Mussel Mortality Research Articles

Bacteriological analysis of unionid hemolymph collected from freshwater mussel populations in the Pacific northwestern United States

AbstractNative freshwater mussel (Unionidae) mortality events have been occurring with increased frequency in recent decades, with few investigations into potential etiological agents. In the western United States, no surveys have been published regarding the bacteria associated with unionid mussels. Herein, we examine locations of known mussel mortality events in the Chehalis River (Washington), in the Crooked River (Oregon), and Owyhee River (Oregon). Mussel populations considered healthy were sampled in the Skookumchuck River (Washington) for comparison. A variety of bacteria were isolated from these populations, and most notably, Acinetobacter spp. were identified from 82% of moribund individuals of Gonidea angulata in the Owyhee River. Future work evaluating whether Acinetobacter spp. are pathogenic to freshwater mussels could be valuable in unraveling the factors associated with these enigmatic mortality events.

Genetic parameters for resistance to field mortality outbreaks and resistance to a pathogenic strain of Vibrio splendidus in Mytilus edulis, Mytilus galloprovincialis and natural hybrid

Since 2014, abnormal mussel mortality (AMM) outbreaks have occurred along the French Atlantic coasts and mortality rates differed among years and sites (30–100%). Still, the etiology of the AMM is undefined. Our study explores the resistance of two mussel species (Mytilus edulis and Mytilus galloprovincialis) and natural hybrids (NH) cultivated in France to AMM. One hundred mussel families were produced using six wild mussel populations in January/February 2017. Mussels were tested in two sites, La Floride (LF) and Maison Blanche (MB), in October 2017, and their mortality/growth was recorded until June 2018. AMM was observed at MB during spring 2018, and reached 70%, 43%, and 63%, for M. edulis, M. galloprovincialis, and NH, respectively, at endpoint. In contrast, mortality was low in LF, reaching 11%, 30%, and 22%, respectively. Heritability for mortality was low to moderate in both sites ranging from 0.12 to 0.37 with higher values in M. edulis, intermediate for NH, and the lowest for M. galloprovincialis. A significant genotype-by-environment interaction was found for mortality between sites, with low genetic correlations not significantantly different from zero (−0.05 to 0.43). In July 2018, mussel families were evaluated under experimental infection using a pathogenic strain of Vibrio splendidus isolated during AMM in 2014. The mean mortality after 72 h post-injection was 53%, 22%, and 31% for M. edulis, M. galloprovincialis, and NH, respectively. The lowest heritability estimates were obtained for NH (0.21) and the highest for M. galloprovincialis (0.41), and the genetic correlation between AMM in MB and experimental infection was not significantly different from 0, suggesting that this Vibrio strain, is not the major cause of AMM outbreak observed in MB in 2018. Finally, significant genetic variation was observed for growth traits with mostly high heritability (> 0.44) for both species. Interestingly, genetic correlation between mortality and growth was weak and non-significant from zero.

Understanding the role of Francisella halioticida in mussel mortalities in France: an integrative approach.

Since 2014, mass mortalities of mussels Mytilus spp. have occurred in production areas on the Atlantic coast of France. The aetiology of these outbreaks remained unknown until the bacterium Francisella halioticida was detected in some mussel mortality cases. This retrospective study was conducted to assess the association between F. halioticida and these mussel mortalities. Mussel batches (n = 45) from the Atlantic coast and English Channel were selected from archived individual samples (n = 863) collected either during or outside of mortality events between 2014 and 2017. All mussels were analysed by real-time PCR assays targeting F. halioticida; in addition, 185 were analysed using histological analysis and 178 by 16S rRNA metabarcoding. F. halioticida DNA was detected by real-time PCR and 16S rRNA metabarcoding in 282 and 34 mussels, respectively. Among these individuals, 82% (real-time PCR analysis) and 76% (16S rRNA metabarcoding analysis) were sampled during a mortality event. Histological analyses showed that moribund individuals had lesions mainly characterized by necrosis, haemocyte infiltration and granulomas. Risk factor analysis showed that mussel batches with more than 20% of PCR-positive individuals were more likely to have been sampled during a mortality event, and positive 16S rRNA metabarcoding batches increased the strength of the association with mortality by 11.6 times. The role of F. halioticida in mussel mortalities was determined by reviewing the available evidence. To this end, a causation criteria grid, tailored to marine diseases and molecular pathogen detection tools, allowed more evidence to be gathered on the causal role of this bacterium in mussel mortalities.

Dislodgement and mortality challenges when restoring shallow mussel beds (Mytilus edulis) in a Danish estuary

In recent decades, mussel beds in the northern Atlantic and Scandinavia have declined rapidly in extent due to anthropogenic impacts, similar to many other marine habitats. In this study, a large‐scale restoration experiment was conducted to identify major challenges that arise during restoration efforts on shallow subtidal mussel beds. Suspension‐grown mussels (Mytilus edulis) were relayed in two different treatments either directly on bare bottom sandy sediments, or on coir nets (Net), used as a proxy for suitable byssal attachment substrate. The treatments were monitored for 1.5 years and coverage (%), biomass (WW), and population dynamics were quantified. Two main challenges of shallow bed restoration were identified: (1) Lack of suitable attachment substrate resulting in dislodgment of individuals during storm events. The Net treatment had significantly higher coverage and biomass of Mytilus at the end of the monitoring period, clearly demonstrating the importance of suitable substrate at physically exposed locations. (2) High mortality of juvenile mussels. Population dynamics revealed a high mortality of juvenile Mytilus, which resulted in almost complete loss of relayed Mytilus individuals less than 30 mm within the first season. This was most likely due to high meso‐predator densities, as a result of declining top‐predator populations. The high mortality of juvenile Mytilus prevented successful annual recruitment, thereby making the population unsustainable long‐term. Both challenges need to be addressed to create stable beds during restoration. Additionally, the experiment demonstrated the viability of using suspension‐grown Mytilus as a seed‐source when restoring mussel beds.

Evaluation of the health of Mediterranean mussels (Mytilus galloprovincialis Lamarck, 1819) distributed in the Çanakkale strait, Turkey

The observation of mortality in Mediterranean mussels (Mytilus galloprovincialis) distributed in the Çanakkale Strait in recent years was influential in developing the research question for this study. In this study, the presence of bacteria (Vibrio spp.) and parasites (Marteilia spp. and Haplosporidium spp.) in mussels collected from Kumkale, Kepez, and Umurbey stations in the Çanakkale Strait was investigated seasonally. Microbiological findings, histopathology, oxidative stress enzymes and their gene expressions, lipid peroxidation, lysosomal membrane stability, and changes in haemolymph were examined. In summer samples, both the defence system and the extent of damage were higher in gill tissue. In winter samples, enzyme activities and lipid peroxidation were found to be predominantly higher in digestive gland tissues. Histological examinations and Hemacolor staining revealed the presence of protozoan cysts, and for bacterial examination, molecular analysis performed after culturing revealed the presence of 7 Vibrio species. While the total numbers of heterotrophic bacteria detected in all samples were at acceptable levels, the predominance of Vibrio spp. numbers among the total heterotrophic bacteria detected in almost all samples were noteworthy. The total hemocyte count was calculated as 5.810(4)±0.58 (cells/mm3) in winter and 7.210(4)±1.03 (cells/mm3) in summer. These factors are considered to be possible causes of mussel mortality.

Quantification of Photobacterium swingsii and characterisation of disease progression in the New Zealand Greenshell™ mussel, Perna canaliculus

Greenshell™ mussels (Perna canaliculus) are endemic to New Zealand and support the largest aquaculture industry in the country. Photobacterium swingsii was isolated and identified from moribund P. canaliculus mussels following a mass mortality event. In this study, a challenge experiment was used to characterise, detect, and quantify P. swingsii in adult P. canaliculus following pathogen exposure via injection into the adductor muscle. A positive control (heat-killed P. swingsii injection) was included to account for the effects of injection and inactive bacterial exposure. Survival of control and infected mussels remained 100% during 72-hour monitoring period. Haemolymph was sampled for bacterial colony counts and haemocyte flow cytometry analyses; histology sections were obtained and processed for histopathological assessments; and adductor muscle, gill, digestive gland were sampled for quantitative polymerase chain reaction (PCR) analyses, all conducted at 12, 24, 48 h post-challenge (hpc). The most profound effects of bacterial injection on mussels were seen at 48 hpc, where mussel mortality, haemocyte counts and haemolymph bacterial colony forming were the highest. The quantification of P. swingsii via qPCR showed highest levels of bacterial DNA at 12 hpc in the adductor muscle, gill, and digestive gland. Histopathological observations suggested a non-specific inflammatory response in all mussels associated with a general stress response. This study highlights the physiological effects of P. swingsii infection in P. canaliculus mussels and provides histopathological insight into the tissue injury caused by the action of injection into the adductor muscle. The multi-technique methods used in this study can be applied for use in early surveillance programs of bacterial infection on mussel farms.

Impact of epizootics on mussel farms: Insights into microbiota composition of Mytilus species

Outbreaks of marine mussel mortality on French farms could have different aetiologies. One of them implies Vibrio splendidus strains. Beyond the involvement of this pathogen, there is considerable evidence that diseases often result from interactions between several microbes and the host. In this study, we explored the bacterial communities associated with mussel species and the surrounding water collected from a mussel farm affected by mortalities. The microbiota of Mytilus edulis, Mytilus galloprovincialis and their hybrids displayed an abnormal abundance of Proteobacteria, in particular the genera Vibrio, Cobetia and Arcobacter. Despite the dysbiosis, the Mediterranean mussel showed a different microbiota profile with a higher richness and presence of the phylum Bacteroidetes. Bipartite network analyses at the level of bacteria families confirmed this finding and showed that the microbiomes of M. edulis and the hybrids tended to cluster together. In addition, injection of mussels with the virulent V. splendidus induced less mortality rate in M. galloprovincialis compared to the other Mytilus sp. suggesting a better resistance of the Mediterranean mussel to infection. Our findings point to a probable aetiology of pathobiome-mediated disease in mussels. To fully understand this phenomenon, more knowledge is needed on the roles of pathobiotic systems and their development during disease establishment.

A facultative mutualism between habitat-forming species enhances the resistance of rocky shore communities to heat waves

Heat waves have increased in frequency, duration, and magnitude in recent decades, causing mass mortality events in terrestrial and aquatic ecosystems. Arguably, mass mortalities of habitat-forming organisms – i.e., dominant sessile organisms that define habitats via their own physical structure – would be amongst the most dramatic impact of heat waves because of their negative, cascading consequences on their associated biodiversity. However, the resistance of habitat-forming organisms to heat waves can be enhanced if they associate with secondary habitat formers able to tolerate and modulate extreme heat levels. Here we show that a seaweed of the Porphyra/Pyropia (P/P) clade can shield primary habitat-forming mussels, Brachidontes rodriguezii, from the impacts of extreme temperatures in a southwestern Atlantic rocky intertidal shore. By means of P/P removal experiments and surveys, we illustrate that P/P cover (a) buffers temperatures in the understory mussel beds during daytime air exposure periods in the summer, (b) reduces mussel mortality and leads to increased mussel body condition during warm summer periods, and (c) can prevent mass mortality of mussels during the course of a heat wave. Additionally, by means of a mussel removal experiment we illustrate that mussel cover is critical for P/P establishment, which is in consonance with the remarkably higher P/P densities and cover observed in mussel beds relative to exposed rock surfaces across a ~70 km coastal range. Collectively, these findings reveal a facultative mutualism where mussels provide a favorable substrate for P/P colonization and P/P attenuates heat mediated mortality on mussels. The ability of P/P to enhance the resistance of mussel beds to extreme heat events and the occurrence of similar P/P-mussel associations during spring-summer at globally dispersed sites suggests a widespread importance of P/P for the stability of mussel beds and their associated communities under warming climates.

The signal crayfish Pacifastacus leniusculus (Dana, 1852) (Crustacea, Decapoda) is threatening the near future of Margaritifera margaritifera Linnaeus, 1758 (Bivalvia, Unionoida) in the Negro River (NW Zamora, Spain)

We studied the incidence of the signal crayfish on a population of Margaritifera margaritifera in the Negro River (Zamora, Spain) during the summers of 2019 to 2022. The incidence of predation was assessed as a mortality factor in relation to floods and the hydrology of the river. The crayfish were trapped in a small plot of high pearl mussels density monitored since 2002, and collected each summer since 2019, during which time their abundance increased (65 % in three years). Simultaneously, we counted the shells carried by the floods to the gravel riverbanks. The incidence of floods was stable between 2019 and 2022 (10.3 % to 19.4 % of mortality), while at the bottom of the river the mortality of pearl mussels increased due to predation from 2.7 % to 43.3 %. During the 2022 dry season, 29 pearl mussels that had recently died and whose shell edges were widely gnawed by crayfishes were collected from the plot. The shells appeared bitten only in the contour exposed above the gravel, ruling out the possibility that the marks could be the effect of the scavenging of dead specimens by the crayfish. Low intensity trapping barely affected the crayfish population, since the following year their abundance in the controlled section had recovered.

Additive multiple predator effects increase prey mortality under experimental warming in a rocky intertidal food web

Climate warming is expected to alter species interactions, which are shaped by components of individual foraging behavior. Yet few studies examine species' behavioral responses to inform predictions about how species interactions will be affected by warming. Multiple predator effects (MPEs) may be mediated by climatic warming, especially in highly diverse coastal ecosystems that are prone to extreme warming events. In a laboratory experiment, we quantified the effects of elevated sea water temperature on mortality rates of mussels (Mytilus californianus, Conrad, 1837), and the behavioral responses of their predators: crabs (Romaleon antennarium, Stimpson, 1856) and sea stars (Pisaster ochraceus, Brandt, 1835), to understand how warming affects MPEs and if predators' behavioral responses to warming are indicative of the outcome of MPEs. Sea star effects on mussel mortality did not vary with temperature, but declined threefold in the presence of crabs, due to behavioral avoidance of physical contact with crabs. In contrast, the effect of crabs on mussel mortality was ten times higher under elevated temperature, but was unaffected by sea stars. In mixed predator treatments, crabs caused significant mussel mortality, but sea stars had weaker impacts, regardless of temperature. Consequently, multiple predators had effects that did not differ from those of the more effective crab predator. Increasing temperature in the eastern North Pacific may create locations where crabs become the dominant predator of mussels, with warming increasing predation risk for mussels in this region. Field studies should examine the effects of warming on crab predation on mussels in this system.

Siting and evaluation of offshore suspended mussel farms through nutritional restriction approach

An individual-based model based on mussel feeding on algae was developed for the first time, and the model was introduced to a three-dimensional water quality model based on nitrogen, phosphorus, phytoplankton cycling and dissolved oxygen balance, describing the ecological influence of mussel feeding. The drag of aquaculture facilities on suspended farms was introduced to a three-dimensional hydrodynamic model. The water quality model was coupled with the three-dimensional hydrodynamic model to simulate the hydrodynamic environment field of a large offshore suspended mussel farm in Gouqi Island throughout a year. The simulation results agree well with observations. Based on the nutritional restriction approach, the cause of mussel mortality in Gouqi Island is insufficient algal concentration and insufficient filtration rate. Accordingly, an optimized site selection of the suspended mussel farms was proposed, and its feasibility was validated in hydrology and water quality by using the model. Furthermore, hydrodynamic and ecological effects of aquaculture facilities and site selection were quantified. Aquaculture facilities in Gouqi Island reduces surface current speed by 86% and increases half-time of water exchange by 64%. P is the limiting nutrient in the suspended mussel farms of Gouqi Island. The siting of offshore suspended mussel farms only changes the local distribution of nutrients, causing minimal changes in nutrient concentrations and having limited ecological influence.

Different isoforms of parabens into marine environment: Biological effects on the bacterium Aliivibrio fischeri and the marine mussel Mytilus galloprovincialis

Different isoforms of alkyl esters of p-Hydroxybenzoic acid, also known as parabens, are of great concern due to their widespread presence into the aquatic environment, their high concentrations in wastewater discharges, as well as their ability to induce adverse effects on aquatic organisms. Considering the imperative need for assessing their fate and risk to aquatic environment, the present study investigated the biological effects of two isoforms of parabens, methyl- (MeP) and propyl- (PrP), on the bacterium Aliivibrio fischeri (using the Bioluminescence Inhibition/Microtox® bioassay) and the mussel Mytilus galloprovincialis (in terms of mussel mortality, cellular, oxidative and genotoxic stress indices). The assessment of MeP and PrP behavior into aquatic media (artificial sea water/ASW and 2 % NaCl), primarily performed by UHPLC-UV-MS analysis, showed only a slight hydrolysis of PrP to 4-Hydrobenzoic acid (4-HBA). Furthermore, exposure of both species to different concentrations of each paraben revealed differences among their toxic potential, as well as their ability to cause cellular, oxidative and genotoxic effects on hemocytes of challenged mussels. Interestingly, the Microtox® bioassay showed that PrP mediated toxicity in A. fischeri were more pronounced than MeP, as revealed by the estimated toxic endpoints (in terms of concentration that promote 50 % of bioluminescence inhibition, EC50). Similarly, in challenged mussels, a significant disturbance of mussel hemocytes' lysosomal membrane integrity, as well as enhanced levels of superoxides, nitric oxides, lipid peroxidation byproducts, and micronuclei formation were observed. These findings are of great interest, since MeP and PrP differential toxic potential, as well their ability to induce pre-pathological alterations in marine species, like mussels, give new evidence for their risk to aquatic biota.

Histopathologic survey of free-living populations of 2 species of freshwater mussels in Indiana.

Freshwater mussels are one of the most endangered groups of animals in Indiana, with nearly half of the native species either extirpated or listed as "state endangered" or of "special concern." Nationally, numerous freshwater mussel species are considered threatened. Freshwater mussel diseases are not well understood and few published accounts of freshwater mussel diseases with detailed histological descriptions exist. Mass mortality events within mussel populations are increasingly recognized, often with undetermined etiology. Our objective was to determine baseline histopathology in free-living populations of freshwater mussels. One-hundred twenty individual mussels representing 2 species-plain pocketbook (Lampsilis cardium) and fatmucket (Lampsilis siliquoidea)-were collected from 3 different locations within the Wildcat Creek watershed in central Indiana during June and July 2019. A cross-section through the visceral mass was obtained and immersed in 10% neutral-buffered formalin, with routine processing and hematoxylin and eosin staining. Branchial acariasis occurred in 43/60 fatmuckets and 22/60 plain pocketbooks. Infection with a bucephalid trematode was recognized in 18/60 fatmuckets, while infection of the gonadal duct with an unidentified trematode species was identified in 4/60 fatmuckets and 18/59 plain pocketbooks. Additional changes associated with unidentified trematodes, bacteria, fungi or oomycetes, and ciliates were observed. Other miscellaneous changes included mineralization, neuronal lipofuscinosis, and gonadal atrophy/atresia. A range of histological changes were observed. These changes likely represented background lesions: incidental findings, spontaneous infectious or endosymbiotic conditions, or normal physiological changes that routinely occur in free-living wild populations. Awareness of baseline lesions should inform future diagnostic investigations of mussel mortality events.

Dynamics of a diffusive mussel-algae system in closed advective environments

In this paper, we are concerned with a diffusive system modeling the interaction of mussel and algae in the water layer overlying the mussel bed, where the algae are the main food source for mussels, and the advection pushes algaes in one direction but not out of the domain. We present a threshold result on the global extinction and persistence of mussels. It shows that the condition for persistence depends on the principal eigenvalue of a scalar boundary value problem, which is related to the diffusion, the speed of the tidal flow, the conversion rate of algae to mussel production, and mussel mortality rate. We further investigate the asymptotic profile of the positive steady state when it exists.

Examination of the potential of refrigerated seawater to improve live transport of the mussel Perna canaliculus: Physiological responses, meat quality and safety implications under different chilled storage conditions

Refrigerated seawater (RSW) based systems for post-harvest storage of green-lipped mussels (Perna canaliculus; also known as Greenshell™ mussels) were assessed. Two post-harvest storage trials were conducted in October 2020 and March 2021 (austral spring and summer respectively) on mussels recovered for 2.5 days after harvest in flow-through seawater. The mussels were randomly assigned to three post-harvest treatments, all conducted at 6.5 °C for 10 days: Chilled in air; immersion in recirculated RSW; and Periodic immersion in RSW (6 min h−1). A range of physiological, morphometric and quality measures were screened to assess their ability to indicate mussel performance. Further to this, mussels in duplicate systems were inoculated with Escherichia coli and Vibrio parahaemolyticus to inform on risks associated with pathogen proliferation. The mussels interacted with the seawater in both the RSW and Periodic RSW systems as seawater ammonia concentrations increased in both. Mussel mortalities counted across the 10-day trials were highest in the Chilled treatment (5.5%), followed by RSW (3.5%) and Periodic RSW (2.4%). Key measurements including haemolymph pH and osmolality, meat and intra-valvular water weight, and textural hardness, were all affected by the post-harvest treatments. For inoculated mussels, E. coli and V. parahaemolyticus most probable numbers (MPN) remained stable in both the Chilled and Periodic RSW treatments with reductions in both bacteria recorded in the RSW treatment. Higher variances in intra-valvular water retention and haemolymph haemocyte numbers and differences in volatile organic compound profiles were seen in the summer mussels, as compared with spring, highlighting mussel condition as a determining factor for post-harvest performance. RSW has therefore been demonstrated to be beneficial, but optimisation (e.g., through further reduction of metabolic activity) may further increase practical post-harvest transport distances and the quality of live/fresh mussel products.

Salinity and pH effects on survival, growth, and reproduction of quagga mussels.

In recent decades, invasive quagga mussels have expanded to the Western United States from the Great Lakes region of North America. Most studies that evaluate the invasion potential of quagga mussels in western water bodies have utilized physiological and life history information from zebra mussels, a related taxon. Few studies have assessed the potential for invasion using specific information from quagga mussel life history or experiments that test for their survival in the fresh and saline waters of the western United States. We investigated quagga mussel survival, growth, and reproduction using semi-natural experiments under temperature and light controlled conditions across a gradient of water salinity (fresh to brackish) and pH (8.4-11). Water from Lake Mead was used as a positive control in our experiment, and water from Pyramid Lake and the Truckee River was used as brackish and freshwater treatments, respectively. The mussels used in the experiments were collected from Lake Mead. After 12 h in brackish water (4 ppt, pH 9.3), we observed 100% mortality of adult mussels. The swelling and disintegration of body tissues and high mortality rates indicated that high potassium, sodium, and chloride concentrations were the likely causes of death in brackish water treatments. In contrast, mussels were able to survive, grow, and reach sexual maturity in freshwater (0.1 ppt) with a low calcium concentration (17 mg L-1) after 57 days. Mussels died after 2 days at pH 11 and after 12 days at pH 10; during the 14-day monitoring period, no mortality was detected at pH 9.0, 9.3, or 9.5 and mussels did not exhibit any visual indications of stress. Understanding quagga mussel physiological and environmental tolerances appears to be essential for assessing their invasion potential in aquatic habitats.

Pathology and infectious agents of unionid mussels: A primer for pathologists in disease surveillance and investigation of mortality events.

Freshwater mussels are one of the most imperiled groups of organisms in the world, and more than 30 species have gone extinct in the last century. While habitat alteration and destruction have contributed to the declines, the role of disease in mortality events is unclear. In an effort to involve veterinary pathologists in disease surveillance and the investigation of freshwater mussel mortality events, we provide information on the conservation status of unionids, sample collection and processing techniques, and unique and confounding anatomical and physiological differences. We review the published accounts of pathology and infectious agents described in freshwater mussels including neoplasms, viruses, bacteria, fungi, fungal-like agents, ciliated protists, Aspidogastrea, Digenea, Nematoda, Acari, Diptera, and Odonata. Of the identified infectious agents, a single viral disease, Hyriopsis cumingii plague disease, that occurs only in cultured mussels is known to cause high mortality. Parasites including ciliates, trematodes, nematodes, mites, and insects may decrease host fitness, but are not known to cause mortality. Many of the published reports identify infectious agents at the light or ultrastructural microscopy level with no lesion or molecular characterization. Although metagenomic analyses provide sequence information for infectious agents, studies often fail to link the agents to tissue changes at the light or ultrastructural level or confirm their role in disease. Pathologists can bridge this gap between identification of infectious agents and confirmation of disease, participate in disease surveillance to ensure successful propagation programs necessary to restore decimated populations, and investigate mussel mortality events to document pathology and identify causality.

Comparison of the Efficiency of Different Eradication Treatments to Minimize the Impacts Caused by the Invasive Tunicate Styela plicata in Mussel Aquaculture.

In 2017, aquaculture producers of the Albufeira lagoon, Portugal, reported an invasion of tunicates that was disrupting mussel production, particularly the tunicate Styela plicata (Lesueur, 1823). A totally effective eradication method still does not exist, particularly for S. plicata, and the effects of the eradication treatments on bivalves' performance are also poorly understood. Our study examined the effectiveness of eradication treatments using three laboratory trials and five treatments (air exposure, freshwater immersion, sodium hypochlorite, hypersaline solution and acetic acid) for S. plicata, as well as their effects on survival and growth of blue mussel Mytilus edulis Linnaeus, 1758. While air exposure and freshwater immersion caused a 27% mortality rate in S. plicata, the acetic acid treatment was the most effective in eliminating this species (>90% mortality). However, a 33-40% mortality rate was registered in mussels. Both species were not affected by the hypersaline treatment in the last trial, but the sodium hypochlorite treatment led to a 57% mortality rate in mussels. Differences in mussels' growth rates were not detected. These trials represent a step forward in responding to the needs of aquaculture producers. However, further studies are needed to investigate the susceptibility of tunicates to treatments according to sexual maturation, as well as to ensure minimum mussel mortality in the most effective treatments, and to better understand the effects on mussel physiological performance in the long-term.

A Bacteriological Comparison of the Hemolymph from Healthy and Moribund Unionid Mussel Populations in the Upper Midwestern U.S.A. Prompts the Development of Diagnostic Assays to Detect Yokenella regensburgei.

Recent bacteriological investigations of freshwater mussel mortality events in the southeastern United States have identified a variety of bacteria and differences in bacterial communities between sick and healthy mussels. In particular, Yokenella regensburgei and Aeromonas spp. have been shown to be associated with moribund mussels, although it remains unclear whether these bacteria are causes or consequences of disease. To further understand the role of bacteria in mussel epizootics, we investigated mortality events that occurred in the upper Midwest in the Embarrass River (Wisconsin) and the Huron River (Michigan). For comparison, we also studied mussels from an unaffected population in the St. Croix River (Wisconsin). Diverse bacterial genera were identified from these sites, including Y. regensburgei from moribund mussels in the Embarrass River (Wisconsin). This bacterium has also been consistently isolated during ongoing mortality events in the Clinch River (Virginia). Subsequently, we developed and validated molecular assays for the detection of Yokenella to use in future investigations of mussel mortality events and to identify environmental reservoirs of this bacterium.

Further Bacteriological Analysis of Annual Pheasantshell (Actinonaias Pectorosa) Mussel Mortality Events in the Clinch River (Virginia/Tennessee), USA, Reveals a Consistent Association with Yokenella Regensburgei

Pheasantshell (Actinonaias pectorosa) mussels in the Clinch River (Tennessee/Virginia, USA) have declined dramatically in recent years. The bacterium Yokenella regensburgei was first isolated with high prevalence from Pheasantshells during the peak of a 2017 mortality event, but it was not identified after mortality subsided a few months later. Since 2017, Pheasantshell mortality in the Clinch River has occurred each autumn. We extended the investigation of culturable bacterial communities in the Clinch River during mussel mortality events in 2018, 2019, and 2020 and examined the spatial and temporal distribution of bacterial genera among Pheasantshells, as well as among other unionid mussels. We identified Y. regensburgei each year, almost exclusively during active mortality events. The significance of Y. regensburgei remains unclear, but the continued association of this bacterium with mussel mortality events warrants further study.