Molecular and bioassay-based detection of Toxoplasma gondii oocyst uptake by mussels (Mytilus galloprovincialis)

Toxoplasma gondii is associated with morbidity and mortality in a variety of marine mammals, including fatal meningoencephalitis in the southern sea otter (Enhydra lutris nereis). The source(s) of T. gondii infection and routes of transmission in the marine environment are unknown. We hypothesise that filter-feeding marine bivalve shellfish serve as paratenic hosts by assimilation and concentration of infective T. gondii oocysts and their subsequent predation by southern sea otters is a source of infection for these animals. We developed a TaqMan PCR assay for detection of T. gondii ssrRNA and evaluated its usefulness for the detection of T. gondii in experimentally exposed mussels (Mytilus galloprovincialis) under laboratory conditions. Toxoplasma gondii-specific ssrRNA was detected in mussels as long as 21 days post-exposure to T. gondii oocysts. Parasite ssrRNA was most often detected in digestive gland homogenate (31 of 35, i.e. 89%) compared with haemolymph or gill homogenates. Parasite infectivity was confirmed using a mouse bioassay. Infections were detected in mice inoculated with any one of the mussel sample preparations (haemolymph, gill, or digestive gland), but only digestive gland samples remained bioassay-positive for at least 3 days post-exposure. For each time point, the total proportion of mice inoculated with each of the different tissues from T. gondii-exposed mussels was similar to the proportion of exposed mussels from the same treatment groups that were positive via TaqMan PCR. The TaqMan PCR assay described here is now being tested in field sampling of free-living invertebrate prey species from high-risk coastal locations where T. gondii infections are prevalent in southern sea otters.     

Heterogeneity and tissue specificity of tropomyosin isoforms from four species of bivalves

Heterogeneity and tissue specificity of tropomyosin isoforms obtained from four species of bivalves (Scapharca broughtonii (ark shell), Mytilus galloprovincialis (mussel), Atrina pectinata (surf clam) and Crassostrea gigas (Pacific oyster)), were examined. Tropomyosins were extracted from translucent and opaque portions of posterior adductor muscle, respectively, and cardiac muscle of each bivalve. There were two tropomyosin isoforms in the ark shell, the surf clam and the Pacific oyster. They were designated as TMa and TMb. In the ark shell, TMa was the common isoform and TMb was specific for the opaque portion of the adductor muscle. In the surf clam, TMb was the common isoform present in all tissues. TMa was found only in the translucent portion of muscle. In the Pacific oyster, TMb was the major component in both portions of adductor muscle and TMa was the major component in cardiac muscle, although both tropomyosins were included in all tissues. The mussel had only one tropomyosin.     

Neurons in a variety of molluscs react to antibodies raised against the VD1/RPD2 α-neuropeptide of the pond snail Lymnaea stagnalis

The VD₁ and RPD₂ neurons of Lymnaea stagnalis innervate other central neurons, certain skin areas, the pneumostome area, and the auricle of the heart. Recently, a set of four (, , , ) neuropeptides produced by these giant neurons and by certain other central neurons has been characterized. Although alternative splicing of the preprohormone of these neurons yields at least 10 different neuropeptides, an affinity-purified antiserum directed against a domain common to all neuropeptides has previously been shown to be highly selective in staining VD₁, RPD₂ and other neurons that produce the preprohormone. Since the gene encoding the neuropeptides is structurally similar to that expressed in R15 of the marine opisthobranch Aplysia californica, we have used the affinity purified antiserum as a marker for VD₁/RPD₂-related systems in other molluscs. Immunopositive neurons and fibers are observed in the central nervous systems of all species studied (Achatina fulica, Anodonta sp., Aplysia brasiliana, A. californica, Bulinus truncatus, Cepea sp., Eobania vermiculata, Helix aspersa, H. pomatia, Limax maximus, Mytilus edulis, Nassarius reticulatus, Viviparus viviparus). Several medium-sized and small neurons and 1–4 giant neurons are found in the pulmonates and opisthobranchs. The giant neurons in pulmonates have locations in the subesophageal ganglion, axonal branching patterns, and terminal arborizations in the auricle of the heart; all these characteristics are similar to those of VD₁ and RPD₂. Double-labelling (Lucifer yellow injection, immunocytochemistry) confirms that the two giant neurons in Helix pomatia are Br and Br. The immunoreactive cells in A. fulica appear to include the VIN and PON neurons. The antiserum also stains cells that appear to be the R15 neurons in two Aplysia species. The small and medium-sized neurons are distributed widely over the central ganglia of opisthobranchs and pulmonates. In prosobranchs and bivalves, small neurons are found in the cerebral and abdominal ganglia. No evidence has been found for innervation of the heart in these latter groups but in M. edulis, immunoreactive terminals can be observed in the gill. The results suggest the evolutionary conservation of immunoreactive peptides and the neurons that produce them, and thus support and extend previous hypotheses regarding the homology of certain giant neurons across molluscan species.     

Feeding activity of scallops and mussels measured simultaneously in the field: Repeated measures sampling and implications for modelling

Many studies on the biology of bivalves have focused on identifying the importance of various environmental factors in regulating feeding behaviour in order to predict ingestion and growth responses in a variety of habitats. When taking into consideration the diversity of studies undertaken and the variety of methodologies employed, it is perhaps not surprising that there have been inconsistencies in both interspecific and intraspecific comparisons. In order to compare and contrast the way in which feeding activities of a mussel species (Mytilus edulis) and a scallop species (Placopecten magellanicus) respond to environmental factors and seston characteristics, we exposed both species simultaneously to a fluctuating assemblage of natural particles found in their local environment. Similar sizes of mussels (40-73 mm) and scallops (40-88 mm) were held in flow through chambers at two different sites in Atlantic Canada on four separate occasions, during which environmental conditions, including seston characteristics, were monitored and individual clearance rates were measured. Because the same individuals were measured over time, repeated-measures statistical procedures were used to analyze the data. Results indicate that weight standardized clearance rates of mussels were significantly higher than those of scallops only at the highest concentration tested (12.8 mg l^− 1), which translates into almost 3 times more material being filtered. Scallops appeared to respond to fluctuations in environment parameters and suspended food particles more consistently than mussels. These data suggest that the feeding behaviour of these two species is mediated differently by the same environmental parameters. Our study also demonstrates the importance of selecting appropriate statistical tests (e.g., repeated measures) for the analyses of data obtained from repeated sampling of the same individuals over time, and the need to consider species-specific feeding responses when developing models that predict the influence of bivalve feeding on ecosystem function.     

黑龙江省东部七虎林组的一些双壳类化石

系统地描述了产于黑龙江省东部龙爪沟群七虎林组的一些双壳类化石,共计6属8种,结合所研究标本对Leionucula与Palaeonucula、MaUetia与Mesosaccella等属之间的区别及Entolium orbiculare的定向等问题进行了探讨,并对七虎林组的时代进行了讨论。     

Cardiac and branchial physiology associated with copper accumulation and detoxication in the mytilid mussel Perna viridis (L.)

Acute, sublethal, copper exposures induced bimodal physiological responses in the mytilid mussel Perna viridis. Whole animal studies revealed that copper increased heart rate (mild tachycardia) whilst clearance rates and hence gill cilia were simultaneously inhibited. The inhibitory bioamine dopamine reversed tachycardia, indicating that elevated heart rates were due to nervous (serotonergic) stimulation of the myocytes. The excitatory bioamine 5-hydroxytryptamine failed to reverse copper-induced inhibition of clearance rates, suggesting that mechanical damage or blockage of the cilia rather than overt neurotoxicity was impairing the function of the gill. The digestive diverticula was the major copper-accumulating organ although rapid metal exocytosis was evident when individuals were allowed to depurate in uncontaminated seawater. Waste products are expelled from the digestive diverticula into the intestine and as the rectum passes through the heart, elevated rates of ventricular contraction may facilitate egestion of metal-rich faeces. Elevated cardiac activity during exposure to copper may serve to increase ventricular pulsatile pressure on the rectum for the expulsion of metal-rich faecal inclusions (coproliths) derived from the digestive diverticula lysosomes. This study describes a relationship between the heart, alimentary canal and digestive diverticula lysosomes as a pathway for metal detoxication in mytilid mussels.     

Effects of digenean trematodes and heterotrophic bacteria on mortality and burying capability of the common cockle Cerastoderma edule (L.)

Population dynamics of marine invertebrates is controlled by a variety of abiotic and biotic factors. Among these, some have received lesser attention from marine ecologists because of their ‘discrete’ nature. This is the case of parasitism and bacterial load. In the present study, we focused on the role that both digenean trematodes and heterotrophic aerobic bacteria might play in the mortality and burying behaviour of cockles, Cerastoderma edule.The bivalves were sampled monthly during 1 year from two sites in Arcachon Bay (French Atlantic coast). Mortality rates were assessed after transferring in the laboratory normally buried and unburied (i.e. found lying at the sediment surface at low tide) cockles. Their digenean and bacterial loads were determined for both positions (normally buried and unburied). Mortality rate was significantly higher for cockles found out of the sediment at low tide, suggesting that this abnormal position was a prelude to cockles' death. Comparison of digenean load of cockles showed no significant difference between buried and unburied bivalves. In contrast, bacterial load was significantly higher in unburied cockles than in normally buried animals. The effect of high concentration of a marine bacterial strain (Pseudomonas fluorescens) on cockles' burying behaviour and mortality was tested in the laboratory. Results showed that these bacteria could trigger the emergence of animals from the sediment but did not cause cockles' death. These field observations and laboratory experiments suggest that bacteria, rather than digenean trematodes, could play a role in the emergence of cockles and, hence, affect their survival in the wild.     

Bivalve borings in Lower Jurassic Lithiotis fauna from northeastern Italy and its palaeoecological interpretation

Random shell sections of the Pliensbachian (Early Jurassic) larger bivalve Opisoma from columns within the Main Post Office building of Ferrara, northern Italy, have been discovered to bear neat clavate-shape boreholes. These boreholes belong to the ichnogenus Gastrochaenolites Leymerie and represent bivalve borings. Opisoma is a subordinate component of the Lithiotis fauna characterised by aberrant shells thriving in tropical lagoonal settings which were widespread throughout the Tethyan and Panthalassa coasts. Although the Lithiotis fauna is well known in the palaeontological literature, no bivalve boring have been so far been formally described. The uniqueness of the morphology, size and substrate of these borings merits the designation of the new species Gastrochaenolites messisbugi ichnosp. nov. which thereby represents the first ichnospecies described from this fauna. The morphology of the boreholes and the included bivalves allows the boring activity to be ascribed to a mytilid bivalve. Palaeoecological and taphonomic analyses allowed the presence of the boreholes to be correlated to the Opisoma mode of life (epifaunal, free-living form) as well as to generally low sedimentation rates and seasonal mesotrophic conditions during an overall oligotrophic regime.

http://zoobank.org/urn:lsid:zoobank.org:act:CCF77B32-D459-4305-BC86-93F228852E50     

The death pathways in mussel larval cells after a freeze-thaw cycle

We analyzed cell viability, caspase activity, plasma membrane alterations and cell ultrastructure morphology to estimate the morphological and biochemical alterations that occur in bivalve molluscan cell cultures during cryopreservation. The use of 5% dymethyl sulfoxide as a cryoprotectant resulted in greater cell survival and a scarcity of destroyed cells lacking cytosol among dead cells. In this case, almost all cells died through necrosis or apoptosis, which appeared to increase in mussel cell cultures after a freeze-thaw cycle. Apoptosis was not a main death pathway in mussel cells, but it was induced in a significant part of these cells (up to 24%) immediately after thawing and depended mostly on the cryoprotectant used. Regardless of the type of the used cryoprotectant, we observed some nuclear aberrations in cells after freezing-thawing, such as few multipolar mitoses or the absence of a division spindle in mitotic cells. After analyzing different methods for assessing cell damage, the best results were obtained from optimal approaches that could provide information regarding the cell disruption level after freezing-thawing and could be considered for future studies.     

How fast do marine invertebrates burrow?

The burrowing of bivalves, arthropods, and echinoderms collected from tidal flats and shallow subtidal sediments of the Ogeechee estuary, Georgia, U.S.A was analyzed using time-elapse, X-ray analysis of thin-walled aquaria. The rate of sediment intrusion was determined for each animal. Burrowing rates ranged between 0.01 and 0.15 cm³/h for suspension-feeding animals. Deposit-feeding animals moved between 1 and 10 cm³ of sand per hour, approximately 10 to 100 times more sediment than the suspension feeders moved over similar times.Neoichnological experiments show that ten filter-feeding individuals could take as long as 115 yr to churn a 1 m² plot of sediment, by indexing the measured burrowing rates to realistic animal population densities. Ten such mobile deposit feeders as irregular echinoderms could bioturbate the same sediment in just 42 days. Under the maximum population densities modeled, the animals could bioturbate the sediment plot in 61 min. Given the reported results, qualitative interpretation of the rock record is possible: highly burrowed examples of the Skolithos Ichnofacies reflect high population densities and at least seasonal time spans. Highly burrowed examples of the Cruziana Ichnofacies may represent moderate population densities and short time spans.