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Even after almost 30 years of Limnoperna fortunei introduction into South America, it is still unclear how the source and propagules are connected. Here, we present genetic evidence of population connectivity and gene flow of L. fortunei propagules from Asia into South America, proposing the main invasion routes into South America. To achieve that we expanded the sampling effort to cover all occurrence points of L. fortunei in South America. We sequenced the mtDNA COI gene and genotyped eight microsatellite loci (ML), and we evaluated the genetic source of the recently introduced population in Sobradinho hydroelectric power plant reservoir in Northeast Brazil. Our results revealed that China is the main genetic source of propagules for the Sobradinho population. We also found COI haplotypes and ML genotypes unique to South American populations, demonstrating a bridgehead effect likely caused by local mutation, adaptation, and admixture patterns that are maintained by high levels of gene flow among them. However, two genetic barriers were also detected. We concluded that L. fortunei is a well-established invader and is still rapidly expanding in Brazil, and the Amazon hydrographic basin is under an alarming threat of invasion.

Limnoperna fortunei, the golden mussel, is a bivalve mollusc native to Southeast Asia, which was introduced to South America via ballast water from commercial cargo ships from China and South Korea. In Brazil, the golden mussel is considered an invasive species, which has caused enormous economic and environmental damage. This is mainly due to the obstruction of pipes carrying water for the cooling systems within hydroelectric power plants. The presence of this invasive species also negatively impacts native fauna and flora. Early detection and rapid response during the early stages of infestation for this invasive species is essential to effectively control populations and prevent new infestations from forming. Techniques within molecular biology allow for more efficient detection of the golden mussel in all stages of development. In addition, the analysis of the genome, transcriptome and proteome of this species makes it possible to identify targets of biotechnological interest, which may be useful for the control of the golden mussel or for the generation of products of technological innovation. This review presents an overview of molecular studies involving this species and the perspectives on the prevention, control and combat of the golden mussel using these approaches.

This work evaluates the tolerance of adult Limnoperna fortunei to aerial exposure at different temperatures. The aim was to contribute knowledge about factors, such as the mussel’s capacity for survival out of water over long periods, allowing the invasion of new environments after terrestrial transport. The analysis of the results revealed that the time of exposure to air influenced mortality at different temperatures throughout the experimental period (P

The study was conducted to compare the distribution and composition of the Molluscs community in 4 ecosystems (4 sites) in southern of Iraq, namely: (Site 1)- in Euphrates River at Al-Madinah city, (Site 2)- in Shatt Al-Arab at Al-Sharsh area, (Site 3)- in the Garmat Ali River at Al-Mashib area and Site 4- in the Al-seba area. Samples were collected from study sites during a period of January to August 2019. (11) species from the class Gastropoda and (4) species from the class Bivalves, species were predominated in most of the study months which are species: Melenodies nodosa, Bellamya bengalensis and Melanopsis turbculata from Gastropoda and two of the species are bivalve: Corbicula fluminalis and Limnoperna fortunei (Dunker, 1857), and the last species, L. fortunei, was recorded in the current study for the first time in freshwater of Iraq. The total biodiversity values of Molluscs at the Euphrates River at Al-Madinah area (Site. 1), Al-Sharsh area (Site 2), Al-Mashib area (Site 3) and Al-seba area (Site 4) were 3.1, 2.9, 2.4, 1.8, respectively, and It was noted that the highest seasonal densities of the species in the current study were recorded in the winter season, reaching 1013, 905, 762 and 639 individuals / m2 in four sites, respectively. Resumo O estudo foi realizado para comparar a distribuição e composição da comunidade de moluscos em quatro ecossistemas (quatro locais) no sul do Iraque, a saber: Local 1- no rio Eufrates, na cidade de Al-Madinah, Local 2- em Shatt Al-Arab na área de Al-Sharsh, Local 3- no rio Garmat Ali na área de Al-Mashib, e Local 4- na área de Al-Seba. As amostras foram coletadas nos locais de estudo durante um período de janeiro a agosto de 2019. Onze espécies da classe Gastropoda e quatro espécies da classe Bivalves predominaram espécies na maioria dos meses de estudo, que são as espécies Melenodies nodosa, Bellamya bengalensis e Melanopsis turbculata, de Gastropoda, e duas das espécies, Corbicula fluminalis e Limnoperna fortunei (Dunker, 1857), de Bivalvia, e L. fortunei, foram registradas pela primeira vez em água doce do Iraque. Os valores totais de biodiversidade de moluscos no rio Eufrates na área de Al-Madinah (Local 1), área de Al-Sharsh (Local 2), área de Al-Mashib (Local 3) e área de Al-Seba (Local 4) foram 3,1, 2,9, 2,4, 1,8, respectivamente, e notou-se que as maiores densidades sazonais da espécie no presente estudo foram registradas na estação de inverno, chegando a 1013, 905, 762 e 639 indivíduos/m2 em quatro locais, respectivamente.

The literature indicates that exotic species have a greater tolerance to environmental stressors compared with native species. In recent decades, the introduction of contaminants into the environment has increased as a result of industrialization. The objective of this study was to verify the resistance of bivalve mollusks from freshwater native (Anodontites trapesialis) and exotic (Limnoperna fortunei) species to chemical contamination using an ex vivo/in vitro approach. Gill and muscle tissues were exposed to two different types of environmental stressors, copper (metal), and Roundup Transorb® (herbicide). The tissues were submitted to a cytotoxicity test in which the lysosomal integrity was assessed, from the adaptation of a method to isolated cells, and multixenobiotic resistance (MXR) test which evaluated cellular defense. In the exotic species, only copper at 9000 μg/L and Roundup Transorb® at 5000 μg/L were cytotoxic. In the native species, copper cytotoxicity at 900 and 9000 μg/L and Roundup Transorb® at 50 and 5000 μg/L were observed. Results were the same in both tissues. The MXR, responsible for the extrusion of contaminants (cell defense), was inhibited in both species when exposed to the contaminants, this cell defense system seems to be more inhibited in the native species, when exposed to both pollutants, indicating greater sensitivity. Therefore, cytotoxicity may be related to the lack of capacity of cellular defense. In relation to lysosomal integrity, the native species was more sensitive to cytotoxic pollutants, where a greater number of experimental conditions of metals and herbicide showed cytotoxicity, as well as more experimental situations inhibited its ability to defend itself.

Although some studies have showed the effects of different crystalline structures of nTiO2 (anatase and rutile) and their applicability in several fields, few studies has analyzed the effect of coexposure with other environmental contaminants such as copper. Thus, the objective of this study was to evaluate if the coexposure to nTiO2 (nominal concentration of 1 mg/L; anatase or rutile) can increase the incorporation and toxic effect induced by Cu (nominal concentration of 56 μg/L) in different tissues of Linmoperna fortunei after 120 h of exposure. Our results showed that the coexposure increased the accumulation of Cu in the gills and adductor muscle independently of the crystalline form and can positively or negatively modulate the antioxidant system, depending on the tissue analyzed. However, exposure only to rutile nTiO2 induced damage in the adductor muscle evidenced by the infiltration of hemocytes in this tissue. Additionally, histomorphometric changes based on fractal dimension analysis showed that coexposure to both forms of nTiO2 induced damage in the same tissue. These results suggest that both crystalline forms exhibited toxicity depending on the analyzed tissue and that coexposure of nTiO2 with Cu may be harmful in L. fortunei, indicating that increased attention to the use and release of nTiO2 in the environment is needed to avoid deleterious effects in aquatic biota.

The Tonle Sap Lake in Cambodia is a crucial freshwater biodiversity hotspot and supports one of the world’s largest inland fisheries. Within the Tonle Sap basin, freshwater molluscs provide vital ecosystem services and are among the fauna targetted for commercial harvesting. Despite their importance, freshwater molluscs of the Tonle Sap basin remain poorly studied. The historical literature was reviewed and at least 153 species of freshwater molluscs have been previously recorded from throughout Cambodia, including 33 from the Tonle Sap basin. Surveys of the Tonle Sap Lake and surrounding watershed were also conducted and found 31 species, 15 bivalves (five families) and 16 gastropods (eight families), in the Tonle Sap basin, including three new records for Cambodia (Scaphula minuta, Novaculina siamensis, Wattebledia siamensis), the presence of globally invasive Pomacea maculata and potential pest species like Limnoperna fortunei. This study represents the most comprehensive documentation of freshwater molluscs of the Tonle Sap basin, and voucher specimens deposited at the Inland Fisheries Research and Development Institute, Cambodia, represent the first known reference collection of freshwater molluscs in the country. In order to combat the combined anthropogenic pressures, including invasive species, climate change and dams along the Mekong River, a multi-pronged approach is urgently required to study the biodiversity, ecology, ecosystem functioning of freshwater molluscs and other aquatic fauna in the Tonle Sap basin.

The intensive use of glyphosate in industrial agriculture may lead to freshwater contamination, encouraging studies of its toxic effect on non-target aquatic organisms. Glyphosate-based commercial formulations contain adjuvants, making them even more toxic than the active ingredient (a.i.) itself. The golden mussel Limnoperna fortunei is a freshwater invasive species which has been found to increase glyphosate dissipation in water and to accelerate eutrophication. The aim of this study is to evaluate the capability of L. fortunei to reduce the concentration of glyphosate in two commercial formulations, Roundup Max® and Glifosato Atanor®. Results were compared with the decay of the a.i. alone and in presence of mussels. Evasive response and toxicity tests were performed in a first set of trials to analyze the response of L. fortunei exposed to Roundup Max® and Glifosato Atanor®. Subsequently, we conducted a 21-day degradation experiment in 2.6-L microcosms applying the following treatments: 6 mg L−1 of technical-grade glyphosate (G), Glifosato Atanor® (A), Roundup Max® (R), 20 mussels in dechlorinated tap water (M), and the combination of mussels and herbicide either in the technical-grade (MG) or formulated form (MA and MR) (all by triplicate). Samples were collected at days 0, 1, 7, 14 and 21. No significant differences in glyphosate decay were found between treatments with mussels (MG: 2.03 ± 0.40 mg L−1; MA: 1.60 ± 0.32 mg L−1; MR: 1.81 ± 0.21 mg L−1), between glyphosate as a.i. and the commercial formulations, and between the commercial formulations, suggesting that the adjuvants did not affect the degrading potential of L. fortunei. In addition to the acceleration of glyphosate dissipation in water, there was an increase in the concentration of dissolved nutrients in water (N–NH4+ and P-PO43-) even higher than that caused by the filtering activity of the mussels, probably resulting from stress or from the degradation of glyphosate and adjuvants. We believe that a larger bioavailability of these nutrients due to glyphosate metabolization mediated by mussels would accelerate eutrophication processes in natural water bodies. The approach used here, where L. fortunei was exposed to two commercial formulations actually used in agricultural practices, sheds light on the potential impact of glyphosate decay on water bodies invaded by this species.

In South America, the presence of the non-native mollusc Limnoperna fortunei ( Dunker, 1857 ) (Mollusca, Bivalvia, Mytilidae) in rivers and reservoirs may result in several impacts on the environment and economic activities. In a continental scale, the factors that most influence the dispersion of the species are anthropogenic vectors such as the movement of boats and people. Due to this, distribution models based solely on climatic variables often fail to predict the areas most subject to invasion, both spatially and temporally. An alternative to overcome this problem is the evaluation assuming the phenomenon as a complex system. In this paper, we have used a cellular automata model to predict the spread of the golden mussel in the Brazilian territory, on a temporal and spatial scale. We assume three parameters of interest: a) altitude b) characteristics of the river, and c) human population density. Transition rules were defined based on various considerations discussed in the article. The algorithm estimated satisfactorily the risk of invasion by L. fortunei to date (year of 2016), and the simulations for the years 2030 and 2050 predicted a high risk of invasion in north and northeastern Brazil. The increased risk of invasion predicted by the model for the next decades indicates that prevention and control measures should be applied immediately and this is of the utmost importance for the country to be able to comply with the #9 Aichi target.

Contamination and biological invasions are important factors that alter the functioning of freshwater systems. We carried out two experiments involving daily measurements of dissolved oxygen (DO) and Limnoperna fortunei mortality: 1) the impact of DO on mussel survival mediated by glyphosate, 2,4-D and their mixture was analysed in a 22-day indoor experiment (IE) under controlled conditions using microcosms with L. fortunei, with and without air supply; and 2) the effect of each herbicide and their mixture on mussel accumulated mortality was compared in a 18-day outdoor experiment (OE) using mesocosms without air supply, with and without L. fortunei. Results showed that glyphosate, alone or mixed affected L. fortunei survival both directly and indirectly. In IE we observed direct toxicity of glyphosate in treatments with air supply, with accumulated mortality of 20.0% for glyphosate and 10.0% for the mixture. In OE, L. fortunei deepened the changes in the patterns of DO fluctuations driven by the herbicides, which led to hypoxia in the system. The accumulated mortality was 46.7, 8.6 and 48.2% for glyphosate, 2,4-D and the mixture, respectively. This study contributes to the understanding of the mechanisms that control the invasion of L. fortunei in freshwater systems influenced by agrochemicals.

The golden mussel, Limnoperna fortunei, is among the most devastating invasive species in freshwater habitats worldwide, leading to severe environmental disturbances and economic losses. Therefore, management efforts would be greatly improved by methods that efficiently detect and quantify the abundance of the golden mussel in freshwater habitats, particularly in early stages of colonization. In this study, we describe a highly-sensitive real-time PCR assay targeting a 100-bp region of the COI mitochondrial gene of the golden mussel. The method was able to detect as little as 0.225 pg of target DNA. This assay represents an important contribution to surveillance methods, as well as to optimize field measures to contain and manage populations of the golden mussel in its introduced range.

The success of Limnoperna fortunei as an invasive freshwater bivalve species is related to its physiological plasticity to endure changes in environmental conditions. The aim of this study was to investigate the physiological responses of L. fortunei after feeding on Microcystis aeruginosa grown at 26 °C (control) and 29 °C during 10 days. At the beginning, we measured biomass, fatty acids (FAs) composition on Cyanobacteria grown at both temperatures at different time intervals. Afterwards, mussels were fed with the thawed M. aeruginosa cells and their FA profile was measured after 15 days of feeding. M. aeruginosa exposed to 29 °C had the highest content of the FAs 18:2ω6 and cis-18:1ω9. The FA profile of the consumer L. fortunei fed with M. aeruginosa cultures grown at 29 °C was also significantly different to those fed with cultures grown at 26 °C, with a significant increased Eicosapentaenoic acid (EPA, 20:5ω3) and Arachidonic acid (ARA, 20:4ω6) concentrations. L. fortunei was already known to be physiologically adapted to live at 29 °C, but our results also shown a high biosynthesis of EPA and ARA (increase of 70 and 40% respectively, compared with 26 °C) and avoided the lipid peroxidation of both FAs. This increased EPA and ARA biosynthesis may be an important source of ω3 and ω6 polyunsaturated FAs (PUFAs) for higher trophic levels, such as the pelagic fishes or birds that mainly prey on these mussels. The transfer of the cyanobacterial response at higher temperature to higher trophic levels will influence the overall functioning of freshwater bodies.

TiO 2 nanoparticles (TiO 2 -NP) have been incorporated into a large range of materials for different applications in the last decades and are very likely to appear in wastewater and effluents, eventually reaching the aquatic environment. Therefore, the assessment of the biological impact of TiO 2 -NP on aquatic ecosystem is of a major concern. The mussels represent a target group for TiO 2 -NP toxicity, as they are filter feeders and are capable of bioaccumulating toxic compounds. Furthermore, the exotic organism Limnoperna fortunei , golden mussel, is a freshwater bivalve that has been used in biomonitoring environmental conditions. In this work, the TiO 2 -NP’s ability to interact with haemocytes of golden mussel was assessed by transmission electron microscopy. The enzymatic and non-enzymatic antioxidant defenses were evaluated by superoxide dismutase (Sod) and catalase (Cat) activities and protein sulfhydryl content, which were measured after the golden mussel was exposed to TiO 2 -NP (1, 5, 10 and 50 μg mL −1 ). Results demonstrate that TiO 2 -NP was internalized by cells, causing alterations in haemocytes membrane. Antioxidant activity of Sod and Cat decreased after 2 h TiO 2 -NP exposure. After 4 h exposure, the enzymatic antioxidant activity was restored. Notably, the protein sulfhydryl content decreased after 2 h to all the TiO 2 -NP concentrations and no alterations were observed after 4 h of TiO 2 -NP exposure. These results demonstrate the potential of golden mussel as sentinel organism to TiO 2 -NP exposure.

Limnoperna fortunei is an aggressive freshwater invader species recorded for the first time in 1991 at Balneario Bagliardi, Río de la Plata Estuary, Argentina. Since then it has spread rapidly within South America. We tested the hypothesis that invasive species need a time span for the development of a reproductive pattern once settled in a new environment. We analyzed the reproductive cycle of a population of L. fortunei from Guaiba Lake (29°55′S; Rio Grande do Sul, Brazil) and compared it with previous studies at Balneario Bagliardi (34°52′S). We studied 834 specimens from Guaiba, following routine histological techniques. At any sampled period, the highest proportion of specimens were mature and gonads showed no resting periods. Female gonad maturation periods and oocyte growth were recorded throughout the year. Five periods of spawning of different intensity were differentiated. The reproductive pattern of the Guaiba population showed more spawning events and a remarkable recovery capacity of the female gonad compared to the Balneario Bagliardi population. The reproductive patterns of L. fortunei from Guaiba two years after invasion shows similarities with the reproductive pattern described previously from Balneario Bagliardi after two and three years of invasion Fil: Giglio, Matías Leonel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentina Fil: Dreher Mansur, María C.. Universidade Federal do Rio Grande do Sul; Brasil Fil: Damborenea, Maria Cristina. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. División de Zoología Invertebrados; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina Fil: Penchaszadeh, Pablo Enrique. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; Argentina Fil: Darrigran, Gustavo Alberto. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. División de Zoología Invertebrados; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina

The invasive bivalve Limnoperna fortunei (Dunker 1857) was introduced in South America in 1991, with the first occurrence in Brazil in 1998. In the Iguassu River, the species was recorded in 2001; however, it is unknown how it was introduced and spread. Adults and larvae were sampled in Iguassu and Parana Rivers, and the genetic profiles were compared. The species was absent in the upper reaches and only larvae were found, in low densities, in intermediary reaches. The L. fortunei populations from the lower Iguassu River presented no genetic differentiation among themselves, suggesting strong connectivity, and were significantly different from the Parana River populations, most likely because of Iguassu Falls. Furthermore, the results suggest that the Parana River represents the source of propagules to the Iguassu River. Generally, no significant differences were observed between the genetic structure inferred from adults and larvae. Only the population from the Iguassu National Park, a lotic environment, differed from the remaining subpopulations. The characterization of genetic profile using larval stages of L. fortunei populations was satisfactory, and represents an important protocol for studying the population genetics of aquatic species with planktonic larval stages.

The widespread use of titanium dioxide nanoparticles (TiO2-NP) in consumer products is the cause of its appearance in wastewater and effluents, reaching the aquatic environment. The evaluation of the biological impact of TiO2-NP and the need to understand its ecotoxicological impact to the aquatic ecosystem are of major concern. Bivalve mollusks may represent a target group for nanoparticle toxicity. Limnoperna fortunei (golden mussel), a freshwater bivalve organism that has been employed in biomonitoring environmental conditions. Comet assay, micronucleus test and oxidative damage to lipids and proteins were performed after the golden mussel was exposed to TiO2-NP (1, 5, 10 and 50 μg mL−1). The results demonstrate that TiO2-NP can damage the DNA of haemocytes after 2 h of exposure and the genotoxic activity significantly increased after 4 h exposure to TiO2-NP, at all the TiO2-NP concentrations. TiO2-NP was ineffective in causing mutagenicity in the haemolymph cells of golden mussel. The increase in the lipid peroxidation levels and carbonyl proteins after the exposure to TiO2-NP indicates the induction of oxidative stress at 2 h exposure with similar results to all TiO2-NP concentrations, but these effects did not occur at 4 h exposure. These results demonstrated that, although TiO2-NP is not mutagenic to golden mussel, it does induce DNA damage and oxidative stress in these organisms.

This study evaluated the Limnoperna fortunei (golden mussel) as a bioindicator of cytotoxicity and genotoxicity in aquatic environments contaminated by heavy metals. Five groups of 50 subjects each were exposed to different concentration of mercuric chloride (HgCl2) (0.001 mg/L, group I; 0.005 mg/L, group II; 0.01 mg/L, group II; 0.02 mg/L, group IV; and 0.1 mg/L, group V). The control group for both chronic and acute treatment did not receive HgCl2. For chronic exposure, the respective groups were placed in aquaria with water contaminated with the above concentrations of HgCl2. For acute exposure, the different concentrations of HgCl2 were injected into the posterior adductor muscle of the individuals belonging to the aforementioned groups. The biological matrix used in the tests was the whole body muscle. Tests (cell viability assay, alkaline comet test; enumeration of micronuclei and necrotic cells, quantification of Hg content in tissues and water, and histopathological analysis of tissues), were carried out on the 7th, 15th, and 30th treatment days or 2 h after injection. Our results demonstrated that L. fortunei showed cell damage in both chronic and acute exposure groups. Significant DNA damage was observed at both the 15th (0.1 mg/L) and 30th (0.01–0.1 mg/L) days of chronic exposure. However, in acute treatment all concentrations induced DNA breaks. The presence of necrosis increased at all concentrations tested for both acute and chronic exposure. Tissue mercury retention on the 15th day was higher than on the 30th day of exposure, while in the same period, there was a decrease in the mercury content of aquarium water. Taking the data together, it is concluded that L. fortunei as a possible bioindicator of the quality of aquatic environments.

Limnoperna fortunei (Dunker 1857) is a freshwater mussel with physiological tolerance to different environmental conditions, which may explain its success as an invasive species. The role of abiotic factors in its establishment, abundance and projections of risk of further spread into several areas has been studied. These mussels may respond to multiple environmental stressors, such as temperature, through physiological mechanisms, behavioral responses, mortality or some combination of these. The aim of this study was to investigate the behavioral responses (valve closing), glycogen concentrations and mortality of L. fortunei under four different temperatures (5°C, 10°C, 20°C and 30°C) during a chronic test (30 days). Two-way analysis of variance (ANOVA) was used to compare glycogen concentrations across days of the experiment and at the different temperatures. Differences in valve-closing behavior and mortality among temperatures were tested using repeated-measures ANOVA. We observed that most of the mussels maintained at 5°C closed their valves (74.7±15.3%), indicating that they remain inactive at low temperatures. The glycogen levels significantly differed among the temperatures tested. These differences occurred mainly due to the high glycogen values observed in mussels exposed to 10°C. Stability in glycogen concentrations was observed within each particular temperature. The cumulative mortality was higher at extreme temperatures (5°C and 30°C). The ideal temperature for laboratory maintenance and tests is approximately 20°C. Our data also show that L. fortunei can survive and maintain their energy reserves (glycogen) for several days at 5°C, an important feature related to its invasion success.

We conducted a study about invertebrates on artificial substrates with different antifouling paints in order to answer the following questions 1) is there lower accumulation of organic matter on substrates with antifouling paints, 2) is invertebrate colonization influenced by the release of biocides from antifouling paints, 3) is the colonization of aquatic invertebrates positively influenced by the material accumulated upon the substrate surface and 4) is the assemblage composition of invertebrates similar among the different antifouling paints? To answer these questions, four structures were installed in the Baía River in February 1st, 2007. Each structure was composed of 7 wood boards: 5 boards painted with each type of antifouling paints (T1, T2, T3, T4 and T5), one painted only with the primer (Pr) and the other without any paint (Cn). After 365 days, we observed a greater accumulation of organic matter in the substrates with T2 and T3 paint coatings. Limnoperna fortunei was recorded in all tested paints, with higher densities in the control, primer, T2 and T3. The colonization of Chironomidae and Naididae on the substrate was positively influenced by L. fortunei density. The non-metric multidimensional scaling (NMDS) of the invertebrate community provided evidence of the clear distinction of invertebrate assemblages among the paints. Paints T2 and T3 were the most similar to the control and primer. Our results suggest that antifouling paints applied on substrates hinder invertebrate colonization by decreasing the density and richness of invertebrates. Resumo O estudo dos invertebrados em substrato artificial com diferentes revestimentos antiincrustantes foi realizado com o intuito de responder as seguintes questões 1) Há menor acumulação de material orgânico nos substratos com revestimentos antiincrustantes? 2) A colonização de invertebrados é afetada pela liberação de biocidas dos revestimentos antiincrustantes? 3) A colonização dos invertebrados aquáticos é facilitada por material acumulado sobre a superfície do substrato? 4) A composição da assembléia é similar entre os diferentes revestimentos? Para realização deste estudo, quatro estruturas foram instaladas em 01 de fevereiro de 2007 no rio Baía. Em cada estrutura foram colocadas sete placas de madeira: cinco placas com aplicação de um tipo de revestimentos antiincrustantes (T1, T2, T3, T4 e T5), uma placa apenas com a aplicação do primer (Pr) e a outra placa permaneceu sem aplicação de nenhum revestimento (Cn). Ao final dos 365 dias em que os substratos ficaram submersos observou-se maior acumulação de material orgânico nos substratos dos revestimentos T2 e T3. Limnoperna fortunei foi registrada em todos os revestimentos testados, com maiores densidades encontradas no controle, primer, T2 e T3. A colonização de Chironomidae e Naididae sobre os substratos foi influenciada pela densidade de L. fortunei. A ordenação (NMDS) evidenciou a separação da assembléia de invertebrados entre os revestimentos. Os revestimentos T2 e T3 foram os mais similares ao controle e ao primer. Os resultados obtidos sugerem que revestimentos antiicrustantes aplicados sobre substratos dificultam a colonização de invertebrados, reduzindo a densidade e riqueza de invertebrados.

The freshwater mussel, Limnoperna fortunei, is adapted to colonize a wide range of aquatic environments, and its ability to contend with environmental stress through phenotypic plasticity has allowed this species to successfully colonize and become established in new regions. Only limited information is currently available on the widely intraspecific variability of this species in response to environmental heterogeneity. Here, we tested the hypotheses that (1) growth and body weight of mussels from a highly polluted environment differed from those from a less polluted habitat, and (2) growth parameters estimated in this study differed from those reported for other invaded ecosystems. We conducted controlled field experiments in two study sites with differing levels of pollution. To compare our results to those reported elsewhere, we considered growth data from studies performed in different locations. We found that mussels from the more polluted habitat showed lower shell growth and body weight than those from the less polluted environment. We also observed differences in the growth performances of the golden mussel between our estimates and those from other invaded habitats. Our findings provide useful information to better understand the striking intraspecific variability of this species in response to stressful conditions. Knowledge on the phenotypic plasticity of L. fortunei is essential for predicting and managing this species. Fil: Bonel, Nicolás. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Laboratorio de Zoología de Invertebrados I; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Lorda, Julio. University of California; Estados Unidos

The golden mussel, Limnoperna fortunei, is an Asian freshwater bivalve introduced in South America in the beginning of the 1990’s, probably through ballast water releases in La Plata River estuary. It dispersed north through Paraná, Uruguay and Paraguay Rivers. The study evaluated the macroscale (18° to 34° S; 45° to 60° W) distribution of L. fortunei (larval stages) in the main rivers and reservoirs of La Plata Basin. Samplings were performed through 68 µm vertical plankton net hauls. Limnological variables were simultaneously determined. Larvae abundance correlated significantly with oxygen (positively) in summer, with temperature (positively) in winter and with total phosphorus and total nitrogen (both negatively) in winter. We expected densities to decrease towards north (latitudinal gradient) and increase in lentic conditions (reservoirs). Despite maximum density was found near the introduction point (La Plata River) similar value was also observed 2,000 km north (Paraguay River). The first hypothesis was refuted. The second hypothesis was partially accepted. Abundances were consistently higher in summer. Higher trophic conditions and fast flow seem to inhibit population growth. Food resources (chlorophyll a) locally influenced temporal variation. Limnoperna fortunei is widely distributed in La Plata basin, reaching high larval densities superior to 10,000 ind. m-3, in all major sub-basins. The species exhibits a high intrinsic dispersal ability (free planktonic larvae), wide tolerance to environmental factors and dispersion is potentialized by natural dispersion processes (e.g. fauna displacement) and human facilitation (e.g. commercial navigation). Resumo O mexilhão dourado, Limnoperna fortunei, é um bivalve de água doce originário da Ásia e introduzido na América do Sul no início de 1990, provavelmente através da água de lastro liberada no estuário do rio La Plata. Dispersando-se para o norte através dos rios Paraná, Uruguai e Paraguai. Esse estudo avaliou a distribuição em macroescala (18° a 34° S; 45° a 60° W) de L. fortunei (estágios larvais) nos principais rios e reservatórios da bacia La Plata. Amostragens foram realizadas através de arrastos verticais com rede planctônica de 68 µm. Variáveis limnológicas foram simultaneamente determinadas. A abundância larval correlacionou-se significativamente com oxigênio (positivamente) no verão, com a temperatura (positivamente) no inverno e com fósforo total e nitrogênio total (ambos negativamente) no inverno. Esperávamos um decréscimo da densidade em direção ao norte (gradiente latitudinal) e aumento em condições lênticas (reservatórios). Apesar de a densidade máxima ter sido encontrada próximo do ponto de introdução (La Plata River) valores similares também foram observados 2.000 km ao norte (rio Paraguai). A primeira hipótese foi refutada. A segunda hipótese foi parcialmente aceita. Abundâncias foram consistentemente maiores no verão. Condições eutróficas e fluxo rápido parecem inibir o crescimento populacional. Recurso alimentar (clorofila a) local influenciou a variação temporal. Limnoperna fortunei está amplamente distribuído na bacia de La Plata, alcançando densidades larvais superiores a 10.000 ind. m-3 em todas as principais sub bacias. A espécie possui alta capacidade de dispersão intrínseca (larva planctônica livre), ampla tolerância aos fatores ambientais e os processos de dispersão são potencializados por processos naturais (e.g. deslocamento de fauna) e facilitação humana (e.g. navegação comercial).

The capacity and mechanism of Limnoperna fortunei to reduce the concentration of forchlorfenuron [or 1-(2-chloropyridin-4-yl)-3-phenylurea (CPPU)] in water has been studied under laboratory conditions. Firstly, the evasive response of mussels to CPPU (10, 20, 40 and 60 mg L(-1)) was evaluated, and a toxicity test was carried out at these concentrations. Secondly, the effect of two different sizes of mussels on CPPU concentrations was investigated in a 24-day experiment. Thirdly, the role of intact mussels and valvae only were respectively evaluated in another 24-day experiment. The CPPU concentration decreased by about 40 % in the presence of large mussels and about 20 % in the presence of valvae only. Finally, nucleic acid extracts from the gut and biofilm microbial communities of L. fortunei were analyzed, and the number of copies of the bacterial genes amoA, nirK and nirS were determined. Based on these results, we propose possible mechanisms for CPPU degradation involving bacteria-associated nitrification and denitrification reactions. In summary, we found that the CPPU half-life depended on the presence of mussels, their size and their associated microorganisms.

We recorded, for the first time, byssal hairs in the Asian freshwater bivalve Limnoperna fortunei from the Paraná River system. We analysed the presence of hairs and their distribution on the shell in relation to habitat and shell size in 12 sites. Hairs were present in lentic habitats associated with macrophytes or organic matter, but were absent in lotic environments. The proportion of mussels with hairs was negatively correlated with current velocity. Hairs were more frequent and abundant in larger mussels. In general, the hairs are a similar length over the bivalve and almost entirely cover the shells in≥60% in lentic habitats. The projections allow L. fortunei to be camouflaged among the roots of macrophytes or coarse organic matter, assisting in avoiding visual predators. The clear-cut separation of L. fortunei populations into two different groups could be associated with phenotypic plasticity in this species. Fil: Montalto, Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto Nacional de Limnología. Universidad Nacional del Litoral. Instituto Nacional de Limnología; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias; Argentina Fil: Rojas Molina, Florencia Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto Nacional de Limnología. Universidad Nacional del Litoral. Instituto Nacional de Limnología; Argentina

The mussel genus Xenostrobus comprises eight named extant species from the Indo-West Pacific, including Xenostrobus inconstans, Xenostrobus pulex and Xenostrobus securis from Australia, the latter two also occurring in New Zealand. Xenostrobus species are predominantly restricted to estuaries or sheltered marine habitats although X. pulex inhabits the rocky intertidal on open coasts. Xenostrobus has recently been suggested to be congeneric with the freshwater invasive species Limnoperna fortunei. Xenostrobus securis is itself invasive in the Far East and Europe. This study employed DNA sequences from cytochrome c oxidase (COI), the D1 expansion region of 28S rRNA and the internal transcribed spacers of the ribosomal cistrons to investigate species relationships and habitat transitions in Australasian Xenostrobus. It is unlikely that Xenostrobus and Limnoperna are congeneric as their COI sequences are very different. There was unexpected complexity in defining Xenostrobus species. Xenostrobus pulex from Ne...

This paper describes the size-selective predation on Limnoperna fortunei (Dunker, 1857) by Pimelodus pintado (Azpelicueta, Lundberg & Loureiro, 2008) from the time it arrived at the Mirim Lagoon basin (2005). Sampling was carried out using bottom trawl in depths of 3-6 m, from January to November 2005, and from October to November 2008. Pimelodus pintado began to prey upon L. fortunei soon after its arrival (austral spring of 2005). On the spring of 2008, L. fortunei was found to be the most important food item of P. pintado. The variation in length of the mussels (0.7-3.2 cm, with a mode of 1.3 cm) indicates that the species is now fully established in the system. Our data indicates that large individuals of P. pintado incorporate more mussels in their diets than small individuals. However, regardless of their size, P. pintado individuals predate only on small (

In the area of Middle Paraná River, Argentina, in the pleuston community associated with the roots of Eichhornia crassipes (Mart.) Solms-Laubach, 1883, an epibiosis of the non-indigenous mussel Limnoperna fortunei (Dunker, 1857) on the native crab Trichodactylus borellianus Nobili, 1896 was occasionally recorded. The organisms were measured, weighed, and photographed. Attached molluscs were larger and heavier than their host crabs. This new interaction in the system was analyzed with regard to several biological and ecological characteristics of both species. En el área del río Paraná Medio, Argentina, en la comunidad pleustónica asociada a raíces de Eichhornia crassipes (Mart.) Solms-Laubach, 1883, se registró ocacionalmente la epibiosis del mejillón invasor Limnnoperna fortunei (Dunker, 1857) sobre el cangrejo nativo Trichodactylus borellianus Nobili, 1896. Los organismos fueron medidos, pesados y fotografiados. Los moluscos adheridos fueron más grandes y pesados que sus cangrejos hospedadores. Se analizó esta nueva interacción en el sistema con respecto a varias características biológicas y ecológicas de ambas especies. Fil: Rojas Molina, Florencia Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto Nacional de Limnología. Universidad Nacional del Litoral. Instituto Nacional de Limnología; Argentina Fil: Williner, Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto Nacional de Limnología. Universidad Nacional del Litoral. Instituto Nacional de Limnología; Argentina

Multiple anthropogenic stressors act simultaneously on the environment, with consequences different from those caused by single-stressor exposure. We investigated how the combination of the invasive mussel Limnoperna fortunei and a widely applied herbicide, Roundup Max®, affected freshwater microscopic communities and water quality. Further, we compared these results with those induced by the combination of the mussel and technical-grade glyphosate. We carried out a 34-day experiment in outdoor mesocosms, applying the following six treatments: 6 mg L−1 of technical-grade glyphosate (G), the equivalent concentration of glyphosate in Roundup Max® (R), 100 mussels (M), the combination of mussels and herbicide either in the technical-grade or formulated form (MG and MR, respectively), and control (C). Herbicides significantly increased total phosphorus in water; R and MR showed greater initial total nitrogen and ammonium. R increased picoplankton abundance and caused an eightfold increase in phytoplankton, with high turbidity values; G had a lower effect on these variables. Herbicide-mussel combination induced an accelerated dissipation of glyphosate in water (MG 6.36 ± 0.83 mg G g DW−1 day−1 and MR 5.16 ± 1.26 mg G g DW−1 day−1). A synergistic effect on ammonium was observed in MR but not in MG. MR and MG had an antagonistic effect on phytoplankton, which showed a drastic reduction due to grazing, as revealed by M. We provide evidence of differential effects of Roundup Max® and technical-grade glyphosate over water quality and microscopic communities, and in combination with mussels. However, in the combination of mussels and herbicides, mussels seem to play a leading role. In the presence of L. fortunei, the effects of higher nutrient availability provided by herbicides addition were counteracted by the filtration activity of mussels, which released nutrients, grazed on picoplankton and phytoplankton, and boosted the development of other primary producers, periphyton and metaphyton. Fil: Gattás, Florencia María. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentina Fil: Vinocur, Alicia Liliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentina Fil: Graziano, Martín. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; Argentina Fil: Dos Santos Afonso, María. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; Argentina Fil: Pizarro, Haydee Norma. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentina Fil: Cataldo, Daniel Hugo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentina

Successful animal rearing under laboratory conditions for commercial processes or laboratory experiments is a complex chain that includes several stressors (e.g., sampling and transport) and incurs, as a consequence, the reduction of natural animal conditions, economic losses and inconsistent and unreliable biological results. Since the invasion of the bivalve Limnoperna fortunei (Dunker, 1857) in South America, several studies have been performed to help control and manage this fouling pest in industrial plants that use raw water. Relatively little attention has been given to the laboratory rearing procedure of L. fortunei, its condition when exposed to a stressor or its acclimation into laboratory conditions. Considering this issue, the aims of this study are to (i) investigate L. fortunei physiological responses when submitted to the depuration process and subsequent air transport (without water/dry condition) at two temperatures, based on glycogen concentrations, and (ii) monitor the glycogen concentrations in different groups when maintained for 28 days under laboratory conditions. Based on the obtained results, depuration did not affect either of the groups when they were submitted to approximately eight hours of transport. The variation in glycogen concentration among the specimens that were obtained from the field under depurated and non-depurated conditions was significant only in the first week of laboratory growth for the non-depurated group and in the second week for the depurated group. In addition, the tested temperature did not affect either of the groups that were submitted to transport. The glycogen concentrations were similar to those of the specimens that were obtained from the field in third week, which suggests that the specimens acclimated to laboratory conditions during this period of time. Thus, the results indicate that the air transport and acclimation time can be successfully incorporated into experimental studies of L. fortunei. Finally, the tolerance of L. fortunei specimens to the stressor tested herein can help us understand the invasive capacity of this mussel during the establishment process. Resumo A criação bem sucedida de animais em condições de laboratório para processos comerciais ou experimentais é uma cadeia complexa que inclui vários fatores de estresse (ex. coleta e transporte) que tem como consequência a redução das condições naturais do animal, prejuízos econômicos e resultados biológicos inconsistentes. Desde a invasão do bivalve Limnoperna fortunei (Dunker, 1857) na América do Sul, vários estudos têm sido realizados para ajudar no controle e gestão dessa praga em plantas industriais que utilizam água. Relativamente pouca atenção tem sido dada ao processo de criação de L. fortunei em laboratório, sua condição quando exposta ao estresse e sua aclimatação a condições de laboratório. Considerando estes aspectos, os objetivos deste estudo foram: (i) investigar as respostas fisiológicas de L. fortunei submetidos ao processo de depuração e subsequente transporte (sem água/condição seca) em duas temperaturas, analisando as diferentes concentrações de glicogênio e (ii) monitorar as concentrações de glicogênio nos diferentes grupos, quando mantidos por 28 dias em condições de laboratório. Com base nos resultados obtidos, a depuração não afetou nenhum grupo quando eles foram submetidos a oito horas de transporte. A variação da concentração de glicogênio entre os espécimes do campo quando depurados e não depurados, foi significativa apenas em relação à primeira semana em laboratório para o grupo não depurado e à segunda semana para o grupo depurado. Além disto, a temperatura testada não afetou os grupos submetidos ao transporte. As concentrações de glicogénio foram semelhantes as dos espécimes do campo a partir da terceira semana, o que sugere que os espécimes estão aclimatados às condições de laboratoriais neste período de tempo. Assim, os resultados indicam que o transporte ao ar e o tempo de aclimatação podem ser incorporados com sucesso aos estudos experimentais com L. fortunei. Finalmente, o conhecimento sobre a tolerância de L. fortunei ao estresse pode ajudar a entender a capacidade invasiva deste durante o processo de estabelecimento.

The success of Limnoperna fortunei as an invasive species is related to its physiological plasticity that allows them to endure adverse environmental conditions. Starvation tolerance is considered to be an important trait associated with bivalve invasiveness. In natural ecosystems, food resources can vary during the year, exposing mussels to variable periods of starvation or limited food availability. Thus, mussels have developed physiological strategies to tolerate fluctuations in food availability. Glycogen concentration has been used in different monitoring studies as an indicator of the nutritional condition of bivalves. The aim of this study was to investigate the physiological responses of L. fortunei based on the glycogen concentrations of specimens under four treatments, comprising differentcombinations of feeding and starvation, during 125 days. The experiment was carried out in two phases. In the phase I, mussels were divided in two treatments: starvation (S) and feeding (F). After 100 days, tissue samples were collected to quantify glycogen concentrations and, each phase I group was divided in two subgroups: starvation (S) and feeding (F), resulting in four treatments. In the phase II, that lasted 25 days, starvation specimens (S) from phase I were allowed to feed (starvation-feeding treatment, or S-F), or continued to undergo starvation (starvation-starvation treatment, or S-S) and the feeding specimens (F) continued feeding (feeding-feeding group, or F-F), or were subjected to starvation (feeding-starvation treatment, or F-S). Behavior (valve-closing) and mortality were recorded in 24 h intervals. After the 25 days (phase II) all specimens were killed, and thei r soft tissue was removed to quantify glycogen concentrations. The glycogen concentration of the S-F treatment was lower than that of the F-S treatment, which was initially allowed to feed (phase I) and then subjected to starvation (phase II). Stability in the glycogen concentrations was observed when the phase II feeding conditions were maintained during the experiments, as observed in the S-S (continued starvation) and F-F (continued feeding) treatments. Based on our glycogen concentrations results, the golden mussel shows a higher tolerance to starvation (125 days) than has previously been published, which suggests that its tolerance strongly influences its invasive behavior., Facultad de Ciencias Naturales y Museo

Limnoperna fortunei, or the golden mussel, invaded South America through the Rio de la Plata estuary in 1991. The size-selective predation on this bivalve by freshwater decapods crustaceans could be an important part of selective trophic behavior because shell sizes are correlated with the flesh contents and their resistance to being broken. The aim of the paper was analyze the size selective predation by the freshwater crab Zilchiopsis collastinensis Pretzmann, 1968 on different sizes of the invasive bivalve L. fortunei. We considered three combinations (AC, AD and BD) of equal numbers of three different shell lengths of mussels: A (small) = 7 to 10.99 mm, B (medium) = 11 to 14.99 mm and C = 15 to 18.99 mm and D = 19 to 22.99 mm (large). These combinations were offered to each adult female intermolt crab individually, and the number of mussels remaining was registered after one, two and 24 hours to evaluate prey-size selection. After this trial, ten female crabs were recorded preying on two different sizes of mussels (A and C) in combination (AC), and we registered the selective feeding behavior (feeding methods and time of predation) with each size of mussel. The results indicated that female crabs ate different sizes of golden mussels, showing an increased consumption of large mollusks after 24 hours, although these sizes demanded higher predation times. Large mussels were encountered at the first time (one and two hours) by the crabs, and these mussels were consumed successfully in the combination AD. These female crabs showed several alternative strategies to access the flesh and were efficient in handling large mussels, presenting an advantageous degree of plasticity for their predatory responses. L. fortunei could be a new food resource for adult Z. collastinensis females, and it could be responsible for a new interaction “native predator-invasive prey” in the system.