Evaluation of Chronic Effects of Potassium Chloride and Nickel on Survival, Growth, and Reproduction of a Unionid Mussel (Lampsilis siliquoidea).

The ASTM International standard test method for freshwater mussels (E2455-13) recommends 4-week toxicity testing with juveniles to evaluate chronic effects on survival and growth. However, concerns remain that the method may not adequately address the sensitivity of mussels to longer term exposures (>4 weeks), particularly in relation to potential reproductive impairments. No standard method directly evaluates toxicant effects on mussel reproduction. The objectives of the present study were to (1) evaluate toxicity endpoints related to reproduction in fatmucket (Lampsilis siliquoidea) using two common reference toxicants, potassium chloride (KCl) and nickel (Ni); (2) evaluate the survival and growth of juvenile fatmucket in standard 4-week and longer term (12-week) KCl and Ni tests following a method refined from the standard method; and (3) compare the sensitivity of the reproductive endpoints with the endpoints obtained from the juvenile mussel tests. Reproductive toxicity tests were conducted by first exposing female fatmucket brooding mature larvae (glochidia) to five test concentrations of KCl and Ni for 6 weeks. Subsamples of the glochidia were then removed from the adults to determine three reproductive endpoints: (1) the viability of brooded glochidia; (2) the viability of free glochidia in a 24-h exposure to the same toxicant concentrations as their mother; and (3) the success of glochidia parasitism on host fish. Mean viability of brooded glochidia was significantly reduced in the high KCl concentration (26 mg K/L) relative to the control, with a 20% effect concentration (EC20) of 14 mg K/L, but there were no significant differences between the control and any Ni treatment (EC20 > 95 µg Ni/L). The EC20s for viability of free glochidia after the additional 24-h exposure and parasitism success were similar to the EC20s of brooded glochidia. The EC20s based on the most sensitive biomass endpoint in the 4-week juvenile tests were 15 mg K/L and 91 µg Ni/L, similar to or greater than the EC20s from the reproductive KCl and Ni tests, respectively. When exposure duration in the juvenile tests was extended from 4 to 12 weeks, the EC20s decreased by more than 50% in the KCl test but by only 8% in the Ni test. Overall, these results indicate that a standard 4-week test with juvenile mussels can prove effective for estimating effects in chronic exposures with different life stages although a longer term 12-week exposure with juvenile mussels may reveal higher sensitivity of mussels to some toxicants, such as KCl. Environ Toxicol Chem 2024;43:1097-1111. © 2024 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Proporción sexual y hermafroditismo del molusco, Anadara tuberculosa (Bivalvia: Arcidae) en Panamá / Sexual proportion and hermaphroditism of the mollusc, Anadara tuberculosa (Bivalvia: Arcidae) in Panama

Introducción: El bivalvo Anadara tuberculosa es un recurso pesquero y económico importante en el Pacífico Este Tropical. La proporción sexual varia a lo largo de su área de distribución, con casos de aparición de hermafroditismo, ocasional para algunas poblaciones, normal en otras. Estas características podrían ser una respuesta a presiones antropogénicas y ambientales. Objetivo: Evaluar la proporción sexual y el hermafroditismo en la especie en el Pacífico de Panamá. Métodos: Se analizó información reproductiva de A. tuberculosa generada entre 1994 y 2020 para el Golfo de Montijo, así como datos de 2017 para el manglar de David. Se aplicaron procedimientos macroscópicos, contrastados con histológicos y se generó un factor de corrección para ajustar la proporción sexual macroscópica, debido al efecto, sobre la proporción sexual, que ocasiona los organismos indeterminados. Resultados: Para el Golfo de Montijo, la histología indicó una proporción sexual hembra a macho de 1:1 hasta 3.2:1, entre 1994 y 2017, mientras que la macroscopía varió de 1.9:1 a 5.1:1 entre 2004 y 2020. Para el manglar de David, la histología indicó una proporción sexual de 3.7:1, y de 7.9:1 por macroscopía en el 2017. En ambas localidades, especímenes más grandes mostraron mayor variación en la proporción sexual, pese a la técnica implementada, así como hermafroditismo protándrico no casual en el 2017. Conclusiones: La aparición del hermafroditismo protándrico y los cambios en la proporción sexual indican que la población de A. tuberculosa, en ambas localidades, está respondiendo a presiones antropogénicas y ambientales.

Introduction: The bivalve Anadara tuberculosa is an important fishery in the Eastern Tropical Pacific. The sex ratio varies along its geographic range and there are cases of hermaphroditism, occasional in some populations, normal in others. These characteristics may be a response to anthropogenic and environmental pressure. Objective: To evaluate sex ratio and hermaphroditism of the species in the Panamanian Pacific. Methods: Data on reproductive features of A. tuberculosa generated between 1994 and 2020 for Gulf of Montijo, as well as 2017 data from mangroves of David was analyzed. Macroscopic diagnosis of individual sex is sensitive to the problem of indeterminate individuals, which in turn can skew the sex ratio analysis; therefore, the macroscopic method was compared with histological sections to generate a correction factor to adjust the macroscopically determined sex ratio. Results: For Montijo, histology indicated that the female to male sex ratio ranged from 1:1 to 3.2:1 between 1994 and 2017, while macroscopy indicated 1.9:1 to 5.1:1 between 2004 and 2020. For David, histology indicated a ratio of 3.7:1, and macroscopy 7.9:1. At both localities, larger specimens had more sex ratio variation, independently of the technique, and there was non-casual protandric hermaphroditism in 2017. Conclusion: The occurrence of protandric hermaphroditism and sex ratio changes suggest anthropogenic and environmental pressure in all populations.

Design and optimization of an experimental maintenance system for yellow clam broodstock Amarilladesma mactroides (Reeve, 1854) / Desenvolvimento e otimização de um sistema de manutenção experimental para reprodutores de marisco branco Amarilladesma mactroides (Reeve, 1854)

The yellow clam is a sand-burrowing bivalve that inhabits the dissipative beaches from southern Brazil to the north coast of Argentina. In the last decades, populations of this species have been impacted by mass mortality events, overfishing and other anthropogenic activities. The production of juveniles in captivity would allow feasibility studies to be carried out to restore the natural stock as well as the production in aquaculture systems. Given the scarcity of studies on the maintenance of this species in captivity, a culture system and a management protocol were developed and tested. Wild-caught clams (total length ≥50 mm) were used in a series of 14 day-long trials. Survival was higher in clams that were allowed to bury into the sand. A permanent ink marker covered with a thin layer of a quick-hardening adhesive proved to be a reliable method to tag clams. The maintenance of yellow clams in this system resulted in high survival and growth, increases in the condition factor and oocyte diameter, and a relative advancement of gonadal development.

O marisco branco é um bivalve de areia que habita as praias dissipativas do sul do Brasil até a costa norte da Argentina. Nas últimas décadas, as populações desta espécie têm sido afetadas por eventos de mortalidade maciça, sobrepesca e outras atividades antropogênicas. A produção de juvenis em cativeiro permitiria a realização de estudos de viabilidade para restaurar o estoque natural, assim como a produção em sistemas de aquicultura. Dada a escassez de estudos sobre a manutenção desta espécie em cativeiro, um sistema de cultivo e um protocolo de manejo foram desenvolvidos e testados. Mariscos branco selvagens (comprimento total ≥50 mm) foram utilizados em uma série de ensaios de 14 dias de duração. A sobrevivência foi maior nos mariscos que podiam ser enterrados na areia. Um marcador de tinta permanente coberto com uma fina camada de adesivo de endurecimento rápido provou ser um método confiável para marcar os mariscos. A manutenção dos mariscos neste sistema resultou em alta sobrevivência e crescimento, aumento do fator de condição e do diâmetro do ovócito, e um relativo avanço do desenvolvimento gonadal.

Possible roles of gonadotropin-releasing hormone (GnRH) and melatonin in the control of gonadal development of clam Ruditapes philippinarum.

Gonadotropin-releasing Hormone (GnRH) is a key reproductive endocrine regulator, and melatonin is considered as a potent candidate in the regulation of photoperiod-related reproductive endocrinology. Nevertheless, their function during gonadal development of molluscs has not been uncovered yet. In the present study, RNAi of GnRH and melatonin injection were conducted on marine bivalve manila clam Ruditapes philippinarum. Tissue section showed that gonadal development was significantly inhibited in male clams injected with GnRH dsRNA for 21 days. For GnRH RNAi treatment group, the expression levels of steroid synthetic enzyme genes 3ß-hydroxysteroid dehydrogenase (3ß-HSD), 17ß-hydroxysteroid dehydrogenase (17ß-HSD), cytochrome P450 (CYP3A) and melatonin receptor homolog (MTNR) gene were significantly down-regulated in female clams while significantly up-regulated in male clams. In melatonin injection group, the expression of GnRH was significantly inhibited and the expression of 3ß-HSD, 17ß-HSD, CYP3A and MTNR genes also increased which was in line with the GnRH dsRNA injection group in male clams. These results suggest that melatonin may affect GnRH expression and both have effects on gonadal development of bivalves. This study provides evidence for understanding the effects of melatonin and GnRH on reproductive endocrinology and gonadal development in bivalve molluscs.

RNAi-mediated knock-down of the dopamine beta-hydroxylase gene changes growth of razor clams.

Dopamine beta-hydroxylase (DßH) plays an essential role in the synthesis of catecholamines (CA) in neuroendocrine networks. In the razor clam, Sinonovacula constricta a novel gene for DßH (ScDßH-α) was identified that belongs to the copper type II ascorbate-dependent monooxygenase family. Expression analysis revealed ScDßH-α gene transcripts were abundant in the liver and expressed throughout development. Knock-down of ScDßH-α in adult clams using siRNA caused a reduction in the growth rate compared to control clams. Reduced growth was associated with strong down-regulation of gene transcripts for the growth-related factors, platelet derived growth factors A (PDGF-A) (P < 0.001) 24 h after ScDßH-α knock-down, vascular endothelial growth factor (VEGF1) (P < 0.001) and platelet derived growth factor B (PDGF-B-2) (P < 0.001) 24 h and 48 h after ScDßH-α knock-down and transforming growth factor beta (TGF-ß1) (P < 0.001) 48 h and 72 h after ScDßH-α knock-down. Taken together the results suggest that the novel ScDßH-α gene through its role in CA synthesis is involved in growth regulation in the razor clam and possibly other bivalves.

Use of sediment dwelling bivalves to biomonitor plastic particle pollution in intertidal regions; A review and study.

We explore the possibility of using the varnish (Nutallia obscurata) and Manila (Venerupis philippinarum) clams as biomonitors of microplastics (MPs) pollution. A short review is first provided on the use of bivalves for biomonitoring MPs in aquatic ecosystems. From the conclusions drawn from our review we determine if the sediment dwelling varnish and Manila clam could possibly be good choices for this purpose. We sampled 8 intertidal sites located within two distinct regions of coastal British Columbia, Burrard Inlet (5 sites) and Baynes Sound (3 sites). Each intertidal region had its own particular use; within Burrard Inlet, BMP a heavily used marine park, CP, EB, J, and AP, popular local beaches, and within Baynes Sound, Met and NHB, two intertidal regions heavily exploited by the shellfish industry and RU an intertidal region with limited aquaculture activity. Microfragments were recovered from bivalves collected from all intertidal regions except for AP. Microspheres were recovered primarily from bivalves sampled from Baynes Sound at NHB where high numbers of spheres within sediments had previously been reported. BMP and Met had the highest number of particles present within individual clams which were predominantly high density polyethylene (HDPE) and a polypropylene composite (PPC). Both polymers are extensively used by the shellfish industry in all gear types, as well as in industrial and recreational marine activities. The spatial distribution of recovered MPs was indicative of the anthropogenic use of the intertidal region suggesting these bivalves, for microfragments and microspheres, may be suitable as biomonitors and could prove to be useful tools for determining whether reduction policies for plastics use are having a positive effect on their release into marine environments.

Copper sulphate impact on the antioxidant defence system of the marine bivalves Cerastoderma edule and Scrobicularia plana.

Anthropogenic activities, such as agriculture and industrial activities, are a main source of pollution contributing for the degradation of water quality and thus affecting the living organisms of aquatic systems. Copper is widely used at these practices being often released into the aquatic systems and may cause negative effects in its communities. This study proposes to determine the effects of copper in the antioxidant defence system of two size classes (big and small sizes) of Scrobicularia plana and Cerastoderma edule, two marine bivalve species with commercial interest. It was observed the behaviour activity of the organisms during the exposure to copper sulphate (CS) and was determined the enzymatic activities of glutathione-S-transferases (GST), glutathione reductase (GR) and glutathione peroxidase (GPx) (both selenium-dependent (SeGPx) and total (tGPx)) in the muscle tissue (foot). Lipid peroxidation (LPO) was evaluated through thiobarbituric acid reactive substances (TBARS) measurement in the foot. Changes in the behaviour and enzymatic activity were observed. Lipid peroxidation was observed at C. edule and S. plana big and small size classes, respectively, according to TBARS levels. The foot showed to be a good tissue to be used in biochemical analysis to detect the presence of toxicants.

Integration of Biochemical, Cellular, and Genetic Indicators for Understanding the Aging Process in a Bivalve Mollusk Chlamys farreri.

The major causal factors for the irreversible decline in physical vitality during organismal aging are postulated to be a chronic state of cellular redox imbalance, metabolic toxicity, and impaired energy homeostasis. We assessed whether the relevant enzyme activity, oxidative stress, and intracellular ATP might be causally involved in the aging of short-lived Chlamys farreri (life span 4~5 years). A total of eight related biochemical and cellular indicators were chosen for the subsequent analysis. All the indicators were measured in seven different tissues from scallops aged one to four years, and our data support that the aging of C. farreri is associated with attenuated tissue enzyme activity as well as a decreased metabolic rate. Through principal component analysis, we developed an integrated vigor index for each tissue for comprehensive age-related fitness evaluation. Remarkably, all tissue-integrated vigor indexes significantly declined with age, and the kidney was observed to be the most representative tissue. Further transcriptional profiling of the enzymatic genes provided additional detail on the molecular responses that may underlie the corresponding biochemical results. Moreover, these critical molecular responses may be attributed to the conserved hierarchical regulators, e.g., FOXO, AMPKs, mTOR, and IGF1R, which were identified as potentially novel markers for chronic fitness decline with age in bivalves. The present study provides a systematic approach that could potentially benefit the global assessment of the aging process in C. farreri and provide detailed evaluation of the biochemical, cellular, and genetic indicators that might be involved. This information may assist in a better understanding of bivalve adaptability and life span.

Effect of sublethal gradient concentrations of nickel on postlarvae of Penaeus monodon, Perna viridis and Terapon jarbua: Enzyme activities and histopathological changes.

The present study evaluates the enzyme activities and histopathological changes in the post larvae (PL) of shrimp (Penaeus monodon), green mussel (Perna viridis) and fingerlings of crescent perch (Terapon jarbua) exposed to sublethal gradient concentrations of Nickel (Ni). The median lethal concentration (LC50) values were 2.49, 66.03 and 43.92 mg Ni L-1 derived for the PL of shrimp, green mussel and fish fingerlings respectively. No Observed Effect Concentration (NOEC), Lowest Observed Effect Concentration (LOEC) and chronic values of the PL of shrimp were 46.5, 73.0 and 58.3 µg Ni L-1 derived for the 21-d survival endpoint. The NOEC, LOEC and chronic values for the 30-d survival endpoint of the green mussels and fish fingerlings were 4.6, 6.32, 5.4 and 1.95, 2.6, 2.25 mg Ni L-1 respectively. The isoforms of esterase, superoxide dismutase and malate dehydrogenase activities in the whole body tissues of test organisms were studied by native polyacrylamide gel electrophoresis after exposure to Ni. Histological examination of compound eye sections of shrimp revealed deformation, compression, fusion and detachement in the corneal cells from the corneal facet of the ommatidia indicating cellular anomalies due to Ni toxicity. Gill sections of the green mussel witnessed reduced haemolymph in sinuses of gill filaments, degenerative changes in interfilamentous junction and necrosis of frontal ciliated epithelial cells with vacuoles after exposure to Ni. Nickel affects the vision of shrimp and fish fingerlings, gills and byssus of green mussels.

Biosseguridade para sistemas de produção de peixes em tanque-rede em função da colmatação agravada por Limnoperna fortunei / Biosecurity for fishfarming cage systems due to the highly clogged screen aggravated by Limnoperna fortunei

Este trabalho avaliou a colmatação por Limnoperna fortunei em diferentes materiais para confecção de telas de arame, a dinâmica da colmatação pelo molusco e a relação custo-benefício dos materiais usados em tanques-rede. Utilizaram-se amostras de telas de simples torção de malha 19mm, de cinco tipos diferentes de revestimentos, colocadas na barragem de Salto Caxias, no Rio Iguaçu, estado do Paraná, entre julho de 2012 e julho de 2014. A cada seis meses, foi verificado o ganho de peso das telas em razão da colmatação, além da contagem do número de mexilhões aderidos. Após as coletas dos dados, foi realizado o teste estatístico de Kruskal-Wallis para se avaliar o ganho de peso das telas devido à colmatação. Realizou-se também uma pesquisa com 21 empresas para se obter o preço comercializado de telas para confecção de tanques-rede e se avaliar a expectativa de durabilidade dos materiais. Verificou-se que os diferentes materiais avaliados para as telas influenciam na colmatação e aderência do mexilhão-dourado; além disso, constatou-se que a tela mais eficiente quanto ao custo/ano é a de arame galvanizado plastificado e, para baixa colmatação, a de arame Bezinal.(AU)

This research evaluated degree of clogging by Limnoperna fortunei in different materials for wire mesh fabrication, the dynamics of mollusk sealing and the cost-benefit ratio of these materials used in tank-nets. Samples of single-twist screens of 19 mm mesh were used, from five different types of coatings were placed in the Salto Caxias dam on the Iguaçu River in the Paraná state, between July 2012 and July 2014. Every six months, it was verified the weight gain of the screens due to clogging and counting of the number of mussels adhered. After the data collection was performed Kruskal-Wallis statistical test to evaluate the weight gain of the screens due to clogging. In addition, a survey was carried out with 21 companies to obtain the commercialized price of screens for the production of net tanks and to evaluate the expected durability of the materials. The different materials evaluated for the screens influenced the sealing and adhesion of the golden mussel. In addition, the most cost-per-year screen is that of plastic-coated galvanized wire and for low clogging it is the Bezinal wire.(AU)

Lipid-protein interactions in mitochondrial membranes from bivalve mollusks: molecular strategies in different species.

The mitochondrial F1FO-ATPase, the key enzyme in cell bioenergetics, apparently works in the same way in mollusks and in mammals. We previously pointed out a raft-like arrangement in mussel gill mitochondrial membranes, which apparently distinguishes bivalve mollusks from mammals. To explore the relationship between the microenvironmental features and the enzyme activity, the physico-chemical features of mitochondrial membranes and the F1FO-ATPase activity temperature-dependence are here explored in the Manila clam (Ruditapes philippinarum). Similarly to the mussel, clam gill mitochondrial membrane lipids exhibit a high sterol content (42 mg/g protein), mainly due to phytosterols (cholesterol only attains 42% of total sterols), and abundant polyunsaturated fatty acids (PUFA) (70% of total fatty acids), especially of the n-3 family. However, the F1FO-ATPase activation energies above and below the break in the Arrhenius plot (22.1 °C) are lower than in mussel and mammalian mitochondria. Laurdan fluorescence spectroscopy analyses carried out at 10 °C, 20 °C and 30 °C on mitochondrial membranes and on lipid vesicles obtained from total lipid extracts of mitochondria, indicate a physical state without coexisting domains. This mitochondrial membrane constitution, allowed by lipid-lipid and lipidprotein interactions and involving PUFA-rich phospholipids, phytosterols (much more diversified in clams than in mussels) and proteins, enables the maintenance of a homogeneous physical state in the range 10-30 °C. Consistently, this molecular interaction network would somehow extend the temperature range of the F1FO-ATPase activity and may contribute to clam resilience to temperature changes.

Sensitivity of multiple life stages of 2 freshwater mussel species (Unionidae) to various pesticides detected in Ontario (Canada) surface waters.

Freshwater mussels contribute important ecological functions to aquatic systems. The water filtered by mussel assemblages can improve water quality, and the mixing of sediments by burrowing mussels can improve oxygen content and release nutrients. However, nearly 70% of North American freshwater mussel species are listed as either endangered, threatened, or in decline. In Ontario, 28 species are in decline or in need of protection. Even though freshwater mussels have a heightened sensitivity to some contaminants, few studies have investigated the risks that various pesticide classes pose to one freshwater mussel species or among life stages. Lampsilis siliquoidea and Villosa iris were the focus of the present study, with the latter currently listed as of "special concern" in Canada. A potential risk to the recovery of freshwater mussel species is the presence and persistence of pesticides in Ontario surface waters. Acute (48 h) toxicity tests were performed with V. iris glochidia to determine the effect on viability (surrogate for survival) following exposure to 4 fungicides (azoxystrobin, boscalid, metalaxyl, and myclobutanil), 3 neonicotinoids (clothianidin, imidacloprid, and thiamethoxam), 2 carbamates (carbaryl and malathion), 1 organophosphate (chlorpyrifos), and 1 butenolide (flupyradifurone). Juvenile and adult L. siliquoidea were also exposed to azoxystrobin, clothianidin, imidacloprid (juvenile only), and carbaryl (adult only). Our study found in general that all life stages were insensitive to the pesticides tested, with median effect and lethal concentrations >161 µg/L. The pesticides tested likely represent a minimal risk (hazard quotients <5.4 × 10-3 ) to freshwater mussel viability and survival in acute (48 h) and subchronic (28 d) exposures, respectively, in Ontario streams where pesticide concentrations were considerably lower than those tested in the present study. Environ Toxicol Chem 2018;37:2871-2880. © 2018 SETAC.

Hot and bothered: effects of elevated Pco2 and temperature on juvenile freshwater mussels.

Multiple environmental stressors may interact in complex ways to exceed or diminish the impacts of individual stressors. In the present study, the interactive effects of two ecologically relevant stressors [increased temperature and partial pressure of carbon dioxide (Pco2)] were assessed for freshwater mussels, a group of organisms that are among the most sensitive and rapidly declining worldwide. The individual and combined effects of elevated temperature (22°C-34°C) and Pco2 (~230, 58,000 µatm) on juvenile Lampsilis siliquoidea were quantified over a 5- or 14-day period, during which physiological and whole animal responses were measured. Exposure to elevated temperature induced a series of physiological responses, including an increase in oxygen consumption rates following 5 days of exposure at 31°C and an increase in carbonic anhydrase ( ca) and heat shock protein 70 mRNA levels following 14 days of exposure at 28°C and 34°C, respectively. Treatment with elevated Pco2 activated acid-base regulatory responses including increases in CA and Na+-K+-ATPase activity and a novel mechanism for acid-base regulation during Pco2 exposure in freshwater mussels was proposed. Thermal and CO2 stressors also interacted such that responses to the thermal stressor were diminished in mussels exposed to elevated Pco2, resulting in the greatest level of mortality. Additionally, larger mussels were more likely to survive treatment with elevated Pco2 and/or temperature. Together, exposure to elevated Pco2 may compromise the ability of juvenile freshwater mussels to respond to additional stressors, such as increased temperatures, highlighting the importance of considering not only the individual but also the interactive effects of multiple environmental stressors.

Reproductive periods of Lucina pectinata (BIVALVE; LUCINIDAE) in the Paranaguá Estuarine Complex, Paraná - Brazil / Período reprodutivo de Lucina pectinata (BIVLAVE; LUCINIDAE) no Complexo Estuarino de Paranaguá, PR - Brasil

Abstract The objectives of the present study is to describe the size for exploitation and gonadal maturation of L. pectinata in Peças Island in Paraná coast. Twenty two individuals were sampled every month, from an intertidal flat of the Peças River, Municipality of Guaraqueçaba, during two periods – summer/autumn and winter of the 2009. Environmental parameter data was obtained at the same time of the biological sampling: temperature, salinity and seawater transparency at the Peças River mouth. The animals’ height, length, width, gross weight, wet weight of meat and dry weight of meat were measured at the lab. The gonad development stages (GDS), condition index (CI) and yield index (Y) were calculated. Sex identification was conducted by direct observation of the gonads and gametes under an optic microscope. Individuals were classified as males (M), females (F) or undetermined (U). The medium height observed was of 49.62 ± 6.84 mm. Period of more expressive gonad repletion were observed in summer when water temperature was higher. Results of GDS in both periods showed a predominance of partially filled gonad stages. This may be related with the fact that L. pectinata has intense gametogenesis activity, that is, the organisms do not have a resting interval, but go back to gonad restructuring rapidly after releasing the gametes. The continuity of the reproductive cycle of the specimens found in both studied periods could demonstrate a possible continuous reproduction of the species and a size for exploitation above 40.00 mm of height.

Resumo O presente estudo tem como objetivo descrever o tamanho de exploração e a maturação gonadal de L. pectinata na Ilha das Peças no litoral do Paraná. Vinte e dois indivíduos foram amostrados mensalmente na região entre-marés no Rio das Peças, Município de Guaraqueçaba, durante 2 períodos – verão/outono e inverno de 2009. Paralelamente às amostragens, foram obtidos dados dos parâmetros ambientais: temperatura, salinidade e transparência da água do mar na foz do Rio das Peças. A altura, comprimento, largura, peso bruto, peso úmido da carne e peso seco da carne dos animais foram medidos no laboratório. Foram calculados os estágios de desenvolvimento gonadal (EDG), índices de condição (IC) e de rendimento (R). A Identificação do sexo foi realizada por observação direta das gônadas e gametas sob microscópio óptico. Os indivíduos foram classificados como: machos (M), fêmeas (F) ou indeterminados (I). A altura média observada foi de 49,62 ± 6,84 mm. O período de maior repleção gonadal foi observado no verão, quando a temperatura da água foi mais elevada. Resultados dos EDG em ambos os períodos mostrou um predomínio de gônadas parcialmente cheias. Isso pode estar relacionado com o fato de que L. pectinata tem atividade gametogênese intensa, ou seja, os organismos não têm um intervalo de repouso, mas voltam a reestruturação das gônadas rapidamente após liberação dos gametas. A continuidade do ciclo reprodutivo dos espécimes encontrados em ambos os períodos estudados, pode demonstrar possível reprodução contínua da espécie e tamanho de exploração superior a 40,00 mm de altura.

Exposure to2,2',4,4'-tetrabromodiphenyl ether (BDE-47) alters thyroid hormone levels and thyroid hormone-regulated gene transcription in manila clam Ruditapes philippinarum.

Polybrominated diphenyl ethers (PBDEs) have the potential to disturb the thyroid endocrine system in vertebrates, but little is known about the disruptive effects of PBDEs on marine bivalves. In this study, we first examined the effects of BDE-47 exposure on growth of juvenile manila clams Ruditapes philippinarum. The result showed that 1.0 and 10 µg L(-1) BDE-47 had adverse effects on 14-d shell-length growth of juvenile clams. Then, one-year-old adult clams were exposed to 0, 0.1 and 1 µg L(-1) BDE-47 for 15 d. BDE-47 (1 µg L(-1)) exposure caused significant decreases of total T4 (thyroxine) by 40% and T3 (3,5,3'-triiodothyronine) by 75% concentrations in haemolymph of the clams. Transcription of genes involved in thyroid hormone synthesis and metabolism were also studied by quantitative RT-PCR. Gene expression levels of sodium iodide symporter (rp-NIS), iodothyronine deiodinase (rp-Deio) and thyroid peroxidase (rp-TPO) were increased in a dose-dependent manner at day 5 and day 10, while monocarboxylate transporter 8 (rp-Mct8) was downregulated at day 5, day 10 and day 15. The effect and preliminary mechanism observed in the present study were consistent with the results from previous studies on rodent and fish, implying that exposure to BDE-47 may pose threat to thyroid hormone homeostasis in bivalves through thyroid synthesis and metabolism pathways. This study may provide a first step towards understanding of the thyroid function disruptive effects of PBDEs on marine bivalves and the underlying mechanism across taxonomic groups and phyla.

Acute effects of non-weathered and weathered crude oil and dispersant associated with the Deepwater Horizon incident on the development of marine bivalve and echinoderm larvae.

Acute toxicity tests (48-96-h duration) were conducted with larvae of 2 echinoderm species (Strongylocentrotus purpuratus and Dendraster excentricus) and 4 bivalve mollusk species (Crassostrea virginica, Crassostrea gigas, Mytilus galloprovincialis, and Mercenaria mercenaria). Developing larvae were exposed to water-accommodated fractions (WAFs) and chemically enhanced water-accommodated fractions (CEWAFs) of fresh and weathered oils collected from the Gulf of Mexico during the Deepwater Horizon incident. The WAFs (oils alone), CEWAFs (oils plus Corexit 9500A dispersant), and WAFs of Corexit alone were prepared using low-energy mixing. The WAFs of weathered oils had no effect on survival and development of echinoderm and bivalve larvae, whereas WAFs of fresh oils showed adverse effects on larval development. Similar toxicities were observed for weathered oil CEWAFs and WAFs prepared with Corexit alone for oyster (C. gigas and C. virginica) larvae, which were the most sensitive of the tested invertebrate species to Corexit. Mean 10% effective concentration values for total polycyclic aromatic hydrocarbons and dipropylene glycol n-butyl ether (a marker for Corexit) in the present study were higher than all concentrations reported in nearshore field samples collected during and after the Deepwater Horizon incident. The results suggest that water-soluble fractions of weathered oils and Corexit dispersant associated with the Deepwater Horizon incident had limited, if any, acute impacts on nearshore larvae of eastern oysters and clams, as well as other organisms with similar sensitivities to those of test species in the present study; however, exposure to sediments and long-term effects were not evaluated. Environ Toxicol Chem 2016;35:2016-2028. © 2016 SETAC.

Allatostatin-type A, kisspeptin and galanin GPCRs and putative ligands as candidate regulatory factors of mantle function.

Allatostatin-type A (AST-A), kisspeptin (KISS) and galanin (GAL) G-protein coupled receptor (GPCR) systems share a common ancestral origin in arthropods and the vertebrates where they regulate metabolism and reproduction. The molluscs are the second most diverse phylum in the animal kingdom, they occupy an important phylogenetic position, and their genome is more similar to deuterostomes than the arthropods and nematodes and thus they are good models for studies of gene family evolution and function. This mini-review intends to extend the current knowledge about AST-A, KISS and GAL GPCR system evolution and their putative function in the mollusc mantle. Comparative evolutionary analysis of the target GPCR systems was established by identifying homologues in genomes and tissue transcriptome datasets available for molluscs and comparing them to those of other metazoan systems. Studies in arthropods have revealed the existence of the AST-A system but the loss of homologues of the KISS and GAL systems. Homologues of the insect AST-AR and vertebrate KISSR genes were found in molluscs but putative GALR genes were absent. Receptor gene number suggested that members of this family have suffered lineage specific evolution during the molluscan radiation. In molluscs, orthologues of the insect AST-A peptides were not identified but buccalin peptides that are structurally related were identified and are putative receptor agonists. The identification of AST-AR and KISSR genes in molluscs strengthens the hypotheses that in metazoans members of the AST-AR subfamily share evolutionary proximity with KISSRs. The variable number of receptors and large repertoire of buccalin peptides may be indicative of the functional diversity of the AST-AR/KISSR systems in molluscs. The identification of AST-A and KISS receptors and ligands in the mantle transcriptome indicates that in molluscs they may have acquired a novel function and may play a role in shell development or sensory detection in the mantle.

Growth and production of Donax striatus(Bivalvia: Donacidae) from Las Balsas beach, Gibara, Cuba / Crecimiento y producción de Donax striatus (Bival-via: Donacidae) en playa Las Balsas, Gibara, Cuba

Abstract:Clams of the genus Donaxare worldwide the dominating group of the invertebrate community on sandy beaches. They are primary consumers that provide a significant abundance and biomass to the ecosystem. In the Caribbean, Donax striatushas an important role for nature and human, nonetheless studies on the population dynamics of this beach clam are scarce and no information exists on secondary production of this species. Growth parameters and secondary production of D. striatuswere estimated from February 2008 to November 2009 at Las Balsas beach, Northeastern Cuba, in order to provide basic information for management purposes. In each month 45 samples were taken by means of a PVC corer of 0.025 m2 area and sieved with a 1 mm mesh. Animals were measured and weighted with and without shell. A total of 5 471 specimens were collected during the sampling period. Shell length ranged from 2.7-33.3 mm. Growth parameters estimated from length frequency data were Lm = 36.1 mm, K= 0.8/yr and t0 = 0.2/yr. The growth performance resulted in values of 0'= 3.02. Life span was 2.4 yrs and mortality rate was 3.07 /yr. In 2008, mean abundance of D. striatusranged between 17.1770.7 ind./m2. In 2009 the lowest mean abundance was 34.4 and the highest was 892.5 ind./m2. During 2009 biomass and production was more than twice higher in comparison with 2008. Individual production showed highest values in the 24 mm shell size (3.74 g/m2.yr) and 25 mm (0.71 g/m2.yr), considering mass with shell and without shell, respectively. During 2009 abundance of individuals with 15 mm shell length or more increased resulting in higher biomass and production, compared to 2008. Using the conversion factor of wet mass to ash free dry mass (AFDM), annual production ranged between 2.87-6.11 g AFDM/m2.yr, resulting in a turnover rate (P/B) between 5.11 and 3.47 in 2008 and 2009, respectively. The rapid growth and high turnover rate of D. striatussuggest a rapid recovery of the population. These results support the idea that this beach clam is an important resource at Las Balsas beach. Thus its exploitation must continue with caution, and only at the level of a recreational fishery.

Resumen:Los estudios sobre la dinámica poblacional de la almeja de playa Donax striatusson escasos y no existe información sobre la producción secundaria de alguna población de esta especie. Los parámetros del crecimiento y la producción secundaria de D. striatusfueron estimados a partir de datos de muestreo obtenidos entre febrero 2008 y noviembre 2009 en playa Las Balsas en la zona norte oriental de Cuba para aportar información básica para su manejo. Cada mes se recolectaron 45 muestras usando un núcleo de PVC de 0.025 m2, el sedimento fue tamizado en malla de 1 mm. Los individuos recolectados fueron medidos y pesados con concha y sin concha. La longitud de la concha fue de 2.7-33.3 mm. Los parámetros de crecimiento estimados a partir de las frecuencias de tallas fueron: Lm= 36.1 mm, K= 0.8/año y t0 = 0.2/año. El índice de crecimiento fue de 0'= 3.02. El tiempo de vida estimado de 2.4 años y la mortalidad 3.07/año. En 2008 la abundancia media de D. striatusfluctuó entre de 17.1-770.7 ind/m2. En 2009 la menor abundancia fue de 34.4 y la mayor de 892.5 ind/ m2. En 2009 la biomasa y la producción fueron más del doble que las estimadas en 2008. La mayor producción somática individual fue aportada por individuos de 24 mm (3.74 g/m2.año) y 25 mm (0.71 g/m2.año), considerando la masa total con concha y sin concha respectivamente. Durante 2009 la abundancia de individuos de 15 mm de longitud o más, se incrementó resultando en mayor bio-masa y producción en comparación con 2008. Usando el factor de conversión de peso húmedo a peso seco libre de cenizas (PSLC), la producción anual en 2008 fue de 2.87 y en 2009 fue 6.11 g PSLC/m2.año, lo que resulta en una tasa de renovación (P/B) de 5.11 en 2008 y 3.47 en 2009. Estos resultados apoyan la idea que la almeja de playa es un recurso importante y su explotación debe continuar con cautela y solo a un nivel de una pesquería recreativa.

Occurrence of lipophilic marine toxins in shellfish from Galicia (NW of Spain) and synergies among them.

Lipophilic marine toxins pose a serious threat for consumers and an enormous economic problem for shellfish producers. Synergistic interaction among toxins may play an important role in the toxicity of shellfish and consequently in human intoxications. In order to study the toxic profile of molluscs, sampled during toxic episodes occurring in different locations in Galicia in 2014, shellfish were analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS), the official method for the detection of lipophilic toxins. The performance of this procedure was demonstrated to be fit for purpose and was validated in house following European guidelines. The vast majority of toxins present in shellfish belonged to the okadaic acid (OA) group and some samples from a particular area contained yessotoxin (YTX). Since these toxins occur very often with other lipophilic toxins, we evaluated the potential interactions among them. A human neuroblastoma cell line was used to study the possible synergies of OA with other lipophilic toxins. Results show that combination of OA with dinophysistoxin 2 (DTX2) or YTX enhances the toxicity triggered by OA, decreasing cell viability and cell proliferation, depending on the toxin concentration and incubation time. The effects of other lipophilic toxins as 13-desmethyl Spirolide C were also evaluated in vitro.

Identification and expression characterization of the myostatin (MSTN) gene and association analysis with growth traits in the razor clam Sinonovacula constricta.

Myostatin (MSTN) is a member of the transforming growth factor-ß superfamily (TGF-ß) and is an important negative regulator of muscle growth in vertebrates. In this study, we cloned and analyzed the MSTN gene (Sc-MSTN) from razor clam (Sinonovacula constricta). The full length of Sc-MSTN cDNA sequence consists of 4226 base pairs (bp), comprising a 522-bp 5' untranslated region (UTR), a 2342-bp 3'UTR, and an open reading frame (ORF) that is 1362 in length. The ORF encodes 453 amino acids with a RXXR proteolytic site and nine conserved cysteines. Quantitative real-time PCR analysis revealed that the Sc-MSTN transcript was expressed in a wide range of tissues but appeared to exhibit the greatest level of expression in the foot. The transcript was widely detected in early developmental stages, showing the highest expression in the trochophore stage. Furthermore, six SNPs were identified in the coding region of the Sc-MSTN gene using direct sequencing. SNP-1 is non-synonymous and involves an amino acid change from Leu to Ser. Association analysis showed that SNP-1 and SNP-6 had significant influences on shell length (SL). The results suggested that MSTN could be selected as a candidate gene for the future molecular breeding of razor clam strains.