Leonardo Sandrini-Neto

Is the distribution of the lancelet Branchiostoma caribaeum affected by sewage discharges? An analysis at multiple scales of variability

Spatial variation in the density and biomass of Branchiostoma caribaeum was analyzed along a sewage contamination gradient identified by fecal steroids in a subtropical estuary, southern Brazil. Sampling, repeated in the austral winter and summer, followed a hierarchical design nested at four spatial scales (sector>1 km; area>100 m; site>10 m; replicate<1 m). Density and biomass were significantly lower at sites characterized by high concentrations of fecal steroids. The best combinations of variables that explained the biological similarities among sites involved contamination indicators. Most of the variation of biological data was found at the smallest scales and could be related with the sediment texture. Our study highlighted the usefulness of a multi-scale perspective to evaluate distribution patterns of benthic invertebrates as a biological indication of environmental pollution. Gradient analyses at larger spatial scales may be invalidated by the patchy distribution of benthic fauna if they do not account for such small scale variability.

Antioxidant responses in estuarine invertebrates exposed to repeated oil spills: Effects of frequency and dosage in a field manipulative experiment.

We have experimentally investigated the effects of repeated diesel spills on the bivalve Anomalocardia brasiliana, the gastropod Neritina virginea and the polychaete Laeonereis culveri, by monitoring the responses of oxidative stress biomarkers in a subtropical estuary. Three frequencies of exposure events were compared against two dosages of oil in a factorial experiment with asymmetrical controls. Hypotheses were tested to distinguish between (i) the overall effect of oil spills, (ii) the effect of diesel dosage via different exposure regimes, and (iii) the effect of time since last spill. Antioxidant defense responses and oxidative damage in the bivalve A. brasiliana and the polychaete L. culveri were overall significantly affected by frequent oil spills compared to undisturbed controls. The main effects of diesel spills on both species were the induction of SOD and GST activities, a significant increase in LPO levels and a decrease in GSH concentration. N. virginea was particularly tolerant to oil exposure, with the exception of a significant GSH depletion. Overall, enzymatic activities and oxidative damage in A. brasiliana and L. culveri were induced by frequent low-dosage spills compared to infrequent high-dosage spills, although the opposite pattern was observed for N. virginea antioxidant responses. Antioxidant responses in A. brasiliana and L. culveri were not affected by timing of exposure events. However, our results revealed that N. virginea might have a delayed response to acute high-dosage exposure. Experimental in situ simulations of oil exposure events with varying frequencies and intensities provide a useful tool for detecting and quantifying environmental impacts. In general, antioxidant biomarkers were induced by frequent low-dosage exposures compared to infrequent high-dosage ones. The bivalve A. brasiliana and the polychaete L. culveri are more suitable sentinels due to their greater responsiveness to oil and also to their wider geographical distribution.

Effect of the invader Boccardia proboscidea (Polychaeta: Spionidae) on richness, diversity and structure of SW Atlantic epilithic intertidal community

In Mar del Plata (Argentine, SW Atlantic), a large seaside resort, the sewage discharges impact the littoral ecosystem. The invader polychaete Boccardia proboscidea has developed reefs since spring of 2008. The effect of this species on the richness, diversity and structure of epilithic intertidal community was assessed through an MBACI design in both sewage-impacted and reference sites, and Before/After the invasion. The presence of reefs of B. proboscidea since spring 2008 has caused a significant reduction of total individuals, total taxa and diversity in sewage-impacted sites regarding the reference ones. The species analyzed showed a high variable response because patterns were dominated by small-scale variability. Occasional peaks in abundance were observed on a single sampling site and time and a large variation among replicates. The associated fauna, formerly rich and diverse in impacted sites, shows a tendency to disappear as the ecosystem engineer Brachidontes rodriguezii is replaced by monocultures of B. proboscidea.

Are changes in the structure of nematode assemblages reliable indicators of moderate petroleum contamination

This study assesses through a multiple before-after-control-impact (MBACI) design the effects of diesel oil on the structure of nematode assemblages in unvegetated tidal flats of a subtropical estuary. Oil-exposed treatments were contrasted with controls for a duration of four successive days before and after an experimental spill in three distinct areas of the Paranaguá Estuarine Complex (Southern Brazil). No significant differences were observed in nematode total density, number of taxa and the overall assemblage structure between the control and impact treatments from before to after the experimental spill. This reinforces the idea that, despite being good indicators of environmental stress, free-living marine nematodes are able to tolerate low concentrations of hydrocarbons and to survive in moderately contaminated areas. We also show that robust experimental designs are useful to avoid confounding expected natural variability with the effects of a mild impact.

Are intertidal soft sediment assemblages affected by repeated oil spill events? A field-based experimental approach ☆

This study investigates the impact of repeated diesel spills on the structure of intertidal macrofaunal assemblages of a subtropical estuary. Three frequencies of exposure events were compared against two dosages of oil in a factorial experiment with asymmetrical controls. Hypotheses were tested to distinguish between (i) the overall effect of oil spills, (ii) the effect of diesel dosage via different exposure regimes, and (iii) the effect of time since last spill. Repeated oil spills dramatically altered the overall structure of assemblages and reduced the total density of macrofauna and densities of dominant taxa. Increasing the frequency of oil spills negatively affected macrofauna. In general, frequent low-dosage oil spills were more deleterious than infrequent high-dosage ones. However, increases in densities of some taxa, mainly the gastropod Heleobia australis, were observed in response to infrequent spills. Our results highlight the importance of repeated exposure events in determining the extent of oil impacts.

Antioxidant defense responses in Mytella guyanensis (Lamarck, 1819) exposed to an experimental diesel oil spill in Paranaguá Bay (Paraná, Brazil).

We evaluated the effects of diesel oil on the bivalve Mytella guyanensis using biomarkers of oxidative stress (glutathione S-transferase, glutathione peroxidase, and reduced glutathione) after an experimental in situ spill in a mangrove area in southern Brazil. A linear model was developed for the Multiple Before-After Control-Impact (MBACI) experimental design to assess the significance of biological responses. Control and impacted sites were sampled seven and two days before as well as two and seven days after the spill. With the exception of a late response of reduced glutathione (GSH) levels on day seven, none of the biomarkers were significantly altered by the impact. This result was attributed to the high environmental variability of the experimental sites combined with a low sensitivity of Mytella guyanensis to diesel oil at short time-scales. The high resistance of M. guyanensis suggests that its antioxidant response is triggered only after a medium- to long-term exposure to contaminants.

How and where to perform biomonitoring studies: different levels of toxic metal pollution are detected in the Alagados Reservoir in Southern Brazil

The Alagados Reservoir located in the Brazilian city of Ponta Grossa is used to supply water for human consumption but is impacted by toxic metals. The current study combined chemical, biochemical, and multivariate analyses to determine the bioavailability of toxic metals at three sites (AL-A, AL-B, and AL-C) within the Alagados Reservoir. Metal bioaccumulation was analyzed in the liver, gills, and muscle tissue of a native fish species (Geophagus brasiliensis), and neurotoxicity, xenobiotic metabolism, and oxidative stress were evaluated using biochemical biomarkers. Additionally, histopathological studies were performed on the gills and the liver using scanning electron microscopy (SEM) and conventional light microscopy (LM), respectively. Overall, the bioaccumulation of metals, biomarkers of oxidative stress, and melanomacrophage counts indicate that the AL-C and the AL-A sites are the most and least affected by metals, respectively. The AL-B site presented the lowest acetylcholinesterase enzyme activity, a finding which was probably associated with the agricultural activities around this area of the reservoir. The biomarkers clearly revealed that toxic metals negatively affect all three sites studied herein and that human activity is the major source of pollutants. Despite the existence of different pollution levels within the Alagados Reservoir, it is still used as a human water supply.

Un nuevo método de análisis digital para la determinación de los niveles de consumo foliar

A protocol using the computer program Adobe Photoshop CS2 (version 9.0) is proposed for determining the level of herbivory on intact and consumed leaves, using mangroves as an example. The usual manual methods consist of drawing the leaf on graph paper, restoring the original shape in cases of damaged edges. The computerized methods are intended to accelerate the leaf measurement time with applications, digitalization equipment, and portable area measurers that are more efficient, faster, and more economically viable than the manual methods. The need to restore the edges of the consumed leaves is an obstacle to all methodologies. Nonetheless, the reconstruction of the consumed foliar edges is shown to be more realistic with Adobe Photoshop, as this allows a better fit to the original shape of the leaf. Moreover, this application allows the use of compacted files such as JPEG, permitting a simple, rapid, and precise method for determining the levels or rates of foliar herbivory.

Diffuse sources of contamination in freshwater fish: Detecting effects through active biomonitoring and multi-biomarker approaches

Aquatic organisms are usually exposed to a mixture of xenobiotics that may exert a large effect even in low concentrations, and when information is obtained exclusively from chemical analyses the prediction of the deleterious effects is potentially hindered. Therefore, the application of complementary monitoring methods is a priority. Here, in addition to chemical analyses, an active biomonitoring study using multiple biomarker responses in Nile tilapia Oreochromis niloticus was conducted to assess the effects of a contamination gradient along four reservoirs in Iguaçu River. Chemical analysis in the muscle showed high levels of metals in fish from the reservoir closest to an industrialized and environmentally degraded area, however fish exposed to all studied reservoirs showed hepatic alterations (necrosis and inflammatory processes). Also, significant variations of biochemical biomarkers were observed with no clear indication of contamination gradient, since an indicative of higher impact was found in an intermediary reservoir, including high concentrations of biliary polycyclic aromatic hydrocarbons (PAHs). However, nuclear morphological alterations (NMA) were less frequent at the same reservoir. Thus, the multi-biomarker approach allied to active biomonitoring is a practical and important tool to assess deleterious effects of contamination in freshwater, providing data for monitoring and conservation protocols.

Effects of hydrocarbon pollution in the structure of macrobenthic assemblages from two large estuaries in Brazil

Changes in the structure of benthic macrofauna and its relationship with hydrocarbon contamination were determined at different spatial scales in sublittoral sediments of two large estuaries in Brazil. Guanabara Bay (GB) is a heavily polluted estuary due to the presence of a large industrial complex and high demographic density. Laranjeiras Bay (LB) lies in an Environmental Protection Area and can still be considered as preserved from human activities. Despite some spatial differences within each bay, the PAHs concentrations were significantly and consistently higher in GB, with values generally above the threshold effect levels. No signs of hydrocarbon contamination were observed in LB. Macrofauna abundance, diversity and overall assemblage structure were largely different between bays. Canonical analysis of principal coordinates (CAP), used to model the relationship between macrofauna and PAHs levels, indicated that this class of hydrocarbons is the main structuring factor of soft-bottom assemblages in both bays.