited a more consistent, less variable response than springtails. Overall results showed that avoidance tests with collembolans and earthworms have the potential to be used as screening tools in ecological risk assessment schemes for contaminated land, to trigger other tests in case of concern. However, further method development is needed to reduce variability in the data, particularly in the Collembola assays, and to gain knowledge about the possible effects of soil properties on the outcome of the tests.

Animals↗

PubMed2003

In situ bioassay chambers and procedures for assessment of sediment toxicity with Chironomus riparius.

The purpose of this study was to develop an in situ sediment bioassay chamber and respective procedures, suitable for performing toxicity bioassays with benthic invertebrates, using the midge Chironomus riparius. It was also our objective to compare the responses obtained under controlled conditions (laboratory 10-day larval growth and survival test) with those obtained in situ. Clean sand and a formulated sediment were incorporated in the in situ bioassay, along with local sediments, as a way of minimizing natural variability due to physicochemical differences among sediments or due to interactions with indigenous organisms. Recovery of organisms was good (80-100% in the control and reference site), indicating that the developed chamber and protocol were suitable for exposing and retrieving C. riparius in situ. Results also showed differences between responses obtained with formulated and natural sediment in situ, as well as between laboratory and in situ.

Animals↗

PubMed2002

An in situ bioassay for estuarine environments using the microalga Phaeodactylum tricornutum.

This study aimed at evaluating the potential of an in situ algal bioassay for routine toxicity estimates of potentially contaminated estuarine environments using the microalga Phaeodactylum tricornutum immobilized in alginate beads. The influence of the initial cell density in the beads and of salinity on algal growth was first investigated. The potential of the proposed bioassay was evaluated by comparing laboratory with in situ results. A good growth performance of P. tricornutum was observed at all starting densities of beads. Although the growth rate of P. tricornutum was significantly affected by salinity, acceptability criteria currently adopted in algal assays were met, indicating the suitability of P. tricornutum as a test species for bioassays in estuaries. The large differences observed between the laboratory and in situ responses of P. tricornutum were most probably due to the temperature and light conditions less favorable for algal growth in the field and to the lack of representativeness of water samples compared to the field fluctuating conditions. These results showed the need for in situ assessments, especially in estuarine environments influenced by tides. To a lesser extent, the bioassay itself may also have been responsible for the laboratory and field differences. Further improvements in the bioassay chambers and procedures were also discussed.

Biological Assay↗