Histological techniques for marine bivalve mollusks and crustaceans, 2nd edition

Investigators at the Cooperative Oxford Laboratory (COL) diagnose and study crustaceans, mollusks, finfish, and a variety of other marine and estuarine invertebrates to assess animal health. This edition updates the Histological Techniques for Marine Bivalve Mollusks manual by Howard and Smith (1983) with additional chapters on molluscan and crustacean techniques. The new edition is intended to serve as a guide for histological processing of shellfish, principally bivalve mollusks and crustaceans. Basically, the techniques included are applicable for histopathological preparation of all marine animals, recognizing however that initial necropsy is unique to each species. Photographs and illustrations are provided for instruction on necropsy of different species to simplify the processing of tissues. Several of the procedures described are adaptations developed by the COL staff. They represent techniques based on principles established for the histopathologic study of mammalian and other vertebrate tissues, but modified for marine and aquatic invertebrates. Although the manual attempts to provide adequate information on techniques, it is also intended to serve as a useful reference source to those interested in the pathology of marine animals. General references and recommended reading listed in the back of the manual will provide histological information on species not addressed in the text.

Disease Risks Associated with Importation of Nonindigenous Marine Animals

Transfers and introductions of marine species have occurred and are occurring on a worldwide basis, largely in response to perceived needs of expanding aquaculture industries. Greatest interest is in salmon (cage rearing and ocean ranching), shrimp, and bivalve mollusks, although other organisms are being considered. Such movements of animals carry an associated risk of moving pathogens into areas where they did not occur previously, possibly resulting in infections in native species. Many case histories of the effects of introduced pathogens and parasites now exist-enough to suggest that national and international action is necessary. Viral pathogens of shrimp and salmon, as well as protozoan parasites of mollusks and nematode parasites of eels, have entered complex "transfer networks" developed by humans, and have been transported globally with their hosts in several well-documented instances. Examining the records of transfers and introductions of marine species, incomplete as they are, permits the statement of emerging principles-foremost of which is that severe disease outbreaks can result from inadequately controlled or uncontrolled movements of marine animals.

Trace metals in common marine foods of the Pacific Coast

Trace metal analysis of 23 species of common Pacific Coast marine foods revealed high cadmium values for Bent-nosed clams (Macoma nasuta), Bay mussels (Mytilus edulis), Bay oysters (Osrtrea lurida), Pacific oysters (Crassostrea gigas), and Littleneck clams (Protothaca staminea). Metals were found to concentrate in the gills, heart, and visceral mass of all 10 species of bivalve mollusks examined. Swordfish (Xiphias gladius) and Salmon (Oncorhynchus tshawytscha) demonstrated the highest cadmium values for fish flesh.

National Status and Trends Bioeffects Program: field methods

Environmental quality indicators provide resource managers with information useful to assess coastal condition and scientifically defensible decisions. Since 1984, the National Oceanic and Atmospheric Administration (NOAA), through its National Status and Trends (NS&T) Program, has provided environmental monitoring data on chemical, physical, and biological indicators of coastal environments. The program has two major monitoring components to meet its goals. The Bioeffects Assessments Program evaluates the health of bays, estuaries, and the coastal zone around the nation using the Sediment Quality Triad technique that includes measuring sediment contaminant concentrations, sediment toxicity and benthic community structure. The Mussel Watch Program is responsible for temporal coastal monitoring of contaminant concentrations by quantifying chemicals in bivalve mollusks. The NS&T Program is committed to providing the highest quality data to meet its statutory and scientific responsibilities. Data, metadata and information products are managed within the guidance protocols and standards set forth by NOAA’s Integrated Ocean Observing System (IOOS) and the National Monitoring Network, as recommended by the 2004 Ocean Action Plan. Thus, to meet these data requirements, quality assurance protocols have been an integral part of the NS&T Program since its inception. Documentation of sampling and analytical methods is an essential part of quality assurance practices. A step-by–step summary of the Bioeffects Program’s field standard operation procedures (SOP) are presented in this manual.

Chemical ecology fatty acids of mollusks dominant species in intertidal zone of Chabahar Bay

Chemical ecology is the science of study and analysis of natural chemical products in result of biochemical processes in organisms and their reactions to variations of ecological and environmental parameters. In marine chemical ecology the existence of natural products in aquatic organisms and their ecological roles in marine animals and their reactions to environmental parameters variations will be studied. Among them, fatty acids are the most various and abundant ones in natural products which had been extracted from many marine organisms such as mollusks and algae. In this study selected animals were the dominant species of mollusks in intertidal zone of chabahar bay including gastropods, bivalves and polyplacophora classes. Nerita textilis and Turbo coronatus species are among gastropoda, Saccostrea cucullata is from bivalve, and Chiton lamyi is from polyplacophora. After seasonal sampling, separation and identification of natural products of these species, fatty acids had been isolated and identified by GC mass chromatography and their seasonal variations had been identified. In addition environmental factors of the location including pH, salinity temperature, dissolved oxygen, chlorophyll a and nutrients were measured monthly. Then the effect of seasonal variations of environmental factors on fatty acids had been studied by applying statistical analysis. GC/MS resulted thirteen fatty acids, which the most importants were myristic, stearic, oleic, palmitoleic, arachidonic and eicosapentaenoic acids. In majority of species palmitic acid was most abundant than the others and saturatedes had the most percentage levels than unsaturated ones. Although seasonal variations of identified fatty acids was not similar in species, but the majority of unsaturated ones had their maximum during winter, while saturated acids reached their maximum in summer. Statistical Analysis showed the strong correlations between Environmental factors and some fatty acids and temperature, nitrate, silicate and pH had strong correlations in all species. The species was studied from the point of lipid content and the results showed a good quality of lipid content in the selected species in the intertidal zone of Chabahar bay.

Baseline aquatic faunal survey of Avon Park Air Force Range, Florida: fishes, mollusks, and crayfishes

This report presents results of the first systematic study of the diversity and distribution of fishes and mussels in Avon Park Air Force Range (APR). We also provide information on crayfishes and aquatic snails taken during our fish and mussel sampling activities. Our surveys documented the presence of 46 species of fishes (43 native and 3 nonindigenous), 9 species of mussels (including 8 native and 1 nonindigenous species), 5 species of aquatic snails, and two crayfish species. (347 page document)

Mollusks taken by Beam Trawl in the vicinity of Gray's Reef National Marine Sanctuary on the Continental Shelf off Georgia, Southeastern U.S.

Mollusks were sorted from samples of shell hash (obtained as bycatch during NOAA-sponsored studies of larval and juvenile fish distribution), and analyzed to gain qualitative insights on species composition, distribution and habitat affinities of the molluscan fauna on the continental shelf off Georgia. Samples came from beam trawls at 37 stations located in the immediate vicinity and offshore of the Gray’s Reef National Marine Sanctuary (GRNMS) at depths of 4.9 to 103 m. Two hundred sixty-three (263) taxa of mollusks (~58% as dead shells only) were collected, and nearly all (~99%) were identified to the species level. Ninety-seven of these taxa appeared in samples from one or more of the four stations established near the corners of the GRNMS. Samples were highly variable in terms of appearance, volume and species composition of mollusks, reflecting the extreme patchiness of benthic habitats within this region of the continental shelf. With very few exceptions, the mollusks were generally characteristic of either the Carolinian or Caribbean faunal provinces. The Georgia continental shelf, however, was outside the previously reported ranges for at least 16 of the species reported here. Most of these extralimital species were known previously from the East Coast of Florida, and represented northerly range extensions of 1-5° Latitude (110-560 km). One species represented a more significant range extension from the Bahamas and the southern Caribbean, and two represented southerly range extensions, known previously from only as close as off North Carolina. The high incidence of range extensions found in this study and the potential for discovery of additional species are discussed in the context of the diversity and patchiness of benthic habitats on the continental shelf of the region, and the sensitivity of species recruitment to variability in Gulf Stream patterns and global climate change. (PDF contains 52 pages)

Farming experiments and transfer of technology of bivalve culture along the southwest coast of India

The Central Marine Fisheries Research Institute (CMFRI) in India developed bivalve farming technologies in the 1970s, but these were not widely adopted at the time. In 1993, CMFRI undertook an action research program to encourage farming of edible oysters (Crassostrea madrasensis), mussels (Perna viridis and Perna indica), clams (Paphia malabarica) and pearls (Pinctada fucata) along the southwest coast of India. Successful demonstration of the viability of bivalve farming led to the initiation of commercial farming of mussels and generated interest among farmers and entrepreneurs in developing production of pearls and farming of edible oysters. Given the high potential for the mollusc aquaculture, both for the local and export market, issues such as demarcation and issuance of lease right on aquaculture zones in public waterbodies by the government, organization of marketing systems and provisions for technical and financial support to farmers need to be addressed.

Histological techniques for marine bivalve molluscs: update

This chapter describes the procedures for determining the reproductive stage of oysters, mytilid mussels, and dreissenid mussels collected for NOAA’s National Status and Trends Mussel Watch Project. Analyses are conducted on paraffin-embedded tissues sectioned at a 5-μm thickness and stained using a pentachrome staining procedure. Each slide is examined microscopically to determine the animal’s sex and stage of gonadal development. A semi-quantitative ranking is assigned.

An environmentally based growth model that uses finite difference calculus with maximum likelihood method: its application to the brackish water bivalve Corbicula japonica in Lake Abashiri, Japan

We present a growth analysis model that combines large amounts of environmental data with limited amounts of biological data and apply it to Corbicula japonica. The model uses the maximum-likelihood method with the Akaike information criterion, which provides an objective criterion for model selection. An adequate distribution for describing a single cohort is selected from available probability density functions, which are expressed by location and scale parameters. Daily relative increase rates of the location parameter are expressed by a multivariate logistic function with environmental factors for each day and categorical variables indicating animal ages as independent variables. Daily relative increase rates of the scale parameter are expressed by an equation describing the relationship with the daily relative increase rate of the location parameter. Corbicula japonica grows to a modal shell length of 0.7 mm during the first year in Lake Abashiri. Compared with the attain-able maximum size of about 30 mm, the growth of juveniles is extremely slow because their growth is less susceptible to environmental factors until the second winter. The extremely slow growth in Lake Abashiri could be a geographical genetic variation within C. japonica.

Characterisation of neurons in the pedal ganglia of the green-lipped mussel, Perna canaliculus, using antibodies raised against neuropeptides and neurotransmitters involved in gastropod egg-laying behaviour and bivalve reproduction and spawning

To characterise central neurons in the pedal ganglia of both male and female green lipped mussel, Perna canaliculus immunohistochemical techniques were used. Mollusc antibodies were used against neuropeptides and neurotansmitters known to control reproduction and spawning. Anti-ELH and anti-APGWamide showed very strong immunoreactivity in small type of neurons. Anti-5-HT and anti-DA immunoreactivity was mostly in large type of neurons. The labelled neurons are consistent with descriptions of neurosecretory cells implicated in the control of reproduction and spawning on the basis of earlier histological staining techniques used in this species. The use of selective immunological markers for peptides and amines appears to be a, promising tool for further characterisation of neurosecretory cells, and to isolate an'tl characterise neuropeptides and other biologically active materials involved in the control of reproduction in Perna canaliculus.

Neurons characterization in the cerebral ganglia of the green-lipped mussel, Perna canaliculus, using antibodies raised against neuropeptides and neurotransmitters involved in gastropod egg-laying behaviour and bivalve reproduction and spawning

Immunohistochemical techniques were used to characterise central neurons in the cerebral ganglia of both male and female Perna canaliculus. We used mollusc antibodies raised against neuropeptides and neurotransmitters known to control reproduction and spawning. Anti-ELH and anti-APGWamide showed very strong immunoreactivity in small type of neurons. Anti-5-HT and anti-DA immunoreactivity was mostly in large type of neurons. The labelled neurons are consistent with descriptions of neurosecretory cells implicated in the control of reproduction and spawning on the basis of earlier histological staining techniques used in this species. The use of selective immunological markers for peptides and amines appears to be a promising tool for further characterisation of neurosecretory cells, and to isolate and characterise neuropeptides and other biologically active materials involved in the control of reproduction in Perna canaliculus.

Agiis: a bivalve cultivated as live shrimp feed

Live 'agiis' have been proven to be a good feed for shrimp for the past ten years by polyculture fish farmers in the province of Capiz, in the island of Panay, west central Philippines. A brief account is given of culture and feeding operations. Its cultivation period is short and seeds are readily available. It can reduce dependence on trash fish which is now getting to be scarce; it also seems much cheaper. Perhaps this fast-growing tiny bivalve can be scientifically investigated by students of aquaculture as feed for other commercial aquaculture species. Its scientific identification can be a good start.

Sea otter predation and the distribution of bivalve prey in the Elkhorn Slough National Estuarine Research Reserve

The California sea otter population is gradually expanding in size and geographic range and is consequently invading new feeding grounds, including bays and estuaries that are home to extensive populations of bivalve prey. One such area is the Elkhorn Slough, where otters have apparently established a spring and summer communal feeding and resting area. In anticipation of future otter foraging in the slough, an extensive baseline database on bivalve densities, size distributions, biomasses, and burrow depths has been established for three potential bivalve prey species, Saxidomus nuttalli, Tresus nutallii, and Zirphaea pilsbryi. In 1986, the Elkhorn Slough otters were foraging predominately at two areas immediately east and west of the Highway 1 bridge (Skipper's and the PG&E Outfall). Extensive subtidal populations of Saxidomus nuttalli and Tresus nuttallii occur in these areas. Shell records collected at these study areas indicated that sea otters were foraging selectively on Saxidomus over Tresus. The reason for this apparent preference was not clear. At the Skipper's study site, 51% of the shell record was composed of Saxidomus, yet this species accounted for only 16% of the in situ biomass, and only 39% of the available clams. Tresus represented 49% of the shell record at Skipper's, yet this species accounted for 84% of the in situ biomass and 61% of the available clams. There was no difference in mean burrow depth between the two species at this site so availability does not explain the disparity in consumption. At the PG&E Outfall, Saxidomus represents 66% of the in situ biomass and 81% of the available clams, while Tresus accounts for 34% of the in situ biomass and 19% of the available clams. Saxidomus accounts for 96% of the shell record at this site vs. 4% for Tresus, again indicating that the otters were preying on Saxidomus out of proportion to their density or biomass. High densities and biomasses of a third species, Zirphaea pilsbryi, occur in areas where sea otters were observed to be foraging, yet no cast-off Zirphaea shells were found. Although it is possible this species was not represented in the shell record because the otters were simply chewing up the shells, it is more likely this species is avoided by sea otters. There were relatively few sea otters in the Elkhorn Slough in 1986 compared to the previous two years. This, coupled with high bivalve densities, precluded any quantitative comparison of bivalve densities before and after the 1986 sea otter occupation. Qualitative observations made during the course of this study, and quantitative observations from previous studies indicate that, after 3 years, sea otters are not yet significantly affecting bivalve densities in the Elkhorn Slough.