Histopathological Lesions Caused by a Digenean Trematode in a Pest Apple Snail, Pomacea canaliculata, in Its Native Geographic Distribution Area.

Pomacea canaliculata is one of the most dangerous invasive species. Morphological and molecular analyses have revealed that a digenean species belonging to the family Echinostomatidae parasitizes this snail at two sites in Buenos Aires Province, Argentina, South America. Molecular results confirmed that the species belongs to a genus closely related to Patagifer. Analysis of the 28S rDNA showed that the sequences of the rediae and metacercariae are identical, indicating that the apple snail acts as the first and second intermediate host. The cercariae may encyst as metacercaria inside the redia and also emerge and re-infect the same snail or another snail. The prevalence of digeneans was higher in one of the sampling locations (15.1% vs. 0.72%), probably because the bird species that acts as the definitive host is more abundant in that area. Histopathological examination showed that the parasite quickly invades multiple host organs (gills, intestines, albumen gland, lung, kidney, and mantle border) besides the gonad and digestive gland, as is usual in digeneans. In addition, the partial or total castration of snails was observed in cases of moderate and high infection intensity. In males, there was loss of integrity in testicular tubules, while in females, the replacement of ovarian tissue by rediae was found.

Cryptosporidium sp. in cultivated oysters and the natural oyster stock of the state of Maranhão, Brazil / Detecção de Cryptosporidium sp. em ostras de cultivo e estoque natural no estado do Maranhão, Brasil

This study detected Cryptosporidium spp. in cultivated oysters and the natural oyster stock of the state of Maranhão and determine the elective tissue(s) to examine this protozoan. For this purpose, 200 cultivated oysters were purchased from the municipality of Raposa and another 100 from Paço do Lumiar. Additionally, 100 oysters were extracted from the natural stock of the municipality of Primeira Cruz, thus making up a total of 400 oysters. They were grouped into 80 pools consisting of 5 oysters each. From each pool, the gills and visceral mass were removed to obtain 160 pools, 80 pools for the gill group and another 80 for the visceral mass group. Then, DNA was extracted from each pool using a commercial kit with modifications. Subsequently, the protozoan DNA was detected using nested polymerase chain reaction. With this technique, the DNA of the protozoan under investigation was detected in 2.5% (n = 2/80) of the pools containing gills, with 1.25% of the pools (n = 1/80) belonging to the cultivation group of oysters and the other 1.25% (n = 1/80) to the natural stock. With the results obtained in this study, it was concluded that the analyzed oysters of the genus Crassostrea, from cultivation and natural stock groups, found in the state of Maranhão, were contaminated by Cryptosporidium spp. and may become potential sources of infection in humans and other animals. In addition, the gills are the elective tissue for the study of Cryptosporidium spp. in oysters.

Objetivou-se com o estudo detectar Cryptosporidium sp. em ostras de cultivo e estoque natural no estado do Maranhão e determinar o(s) tecido(s) eletivo(s) para pesquisa desse protozoário. Para a realização do estudo foram adquiridas 200 ostras de cultivo do município de Raposa e 100 de Paço do Lumiar, além de 100 ostras extraídas de estoque natural do município de Primeira Cruz, totalizando 400 ostras. Estas foram agrupadas em 80 pools constituídos por cinco animais. De cada pool, as brânquias e a massa visceral foram removidas totalizando 160 pools, sendo 80 para o grupo das brânquias e 80 para o grupo de massa visceral. Na sequência, procedeu-se à extração de DNA de cada pool com a utilização de kit comercial com modificações. Posteriormente, realizou-se a detecção do DNA do protozoário por meio da técnica de Nested-PCR. Com a técnica utilizada, foi detectado o DNA do protozoário pesquisado em 2,5% (n=2/80) pools apenas de brânquias, sendo 1,25% pools (n=1/80) oriundos de cultivo e os outros 1,25% (n=1/80) de estoque natural. Com os resultados obtidos nesse estudo, conclui-se que as ostras analisadas do gênero Crassostrea sp., oriundas de cultivo e estoque natural no estado do Maranhão, estavam contaminadas por Cryptosporidium sp. e podem se reverter em fontes potenciais para seres humanos e outros animais. Para a pesquisa de Cryptosporidium sp. em ostras, as brânquias são o tecido eletivo.

Cryptosporidium sp. in cultivated oysters and the natural oyster stock of the state of Maranhão, Brazil / Detecção de Cryptosporidium sp. em ostras de cultivo e estoque natural no estado do Maranhão, Brasil

ABSTRACT: This study detected Cryptosporidium spp. in cultivated oysters and the natural oyster stock of the state of Maranhão and determine the elective tissue(s) to examine this protozoan. For this purpose, 200 cultivated oysters were purchased from the municipality of Raposa and another 100 from Paço do Lumiar. Additionally, 100 oysters were extracted from the natural stock of the municipality of Primeira Cruz, thus making up a total of 400 oysters. They were grouped into 80 pools consisting of 5 oysters each. From each pool, the gills and visceral mass were removed to obtain 160 pools, 80 pools for the gill group and another 80 for the visceral mass group. Then, DNA was extracted from each pool using a commercial kit with modifications. Subsequently, the protozoan DNA was detected using nested polymerase chain reaction. With this technique, the DNA of the protozoan under investigation was detected in 2.5% (n = 2/80) of the pools containing gills, with 1.25% of the pools (n = 1/80) belonging to the cultivation group of oysters and the other 1.25% (n = 1/80) to the natural stock. With the results obtained in this study, it was concluded that the analyzed oysters of the genus Crassostrea, from cultivation and natural stock groups, found in the state of Maranhão, were contaminated by Cryptosporidium spp. and may become potential sources of infection in humans and other animals. In addition, the gills are the elective tissue for the study of Cryptosporidium spp. in oysters.

RESUMO: Objetivou-se com o estudo detectar Cryptosporidium sp. em ostras de cultivo e estoque natural no estado do Maranhão e determinar o(s) tecido(s) eletivo(s) para pesquisa desse protozoário. Para a realização do estudo foram adquiridas 200 ostras de cultivo do município de Raposa e 100 de Paço do Lumiar, além de 100 ostras extraídas de estoque natural do município de Primeira Cruz, totalizando 400 ostras. Estas foram agrupadas em 80 pools constituídos por cinco animais. De cada pool, as brânquias e a massa visceral foram removidas totalizando 160 pools, sendo 80 para o grupo das brânquias e 80 para o grupo de massa visceral. Na sequência, procedeu-se à extração de DNA de cada pool com a utilização de kit comercial com modificações. Posteriormente, realizou-se a detecção do DNA do protozoário por meio da técnica de Nested-PCR. Com a técnica utilizada, foi detectado o DNA do protozoário pesquisado em 2,5% (n=2/80) pools apenas de brânquias, sendo 1,25% pools (n=1/80) oriundos de cultivo e os outros 1,25% (n=1/80) de estoque natural. Com os resultados obtidos nesse estudo, conclui-se que as ostras analisadas do gênero Crassostrea sp., oriundas de cultivo e estoque natural no estado do Maranhão, estavam contaminadas por Cryptosporidium sp. e podem se reverter em fontes potenciais para seres humanos e outros animais. Para a pesquisa de Cryptosporidium sp. em ostras, as brânquias são o tecido eletivo.

Chemical contamination in coastal areas alters shape, resistance and composition of carnivorous gastropod shells.

Morphological, structural and compositional alterations in shells of molluscs have been proposed as putative biomarkers of chemical contamination in coastal zones. Despite this, few studies were carried out using top predator gastropods which tend to be more susceptible to contamination exposure. Thus, the present study assessed disturbances on shells of Stramonita brasiliensis considering compression resistance and organic and mineralogical matrix composition, related to morphometric alterations. Results showed reductions in compression resistance and organic matrix content associated with higher contaminated sites. In addition, a predominance of calcite polymorphs was seen in shells obtained in polluted areas. Such outputs were consistent with local contamination levels which may have induced the observed alterations. Thus, changes in mollusc shells showed good performance as potential biomarkers of coastal contamination, being probably observed in other species of carnivorous gastropods around the world.

Influence of pH on physiological and behavioral responses of Pomacea canaliculata.

The physiological and behavioral responses of Pomacea canaliculata exposed to different pH values (4, 5, 6, 7, 8, 9, and 10) were evaluated. Survival, behavior (avoidance), metabolites (mantle), net ion fluxes (Na+ and K+), and ATPase activity (gills) were the parameters analyzed. The final survival rates were 100% (pH 4-9) and 90% (pH 10), and the groups did not differ significantly. Avoidance behavior was not identified in animals exposed to an extreme pH compared to pH 7. The main changes observed in the metabolites were in those exposed to an alkaline pH. Glucose (pH 9) and total protein (pH 9 and 10) levels increased, and lactate decreased (pH 9 and 10) compared to a neutral pH. There was an increase in Na+ efflux at pH 4, 5, and 8 and an influx at pH 9 and 10. Extreme pH values (4 and 10) also caused an increase in K+ efflux. At pH values outside the neutrality range (pH 7), there was a significant decrease in the activities of Na+/K+-ATPase (4, 5, 6, 9, and 10) and H+-ATPase (pH 4, 5, 9, and 10). Variations in environmental pH did not cause statistically significant mortality or avoidance behavior in P. canaliculata at the analyzed times. However, due to changes in energy metabolism (glucose and lactate, mainly) and ionoregulation, these can be considered sensitive biomarkers of stress in this species.

Artisanal mollusc fisheries co-management in Brazil and Italy: Institutional innovations to address environmental crisis.

Initiatives of artisanal fisheries co-management and the construction of differentiated markets for seafood products have been emerging in different parts of the world, as an institutionalized way of coping with a global fishery crisis. This paper analyses some institutionalization processes of artisanal mollusc fisheries, considering the role of co-management in two Brazilian and Italian protected areas (Resex Pirajubaé and Conero Regional Park). Within a theoretical framework aiming at moving beyond the dualism between nature and society, the methodology of multiple-case-study has been used to carry on research about mollusc artisanal fisheries co-management networks in their constitution and development. The paper analyses how these networks are organised in the two contexts and the relations social actors have been developing for a sustainable fishery as a possible way to influence and increase their capacity to address environmental crisis. In the artisanal mollusc fishery co-management experiences, fishers' participation may favour institutional innovations and the co-management networks stability may be generated by the institutions legitimacy. Furthermore, the case studies offer complementary insights to better understand the linkage between artisanal fishery institutionalization processes, common natural resources co-management and value aggregation for traditional seafood. Artisanal mollusc fishery co-management experiences should be stimulated and investigated since they can help in diagnosing early climate and environmental changes in the oceans.

Opaepupu, a new genus and species of bivalve-associated shrimp (Decapoda: Caridea: Palaemonidae) from Hawai'i.

A new palaemonid shrimp genus, Opaepupu gen. nov., is established to accommodate a new species of bivalve-associated shrimp, Opaepupu huna sp. nov. from Hawaii. A single mated pair, the female holotype and the male allotype, were found inside the trapezid bivalve Trapezium oblongum (Linnaeus, 1758) at a depth of 14 m in Kâne'ohe Bay, Hawai'i. The new genus is characterised by the rostrum being proximally broad, distally pointed, mid-dorsally carinate, and non-dentate; the anterolateral margin of the carapace without supraorbital, hepatic or epigastric teeth, but with a strong sharp antennal tooth; the sixth pleonite posteriorly unarmed; the telson medially depressed, with the dorsal surface armed with two pairs of submarginal cuspidate setae and with the posterior margin armed with two pairs of spiniform setae; the distolateral angle of the first article of the antennular peduncle without a sharp tooth; the mandible without a palp; the maxillular palp furnished with one long stiff seta dorsal to a small tooth-like extension; the first maxilliped without a palp; the third maxilliped not being operculate; the second pereiopods moderately robust, relatively slender, subequal, subsymmetrical, with simple teeth on the cutting edges of the fingers; the ambulatory pereiopods being slender, each ending in an elongate biunguiculate dactylus; and the uropodal exopod with a faint diaeresis and greatly reduced distolateral spiniform seta. The phylogenetic position of Opaepupu gen. nov. remains unclear, although it does not appear to be closely related to other bivalve-associated palaemonid genera.

Functional and evolutionary perspectives on gill structures of an obligate air-breathing, aquatic snail.

Ampullariids are freshwater gastropods bearing a gill and a lung, thus showing different degrees of amphibiousness. In particular, Pomacea canaliculata (Caenogastropoda, Ampullariidae) is an obligate air-breather that relies mainly or solely on the lung for dwelling in poorly oxygenated water, for avoiding predators, while burying in the mud during aestivation, and for oviposition above water level. In this paper, we studied the morphological peculiarities of the gill in this species. We found (1) the gill and lung vasculature and innervation are intimately related, allowing alternation between water and air respiration; (2) the gill epithelium has features typical of a transporting rather than a respiratory epithelium; and (3) the gill has resident granulocytes within intraepithelial spaces that may serve a role for immune defence. Thus, the role in oxygen uptake may be less significant than the roles in ionic/osmotic regulation and immunity. Also, our results provide a morphological background to understand the dependence on aerial respiration of Pomacea canaliculata. Finally, we consider these findings from a functional perspective in the light of the evolution of amphibiousness in the Ampullariidae, and discuss that master regulators may explain the phenotypic convergence of gill structures amongst this molluscan species and those in other phyla.

Detection of Toxoplasma gondii in Crassostrea spp. oysters cultured in an estuarine region in eastern Amazon.

The aim of the present study was to detect DNA of Toxoplasma gondii in Crassostrea spp. oysters cultured in the state of Pará, Brazil. A total of 400 oysters were directly collected from a fixed rack system. Gills, gastrointestinal tract (GIT) and intervalvular liquid were separated and grouped into pool samples of 10 animals, resulting in 40 samples each of gills, GIT and intervalvular liquid. DNA extraction was performed using a commercial kit, and T. gondii DNA was detected by nested PCR using the primers Toxo3 and Toxo4, which produced an amplification product of 155 bp of the T. gondii gene B1. Nucleotide sequencing was performed for positive samples, and the obtained sequences were identified by comparison with sequences in GenBank. The DNA of T. gondii was detected in 5.8% (7/120) of the pool samples, of which 7.5% (3/40) was in the GIT, 5% (2/40) in the gills, and 5% (2/40) was in the intervalvular liquid. The obtained sequences presented 100% identity and overlap with T. gondii DNA sequences. This is the report of detection of T. gondii DNA in oysters from genus Crassostrea spp. originating from the state of Pará, eastern Amazon.

Nanoemulsion containing essential oil from Xylopia ochrantha Mart. produces molluscicidal effects against different species of Biomphalaria (Schistosoma hosts)

BACKGROUND This work describes a chemical study of the essential oil from leaves of Xylopia ochrantha, an endemic Annonaceae species from Brazil, and its activity against Biomphalaria species. Considering its poor solubility in aqueous medium, the essential oil was nanoemulsified to evaluate its action on controlling some mollusc species of genus Biomphalaria, snail hosts of Schistosoma mansoni that causes schistosomiasis, which mainly affects tropical and subtropical countries. OBJECTIVES The main aims of this work were to analyse the chemical composition of essential oil from X. ochrantha, and to evaluate the effect of its nanoemulsion on molluscs of genus Biomphalaria and their oviposition. METHODS Chemical analysis was performed by gas chromatography coupled to mass spectrometry. Nanoemulsions were prepared by a low energy method and characterised by particle size and polydispersity index. Biological assays evaluating the mortality of adult species of B. glabrata, B. straminea and B. tenagophila and their ovipositions upon contact with the most stable nanoemulsion during 24 and 48 h were performed. FINDINGS Chemical analysis by mass spectrometry revealed the majority presence of bicyclogermacrene and germacrene D in the essential oil. The formulation with a hydrophilic-lipophilic balance (HLB) of 9.26 was the most suitable for the oil delivery system. This nanoemulsion caused the mortality in B. tenagophila, B. straminea and B. glabarata of different sizes at levels ranging from 50 to 100% in 48 h. Additionally, the formulation could inhibit the development of deposited eggs. CONCLUSION Thus, these results suggest the use of nanoemulsified essential oil from X. ochrantha as a possible alternative in controlling some Biomphalaria species involved in the schistosomiasis cycle.

Differential response between histological and biochemical biomarkers in the apple snail Pomacea canaliculata (Gasteropoda: Amullariidae) exposed to cypermethrin.

To develop effective programs to monitor water quality is necessary to identify sensitive biomarkers in indicator species. The aim of this study was to evaluate different biomarkers in the apple snail Pomacea canaliculata exposed to the insecticide Cypermethrin (CYP). Adult male and female snails were exposed to sublethal CYP concentrations (10, 25 and 100µgl-1) for 1, 4, 7 and 14days. The recovery of the exposed snails was also studied by a post-exposure assay. The activities of the enzymes superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST), the levels of lipid peroxidation (LPO) and protein oxidation (PC) in digestive gland and gills were studied as biomarkers of exposure. Histopathological changes in target tissues were also evaluated. In digestive gland, CYP caused a significant increase in SOD, CAT and GST activities compared to control (p<0.05) as well as in LPO and PC levels (p<0.05). However, such biochemical effects were neither concentration nor time dependent. Histopatological changes were observed in the exposed groups, such as an increase in the number of basophilic cells, hemocytic infiltration and epithelia atrophy. Additionally, a positive correlation between the surface occupied by pigmented corpuscles and CYP concentrations was observed at all exposure periods. Gills showed greater sensitivity to oxidative damage than digestive gland. CYP caused an acute toxic effect in LPO levels in this respiratory organ. The gill filament of exposed snails, exhibited a reduction or loss of cilia, vacuolization of the columnar cells and an increase in haemocyte content irrespective of the concentration. High concentrations of CYP caused disruptions in the columnar muscle fibers. In general, snails did not show an improvement in their basal state during post-exposure treatment. Apparently, males and females do not have differential sensitivity to the pesticide. The results of this study suggest that histopathological changes are the most sensitive time- and dose-dependent biomarkers of toxicity induced by CYP in P. canaliculata.

Maintenance of Octopus vulgaris Type II paralarvae in an estuarine area

We assessed the survival of paralarvae kept in a floating wooden box attached to an oyster extensive cultivation system with no extra food supply. A total of 7700 newly hatched paralarvae were maintained in a 10.5-L floating box (7 cm height × 30 cm width × 50 cm length) covered with a 180-µm mesh net for 14 days with no extra food supply. Skin damages and tentacle deformities were observed in 43% of the paralarvae at 14 days after hatching (DAH). The survival rate was 64.7% at 7 DAH and 42.8% at 14 DAH. The floating box is a promising structure for culturing O. vulgaris paralarvae in an extensive system.(AU)

Methanolic extract of Euchelus asper exhibits in-ovo anti-angiogenic and in vitro anti-proliferative activities.

BACKGROUND: The marine environment is a rich source of bioactive natural products. Many of the marine bioactive compounds have been derived successfully from molluscs. Euchelus asper is a marine mollusc which is commonly found in the intertidal rocky regions of the Mumbai coast. The present study was focused on evaluating the anti-angiogenic and anti- proliferative activities of methanolic extract of Euchelus asper (EAME). METHODS: The anti-angiogenic activity of EAME (50-800 µg/mL) was assessed by chick chorio-allantoic membrane (CAM) model wherein multiple parameters in the CAM blood vessels were analysed through morphometric and histological investigations. In vitro testing of EAME (5-20 µg/mL) included its cytotoxicity against three different cancer cell lines, its effect on cell proliferation by wound healing assay as well as their relevant molecular mechanisms. Statistical analysis was carried out by two-tailed student's t test for two unpaired groups. RESULTS: Analysis of CAM revealed that the extract is effective in reducing the branching points of the 1st order blood vessels or capillaries of CAM. Histological analysis of CAM showed significant decrease in capillary plexus and compartmentalization along with increase in mesodermal blood vessels, thus establishing its anti-angiogenicity. Further, EAME exhibited moderate but significant cytotoxicity against A549 non-small cell lung carcinoma cell line. We also demonstrated that the cytotoxicity of EAME in A549 was associated with its apoptotic activity by subG1 phase arrest. Lastly, EAME significantly reduced A549 proliferation by reducing the expression of Matrix metalloproteinase-2 (MMP-2) and Matrix metalloproteinase-9 (MMP-9). CONCLUSION: Overall, our study suggested that EAME has potential to inhibit tumour angiogenic and proliferative activity and may be a potential source for development of new anti-cancer pharmaceuticals.

Bioinvasion Triggers Rapid Evolution of Life Histories in Freshwater Snails.

Biological invasions offer interesting situations for observing how novel interactions between closely related, formerly allopatric species may trigger phenotypic evolution in situ. Assuming that successful invaders are usually filtered to be competitively dominant, invasive and native species may follow different trajectories. Natives may evolve traits that minimize the negative impact of competition, while trait shifts in invasives should mostly reflect expansion dynamics, through selection for colonization ability and transiently enhanced mutation load at the colonization front. These ideas were tested through a large-scale common-garden experiment measuring life-history traits in two closely related snail species, one invasive and one native, co-occurring in a network of freshwater ponds in Guadeloupe. We looked for evidence of recent evolution by comparing uninvaded or recently invaded sites with long-invaded ones. The native species adopted a life history favoring rapid population growth (i.e., increased fecundity, earlier reproduction, and increased juvenile survival) that may increase its prospects of coexistence with the more competitive invader. We discuss why these effects are more likely to result from genetic change than from maternal effects. The invader exhibited slightly decreased overall performances in recently colonized sites, consistent with a moderate expansion load resulting from local founder effects. Our study highlights a rare example of rapid life-history evolution following invasion.

A new terrestrial snail species (Gastropoda: Bulimulidae) from the Región de Antofagasta, northern Chile.

A new species of Scutalus Albers, 1850 (Gastropoda: Bulimulidae), Scutalus chango sp. n., is described from a coastal area of northern Chile. Empty shells of this new species were found buried in sand and under boulders and rocks in the foothills of the Chilean Coastal Range at Paposo, Región de Antofagasta. This new species is distinguished from all other Chilean terrestrial snails by its slender shell with a flared and reflected aperture, and by the presence of a columellar fold. This is the first record of Scutalus in Chile, and the southernmost record for this endemic South American bulimulid genus. The presence of this species in Paposo highlights the need for further research and for conservation guidelines in coastal areas of northern Chile, which have comparatively high levels of biodiversity and endemism.

Analysis of transcriptional responses of normalizing genes on Crassostrea brasiliana under different experimental conditions.

Bivalves show remarkable plasticity to environmental changes and have been proposed as sentinel organisms in biomonitoring. Studies related to transcriptional analysis using quantitative real-time polymerase chain reaction (qRT-PCR) in these organisms have notably increased, imposing a need to identify and validate adequate reference genes for an accurate and reliable analysis. In the present study, 9 reference genes were selected from transcriptome data of Crassostrea brasiliana to identify their suitability as qRT-PCR normalizer genes. The transcriptional patterns were analyzed in gills of oysters under 3 different conditions: different temperatures (18, 24, or 32 °C) and phenanthrene (100 µg L-1 ) combined exposure; different salinities (10, 25, or 35‰) and phenanthrene combined exposure; and 10% of diesel fuel water-accommodated fraction (diesel-WAF) exposure. Reference gene stability was calculated using 5 algorithms (geNorm, NormFinder, BestKeeper, ΔCt, RefFinder). Transcripts of ankyrin-like (ANK), glyceraldehyde 3-phosphate dehydrogenase-like (GAPDH), and α-tubulin-like (TUBA) genes showed minor changes in different temperature/phenanthrene treatment. Transcripts of ANK, ß-actin-like, and ß-tubulin-like genes showed better stability at salinity/phenanthrene treatment, and ANK, TUBA, and 28S ribosomal protein-like genes showed the most stable transcription pattern in oysters exposed to diesel-WAF exposure. The present study constitutes the first systematic analysis of reference gene selection for qRT-PCR normalization in C. brasiliana. These genes could be employed in studies using qRT-PCR analysis under similar experimental conditions. Environ Toxicol Chem 2017;36:2190-2198. © 2017 SETAC.

Methanolic extract of Euchelus asper exhibits in-ovo anti-angiogenic and in vitro anti-proliferative activities

BACKGROUND: The marine environment is a rich source of bioactive natural products. Many of the marine bioactive compounds have been derived successfully from molluscs. Euchelus asper is a marine mollusc which is commonly found in the intertidal rocky regions of the Mumbai coast. The present study was focused on evaluating the anti-angiogenic and anti- proliferative activities of methanolic extract of Euchelus asper (EAME). METHODS: The anti-angiogenic activity of EAME (50-800 µg/mL) was assessed by chick chorio-allantoic membrane (CAM) model wherein multiple parameters in the CAM blood vessels were analysed through morphometric and histo-logical investigations. In vitro testing of EAME (5-20 µg/mL) included its cytotoxicity against three different cancer cell lines, its effect on cell proliferation by wound healing assay as well as their relevant molecular mechanisms. Statistical analysis was carried out by two-tailed student's t test for two unpaired groups. RESULTS: Analysis of CAM revealed that the extract is effective in reducing the branching points of the 1st order blood vessels or capillaries of CAM. Histological analysis of CAM showed significant decrease in capillary plexus and compartmentalization along with increase in mesodermal blood vessels, thus establishing its anti-angiogenicity. Further, EAME exhibited moderate but significant cytotoxicity against A549 non-small cell lung carcinoma cell line. We also demonstrated that the cytotoxicity of EAME in A549 was associated with its apoptotic activity by subG1 phase arrest. Lastly, EAME significantly reduced A549 proliferation by reducing the expression of Matrix metalloproteinase-2 (MMP-2) and Matrix metalloproteinase-9 (MMP-9). CONCLUSION: Overall, our study suggested that EAME has potential to inhibit tumour angiogenic and proliferative activity and may be a potential source for development of new anti-cancer pharmaceuticals.

Leachate from microplastics impairs larval development in brown mussels.

Microplastic debris is a pervasive type of contaminant in marine ecosystems, being considered a major threat to marine biota. One of the problems of microplastics is that they can adsorb contaminants in extremely high concentrations. When released from the particle, these contaminants have the potential to cause toxic effects in the biota. So far, reports of toxic effects are mostly linked with the direct exposure of organisms through ingestion of contaminated microplastics. There is little information on the toxicity of leachates from microplastics to marine organisms. In this study, we conducted experiments to evaluate the toxicity of leachates from virgin and beached plastic pellets to embryo development of the brown mussel (Perna perna). We compared the efficiency of two test procedures, and evaluated the toxicity of beached pellets collected in a coastal marine protected area. We observed that mussel embryo is sensitive to leachate from both virgin and beached pellets. However, the toxicity of the leachate from beached pellets was much higher than that of virgin pellets. We suggest contaminants adsorbed onto the surface of beached pellets were responsible for the high toxicity of leachate from beached pellets, while the toxicity of leachate from virgin pellets was mainly due to plastic additives. Our results suggest microplastic debris may be harmful even if ingestion is not the only or main pathway of interaction of marine organisms with contaminated plastic debris.

O mexilhão Perna perna no Brasil: nativo ou exótico? / The brown mussel Perna perna in Brazil: native or exotic?

A mitilicultura no Brasil se baseia no mexilhão Perna perna (Linnaeus, 1758), que é encontrado em todo o litoral brasileiro, sendo especialmente abundante do Espírito Santo a Santa Catarina. Nos últimos anos, foi lançada a hipótese de que P. perna seja uma espécie exótica no litoral brasileiro. A hipótese se baseia na análise da malacofauna presente em sítios arqueológicos. Todas as contestações levantadas não remetem à uma conclusão, pois precisam de estudos específicos, com metodologias claras, aliando arqueologia, ecologia e biologia molecular. O objetivo deste trabalho foi estudar a condição de nativo ou exótico do mexilhão Perna perna no Brasil, a partir de levantamento dos resultados em sítios arqueológicos, de técnicas moleculares e de datação de conchas com C14. A datação indicou que as amostras de Perna perna do sítio arqueológico Rio do Meio/Jurerê, Florianópolis/SC, têm idade de 720±30 e 780±30 anos. O cálculo do tempo de divergência indicou que a separação das populações brasileiras e africanas ocorreu por volta de 200 mil anos. Os resultados apontam a presença da espécie no território brasileiro muito antes do descobrimento do Brasil pelos portugueses no ano de 1500, indicando que P. perna é de fato uma espécie nativa.(AU)

Mussel farming in Brazil is based on brown mussel Perna perna (Linnaeus, 1758), which is found throughout the Brazilian coast, especially abundant from Espírito Santo to Santa Catarina coast. In recent years it was suggested that Perna perna is an exotic species for the Brazilian coast. The hypothesis is based on the analysis of the zooarchaeology studies in archaeological sites in Brazil. All objections raised do not offer a conclusion, because they need specific studies with clear methodology, combining archeology, ecology and molecular biology. The aim of this work was to study the condition of the brown mussel Perna perna in Brazil if native or exotic, from survey results in archaeological sites, molecular techniques and dating of shells with C14. The dating indicated that the shells were 720±30 and 780±30 years, respectively. The calculation of divergence time indicated that the separation of the African and Brazilian mussel populations occurred around 200 thousand years ago. The results indicate the presence of the P. perna species in Brazilian territory long before the discovery of Brazil by the Portuguese in 1500, indicating that P. perna is actually a native species of Brazil.(AU)

O mexilhão Perna perna no Brasil: nativo ou exótico? / The brown mussel Perna perna in Brazil: native or exotic?

A mitilicultura no Brasil se baseia no mexilhão Perna perna (Linnaeus, 1758), que é encontrado em todo o litoral brasileiro, sendo especialmente abundante do Espírito Santo a Santa Catarina. Nos últimos anos, foi lançada a hipótese de que P. perna seja uma espécie exótica no litoral brasileiro. A hipótese se baseia na análise da malacofauna presente em sítios arqueológicos. Todas as contestações levantadas não remetem à uma conclusão, pois precisam de estudos específicos, com metodologias claras, aliando arqueologia, ecologia e biologia molecular. O objetivo deste trabalho foi estudar a condição de nativo ou exótico do mexilhão Perna perna no Brasil, a partir de levantamento dos resultados em sítios arqueológicos, de técnicas moleculares e de datação de conchas com C14. A datação indicou que as amostras de Perna perna do sítio arqueológico Rio do Meio/Jurerê, Florianópolis/SC, têm idade de 720±30 e 780±30 anos. O cálculo do tempo de divergência indicou que a separação das populações brasileiras e africanas ocorreu por volta de 200 mil anos. Os resultados apontam a presença da espécie no território brasileiro muito antes do descobrimento do Brasil pelos portugueses no ano de 1500, indicando que P. perna é de fato uma espécie nativa.

Mussel farming in Brazil is based on brown mussel Perna perna (Linnaeus, 1758), which is found throughout the Brazilian coast, especially abundant from Espírito Santo to Santa Catarina coast. In recent years it was suggested that Perna perna is an exotic species for the Brazilian coast. The hypothesis is based on the analysis of the zooarchaeology studies in archaeological sites in Brazil. All objections raised do not offer a conclusion, because they need specific studies with clear methodology, combining archeology, ecology and molecular biology. The aim of this work was to study the condition of the brown mussel Perna perna in Brazil if native or exotic, from survey results in archaeological sites, molecular techniques and dating of shells with C14. The dating indicated that the shells were 720±30 and 780±30 years, respectively. The calculation of divergence time indicated that the separation of the African and Brazilian mussel populations occurred around 200 thousand years ago. The results indicate the presence of the P. perna species in Brazilian territory long before the discovery of Brazil by the Portuguese in 1500, indicating that P. perna is actually a native species of Brazil.