Use of Competitive PCR to Detect and Quantify Haplosporidium nelsoni Infection (MSX disease) in the Eastern Oyster (Crassostrea virginica)

This study was undertaken to develop a quantitative polymerase chain reaction assay that would improve the utility of PCR for detecting Haplosporidium nelsoni (MSX), a serious parasite of the eastern oyster Crassostrea virginica. A competitive PCR sequence was generated from the H. nelsoni small subunit ribosomal DNA fragment, originally described by Stokes and colleagues, that was amplified by the same PCR primers and had similar amplification performance. Assays performed using competitor dilutions ranging from 0.05 to 500 pg/μl DNA were used to test oyster samples designated using histological techniques as having ``light' or ``heavy' MSX infections. Visual diagnoses were confirmed equally well with three methods: densitometry of ethidium-bromide-stained agarose, densitometry of SYBRGreen-stained polyacrylamide gels, and analysis by GeneScan 3.0 of fluorescent products detected in ultrathin gels. Oysters diagnosed as negative for MSX tested as negative or light by PCR. Oysters with light MSX infections generally had less than 5 pg/μl infectious DNA. Oysters with heavy infections generally corresponded to 5 pg/μl or greater competitor dilutions. Received September 3, 1999; accepted March 3, 2000.     

Vegetation differentiation in the patterned landscape of the central Everglades: importance of local and landscape drivers

Aim We present a model to account for self‐assembly of the slough–ridge–tree island patterned landscape of the central Everglades in southern Florida via feedbacks among landforms, hydrology, vegetation and biogeochemistry. We test aspects of this model by analysing vegetation composition in relation to local and landscape‐level drivers. Location We quantified vegetation composition and environmental characteristics in central Water Conservation Area (WCA) 3A, southern WCA‐3A and southern WCA‐3B in southern Florida, based on their divergence in water management and flow regimes over the past 50 years. Methods In 562 quadrats, we estimated species coverages and quantified maximum, minimum and average water depth, soil depth to bedrock, normalized difference vegetation index (NDVI) and proximity to the nearest tree island. We used non‐metric multi‐dimensional scaling (NMS) to relate compositional variation to local and landscape‐level factors, and evaluated environmental differences among eight a priori vegetation types via anova . Results Water depth and hydroperiod decreased from sloughs to ridges to tree islands, but regions also differed significantly in the abundance of several community types and the hydroregimes characterizing them. NMS revealed two significant axes of compositional variation, tied to local gradients of water depth and correlated factors, and to a landscape‐scale gradient of proximity to tall tree islands. Sawgrass height and soil thickness increased toward higher ridges, and NDVI was greatest on tree islands. Main conclusions This study supports four components of our model: positive feedback of local substrate height on itself, woody plant invasion and subsequent P transport and concentration by top predators nesting on taller tree islands, compositional shifts in sites close to tree islands due to nutrient leakage, and flow‐induced feedback against total raised area. Regional divergence in the relationship of community types to current hydroregimes appears to reflect a lag of a few years after shifts in water management; a longer lag would be expected for shifts in landscape patterning. Both local and landscape‐level drivers appear to shape vegetation composition and soil thickness in the central Everglades.     

Declining Diversity in Natural and Restored Salt Marshes: A 30-Year Study of Tijuana Estuary

Between 1974 and 2004, Tijuana Estuary's natural salt marsh underwent pulse disturbance (an 8-month nontidal period in 1984), which caused the sudden loss of two short-lived halophytes ( Salicornia bigelovii [Sb] and Suaeda esteroa [Se]) and rapid dominance of a productive native succulent ( Sa. virginica [Sv]), plus ramp disturbance that led to gradual codominance by Jaumea carnosa (Jc) (another productive succulent) by 1994. Species richness was high in 1974 (4.2 species/0.25-m² plot), low in 1984 (1.4 species), and not fully recovered by 1994 (3.7 species) or 2004 (3.9 species). Restoration efforts (reseeding former habitat and excavating and planting new sites) did not recover the populations of Sb or Se. In a 1997 project, plantings of these and six other native halophytes survived initially, but by 2005, short-lived species were lost and Sv and Jc dominated, as in the natural marsh. In a 2000 restoration site, planting mortality was high for five species, but Sv recruited voluntarily and dominated by 2005. We attribute recent vegetation changes to frequent catastrophic storms, flooding, and sedimentation, which contrasted strongly with the benign conditions of decades prior to 1974. Sediment blocked tidal channels in 1984 and gradually elevated the marsh plain, degrading the diverse salt marsh and hindering efforts to restore it. Future restoration efforts will require even greater control over sediment inflows plus contouring sites to include natural topographic features that appear critical to sustaining high species richness and evenness.     

Extending the host range of the bioherbicidal fungus Colletotrichum gloeosporioides f. sp. aeschynomene

Spore formulations of the bioherbicidal fungus Colletotrichum gloeosporioides f. sp. aeschynomene (ATCC No. 20358) (CGA) were evaluated for control of three weed species: northern jointvetch (Aeschynomene virginica), Indian jointvetch (A. indica), and hemp sesbania (Sesbania exaltata) in greenhouse experiments. Mortality, dry weight reduction, and plant height reduction of A. virginica seedlings ranged from 98% to 100%, 15 days after inoculation with CGA in water, in an invert emulsion, or in Silwet L-77 surfactant. However, CGA in water caused no effects of these parameters on S. exaltata, and only slight effects on A. indica. A. indica and S. exaltata were also severely injured (mortality, dry weight and plant height reduction, 98–100%) by CGA in the invert emulsion or in Silwet L-77. The CGA in Silwet formulation incited severe disease development more rapidly than the invert emulsion or water formulations of CGA in all species. These results suggest that the host range of CGA can be expanded though formulation modification to enable this bioherbicide to control multiple weeds, thus making this product more economically acceptable.     

Latitudinal compensation in oyster ciliary activity

1. Theories of latitudinal compensation predict that individuals living in colder temperature regimes should physiologically compensate for the slowing of standard physiological rates, owing to the relatively low temperature of their local environments, by increasing their metabolic rate in colder water temperatures relative to individuals living in warmer water temperature regimes.
2. This hypothesis was tested with oyster strains originally from geographically separated populations that were raised in a common environment for seven generations. The physiological parameter measured was ciliary activity across a temperature gradient.
3. Support for the latitudinal compensation hypothesis was found: the strain originally from the colder temperature regime had more active cilia at lower experimental temperatures than individuals originally from the warmer temperature regime. Ciliary activity of the more northern Long Island Sound oysters was significantly greater than activity in the more southern Delaware Bay oysters at temperatures of –1, 2 and 6 °C.
4. These results suggest that there is genetically based physiological differentiation between these populations of oysters consistent with the latitudinal compensation for local temperature regime.     

Fatty acid composition of Goniobasis virginica, Physa sp. and Viviparus malleatus (Mollusca: Gastropoda) from Lake Musconetcong, New Jersey

Gas—liquid chromatographic studies were done to determine the fatty acid composition of Goniobasis virginica Physa sp., and Viviparus malleatus (Mollusca: Gastropoda), from Lake Musconetcong, NJ, U.S.A. Palmitic acid was the predominant saturated fatty acid, followed by stearic acid, in all three molluscan species. The chief monoenes were 16:n−7, 18:1n−7; 20:1n−11+9, and 22:1n−11+13, which together accounted for all monoene fatty acids and one-quarter of the total fatty acids. Considerable amounts of linolenic acid (2.7−4.1%) and arachidonic acid (7.8−12%) were found in all three species. The percentage composition of docosahexaenoic acid (22:6n−3) was low compared to that of eicosapentaenoic acid (22:5n−3). Non-methylene interrupted dienes (20:2 NMID), characteristic of marine molluscs, ranged from 2 to 3% in the three species of freshwater snails.