Description of the glochidium of Margaritifera auricularia (Spengler 1793) (Bivalvia,Unionoidea)

The glochidium of Margaritifera auricularia is described for the first time by using light microscopy, scanning electron microscopy and histological techniques. The larval mantle is formed by only two layers of cells; the inner one being much thicker, with microvilli. All cell masses of the glochidium are temporary aggregations that are the rudiments of organs of the subsequent juveniles which will be released after metamorphosis in the host tissues. In the glochidium there are three main masses of cells: (i) the muscle, which is in an anterior position; (ii) the oral plate in the centre of the larva; and (iii) the more ventrally and posteriorly situated ventral plate, or foot rudiment, flanged with lateral pits all bearing dense cilia. No rudimentary organs such as the pericardium, the kidney, the heart or nerve ganglia have developed. There are no visible hooks in the valve margins, but by using light microscopy we observed minute teeth covered by a rim of the periostracum. Near the margin of the shell there are two pairs of sensory hair tufts only observable by scanning electron microscopy. The glochidium of M. auricularia is the largest of the family Margaritiferidae and intermediate between the glochidium of the known species of this family and those of Unionidae.     

Who wins in the weaning process? Juvenile feeding morphology of two freshwater mussel species

The global decline of freshwater mussels can be partially attributed to their complex life cycle. Their survival from glochidium to adulthood is like a long obstacle race, with juvenile mortality as a key critical point. Mass mortality shortly after entering into a juvenile state has been reported in both wild and captive populations, thus weakening the effective bivalve population. A similar phenomenon occurs during metamorphosis in natural and hatchery populations of juvenile marine bivalves. Based on a morphological analysis using scanning electron microscopy of newly formed juveniles of the freshwater species Margaritifera margaritifera (L.) (Margaritiferidae) and Unio mancus Lamarck (Unionidae), we show that a second metamorphosis, consisting of drastic morphological changes, occurs that leads to suspension feeding in place of deposit feeding by the ciliated foot. We hypothesize that suspension feeding in these two species improves due to a gradual development of several morphological features including the contact between cilia of the inner gill posterior filaments, the inner gill reflection, the appearance of the ctenidial ventral groove and the formation of the pedal palps. Regardless of the presence of available food, a suspension feeding mode replaces deposit feeding, and juveniles unable to successfully transition morphologically or adapt to the feeding changes likely perish.     

Scanning electron microscopy of glochidia and juveniles of the freshwater mussel,Hyriopsis myersiana

Summary

The ultrastructure of early stages of the mussel, Hyriopsis (Limnoscapha) myersiana (Lea, 1856), was observed by scanning electron microscopy from the glochidial period until the onset of the juvenile stage 10 days later. Further observations were performed for an additional 13 days to assess juvenile development. Glochidia extracted from the brood chambers have a hookless, semi-oval and equivalve calcareous shell with numerous pores in the internal surface, pits in the external surface and cuticular spines in the ventral region. Keratin fibers with a random arrangement in the cuticle of the glochidial shell were also detected. The appearance of the foot within 10 days of in vitro glochidial culture was considered the main feature of metamorphosis to the juvenile stage. Another change during the following 13 days was the formation of a new periostracum exhibiting growth lines under the old glochidial shell. This development occurs mainly in the anterior region and is followed by hardening of the periostracum matrix by calcium deposition. Periostracum growth gradually became apparent in the lateral and posterior regions at the end of this period. The retraction of spines and the alteration of the external surface of the old shell are also described. It is speculated that transcuticular filaments identified in the juvenile stage may have sensory or metabolic exchange functions. The prominent foot, gradually covered by long dense cilia, shows rhythmical movements which suggest a role in feeding. Similarly, cilia present in the mantle may also be involved in the capture of food, while microvilli may facilitate absorption of dissolved materials. Longer cilia, sparsely distributed in the mantle, may function as chemo- or tactile sensors.     

Evolution of bilaterally asymmetrical larvae in freshwater mussels (Bivalvia: Unionoida: Unionidae)

Bilaterally asymmetrical glochidia (i.e. bivalved parasitic larvae bearing a large marginal appendage on a single valve) have been reported from five Asian freshwater mussel genera belonging to two separate subfamilies, the Gonideinae (i.e. Pseudodon, Solenaia, and Physunio) and Rectidentinae (i.e. Contradens and Trapezoideus). This classification requires that the bilaterally asymmetrical glochidium‐bearing mussels are not monophyletic, and suggests that this atypical larval morphology evolved twice in the same geographic region. Although homoplastic glochidium characters are known (e.g. marginal appendages and size), we hypothesized that bilaterally asymmetrical glochidia represent a novel morphological synapomorphy. We tested the monophyly of the mussels bearing bilaterally asymmetrical glochidia using a molecular matrix consisting of representatives from all six freshwater mussel families and three molecular markers (28S, 16S, and COI). Bayesian inference, maximum likelihood, and ancestral state reconstruction were employed to estimate the phylogeny and larval trait transformations. The reconstructed phylogeny rejects the monophyly of the asymmetrical glochidium‐bearing mussels and resolves two putative origins of asymmetrical glochidia; however, ancestral state reconstruction supports asymmetrical glochidia as a synapomorphy of only one supraspecific taxon of the Rectidentinae. In the Gonideinae, asymmetrical glochidia were autapomorphic of Pseudodon cambodjensis (Petit, 1865). That is, no other taxa resolved among the Gonideinae had bilaterally asymmetrical glochidia, including other Pseudodon species. We describe how the alleged intrageneric glochidial variation in Pseudodon, and in the other genera of the Gonideinae reported to have asymmetrical glochidia (i.e. Solenaia and Physunio), challenge the resolved convergence of asymmetrical glochidia. Our results are discussed in the context of freshwater mussel larval evolution, patterns in life‐history traits, and the classification of freshwater mussels generally. © 2015 The Linnean Society of London     

Investigation of early mussel (Perna canaliculus) development using histology,SEM imaging,immunochemistry and confocal microscopy

A comprehensive study, incorporating histology, light microscopy, scanning electron microscopy, immunochemistry and confocal microscopy, was performed to investigate embryogenesis and larval development of the New Zealand Greenshell? mussel, Perna canaliculus. Detailed observations with this multi-technique approach revealed a gastrula stage at 18 hours post-fertilization, with the appearance of a blastopore, apical sense organ and enclosing vegetal pole. Early D-stage larvae showed limited alimentary organogenesis and clear initiation of a developing nervous system. Shell morphology of D-larvae was characterized by a flat, hinged, pitted–punctate prodissoconch I shell, followed closely by commarginal growth lines within the prodissoconch II shell. Early umbo larvae had a protruding functioning velum, and well-developed posterior adductor and velar retractor muscles. Significant progression in neuronal development occurred just before the umbo stage with noticeable paired cerebral, pedal and visceral ganglia. Shell morphology was characterized by further prodissoconch II secretion with a more rounded umbonate appearance. During the transition through the pediveliger stage, rapid development of the gill rudiment, eye spot and functioning foot was observed with ongoing neuronal development. The first appearance of the dissoconch shell layer took place during this transition, at which point the nervous system was highly distinct with innervations extending throughout muscle regions and between ganglia. This study provides the first comprehensive documentation of the developmental stages of P. canaliculus larvae from fertilization to settlement. The study highlights the advantages of using a combination of techniques to understand larval development and provides crucial information to identify larval performance during larval rearing.     

Molecular characteristics and expression of calmodulin cDNA from the freshwater pearl mussel, Hyriopsis schlegelii

Calmodulin (CaM) is a multifunctional intracellular calcium ion receptor protein that participates in a range of cellular processes, including calcium metabolism in mussels. To investigate the role of CaM in freshwater mollusk shell calcium metabolism, the full-length CaM cDNA was isolated from the freshwater pearl mussel, Hyriopsis schlegelii (referred to as hsCaM) using SMART RACE technology. The full-length hsCaM was 855 bp in size, containing a 70-bp 5'-untranslated sequence, a 447-bp open reading frame, a 309-bp 3'-untranslated sequence, and a 26-nucleotide long poly(A) tail. The hsCaM mRNA expression in different mussel tissues was examined using real-time PCR. The hsCaM mRNA was found to be ubiquitously expressed, but far more abundant in the gill, foot, and mantle than in the posterior adductor muscle. Real-time PCR was also used to determine hsCaM mRNA expression levels in mantle tissues of H. schlegelii at different ages. No significant differences between one-, two-, and three-year-old mussels were detected, but expression increased in four-year-old mussels and then decreased in five-year-old mussels. CaM appears to be involved in calcium regulation of the mantle in four-year-old mussels, which may secrete more mother of pearl during pearl culture.     

Vascular organization of the head and respiratory organs of the air-breathing catfish,Heteropneustes fossilis

Light and scanning electron microscopy of vascular replicas from the facultative air-breathing fish Heteropneustes fossilis show modifications in the macrocirculation of the respiratory organs and systemic circulation, whereas, gill microcirculation is similar to that found in typical water-breathing fish. Three and sometimes four ventral aortae arise directly from the bulbus. The most ventral vessel supplies the first pair of arches. Dorsal to this another aorta supplies the second gill arches, and a third, dorsal to, and larger than the other two, supplies the third and fourth arches and the air sacs. Occasionally a small vessel that may be the remnant of a primitive aortic arch arises from the first ventral aorta and proceeds directly to the mandibular region without perfusing gill tissue. The air sac is perfused by a large-diameter extension of the afferent branchial artery of the fourth gill arch and its circulation is in parallel with the gill arches. Blood drains from the air sac into the fourth arch epibranchial artery. A number of arteries also provide direct communication between the efferent air sac artery and the dorsal aorta. All four gill arches are well developed and contain respiratory (lamellar) and nonrespiratory (interlamellar and nutrient) networks common to gills of water-breathing fish. Air sac lamellae are reduced in size. The outer 30% of the air sac lamellar sinusoids are organized into thoroughfare channels; the remaining vasculature, normally embedded in the air sac parenchyma, is discontinuous. A gill-type interlamellar vasculature is lacking in the air sac circulation. Despite the elaborate development of the ventral aortae, there is little other anatomical evidence to suggest that gill and air sac outflow are separated and that dorsal aortic oxygen tensions are maintained when the gills are in a hypoxic environment. Physiological adjustments to hypoxic water conditions probably include temporal regulation of gill and air sac perfusion to be effective, if indeed they are so.     

Byssal proteins of the freshwater zebra mussel,Dreissena polymorpha

The freshwater zebra mussel (Dreissena polymorpha) is a notorious biofouling organism. It adheres to a variety of substrata underwater by means of a proteinaceous structure called the byssus, which consists of a number of threads with adhesive plaques at the tips. The byssal proteins are difficult to characterize due to extensive cross-linking of 3,4-dihydroxyphenylalanine (DOPA), which renders the mature structure largely resistant to protein extraction and immunolocalization. By inducing secretion of fresh threads and plaques in which cross-linking is minimized, three novel zebra mussel byssal proteins were identified following extraction and separation by gel electrophoresis. Peptide fragment fingerprinting was used to match tryptic digests of several gel bands against a cDNA library of genes expressed uniquely in the mussel foot, the organ which secretes the byssus. This allowed identification of a more complete sequence of Dpfp2 (D. polymorpha foot protein 2), a known DOPA-containing byssal protein, and a partial sequence of Dpfp5, a novel protein with several typical characteristics of mussel adhesive proteins.     

Development of cranial muscles in the actinopterygian fish Senegal bichir,Polypterus senegalus Cuvier, 1829

Polypterus senegalus Cuvier, 1829 is one of the most basal living actinopterygian fish and a member of the Actinopterygii. We analyzed the spatial and temporal pattern of cranial muscle development of P. senegalus using whole‐mount immunostaining and serial sectioning. We described the detailed structure of the external gill muscles which divided into dorsal and ventral parts after yolk exhaustion. The pattern of the division is similar to that of urodeles. We suggest that, the external gill muscles of P. senegalus are involved in spreading and folding of the external gill stem and the branches. The fibers of the external gill muscles appear postero‐lateral to the auditory capsule. In addition, the facial nerve passes through the external gills. Therefore, the external gill muscles are probably derived from the m. constrictor hyoideus dorsalis. In contrast to previous studies, we described the mm. interhyoideus and hyohyoideus fibers as independent components in the yolk‐sac larvae. The m. hyohyoideus fibers appear lateral to the edge of the ventral portion of the external gill muscles, which are probably derived from the m. constrictor hyoideus dorsalis. These findings suggest that the m. hyohyoidues is derived from the m. constrictor hyoideus dorsalis in P. senegalus . In other actinopterygians, the m. hyohyoideus is derived from the m. constrictor hyoideus ventralis; therefore, the homology of the m. hyohyoidues of P. senegalus and other actinopterygians remains unclear. J. Morphol. 278:450–463, 2017. © 2017 Wiley Periodicals, Inc.     

The morphology and evolution of the ventral gill arch skeleton in batoid fishes (Chondrichthyes: Batoidea)

The ventral gill arch skeleton was examined in some representatives of batoid fishes. The homology of the components was elucidated by comparing similarities and differences among the components of the ventral gill arches in chondrichthyans, and attempts were made to justify the homology by giving causal mechanisms of chondrogenesis associated with the ventral gill arch skeleton. The ceratohyal is present in some batoid fishes, and its functional replacement, the pseudohyal, seems incomplete in most groups of batoid fishes, except in stingrays. The medial fusion of the pseudohyal with successive ceratobranchials occurs to varying degrees among stingray groups. The ankylosis between the last two ceratobranchials occurs uniquely in stingrays, and it serves as part of the insertion of the last pair of coracobranchialis muscles. The basihyal is possibly independently lost in electric rays, the stingray genus Urotrygon (except U. daviesi) and pelagic myiiobatoid stingrays. The first hypobranchial is oriented anteriorly or anteromedially, and it varies in shape and size among batoid fishes. It is represented by rami projecting posterolaterally from the basihyal in sawfishes, guitarfishes and skates. It consists of a small piece of cartilage which extends anteromedially from the medial end of the first ccratobranchial in electric rays. It is a large cartilaginous plate in most of stingrays. It is absent in pelagic myliobatoid stingrays. The remaining hypobranchial cartilages also vary in shape and size among batoid fishes. Torpedo and possibly the Jurassic Belemnobalis and Spathobatis possess the generalized or typical chondrichthyan ventral gill arch structure in which the hypobranchials form a Σ-shaped pattern. In the electric ray Hypnos and narkinidid and narcinidid electric rays, the hypobranchial components are oriented longitudinally along the mid-portion of the ventral gill arches. They form a single cartilaginous plate in the narkinidid electric rays, Narcine and Diplobatis. In guitarfishes and skates, the second hypobranchial is unspecialized, and in skates, it does not have a direct contact with the second ceratobranchial. In both groups, the third and fourth hypobranchials are composed of a small cartilage which forms a passage for the afferent branches of the ventral aorta and serve as part of the insertion of the coracobranchialis muscle. In sawfishes and stingrays, the hypobranchials appear to be included in the medial plate. In sawfishes, the second and third components separately chondrify in adults, but the fourth component appears to be fused with the middle medial plate. In stingrays, a large medial plate appears to include the second through to the last hypobranchial and most of the basibranchial copulae. The medial plate probably develops independently in sawfishes and stingrays. Because the last basibranchial copula appears to be a composite of one to two hypobranchials and at least two basibranchial copulae, the medial plate may be formed by several developmental processes of chondrogenesis. More detailed comparative anatomical and developmental studies are needed to unveil morphogenesis and patternings of the ventral gill arch skeleton in batoid fishes.     

Distribution of chloride cells in teleost larvae

Distribution and density of the chloride cells in the newly hatched larvae of teleosts vary depending on species and environmental salinity at hatching. In the euryhaline freshwater ayu (Plecoglossus altivelis), chloride cells are concentrated in the skin posterior to the pectoral fins and gradually decrease in number toward the head and tail. In the stenohaline sea water flounder (Kareius bicoloratus), most chloride cells are localized at the inner membrane of gill chambers and in the skin near the openings of gill chambers, but only a few cells appear in the skin of the yolk sac. In the stenohaline freshwater carp (Cyprinus carpio), only a few small chloride cells are scattered in the body skin. The density and abundance of chloride cells appears to be correlated with the different requirements for osmoregulation in teleost larvae.     

Methods of DNA extraction and PCR amplification for individual freshwater mussel (Bivalvia: Unionidae) glochidia,with the first report of multiple paternity in these organisms

Freshwater mussels (Unionidae) are among North America's most imperilled organisms. Mussels produce small larvae (glochidia) that parasitize aquatic vertebrates. We modified the Epicentre QuickExtract protocol to extract DNA from a single glochidium, collected directly from the marsupium of a female mussel, to use as template in polymerase chain reactions (PCRs). Yield per glochidium in a 40 µL extraction volume provided enough DNA for ≥ 15 PCRs per individual. We were successful in using this DNA for microsatellite analysis of up to three loci per individual. Offspring from one female showed evidence for multiple paternity within her brood. Our results are the first documentation of this phenomenon in freshwater mussels.     

褶纹冠蚌钩介幼虫非寄生变态发育观察及早期稚蚌的生长

为揭示褶纹冠蚌钩介幼虫变态发育特征及过程,采用体外培养方法实现了褶纹冠蚌钩介幼虫的非寄生变态发育。运用光学显微镜和扫描电子显微镜对变态发育过程中幼虫外部形态、内部器官发育进行了系列观察,对非寄生变态发育的稚蚌后期生长发育进行跟踪研究,并分析了底泥和光照两种环境因子对稚蚌存活及生长的影响。结果显示:在整个培养过程中,钩介幼虫的外部形态及大小未出现显著性变化,而斧足、鳃丝、外套膜及内脏团等组织器官逐步形成;在培养第3天,幼虫可见斧足雏形,鳃丝、外套膜纤毛尚未发现;在培养第6天,斧足成形,可见斧足侧沟,外套膜纤毛稀疏,鳃丝出现;培养第9天,斧足纤毛、外套膜纤毛增多,鳃丝密集。稚蚌投喂30d后,鳃丝基本成形。养殖试验结果表明:底泥对稚蚌存活和生长具有显著影响（P < 0.01）,而光照无显著性影响（P>0.05）。该结果为蚌科钩介幼虫变态发育生物学研究积累了基础资料,也通过对稚蚌生长的评估证实了体外培养是蚌类人工繁育及保护的有效技术途径。     

Molecular properties and immune defense of two ferritin subunits from freshwater pearl mussel,Hyriopsis schlegelii

Ferritin is a conserved iron-binding protein involved in cellular iron metabolism and host defense. In the present study, two distinct cDNAs for ferritins in the freshwater pearl mussel Hyriopsis schlegelii were identified (designated as HsFer-1 and HsFer-2) by SMART RACE approach and expressed sequence tag (EST) analysis. The full-length cDNAs of HsFer-1 and HsFer-2 were of 760 and 877 bp, respectively. Both of the two cDNAs contained an open reading frame (ORF) of 522 bp encoding for 174 amino acid residues. Sequence characterization and homology alignment indicated that HsFer-1 and HsFer-2 had higher similarity to H-type subunit of vertebrate ferritins than L-type subunit. Analysis of the HsFer-1 and HsFer-2 untranslated regions (UTR) showed that both of them had an iron response element (IRE) in the 5′-UTR, which was considered to be the binding site for iron regulatory protein (IRP). Quantitative real-time PCR (qPCR) assays were employed to examine the mRNA expression profiles. Under normal physiological conditions, the expression level of both HsFer-1 and HsFer-2 mRNA were the highest in hepatopancreas, moderate in gonad, axe foot, intestine, kidney, heart, gill, adductor muscle and mantle, the lowest in hemocytes. After stimulation with bacteria Aeromonas hydrophila, HsFer-1 mRNA experienced a different degree of increase in the tissues of hepatopancreas, gonad and hemocytes, the peak level was 2.47-fold, 9.59-fold and 1.37-fold, respectively. Comparatively, HsFer-2 showed up-regulation in gonad but down-regulation in hepatopancreas and hemocytes. Varying expression patterns indicate that two types of ferritins in H. schlegelii might play different roles in response to bacterial challenge. Further bacteriostatic analysis showed that both the purified recombinant ferritins inhibited the growth of A. hydrophila to a certain degree. Collectively, our results suggest that HsFer-1 and HsFer-2 are likely to be functional proteins involved in immune defense against bacterial infection.     

The role of host specificity in explaining the invasion success of the freshwater mussel Anodonta woodiana in Europe

Several freshwater mussel species represent some of the most problematic invasive species and have considerably altered ecosystems worldwide. Their invasion potential has been partially attributed to their free-living larvae, which have a high dispersal capability. We investigated the invasion potential of Anodonta (Sinanodonta) woodiana, a species of East Asian unionid mussel established worldwide despite having an obligatory parasitic stage (glochidium), which must encyst on host fish. The invasion success of A. woodiana has been attributed to the success of worldwide introductions of its sympatric fish hosts. We experimentally found, however, that A. woodiana is a broad host generalist, which can complete its development on all eight fish species tested, both coinvasive and native. Subsequently, we used a data on the occurrence and relative abundance of potential hosts in river habitats in the Czech Republic to project scenarios of the effect of host availability on A. woodiana invasion. We found that host availability does not constitute a major limit for A. woodiana to colonise most aquatic habitats in Central Europe. In addition, we investigated seasonal dynamics of A. woodiana reproduction and did not detect any limitations of its reproduction by ambient water temperatures typical of a Central European lowland river. Consequently, we predict that A. woodiana may further increase the speed and range of its invasion and we discuss possible consequences to native habitats and communities, especially to the endangered species of unionid mussels.     

Molecular cloning and characterization of perlucin from the freshwater pearl mussel, Hyriopsis cumingii

Perlucin is an important functional protein that regulates shell and pearl formation. In this study, we cloned the perlucin gene from the freshwater pearl mussel Hyriopsis cumingii, designated as Hcperlucin. The full-length cDNA transcribed from the Hcperlucin gene was 1460 bp long, encoding a putative signal peptide of 20 amino acids and a mature protein of 141 amino acids. The mature Hcperlucin peptide contained six conserved cysteine residues and a carbohydrate recognition domain, similar to other members of the C-type lectin families. In addition, a “QPS” and an invariant “WND” motif near the C-terminal region were also found, which are extremely important for polysaccharide recognition and calcium binding of lectins. The mRNA of Hcperlucin was constitutively expressed in all tested H. cumingii tissues, with the highest expression levels observed in the mantle, adductor, gill and hemocytes. In situ hybridization was used to detect the presence of Hcperlucin mRNA in the mantle, and the result showed that the mRNA was specifically expressed in the epithelial cells of the dorsal mantle pallial, an area known to express genes involved in the biosynthesis of the nacreous layer of the shell. The significant Hcperlucin mRNA expression was detected on day 14 post shell damage and implantation, suggesting that the Hcperlucin might be an important gene in shell nacreous layer and pearl formation. The change of perlucin expression in pearl sac also confirmed that the mantle transplantation results in a new expression pattern of perlucin genes in pearl sac cells that are required for pearl biomineralization. These findings could help better understanding the function of perlucin in the shell and pearl formation.     

Light and scanning electron microscopy of the embryos and glochidia larvae of the Australian freshwater bivalve Hyridella depressa (Hyriidae)

Summary

As is characteristic of freshwater Unionacea, the Australian species Hyridella depressa incubates its young within the demibranchs of its modified gills. The development of H. depressa was documented through light and scanning electron microscopic examination of the marsupial pouch in the inner demibranchs of brooding females. Transition from the gastrula to the glochidial stage was accompanied by a split in the larval integument. The glochidia of H. depressa have subtriangular shell valves each having a blunt tooth at the ventral margin. Each glochidium has a pair of hooks, one on each valve internal to the tooth. These hooks are used to attach to its fish host and are structured to interlock when the valves snap shut. One or two tufts of sensory hairs are located on the internal surface of the mantle dorsal to the hook, while a single tuft, encircled by a collar-like structure, is located centrally. The valves are pitted by pores and have concentric lines along the margins. Due to the similarity and phenotypic plasticity of adult Hyridella species, the morphology of their glochidia has potential for use as a taxonomic tool. In comparison to related species, the glochidia of H. depressa are medium-sized with a mean length of 243 μm and a mean height of 249 μm. The number of glochidia present in the gills was measured to determine the reproductive output of H. depressa. The embryos of an unknown mite, Unionicola sp., form cysts within the gill tissue of H. depressa and the adult mites were observed on the surface of the gills.     

Laboratory evidence suggests glochidia metamorphosis in Sinanodonta japonica (Bivalvia: Unionidae) is supported by gills,but no other tissues of the host Gymnogobius urotaenia (Perciformes: Gobiidae)

We quantitatively assessed the ability of the gills, caudal fin and scales of the floating goby Gymnogobius urotaenia (Hilgendorf, 1879) (Perciformes: Gobiidae) to serve as substrates for the larvae (glochidia) of the freshwater mussel Sinanodonta japonica (Clessin, 1874) (Unionida: Unionidae) by comparing parasitism success and metamorphosis success. We established three experimental treatments with 10 fish per treatment. Twenty glochidia were introduced onto one of the three body parts of each test fish by direct pipette infestation. Glochidia in the gill group had higher parasitism success than those in the fin and scale groups. Juvenile mussels were obtained only in the gill group. We quantitatively assessed the appropriateness of the three body parts as substrates for glochidia on the basis of three indicators: parasitism success; metamorphosis success; and parasitism and metamorphosis success. We conclude from our laboratory experiment that the artificial introduction of S. japonica glochidia onto G. urotaenia gills is a better procedure for obtaining juvenile mussels than the introduction onto fin or scales.     

Evaluation of potential health risks to Eastern Elliptio (Elliptio complanata) (Mollusca: Bivalvia: Unionida: Unionidae) and implications for sympatric endangered freshwater mussel species

Conservation efforts for freshwater musselspecies require identification and evaluationof potential health risks to populations. Sampling large numbers of individual mussels,however, could damage the small, fragile,extant populations of imperiled mussel species. In this study, a small sample size was designedto reduce lethal sampling, costs, andenvironmental disruption, while still allowing95% confidence in detecting health risks of25% or greater prevalence in the population. Health assessments were conducted on twentyspecimens of the Eastern Elliptio, Elliptio complanata, collected from two NorthCarolina sites as part of a survey to evaluatepotential disease threats to mussels in theregion. Bacteriological sampling of thegastrointestinal tracts yielded 18 aerobicbacterial species, of which Aeromonashydrophila (55.0%), Enterobacter spp.(40.0%), and Bacillus spp.(30.0%) were predominant. Histologicallesions of internal organs included mild tomoderate digestive gland atrophy andinflammation in one mussel, and mild tomoderate parasitism in several individuals. Adistinct difference in parasite prevalence wasevident between infections in E.complanata from the two collection sites. Thetrematode metacercaria of Homalometronarmatum and what appears to be three gillciliate species, the most abundant beingtentatively identified as the scyphidiidperitrich Mantoscyphidia sp., were foundin mussels from one site only. This studydemonstrates a comprehensive diagnosticapproach incorporating multiple modalities toassess the health status of mussel populations,while minimizing the sample size required toobtain valuable information. Furthermore, thisstudy provides baseline health data of E.complanata at two sites in south-central NorthCarolina and suggests the potential usefulnessof E. complanata as an environmentalbioindicator of health risks to sympatricthreatened freshwater mussel populations.