Cytotoxicity and enzymatic biomarkers as early indicators of benthic responses to the soluble-fraction of diesel oil.

Xenobiotics from oil tanker leaks and industrial discharges are amongst the main human impacts to confined coastal areas. We assessed the genotoxic responses to the water-soluble fraction of diesel oil in the polychaete Laeonereis culveri and the bivalve Anomalocardia flexuosa, two widespread benthic species in subtropical estuaries from the Southwestern Atlantic. We hypothesized that the highest responsiveness would be expressed by significantly different biomarkers responses between control and oil-impacted treatments. Responsiveness to diesel oil was investigated using an experimental design with two fixed factors (contaminant percentages and times of exposure). After exposure, we monitored the responses of the oxidative stress enzymes and performed micronuclei tests. Results were congruent for both species. Antioxidant defense of glutathione S-transferase and the induction of micronuclei and nuclear buds, the latter just for the bivalve, were significantly affected by polycyclic aromatic hydrocarbons, with significant increases on the seventh day of exposure and in the higher concentrations, compared to controls groups. We assessed the benefits and drawbacks of using each biomarker in laboratory experiments. Both species are indicators of early, and rapid responses to genotoxic contaminants in subtropical estuarine habitats. We suggest that the micronuclei frequency in A. flexuosa is a simple, fast and cheap test for genotoxicity in oil-impacted areas. Such early biomarkers are needed to develop better protocols for impact assessment and monitoring under real field conditions.

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.

A new empirical index for assessing the vulnerability of peri-urban mangroves.

Environmental vulnerability can be understood as a function of exposure to impacts and the sensitivity and adaptive capacity of ecological systems towards environmental tensors. The present study empirically evaluated the vulnerability of forest stands that make up the subtropical Perequê River mangrove (Paranaguá Bay, S Brazil) as the basis for a more appropriate management of its resources. This mangrove is located in the coastal municipality of Pontal do Paraná (Paraná, Brazil) and is part of the Perequê River Mangrove Natural Park, which is still not under a management plan. This study aimed to contribute to the future management plan of this conservation unit through an environmental diagnosis and an empirical evaluation of its vulnerability. The primary data, collected from 51 plots of 100 m(2) each, involved the evaluation of: 1) the exposure to solid waste, deforestation, trails, landfills, and constructions; 2) the degree of sensitivity according to the environmental conditions of each plot; and 3) the adaptive capacity according to the recovery and reproductive potential and structural complexity of the forests. These data were integrated to compose the empirical index of environmental vulnerability (VI) expressed as VI = (SI + EI) - ACI where SI is the sensitivity sub-index, EI is the exposure sub-index, and ACI is the adaptive capacity sub-index. Empirical indices calculated for local forests varied significantly due to differences in values of SI, EI and ACI. This variability defines the local mangrove as a mosaic of vulnerability conditions and suggests that the planning and implementation of local management actions consider such environmental heterogeneity. Applying or extending these actions should be based on local forest conditions at short spatial scales and should not be applied to the mangrove as a whole.

First Insights into Body Localization of an Osmoregulation-Related Cotransporter in Estuarine Annelids.

The expression of the Na+-K+-2Cl- cotransporter (NKCC), widely associated with cell volume regulation, has never been directly demonstrated in annelids. Its putative presence was firstly recovered in silico, and then using immunofluorescence, its signal was retrieved for the first time in different tissues of four species of estuarine annelids from southern Brazil that are regularly subjected to salinity fluctuations. We tested two euryhaline species (wide salinity tolerance), the nereidids Alitta yarae and Laeonereis acuta (habitat salinity: ~10-28 psu), and two stenohaline species (restricted salinity tolerance), the nephtyid Nephtys fluviatilis (habitat salinity: ~6-10 psu), and the melinnid Isolda pulchella (habitat salinity: ~28-35 psu). All four species showed specific immunofluorescent labelling for NKCC-like expression. However, the expression of an NKCC-like protein was not homogeneous among them. The free-living/burrowers (both euryhaline nereidids and the stenohaline nephtyid) displayed a widespread signal for an NKCC-like protein along their bodies, in contrast to the stenohaline sedentary melinnid, in which the signal was restricted to the branchiae and the internal tissues of the body. The results are compatible with NKCC involvement in cell volume, especially in annelids that face wide variations in salinity in their habitats.

Phycotoxins and marine annelids - A global review.

Several species of microalgae can produce potent phycotoxins that negatively affect aquatic organisms and their consumers following different exposure routes, as well as toxicokinetic (TK) and toxicodynamic (TD) processes. Benthic organisms are especially vulnerable as they are exposed to both benthic and planktonic species causative of harmful algal blooms (HABs). While benthic algae can come into direct contact with annelids during substrate remobilization, planktonic cells can settle to the bottom mostly during senescence and/or encystment stages, and in shallow and calm waters. We performed a systematic, qualitative review of the literature on the phycotoxin TK and TD processes in marine annelids, summarizing the most relevant findings and general trends. Besides, by using innovative analytical/statistical approaches, we were able to detect patterns and gaps in the current literature, thus pointing to future research directions. We retrieved and analyzed studies involving diarrhetic shellfish toxins (DSTs), paralytic shellfish toxins (PSTs), brevetoxins (PbTXs), domoic acid (DA), as well as palytoxin and its congeners, the ovatoxins (treated together as PLTXs). It is worth mentioning that studies evaluating other phycotoxins (e.g., ciguatoxins, yessotoxins) were not found in the literature. The absence of data on PbTXs, PSTs and DA is the largest gap hampering TK assessment in annelids, although some relevant information on TD is already available. Whereas lethal effects from DSTs have not been reported, more potent toxins like PbTXs, PSTs, DA and those grouped as PLTX-like compounds can cause mortality and/or marked decrease in annelid abundance. In addition, phycotoxins have been linked to sublethal effects on annelid cells. Although very sparse, field and laboratory studies offer strong evidence that annelids may be reliable indicators of toxin exposure and their negative effects during both early and later stages of HABs in marine environments. Besides quickly responding to these compounds at both organismic and suborganismic levels, annelids are easily found in areas affected by HABs. The use of annelids in future investigations evaluating the action mechanisms of toxic microalgae on marine invertebrates should be thus encouraged. In this case, the choice for widely dispersed and numerically dominant species of annelids would strengthen the validation and extrapolation of results from risk assessments in areas affected by HABs worldwide.

Mangrove interaction with saltmarsh varies at different life stages.

Mangroves and saltmarshes are two of the most relevant coastal habitats for humans. These ecosystems offer several services like coastal protection, climate mitigation, and nursery habitats for many artisanal and commercially exploited fish, crabs, and shellfish. They mostly dominate different latitudinal ranges but in several places around the world they co-occur and interact. Here, we summarize the current scientific knowledge on mangrove-saltmarsh ecological interactions and propose a conceptual model. We screened 1410 articles from 1945 to 2022 and selected 29 experiments that assessed mangrove-saltmarsh ecological interactions. Both positive and negative interactions are observed but there is variation along different mangrove life stages. Higher retention and establishment of mangrove propagules are found inside saltmarshes than on bare flats, i.e. facilitation, and these effects are higher at grass than at succulent saltmarsh species. Mangrove seedlings, saplings, or trees mostly compete with saltmarshes, negatively affecting mangrove growth. We propose a model with different outcomes considering the interaction between different mangrove's life stages and saltmarsh forms and discussed these interactions in the light of anthropogenic threats and climate change.

A new look at the potential role of marine plastic debris as a global vector of toxic benthic algae.

Marine plastic debris provides a significant surface area for potential colonization by planktonic and benthic harmful microalgae and for the adsorption of their toxins. Furthermore, floating plastics may substantially expand the substrate area available for benthic algae in the ocean, intensifying the transfer of potent toxins through pelagic food webs. In this study, we quantify the available surface area of micro- and macroplastics in different oceanic regions and assess the potential role of floating plastics as vectors for the transfer of toxins from three widespread benthic dinoflagellates, Gambierdiscus spp., Ostreopsis cf. ovata and Prorocentrum lima. To avoid bias associated to the occurrence of benthic algae in deep waters, we selected only records from 0 to 100 m depths. We estimate that 26.8 × 1010 cm2 of plastic surface area is potentially available in surface waters of the global ocean, mostly in the size range of large microplastics (1.01-4.75 mm). Based on the distribution of floating plastics and the habitat suitability of the selected microalgal species, the plastic relative colonization risks will be greater in the Mediterranean Sea and in the subtropical and temperate western margins of the oceans, such as the North American and Asian eastern coasts and, to a lesser extent, southern Brazil and Australia. In places where the colonization of O. cf. ovata cells on floating plastic debris has been properly quantified, such as the Mediterranean and southern Brazil, we estimate a colonization potential of up to 2 × 106 cells km-2 of ocean surface during the regular occurrence period and up to 1.7 × 108 cells km-2 during massive blooms of this species. As plastic pollution and harmful benthic algal blooms have both increased substantially over the past decades, we suggest that their interactive effects can become a major and novel threat to marine ecosystems and human health.

Reviewing the effects of contamination on the biota of Brazilian coastal ecosystems: Scientific challenges for a developing country in a changing world.

Pollution is a major worldwide problem that is increasing with urban growth, mainly along coastal areas. Pollution is often worse, governance is poorer and managerial strategies to improve environmental quality are less advanced in developing than developed countries. Here, we present an overview of the current scientific knowledge of the impacts of contamination on the biota of coastal ecosystems of Brazil and evaluate the scientific challenges to provide baseline information for local managerial purposes. We compiled data from 323 peer-reviewed published papers from the extensive Brazilian coast. We critically evaluated the produced knowledge (target contaminants, sources, ecosystems, taxa, response variables) and the science behind it (rigour and setting) within its socioenvironmental context (land occupation, use of the coast, sanitation status, contamination history). Research was driven largely by environmental outcomes of industrial development with a focus on the single effects of metals on the biota. The current knowledge derives mainly from laboratory manipulative experiments or from correlative field studies of changes in the biota with varying levels of contamination. Of these, 70% had problems in their experimental design. Environmental impacts have mainly been assessed using standard indicators of populations, mostly in ecotoxicological studies. Benthic assemblages have mostly been studied using structural indicators in field studies. Future assessments of impacts should expand research to more taxonomic groups and ecosystem compartments, adding combined functional and structural responses. Furthermore, further investigations need to consider the interactive effects of contaminants and other environmental stressors. By doing so, researchers would deliver more robust and effective results to solve problems of pollution.

Ten new species of Phyllodoce Lamarck, 1818 (Phyllodocidae, Annelida) from Brazil.

Herein we provide a taxonomic revision of Phyllodoce species from Brazil, describing 10 new species in addition to two new records, Phyllodoce longipes Kinberg, 1866 and Phyllodoce cf. madeirensis Langerhans, 1880. Phyllodoce sp. A. and Phyllodoce sp. B. are probably new but the number and condition of available specimens do not provide adequate and reliable diagnostic features for a formal description. These species have been collected in diverse marine and estuarine environments from shallow estuarine bottoms to continental shelf and slope sediments and submarine canyons from off southern and southeastern Brazil. A key to the fourteen species of Phyllodoce occurring in Brazil is also given.

Who's who in Magelona: phylogenetic hypotheses under Magelonidae Cunningham & Ramage, 1888 (Annelida: Polychaeta).

Known as shovel head worms, members of Magelonidae comprise a group of polychaetes readily recognised by the uniquely shaped, dorso-ventrally flattened prostomium and paired ventro-laterally inserted papillated palps. The present study is the first published account of inferences of phylogenetic hypotheses within Magelonidae. Members of 72 species of Magelona and two species of Octomagelona were included, with outgroups including members of one species of Chaetopteridae and four of Spionidae. The phylogenetic inferences were performed to causally account for 176 characters distributed among 79 subjects, and produced 2,417,600 cladograms, each with 404 steps. A formal definition of Magelonidae is provided, represented by a composite phylogenetic hypothesis explaining seven synapomorphies: shovel-shaped prostomium, prostomial ridges, absence of nuchal organs, ventral insertion of palps and their papillation, presence of a burrowing organ, and unique body regionation. Octomagelona is synonymised with Magelona due to the latter being paraphyletic relative to the former. The consequence is that Magelonidae is monotypic, such that Magelona cannot be formally defined as associated with any phylogenetic hypotheses. As such, the latter name is an empirically empty placeholder, but because of the binomial name requirement mandated by the International Code of Zoological Nomenclature, the definition is identical to that of Magelonidae. Several key features for future descriptions are suggested: prostomial dimensions, presence/absence of prostomial horns, morphology of anterior lamellae, presence/absence of specialised chaetae, and lateral abdominal pouches. Additionally, great care must be taken to fully describe and illustrate all thoracic chaetigers in descriptions.

Antioxidant responses and okadaic acid accumulation in Laeonereis acuta (Annelida) exposed to the harmful dinoflagellate Prorocentrum cf. lima.

We evaluated the accumulation of okadaic acid (OA), a diarrhetic toxin, and the antioxidant responses in the marine annelid Laeonereis acuta exposed to the benthic toxigenic dinoflagellate Prorocentrum cf. lima. Nontoxic Tetraselmis sp. was used as a control diet. Living cells of the two algae were supplied as food to animals kept in agar medium for 72 h. To assess the significance of the observed effects, our experimental design treated the algal species (diet), algal cell densities, and exposure time as fixed factors. Responses of the organisms were assessed through oxidative stress biomarkers (glutathione-S-transferase [GST], catalase [CAT], reduced glutathione [GSH] and lipid peroxidation [LPO]). Toxin accumulation was measured by LC-MS/MS in whole-body homogenates after 12, 24 and 72 h of exposure. Worms exposed to the toxigenic dinoflagellate gradually accumulated OA, with toxin levels directly related to the cell density of Prorocentrum cf. lima. Worms fed with Prorocentrum cf. lima exhibited decreased CAT activity, increased LPO levels - both interactively affected by algal species and time - and decreased GSH levels, which were interactively affected by algal species and cell density. Higher LPO levels, along with the inhibition of CAT and GSH, clearly indicated an oxidative stress situation in worms exposed to the toxigenic dinoflagellate. Laeonereis acuta accumulated moderate OA levels and may act as a vector of OA to food webs in estuarine areas under high Prorocentrum cf. lima abundance.

Finding Aquaporins in Annelids: An Evolutionary Analysis and a Case Study.

Aquaporins (AQPs) are a family of membrane channels facilitating diffusion of water and small solutes into and out of cells. Despite their biological relevance in osmoregulation and ubiquitous distribution throughout metazoans, the presence of AQPs in annelids has been poorly investigated. Here, we searched and annotated Aqp sequences in public genomes and transcriptomes of annelids, inferred their evolutionary relationships through phylogenetic analyses and discussed their putative physiological relevance. We identified a total of 401 Aqp sequences in 27 annelid species, including 367 sequences previously unrecognized as Aqps. Similar to vertebrates, phylogenetic tree reconstructions clustered these annelid Aqps in four clades: AQP1-like, AQP3-like, AQP8-like and AQP11-like. We found no clear indication of the existence of paralogs exclusive to annelids; however, several gene duplications seem to have occurred in the ancestors of some Sedentaria annelid families, mainly in the AQP1-like clade. Three of the six Aqps annotated in Alitta succinea, an estuarine annelid showing high salinity tolerance, were validated by RT-PCR sequencing, and their similarity to human AQPs was investigated at the level of "key" conserved residues and predicted three-dimensional structure. Our results suggest a diversification of the structures and functions of AQPs in Annelida comparable to that observed in other taxa.

Euryhalinity of subtropical marine and estuarine polychaetes evaluated through carbonic anhydrase activity and cell volume regulation.

Polychaete worms are widespread and diverse in marine and estuarine habitats subject to varying salinity, in areas influenced by tides, demanding physiological adjustment for internal homeostasis. They are typically considered and reported to be osmoconformers, but they are not often studied for their osmoregulation. Here, three species of polychaete worms from distinct coastal habitats have been investigated: the spionid Scolelepis goodbody (intertidal in saline, exposed sandy beaches), the nereidid Laeonereis culveri (estuarine polyhaline), and the nephtyid Nephtys fluviatilis (estuarine oligohaline). The general objective here was to relate ecological aspects and physiology of the studied species. Constitutive whole body osmolality and carbonic anhydrase activity (CAA, relevant for osmoregulation, acid-base balance and respiration) have been assayed. In addition, cell volume regulatory capacity (from whole body cell dissociation) was challenged under hypoosmotic and hyperosmotic shocks (50% intensity), with respect to isosmotic control. S. googdbody and L. culveri, the two species from most saline environments (marine/estuarine), showed higher CAA than N. fluviatilis, which, in turn, displayed a hyperosmotic gradient to water of salinity 15. Cells from S. goodbody and L. culveri showed regulatory volume decrease upon swelling, with S. goodbody showing the largest volume increase. As in other more studied marine invertebrate groups, polychaetes also show variability in their osmoregulatory physiology, related to distinct saline challenges faced in their coastal habitats.

Seasonal variation of oxidative biomarkers in gills and digestive glands of the clam Anomalocardia flexuosa and the mangrove oyster Crassostrea rhizophorae.

We determined values of antioxidant biomarkers catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione S-transferase (GST) and malondialdehyde (MDA) in two bivalve species from a subtropical system in Brazil. Differences in the biomarkers activities measured in digestive glands and gills sampled at two seasons and in sites with different levels of contamination were tested using univariate and multivariate analyses. Seasonality was the main factor accounting for overall variability. Significant SOD and GPx activities and increased levels of lipid peroxides were detected in summer, while CAT activity was higher in winter. However, biomarkers respond to complex environmental settings as indicated by the significant interaction between season and contamination level. We propose A. flexuosa as a candidate for monitoring studies with sampling occurring at a yearly timescale, and using C. rhizophorae is better suited for designs that include both seasons.

Metataxonomic and metagenomic analysis of mangrove microbiomes reveals community patterns driven by salinity and pH gradients in Paranaguá Bay, Brazil.

While environmental drivers regulate the structure of mangrove microbial communities, their exact nature and the extent of their influence require further elucidation. By means of 16S rRNA gene-based sequencing, we determined the microbial taxonomic profiles of mangroves in the subtropical Paranaguá Bay, Brazil, considering as potential drivers: salinity, as represented by two sectors in the extremes of a salinity gradient (<5 PSU and >30 PSU); proximity to/absence of the prevailing plants, Avicennia schaueriana, Laguncularia racemosa, Rhizophora mangle, and Spartina alterniflora; and the chemical composition of the sediments. Salinity levels within the estuary had the strongest influence on microbial structure, and pH was important to separate two communities within the high salinity environment. About one fourth of the total variation in community structure resulted from covariation of salinity and the overall chemical composition, which might indicate that the chemical profile was also related to salinity. The most prevalent bacterial phyla associated with the mangrove soils analyzed included Proteobacteria, Actinobacteria, Chloroflexi, Bacteroidetes, Acidobacteria, and Cyanobacteria. Taxonomic and functional comparisons of our results for whole-genome sequencing with available data from other biomes showed that the studied microbiomes cluster first according to biome type, then to matrix type and salinity status. Metabolic functions were more conserved than organisms within mangroves and across all biomes, indicating that core functions are preserved in any of the given conditions regardless of the specific organisms harboring them.

Petroleum biomarkers as tracers of low-level chronic oil contamination of coastal environments: A systematic approach in a subtropical mangrove.

Petroleum biomarkers (hopanes, terpanes and steranes) are frequently assessed in estuarine sediments as tracers of oil input. In order to compare distinct patterns of hydrocarbon accumulation in mudflats, salt marsh and mangrove, sediments from two transects (control and impacted areas) were sampled in Paranaguá Bay, SW Atlantic. Concentrations of n-alkanes, polycyclic aromatic hydrocarbons (PAHs) and petroleum biomarkers (hopanes, terpanes and steranes) were determined, as well as bulk parameters (TOC, grain size and δ13C). N-alkanes concentrations were similar between control and impacted sites (respectively, 3.03 ±â€¯1.20 µg g-1 and 4.11 ±â€¯3.02 µg g-1) and reflected a high biogenic input. Conversely, PAHs and petroleum biomarker concentrations were three to six times higher in impacted site than the control site (respectively, 60.4 ±â€¯23.3 ng g-1 and 22.0 ±â€¯25.0 ng g-1 for PAHs and 197.7 ±â€¯51.8 ng g-1 and 40.2 ±â€¯32. ng g-1 for hopanes). Despite these differences, concentrations were lower than those reported for highly impacted areas worldwide. Diagnostic ratios and hydrocarbon parameters (e.g. total PAHs and total petroleum biomarkers) helped to distinguish human impact in the ecological zones, suggesting different sources and/or levels of weathering, confirmed by ANOVA tests. TOC played a fundamental role to the concentration of hydrocarbons, showing similar distributions along the transects. Petroleum biomarkers could clearly indicate the preferential sites of deposition and assign different levels of anthropic contamination by hydrocarbons, thus providing clear information about the chronic petroleum pollution in coastal sediments.

Strategies for tube construction in Owenia caissara (Oweniidae, Annelida) from southern Brazil.

In this paper, we have experimentally assessed tube-building strategies of Owenia caissaraSilva & Lana, 2017, including the particle size preferences. After acclimation, individual tubes were broken by their mid-region, and placed in experimental aquaria with four types of homogeneous substrates (from silt-clay to coarse sand) and four types of mixed substrates. Animals completely removed from their tubes were unable to build new tubes. Adults in broken tubes were able to use a wide range of particles, from fine to coarse sand, but not silt-clay. Owenia caissara showed a clear preference for coarse sediment particles. In extreme conditions, individuals were able to rebuild their tubes using only filamentous debris. Owenia caissara may display higher selection ability and preference for larger particles when growing, although adults can also adapt to bottoms with predominantly smaller particles such as fine sand.

Four new species of Paranaitis Southern, 1914 (Phyllodocidae, Annelida) from southern and southeastern Brazil.

Four new species of Paranaitis are described from estuarine and continental shelf bottoms from southern and southeastern Brazil: Paranaitis chitinosa sp. nov., P. cirriformata sp. nov., P. assimetrica sp. nov., and P. cordiformata sp. nov. Paranaitis chitinosa sp. nov. is diagnosed by a proboscis with rows of conical and chitinous papillae alternating with dorsal chitinous bars, and the distal extremity covered with micropapillae, and dorsal cirri rounded and asymmetrical. Paranaitis cirriformata sp. nov. is similar to P. speciosa (Webster, 1879), previously recorded from Brazil, but differs by the presence of long and slender anal cirri. Paranaitis assimetrica sp. nov. differs from P. cordiformata sp. nov. by the presence of supra-acicular lobes that are considerably longer than the subacicular ones, and oval dorsal cirri. Paranaitis cordiformata sp. nov. has cordiform dorsal cirri and symmetrical supra- and sub-acicular lobes.

Testing biomarker feasibility: a case study of Laeonereis culveri (Nereididae, Annelida) exposed to sewage contamination in a subtropical estuary.

The feasibility of biomarkers to accurately detect pollution in estuaries and coastal areas appears to be overestimated in the scientific literature, and is likely stemming from a bias towards publishing only positive results. Herein, we test the applicability of antioxidant capacity and lipid peroxidation to detect potential in situ sewage impacts on a subtropical estuary. We experimentally assessed variation in malondialdehyde (MDA, an indicator of lipid peroxidation) and total antioxidant capacity against peroxyl radicals (ACAP) through transplantations of the worm Laeonereis culveri from control to sewage-polluted areas. Variations in biomarkers were more related to natural environmental variability and small-scale heterogeneity among areas than to experimental exposure to sewage. Use of oxidative stress biomarkers is promising but further experimental work under real-life conditions is necessary to further validate and establish their accuracy and feasibility for practical purposes.