Pathological conditions of the sentinel bivalve, the little mussel Brachidontes rodriguezii, from contaminated intertidal sites in the Southwestern Atlantic coast.

A survey of pathological conditions affecting the natural beds of the dominant bivalve species of the intertidal rocky coasts of the Southern Atlantic Ocean, was performed. Samples of the little mussel (n = 992), Brachidontes rodriguezii, which is considered a sentinel species, were collected from seven sites that present different anthropogenic activity (low anthropogenic activity, harbor, and sewage waste) along the Argentinean coast, and processed by standard histological techniques. Our results showed for the first time in a B. rodriguezii population from Mar del Plata harbor a bacterium causing cellular lysis of the spermatogenic cells (6.5% of prevalence) and in a population from Villa Gesell, a site with low anthropogenic activity, one little mussel presented cytoplasmic vacuoles in the oocytes's cytoplasm similar to a microsporidian (0.2% of prevalence). A ciliated protozoan parasitizing the cytoplasm of the epithelial digestive tubules and larval stages of two different digenean parasites, was found infecting the little mussel from both contaminated and uncontaminated sites. Sporocysts of the family Bucephalidae invaded the gonad, causing severe gonadal damage including castration, and gymnophallid metacercariae located between the mantle and shell or embedded in the mantle tissues, occupied the gonadal space. In Punta Carballido, a site located next to a sewage outfall, the highest prevalence of infection was found, for the ciliated protozoan in the digestive tubules (5.1%) and for the digenean parasites (bucephalid sporocyst 24.4%, and gymnophallid metacercariae 50%). Moreover, we found a positive correlation between shell length and overall prevalence of parasites.

Successful recruitment, survival and long-term persistence of eastern oyster and hooked mussel on a subtidal, artificial restoration reef system in Chesapeake Bay.

Restoration efforts with native eastern oyster, Crassostrea virginica, in Chesapeake Bay and elsewhere have been limited by shell availability, necessitating the use of alternative structures as subtidal reefs, yet these have rarely been evaluated quantitatively. We quantified population structure, density, abundance and biomass of eastern oyster and hooked mussel, Ischadium recurvum, on a concrete modular reef (75 m2 surface area over 5 m2 of river bottom) deployed subtidally at 7 m depth in the Rappahannock River, Virginia during October, 2000. After nearly 5 y (May 2005), we took 120 stratified random samples over the reef. The reef was heavily colonized by 28-168 oysters and 14-2177 mussels m-2 surface area. These densities translate to 1085 oysters and 8617 mussels m-2 river bottom, which are the highest recorded for artificial oyster reefs. Size structure of oysters reflected four year classes, with over half of oysters more than 1 y old and of reproductive age. Oyster biomass (1663 g dry mass m-2 river bottom) and condition index were equally high, whereas parasite prevalence and intensity were low. Oyster density correlated positively in a sigmoid fashion with mussel density up to high densities, then declined. This modular reef is one of the most successful artificial reefs for eastern oyster and hooked mussel restoration, and details features that are conducive for successful settlement, growth and survival in subtidal habitats.

Permanent culture and parasitic impact of the microalga Coccomyxa parasitica, isolated from horse mussel Modiolus kurilensis.

Animals with deformed shells and microalgal invasion have been identified in the natural population of the horse mussel species Modiolus kurilensis of Peter the Great Bay in the Sea of Japan. The haemolymph is initially infested with algae, followed by the rectum, siphons, mantles and gonads located in the posterior body areas. Mantles, which are primarily exposed to light, are major depots for algae. The microscopic analysis of algal cells has revealed the absence of flagella and pyrenoids, the presence of single chloroplast, and reproduction by autosporulation, with dispores prevailing over tetraspores. These results, together with the nearly complete sequence analysis of small subunit (SSU) 18S rDNA (1728bp), have confirmed that these cells are Coccomyxa parasitica. A newly developed method of isolating microalgae from mollusk tissues has facilitated the continuous pure - probably axenic - culture of C. parasitica, thereby providing a description of the time course of each life stage. Histological analyses have revealed significant haemocyte infiltration into the mantles, gonads, kidneys and digestive gland tissues infested with microalgae and the gill tissues, in which the intruder was not identified. Algal encapsulation with major focal areas of fibrosis and amorphic necrosis has been revealed in these infested organs. The spaces between the gonad follicles and digestive gland tubules were significantly widened as these areas were filled with a mass of algae and phagocytic haemocytes, showing acini with a thickened basement membrane. The mantles and kidneys of Modiolus displayed significant morphological deviations of different cells in epithelial, connective and muscle tissues, resulting in the dysfunction of the infested organs. Therefore, C. parasitica, which reproduces in the culture, regardless of the host, is a facultative parasite, causing major pathological alterations, such as anomalous histomorphological patterns and infested organ dysfunctions.

Qualidade parasitológica e condições higiênico-sanitárias de sururu (Mytella charruana) e alface (Lactuca sativa) comercializados em um mercado público de Maceió-AL / Parasitological quality and hygienic-sanitary conditions of mussels (Charruana mytella) and lettuce (Lactuca sativa) sold at a public market Maceió-AL

As Doenças Transmitidas por Alimentos são síndromes originadas pela ingestão de alimentos e/ou água, que contenham agentes etiológicos em quantidades tais que afetem a saúde do consumidor. Assim, o estudo teve como objetivo avaliar a ocorrência de parasitos, bactérias do grupo coliformes totais e fecais em amostras de alfaces e sururu comercializadas em um mercado público de Maceió-AL. Foram avaliados 13 pés de alfaces e 20 amostras de sururu que foram obtidas de diferentes boxes e acondicionadas individualmente em sacos plásticos estéreis. Para a análise parasitológica, as amostras de sururu e alface foram pesadas (50g), posteriormente submetidas à lavagem manual com 300 mL de água destilada. Em seguida, a água da lavagem foi recolhida em recipiente apropriado e esta foi analisada por duas técnicas de detecção de parasitos: sedimentação espontânea e centrifugação simples. Para analise microbiológica das amostras de alface e sururu foi utilizada a técnica do número mais provável, utilizando caldo Lauril Sulfato Triptose para o teste presuntivo, caldo verde brilhante e caldo Escherichia coli para o teste confirmativo. Dentre as amostras, 84,39% apresentaram larvas da família Rhabdiasidae, e 15,61% das amostras foram negativas, não foram identificadas estruturas parasitárias no sururu avaliado. Na análise microbiológica todas as amostras de alface e sururu apresentaram-se com resultados de >1100 NMP/g para bactérias do grupo coliformes indicando que esses produtos possuem contaminação microbiológica alta, não devendo ser ingeridos por crianças, idosos e pacientes imunocomprometidos. Estes dados indicam a necessidade da orientação dos manipuladores quanto à higienização no preparo de alfaces e sururu pelos consumidores.

The Foodborne Diseases syndromes are caused by the ingestion of food and / or water containing etiologic agents in such quantities that affect the health of the consumer. Thus, the study aimed to evaluate the occurrence of parasites, bacteria total coliform and fecal samples of lettuce and mussels sold in a public market Maceió-AL. 13 feet of lettuce and 20 samples of mussels that were collected from different boxes and individually wrapped in sterile plastic bags were evaluated. For parasitological analysis, the samples were weighed mussels and lettuce (50g) subsequently subjected to manual washing with 300 ml of distilled water. Then, the wash water was collected in an appropriate container, and this was analyzed by two techniques for detection of parasites: single spontaneous sedimentation and centrifugation. For microbiological analysis of samples of lettuce and mussels to the most probable number was used, using broth Lauryl Sulfate Tryptose presumptive test, bright green soup and broth Escherichia coli for the confirmatory test. Among the samples, 84.39 % were larvae Rhabdiasidae family, and 15.61 % of samples were negative, parasitic structures were not identified in the assessed mussels. Microbiological analysis in all samples of lettuce and mussels presented with results of > 1100 MPN / g for coliform bacteria indicates that these products have high microbiological contamination and should not be ingested by children , elderly and immunocompromised patients. These data indicate the need for guidance as to the hygiene of food handlers in the preparation of lettuces and mussels by consumers.

Endoliths in Lithophaga lithophaga shells--Variation in intensity of infestation and species occurrence.

Pronounced differences with respect to the extent of infestation and the degree of Lithophaga lithophaga shell damage inflicted by euendolithic taxa at two sites in the Adriatic Sea representing different productivity conditions, are described. Shells collected from the eastern part of Kastela Bay, which is characterized by higher primary productivity, have significantly more shell damage then the shell collected from a site on the outer coast of the island of Ciovo exposed to the oligotrophic Adriatic Sea. The presence of endoliths and their perforations were detected in different layers of the shell, including solidly mineralized parts of the skeleton and within the organic lamellae incorporated into the shell. Phototrophic endoliths were not observed in the specimens. The most serious damage to L. lithophaga shells was the boring clionaid sponge Pione vastifica, which was more common in shells collected from Kastela.

The life cycle of Prosorhynchoides carvajali (Trematoda: Bucephalidae) involving species of bivalve and fish hosts in the intertidal zone of central Chile.

We describe the life cycle of the bucephalid Prosorhynchoides carvajali from the intertidal rocky zone of central Chile. To elucidate the life cycle of this digenean, two mytilid bivalves, Semimytilus algosus and Perumytilus purpuratus, and ten intertidal fish species belonging to the families Blenniidae, Tripterygiidae, Labrisomidae, Kyphosidae and Gobiesocidae were analysed for natural infections. In addition, experimental infections of fish were undertaken and molecular analyses were performed of several developmental stages of the digeneans in various host species. Experimental infections of fish were made from infected mytilids to determine which fish species were suitable for the metacercarial stage of Prosorhynchoides. We also determined the abundance and prevalence of metacercariae in natural infections in fish and found that they were lower than in the experimental infections. A molecular analysis showed that sporocysts from S. algosus were identical to metacercariae from five fish species and P. carvajali adults. Sporocysts isolated from P. purpuratus were similar to metacercaria found in one fish species only (G. laevifrons) but were different from P. carvajali, with 1.9-2.0% genetic divergence. Therefore, the complete life cycle of P. carvajali consists of the mytilid species S. algosus as the first intermediate host, at least five intertidal fish species as second intermediate hosts (Scartichthys viridis, Auchenionchus microcirrhis, Hypsoblennius sordidus, Helcogrammoides chilensis and Gobiesox marmoratus), two carnivorous fish as definitive hosts (Auchenionchus microcirrhis and A. variolosus) and one occasional definitive host (Syciases sanguineus). This is the second description of a life cycle of a marine digenean from Chile.

Spatio-temporal variation in the prevalence of trematodes in the bivalve Perumytilus purpuratus.

Perumytilus purpuratus is an abundant bivalve located in the intertidal rocky zone of South America that has been considered as a key species of the ecosystem. There are few studies of the host-parasite relationship of this bivalve; thus, this research aims to analyse the spatial and temporal variation in the prevalence of trematodes in P. purpuratus. Bivalves were collected from three localities (El Tabo, Las Cruces and Montemar) of central Chile (33°S, 71°W) during different seasons of 2010. The bivalves were also collected every metre, from the lowest to the highest level of the intertidal rocky zone, to determine the parasite distribution within the localities. Three species of trematodes as sporocyst stages were found: Prosorhynchoides carvajali, Proctoeces sp. and an undetermined fellodistomid species. Of the 37,692 bivalve specimens collected, 2.68% were parasitised. The undetermined fellodistomid species was the most prevalent parasite observed (1.69%). There were little detected differences in the prevalence of some trematode species between seasons. The prevalence of P. carvajali varied between localities, being most prevalent at Montemar. The distribution of trematodes along the rocky zone within the localities was variable, with P. carvajali being more prevalent in the mid-lowest level of the intertidal zone and the undetermined fellodistomid species being more prevalent in the mid-highest level. Both the abundance of definitive hosts and the environmental conditions likely result in different levels of infection by trematodes in P. purpuratus between and within the localities.

Reproduction-related effects of green alga Coccomyxa sp. infestation in the horse mussel Modiolus modiolus.

The effects of Coccomyxa sp. infestation on the reproductive characteristics of the horse mussel Modiolus modiolus from the north-western Pacific (Peter the Great Bay, Sea of Japan) are demonstrated in this study. The mussels were collected monthly from May to September of 2009, and the prevalence of females and males with green tissues was 39% and 47%, respectively. Overall, the green alga infection caused a mild effect on gametogenesis in the horse mussel. The dynamics of gonad development in the healthy and infected mussels during the study period was generally similar, with the spawning being partial and occurring from the beginning of June to the first half of September; total spawning was recorded at the end of this period. However, several negative reproduction-related effects of the green alga infestation were observed, i.e., general gonadal underdevelopment, which was apparent from significant decrease in the acinus areas of the ovaries and testes and an increase in the connective tissue layer between the acini, a delay in some stages of the reproductive cycle and production of decreased number of spermatozoa and large vitellogenic oocytes, especially in the early spawning period (June). All of these results suggest a reduced reproductive output for the infected mussels.

Morphological and molecular analyses of larval trematodes in the intertidal bivalve Perumytilus purpuratus from central Chile.

The bivalve Perumytilus purpuratus is a common species that is widely distributed throughout rocky intertidal zones in Chile. This bivalve is the first intermediate host for three trematode species: one bucephalid (an undetermined species) and two fellodistomids (Proctoeces lintoni and one undetermined species). A few studies based on morphological comparisons, experimental infection and molecular analyses have been performed to ascertain the taxon (at least at the family level) to which these trematodes belong; yet, there remains no clarification about the specific identity of these trematodes. Therefore, in this study, we compared the V4 region nucleotide sequences of the 18S rRNA of these three sporocyst species, classified as morphotypes, found in P. purpuratus and nine adult trematode species from intertidal fishes that are likely definitive hosts for these parasites. The sequences from two of the sporocyst morphotypes matched with adult trematodes from the intertidal fish: type 1 sporocyst was similar to Prosorhynchoides carvajali (Bucephalidae), with a mean genetic divergence of 0.78%, and type 2 sporocyst was similar to Proctoeces sp. (but not P. lintoni), with 0% genetic divergence. The third species (type 3 sporocyst) was classified to the family Fellodistomidae; however, the sequence from this species differed greatly from the three other fellodistomid species documented in the marine fish of Chile and from other fellodistomids in public databases. Moreover, this morphotype has a particular cercarial morphology that greatly differs from other fellodistomid species described thus far. Therefore, this intriguing trematode remains a mystery.

Parasites of the mangrove mussel Mytella guyanensis (Bivalvia: Mytilidae) in Camamu Bay, Bahia, Brazil.

This contribution reports the parasites found in the mangrove mussel Mytella guyanensis in Camamu Bay, Bahia, Brazil. Samples were collected monthly from September 2006 through October 2007. A total of 460 individuals were collected, fixed in Davidson's solution, and processed by standard histological techniques, and the sections were stained with Harris hematoxylin and eosin (H&E). The water temperature ranged from 23.5 to 31.6 ºC, and the salinity from 25 to 37‰. Microscopic analysis showed Rickettsia-like organisms (RLOs), Nematopsis sp. (Apicomplexa), and Platyhelminthes, including a turbellarian, sporocysts of Bucephalus sp., metacercariae, and metacestodes of Tylocephalum sp. Parasites were observed mainly in the gills, mantle, and digestive gland. The prevalence of Nematopsis sp. was 100%, and in heavily infected mussels the tissues of the labial palps were damaged. RLOs occurred in high prevalence and intensity of infection in some periods. The digenean sporocysts showed moderate prevalence but high intensity of infection, and caused parasitic castration. In general, there was no significant spatial or temporal variation (p > 0.05) of the parasites, which is probably attributable to the small variations of temperature and salinity in the region.

Phylogenetic and morphological characterization of the green alga infesting the horse mussel Modiolus modiolus from Vityaz Bay (Peter the Great Bay, Sea of Japan).

In this work, the ultrastructural features and taxonomic position of the green microalga infesting the horse mussel Modiolus modiolus from the north-western Pacific (Vityaz Bay, Peter the Great Bay, Sea of Japan) are reported. Mussels were collected monthly from May to September of 2009. In different months, the prevalence of mussels with green tissues was 16.6-62.5% (mean 43%). The most affected organs were the mantle, digestive gland and gonad. Histological analysis revealed severe infiltration of the connective tissue by hemocytes containing the alga cells. Electron microscopy showed that the alga was morphologically similar to the green algae from the genus Coccomyxa (Chlorophyta: Chlorococcales). Two new primers were designed to generate partial small subunit (SSU) rRNA sequences of the green alga from M. modiolus. Phylogenetic analysis based on the comparison of the SSU rRNA sequences of the trebouxiophyceans confirmed an affiliation of the green alga with the genus Coccomyxa. The sequence (1296 bases) of the green alga from M. modiolus was most closely related to the sequence CPCC 508 (AM981206) (identity 100%), obtained from an acid-tolerant, free-living chlorophyte microalga Coccomyxa sp. and to the sequences EU127470 (identity 99.3%) and EU127471 (identity 99.7%) of the green alga, presumably the true Coccomyxa parasitica, infecting the blue mussel Mytilus edulis from the Flensburg Fjord (North Atlantic).

Parasites of the mangrove mussel Mytella guyanensis (Bivalvia: Mytilidae) in Camamu Bay, Bahia, Brazil / Parasitos do marisco-do-mangue, Mytella guyanensis (Bivalvia: Mytilidae), na Baía de Camamu, Bahia

This contribution reports the parasites found in the mangrove mussel Mytella guyanensis in Camamu Bay, Bahia, Brazil. Samples were collected monthly from September 2006 through October 2007. A total of 460 individuals were collected, fixed in Davidson's solution, and processed by standard histological techniques, and the sections were stained with Harris hematoxylin and eosin (H&E). The water temperature ranged from 23.5 to 31.6 ºC, and the salinity from 25 to 37‰. Microscopic analysis showed Rickettsia-like organisms (RLOs), Nematopsis sp. (Apicomplexa), and Platyhelminthes, including a turbellarian, sporocysts of Bucephalus sp., metacercariae, and metacestodes of Tylocephalum sp. Parasites were observed mainly in the gills, mantle, and digestive gland. The prevalence of Nematopsis sp. was 100%, and in heavily infected mussels the tissues of the labial palps were damaged. RLOs occurred in high prevalence and intensity of infection in some periods. The digenean sporocysts showed moderate prevalence but high intensity of infection, and caused parasitic castration. In general, there was no significant spatial or temporal variation (p > 0.05) of the parasites, which is probably attributable to the small variations of temperature and salinity in the region.

Neste estudo, foram investigados os parasitos de Mytella guyanensis da Baía de Camamu (BA). As amostragens foram feitas mensalmente entre setembro de 2006 e outubro de 2007 e totalizaram 460 animais, que foram fixados em solução de Davidson e processados por técnica rotineira de histologia, com inclusão em parafina e coloração por hematoxilina de Harris e eosina. Foram evidenciados organismos assemelhados a Rickettsia (RLOs); Nematopsis sp. (Apicomplexa) e Platyhelminthes, incluindo um turbelário, esporocistos de Bucephalus sp. e o cestoide Tylocephalum sp. Os parasitos foram encontrados principalmente nas brânquias, no manto e na glândula digestiva. Todos os animais apresentaram Nematopsis sp., o que causou alteração nos palpos labiais em exemplares com alta infecção. RLOs ocorreram em alta prevalência e intensidade de infecção em alguns períodos. Esporocistos de trematódeos digenéticos, com prevalência moderada, mostraram-se em alta intensidade de infecção e causaram castração parasitária. Não houve variação temporal e espacial significativa (p > 0,05) dos parasitos, o que provavelmente se deveu à pequena amplitude de variação da temperatura e da salinidade na região.

First intermediate host of the digenean trematode Proctoeces lintoni (Fellodistomidae) in Chile.

The digenetic trematode, Proctoeces lintoni, is a parasite of the rocky intertidal ecosystems of the Chilean Pacific coastline. Although this species is relatively well known, the first intermediate host has not yet been described. In the present study, we used experimental protocols and field studies to identify the host that harbors the sporocysts and cercariae stages of the parasite. The first intermediate host was shown to be the dominant mussel of the mid-littoral zone, Perumytilus purpuratus.

Evidence of plasticity in the reproduction of a trematode parasite: the effect of host removal.

The parasitic trematode Proctoeces lintoni requires 3 hosts (intertidal mussels, keyhole limpets, and clingfish) to complete its life cycle. The densities and size structure of host communities are modified by selective human harvesting. This study examined clutch and egg size of P. lintoni in 3 adjacent sites in rocky intertidal areas of central Chile presenting differences in the levels of human disturbance (i.e., from a fully protected marine reserve to free open-access areas). We found significant differences in parasite fecundity among sites. An increase in number of eggs was observed inside protected marine areas compared with open-access areas, suggesting a plastic response of the parasite reproductive strategies to the host community modification. These results show that host removal by humans in coastal ecosystems can strongly influence parasite life history traits.