Bioassay responses of micro-organisms to sediment elutriates from the St. Lawrence River (Lake St. Louis)

A sediment study, involving both chemical and biological analyses, was carried out in the St. Lawrence River near Montreal (Lake St. Louis). About 60 stations were sampled during 1984–85, and the sediments were analyzed for support variables, heavy metals, and organochlorinated compounds. Subsamples were elutriated using a 1 to 4 sediment/water ratio. The resulting elutriates were analyzed for several chemical variables, while toxicity was measured using the Microtox test, algal ¹⁴C assimilation, and lethality/developmental inhibition in cladocerans, rotifers, and nematodes. The results showed a great variety of responses and sensitivity, and correlations between the tests were non-significant. In terms of toxic responses, the algal and Microtox tests were the most sensitive. Toxic responses could not be explained in simple terms of contaminant concentrations. Therefore, the chemistry of elutriates is not predictive of the toxic potential of contaminated sediments. Biotests can give an insight into the hazard assessment of sediments, but no single test will be sufficient; the use of a battery of standardized biotests, representing different levels of organization/food chain, including representative natural species, is highly recommended.     

Heterogeneity in levels of serum neutralizing antibodies against viral hemorrhagic septicemia virus genotype IVB among fish species in Lake St. Clair, Michigan, USA

The presence of neutralizing antibodies against viral hemorrhagic septicemia virus (VHSV-IVb) was investigated in sera of 13 fish species collected from Lake St. Clair, Michigan, USA, a VHSV-endemic water body. We tested 297 sera collected May 2004-June of 2010, using a complement-dependent 50% plaque neutralization test (50% PNT). Neutralizing antibodies were detected in 23% (67/297) of the samples. The highest overall antibody prevalence (85%, 34/40) and mean positive antibody titer (12,113 ± 11,699 SD) were detected in muskellunge (Esox masquinongy). Antibodies were also detected in 50% (15/30) of sampled northern pike (E. lucius), 25% (15/61) of freshwater drum (Aplodinotus grunniens), and 7% (3/41) of smallmouth bass (Micropterus dolomieu). All sera from channel catfish (Ictalurus punctatus), lake sturgeon (Acipenser fulvescens), quillback (Carpiodes cyprinus), rock bass (Ambloplites rupestris), shorthead redhorse (Moxostoma macrolepidotum), silver redhorse (M. anisurum), walleye (Sander vitreus), white perch (Morone americana), and yellow perch (Perca flavescens) were negative. Antibodies in one or more fish species were detected in all sampling years (2004, 2006, 2007, 2009, and 2010), whereas in parallel sampling periods, VHS virus was detected only in 2006 and 2009. Our results suggest the continued presence of VHSV-IVb in the Lake St. Clair ecosystem, and underscore the importance of assessing immune responses of fish populations to determine prior virus exposure.     

Lake sturgeon spawning on artificial habitat in the St Lawrence River

In 1996, lake sturgeon (Acipenser fulvescens) spawning was documented for the third consecutive year on an artificially placed gravel bed in the St Lawrence River. Two distinct spawning periods were observed in 1996. Spawning initially commenced on 17 June, when water temperature reached 15°C. A second spawning event was documented from 28 June to 1 July (16°C). Sturgeon egg densities were monitored in three transects on egg trays, on the gravel surface, and within interstitial spaces in the gravel. Counts of developing eggs in the gravel bed during both spawning periods were used to estimate a total of 275 000 eggs on the study area (0.075 ha). Average egg density was highest in the transect with the highest water velocities. Lake sturgeon fry were first observed in the gravel on 24 June (15.5°C), and first emergence from the gravel was documented on 28 June. Hatching following the second spawning event commenced on 3 July. Based on assessment of average embryo viability (61.6%) and egg‐to‐emergent fry survival (17.6%) an estimate of about 171 000 sturgeon eggs hatched, producing over 49 000 emergent fry. Current velocity, substrate particle size, depth of substrate, and maintenance of sediment‐free interstitial spaces are important considerations in planning future spawning habitat enhancement projects.     

Distribution and abundance of young fish in the St. Clair River and associated waters,Ontario

Fish larvae were sampled in 1986 in the St. Clair River, and adjacent waters. Species richness (9 taxa as larvae; 4 others as juveniles) and abundance was lowest in the river, where many larvae (e.g., burbot, rainbow smelt, and yellow perch) were in transit from Lake Huron. The most abundant, and localized, species was gizzard shad, which reached a peak mean density of 4600 larvae 100 m^-3 in an agricultural canal. Adjacent waters contribute greatly to the fish communities of the river and adjoining Lakes Huron and Erie, especially in terms of the number and quantity of forage species.     

Vascular plant species richness and rarity across a minerotrophic gradient in wetlands of St. Lawrence County,New York,USA

Thirteen wetlands in St. Lawrence County, NY were sampled to examine the effect of a minerotrophic gradient on vascular plant species richness and rarity. Wetlands ranged from organic soil based poor fens (average conductivity 46.40 microsemens, average Ca 3.55 ppm) to mineral soil based rich fens (average conductivity 342.10 microsemens, Ca 23.00 ppm). Vascular plant species richness was sampled during 1990 in randomly located 1.0 m² quadrats. Specific conductivity, presence or absence of hummocks, and water depth predicted 62% of the variation in richness. Richness increased as conductivity increased until 413 microsemens at which a down trend became obvious. The negative curvilinear relation between conductivity and richness is in accordance with the hump-backed model of Grime but occurs at high rather than intermediate conductivity values. State-listed rare species were found in species-rich wetlands only and had a mean associated richness value of 14.50 species m^-2. This relationship should be taken into consideration when selecting wetlands for protection or managing wetlands for maximum plant diversity.     

Two viruses isolated from rodents (Clethrionomys gapperi and Microtus pennsvlvanicus) trapped in St. Lawrence County, New York

Four strains of C. gapperi virus were isolated from 3 Clethrionomys gapperi and 47 strains of Microtus virus from 15 Microtus pennsylvanicus and 1 Mus musculus. One of the Microtus strains was isolated from a pool of 20 mites while the others were from rodent tissues. These agehts were insensitive to ether and sodium desoxycholate, withstood freezing at -70 C for 3 years and lyophilization without loss of titer, and were not killed when heated at 60 C for 1 hour. Their size as determined by filtration was less than 50 mg and greater than 20-35 mmicro. The strains within each group appear to be similar. The illness induced in suckling mice by the C. gapperi agents had a 5-day incubation period followed by prostration and death with a histologic picture of extensive encephalomalacia. The incubation period in mice for the Microtus agents was 9 to 11 days followed by convulsions and death. Histopathology showed meningeal infiltration and necrosis of the molecular layer. No antigenic similarity was detected between the C. gapperi and Microtus viruses by cross complement-fixation test.     

Spatiotemporal and teratological analyses of diatom assemblages from sediments contaminated with industrial effluents in the St. Lawrence River near Cornwall (Ontario,Canada)

Hydrobiologia - Throughout the twentieth century, waterfront industries in Cornwall (Ontario, Canada) discharged significant quantities of mercury (Hg) and other industrial effluents to the St....     

Historical changes in herbaceous wetland distribution induced by hydrological conditions in Lake Saint-Pierre (St. Lawrence River,Quebec, Canada)

Historical changes (1961–2002) in the distribution of herbaceous wetland plant associations were inferred from the hydrological regime of Lake Saint-Pierre, a 312 km² broadening of the St. Lawrence River (Quebec, Canada), to assess the cumulative effects of human interventions and climatic variability. Relative abundance index (height × percent cover) of wetland plants in 630 field quadrats sampled at 13 sites (1999–2002) were used to derive a model predicting the occurrence of nine herbaceous plant classes with a 71% (24–84%) accuracy. Wetland types included seasonally dry (meadows), mudflats and wet (low marshes and submerged) assemblages. Over the 1961–2002 period, the total surface area of Lake Saint-Pierre herbaceous wetlands ranged between 11 (in 1972) and 128 (in 2001) km² and was negatively correlated (Spearman r = –0.86, p < 0.0001) to average water level during the current growing season. Within-season variability and level conditions over the previous season defined 5 marsh assemblages characterized by different species composition, relative abundance and diversity. Significant hydrological variables included quadrat elevation, water depth, number of days flooded and depth variability experienced over the current and/or previous growth seasons. The hydrological model suggests that for a given level, wetland plant assemblages differed markedly whether the multi-year sequence of water levels was rising or falling. Lake Saint-Pierre alternated between three broad-scale wetland configurations, dominated by meadows and open marsh with floating-leaved vegetation (in the 1960s), scattered tall Scirpus marshes (in the 1970s and early 1980s) and closed marsh with aggressive emergents (since 1996). The strong response of Lake Saint-Pierre wetlands to hydrological conditions in the current and previous growth seasons underlines their vulnerability to future water level variations resulting from regulation and climate variability.     

Factors affecting the structure of epiphytic gastropod communities in the St. Lawrence River (Quebec,Canada)

A study of epiphytic gastropods associated with two submerged macrophytes (Myriophyllum spicatum and Vallisneria americana) was conducted at a soft and at a hard water site in the St. Lawrence River during two non-consecutive years in order to compare effects of macrophyte species and biomass, site and year in gastropod community structure. There was no effect of macrophyte species on total gastropod abundance nor on gastropod diversity, and few gastropod species showed a marked preference for either macrophyte species. Inter-site and inter-annual variations in diversity, total gastropod abundance and gastropod community structure were greater than variations among macrophytes. However, analysis of variations of epiphytic gastropod communities in one of the two sites shows that abiotic factors are important in explaining epiphytic gastropod distribution. Our results and results of other studies on gastropod population dynamics in the St. Lawrence River indicate that intra- and interspecific competition between gastropods is important on macrophytes and that they must be carefully considered in order to understand epiphytic community structure and population dynamics.     

Experimental horizontal transmission of viral hemorrhagic septicemia virus (VHSV) in Japanese flounder Paralichthys olivaceus

Infection by viral hemorrhagic septicemia virus (VHSV) has recently occurred among wild and farmed Japanese flounder Paralichthys olivaceus in Japan. In the present study, horizontal transmission of VHSV among Japanese flounder was experimentally demonstrated by immersion challenge. Exposure to a flounder isolate (Obama25) of VHSV revealed a dose-response, with higher mortality (81 and 70%) at the 2 higher exposure levels (6.0 and 4.0 log10 TCID50 ml(-1)). In a second experiment, high titers of VHSV were expressed from moribund and dead flounder based on virus detection in holding-tank waters 2 to 3 d prior to death of the fish and 1 d after death. The virus could not be detected in tank waters 2 d after death. Finally, a third cohabitation experiment in small tanks demonstrated horizontal transmission of VHSV from experimentally infected to uninfected fish.     

Generation and characterization of NV gene-knockout recombinant viral hemorrhagic septicemia virus (VHSV) genotype IVa

A recombinant viral hemorrhagic septicemia virus (rVHSV-deltaNV-EGFP) containing the enhanced green fluorescent protein (EGFP) gene instead of the NV gene was produced using the reverse-genetics method. For use as a positive control, another recombinant virus (rVHSV-wild) was also generated, which had an identical nucleotide sequence to the wild-type VHSV genome except for a few artificially replaced nucleotides. The rVHSVs were rescued using a system controlled by T7 RNA polymerase supplied by a retroviral vector. Generation of rVHSV-deltaNV-EGFP and rVHSV-wild was confirmed by sequencing of RT-PCR products, and rescue of infectious rVHSVs was confirmed by observation of plaque formation. Replication efficiency of rVHSV-wild was distinctly lower than that of wild-type VHSV, suggesting that the artificially replaced nucleotides, especially when immediately preceding the G or NV gene start codons, might affect the replication of the virus. Replication of rVHSV-deltaNV-EGFP was slightly lower than that of rVHSV-wild when epithelioma papulosum cyprini cells were infected with multiplicity of infection (MOI) 1.0, but much lower when cells were infected with MOI 0.00001. These results suggest that the NV gene plays an important role in VHSV replication through interactions with host-cell responses, and the lower replication ability of rVHSV-wild compared to wild-type VHSV might be caused by replaced nucleotides just before the NV gene open reading frame (ORF) rather than the G gene ORF. In olive flounder Paralichthys olivaceus, rVHSV-wild produced slower-progressing mortalities than wild-type VHSV, whereas rVHSV-deltaNV-EGFP pathogenesis was highly attenuated. These results suggest that the NV protein of VHSV may play an important role not only in viral replication but also in viral pathogenesis.     

An outbreak of VHSV (viral hemorrhagic septicemia virus) infection in farmed Japanese flounder Paralichthys olivaceus in Japan.

A rhabdoviral disease occurred in farmed populations of market sized Japanese flounder (hirame) Paralichthys olivaceus in the Seto Inland Sea of Japan in 1996. The causative agent was identified as viral hemorrhagic septicemia virus (VHSV) based on morphological, immunological, and genetic analyses. Diseased fish that were artificially injected with a representative virus isolate showed the same pathological signs and high mortality as observed in the natural outbreak. This is the first report of an outbreak of VHSV infection in cultured fish in Japan. Clinical signs of diseased fish included dark body coloration, an expanded abdomen due to ascites, congested liver, splenomegaly, and a swollen kidney. Myocardial necrosis was most prominent and accompanied by inflammatory reactions. Necrotic lesions also occurred in the liver, spleen and hematopoietic tissue, and were accompanied by circulatory disturbances due to cardiac failure. Hemorrhagic lesions did not always appear in the lateral musculature. Transmission electron microscopy revealed many rhabdovirus particles and associated inclusion bodies containing nucleocapsids in the necrotized myocardium. The histopathological findings indicated that the necrotizing myocarditis could be considered a pathognomonic sign of VHSV infection in Japanese flounder.     

Immune response of heterologous recombinant antigenic protein of viral hemorrhagic septicemia virus (VHSV) in mice

Viral hemorrhagic septicemia (VHS) is an important infectious disease in fish worldwide caused by viral hemorrhagic septicemia virus (VHSV). VHSV is the causative agent of serious systemic diseases in fish, affecting a number of teleost fish species. In this study, VHSV glycoprotein (G), including its epitope, as a subunit vaccine candidate, was expressed in tobacco plant (Nicotiana tabacum). The recombinant gene, VHSVG, was fused to the immunoglobulin Fc fragment and extended with the endoplasmic reticulum (ER) retention signal (KDEL) to generate VHSVG-FcK. The recombinant expression vector for VHSVG-FcK was transferred into Agrobacterium tumefaciens (LBA4404), and plant transformation was conducted N. tabacum. Polymerase chain reaction (PCR) was performed to confirm gene insertion and VHSVG-FcK protein expression was confirmed by immunoblot analysis. VHSVG-FcK protein was successfully purified from tobacco plant leaves. Furthermore, ELISA analysis showed that mice serum immunized with the plant-derived VHSVG-FcK (VHSVGP-FcK) had a high absorbance against VHSVG-FcK, indicating that the plant-derived recombinant subunit vaccine protein VHSVG-FcK can induce immune response. Taken together, this recombinant vaccine protein can be expressed in plant expression systems and can be appropriately assembled to be functional in immunogenicity.     

Diet composition of the smooth dogfish (Mustelus canis) in the waters of Long Island,New York,USA

Mustelus canis, one of two species in the genus Mustelus found along the Northwest Atlantic coast and the second most abundant shark in the region, is beginning to comprise large portions of by-catch in various commercial fishing industries. This study analysed the gut contents of sharks taken from three locations in Long Island, NY (Long Beach, Shelter Island and Gardiners Bay) to examine sex- and site-specific diet composition. Our study indicates that smooth dogfish in these waters are opportunistic predators, showing a preference for crustaceans (i.e. Cancer irroratus, Libinia spp., Squilla empusa and Upogebia affinis). Dietary niche breadth was found to vary significantly and niche overlap was moderately low between sexes and between sites. While dietary components of M. canis discovered in this study corroborate other studies performed within the Northwestern Atlantic migratory range of M. canis, smooth dogfish diet varies considerably with region and time of year. Thus, future studies should include surveys of distribution patterns for common prey items and studies of caloric components of these prey items.     

Land-use proximity as a basis for assessing stream water quality in New York State (USA)

The influence of the proximity of urbanization and agriculture to stream water quality is often difficult to quantify. The objectives of this study were to (1) compare the influence of far-field land-use, encompassing a watershed drainage area, to a near-field, 200-m buffer on each side of the stream in an attempt to determine on which zone of influence land-use has the largest impact on water quality, and (2) incorporate the EPA's Rapid Habitat Assessment Protocol (Barbour et al., 1999) to characterize the riparian and channel characteristics of a stream that influence water quality, which can improve New York State's monitoring protocols. Impacts were assessed through biological, chemical, and physical-habitat data from 29 streams located within a variety of land-use categories. Land-use was identified through USGS National Land Cover Data (NLCD). Principal components analysis (PCA) indicated that land-use and water quality variables were associated with non-point source contaminants (e.g. nutrients and specific conductance). Using Spearman's rank correlation coefficient, significant relationships between all three land-use types and stream water quality were determined at the 200-m buffer zone of influence. At the watershed zone of influence, water quality indicators did not correlate significantly with land cover type. DO and BAP values within the 200-m buffer zone varied inversely with the percentage of urban-land cover. The stronger correlation between land cover and stream water quality at the 200-m proximity than that of the watershed suggests that the presence of a riparian buffer zone between streams and agricultural and urban areas is a significant factor in reducing contamination from non-point source loading.     

Modeling the secondary spread of viral hemorrhagic septicemia virus (VHSV) by commercial shipping in the Laurentian Great Lakes

Researchers have only begun to study the role of shipping in the spread of invasive species in the Laurentian Great Lakes despite a well-documented history of introductions in these lakes due to ballast water release. Here, we determine whether ballast water discharge was a likely vector of spread of the fish disease, viral hemorrhagic septicemia virus genotype IVb (VHSV-IVb), throughout the Great Lakes and St. Lawrence Seaway. Three models were developed to assess whether the spread of VHSV was due to (1) chance (random model), or (2) ballast water discharge (location model), and whether (3) increased propagule pressure, as measured by the number of visitations by ships carrying ballast water from VHSV infected areas, increased the likelihood of a discharge location becoming infected with VHSV (propagule pressure model). The third model was also used to assess the probable point of initial introduction of VHSV. Presence and absence accuracies and weighted Cohen’s kappa were calculated to determine which models best predicted observed presences and absences of VHSV. Location models explain the patterns of VHSV detections better than random models, and inclusion of “propagule pressure” often improved model fit; however, the relationship is weak likely because of a long lag time between introduction and detection, a high rate of false negatives in reporting, and the possible contribution of other vectors of spread. Montreal was also identified as the more likely introduction site of VHSV, rather than Lake St. Clair, the site where the virus was first detected.     

Integrative ecosystem analyses of phytoplankton dynamics in the York River estuary (USA)

The relative importance of biotic (top-down) vs. abiotic (bottom-up) controls on phytoplankton dynamics was investigated in the York River estuary, Virginia (USA) by a combination of extensive analyses of long-term data sets collected by the U.S. Environmental Protection Agency (EPA) monitoring program over 17 years (1984–2001), field studies (1996–1997) and ecosystem modeling analyses. Results from the analysis of long-term data records collected at three stations along the salinity gradient suggested that phytoplankton are more likely controlled by abiotic mechanisms such as resource limitation than biotic mechanisms such as grazing since annual cycles of primary production and phytoplankton biomass were similar and no grazing effects were evident based on the observed relationship between phytoplankton and zooplankton biomass (R ² < 0.2, p > 0.1). This scenario was supported by short-term field observations made over an annual cycle at three stations in the mid-channel of the estuary where both chlorophyll a and primary production demonstrated similar patterns of seasonal variation. Ratios of fluorescence before and after acidification at all study sites were relatively high suggesting low grazing pressure in the estuary. A tidally-averaged, size-structured plankton ecosystem model was previously developed and verified for the lower York River estuary. The validated ecosystem model was also used to examine this issue and simulation results supported the importance of bottom-up control in the York River estuary.