Ultrastructure of the gonad and gametogenesis in the eastern oyster,Crassostrea virginica. I. Ovary and oogenesis

The ultrastructural features of the ovary and oogenesis are described in the eastern oyster,Crassostrea virginica (Gmelin, 1791). The ovary is a diffuse organ consisting of highly branching acini in which oocytes develop. The acini are surrounded by a matrix of vesicular connective tissue (VCT tells) which serves a nutrient storage function. Each acinus is bathed by fluid within a surrounding connective tissue compartment, the hemocoel, which likely serves as a means of transporting nutrients to the oocytes. Oocytes begin growth while positioned near to the inner acinus wall. As differentiation proceeds and they enter the late stages of vitellogenesis, they become stalle-shaped and project into the acinus lumen. Follicle tells are closely associated with oocytes during the early and middle stages of vitellogenesis but they are largely confined to the basal, stalked region of late-stage oocytes. Vitellogenesis occurs through a process of autosynthesis, involving the combined activity of the Golgi complex and rough endoplasmic reticulum, and heterosynthesis in which extraovarian precursors are incorporated into oocytes via receptor-mediated endocytosis involving the basal surface of the oocytes. It is suggested that the follicle tells play some important role during oogenesis but probably are not the major source of yolk precursors. The VCT celas are probably the main source of nutrients for vitellogenesis.     

Alkaloids and ascidian chemical defense: Evidence for the ecological role of natural products fromEudistoma olivaceum

The surface of the colonial ascidianEudistoma olivaceum (Van Name) is almost completely free of fouling organisms. I provide evidence that this epibiont-free surface is maintained by the alkaloids, Eudistomins G and H. These alkaloids were extracted from colonies collected in the shallow subtidal of Indian River Lagoon, Florida. USA, in November 1987. Concentrations of less than one-fifth of those found in the living ascidian inhibited the settlement of the larvae of five invertebrate species relative to appropriate controls in laboratory and field trials. Standard pharmacological assays have revealed that Eudistomins G and H were not the most biologically active of the eudistomins, indicating that these standard assays are not necessarily good predictors of antifouling activity. In an examination of possible alternate roles for these eudistomins, they proved ineffective as a fish antifeedant. PinfishLagadon rhomboides consumed agar pellets to which Eudistomins G and H had been added at concentrations 10- to 100-fold higher than those effective against settling larvae. These findings indicate that biologically active marine natural products may serve specific ecological roles.     

Preimplantation genetic diagnosis for single gene disorders: experience with five single gene disorders

We report our experience of 14 preimplantation genetic diagnosis (PGD) cycles in eight couples carrying five different single gene disorders, during the last 18 months. Diagnoses were performed for myotonic dystrophy (DM), cystic fibrosis (CF) [ΔF508 and exon 4 (621+1 G>T)], fragile X and CF simultaneously, and two disorders for which PGD had not been previously attempted, namely neurofibromatosis type 2 (NF2) and Crouzon syndrome. Diagnoses for single gene disorders were carried out on ideally two blastomeres biopsied from Day 3 embryos. A highly polymorphic marker was included in each diagnosis to control against contamination. For the dominant disorders, where possible, linked polymorphisms provided an additional means of determining the genotype of the embryo hence reducing the risk of misdiagnosis due to allele dropout (ADO). Multiplex fluorescent polymerase chain reaction (F-PCR) was used in all cases, followed by fragment analysis and/or single-stranded conformation polymorphism (SSCP) for genotyping. Embryo transfer was performed in 13 cycles resulting in one biochemical pregnancy for CF, three normal deliveries (a twin and a singleton) and one early miscarriage for DM and a singleton for Crouzon syndrome. In each case the untransferred embryos were used to confirm the diagnoses performed on the biopsied cells. The results were concordant in all cases. The inclusion of a polymorphic marker allowed the detection of extraneous DNA contamination in two cells from one case. Knowing the genotype of the contaminating DNA allowed its origin to be traced. All five pregnancies were obtained from embryos in which two blastomeres were biopsied for the diagnosis. Our data demonstrate the successful strategy of using multiplex PCR to simultaneously amplify the mutation site and a polymorphic locus, fluorescent PCR technology to achieve greater sensitivity, and two-cell biopsy to increase the efficiency and success of diagnoses. Copyright © 2002 John Wiley & Sons, Ltd.     

Electrolytic transformation of ordinance related compounds (ORCs) in groundwater: laboratory mass balance studies

Electrolytic reactive barriers (e(-) barriers) consist of closely spaced permeable electrodes installed across a groundwater contaminant plume in a permeable reactive barrier format. Application of sufficient potential to the electrodes results in sequential oxidation and reduction of the target contaminant. The objective of this study was to quantify the mass distribution of compounds produced during sequential electrolytic oxidation and reduction of ordinance related compounds (ORCs) in a laboratory analog to an e(-) barrier. In this study, a series of column tests were conducted using RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine) and TNT (2,4,6-trinitrotoluene) as representative ORCs. The experimental setup consisted of a plexiglass column packed with quartz-feldspar sand to simulate aquifer conditions. A single set of porous electrodes consisting of expanded titanium-mixed metal oxide mesh was placed at the midpoint of the sand column as a one-dimensional analog to an e(-) barrier. Constant current of 20mA (variable voltage) was applied to the electrode set. Initial studies involved quantification of reaction products using unlabeled RDX and TNT. Approximately 70% of the influent concentration was transformed, in one pass, through sequential oxidation-reduction for both contaminants. Following the unlabeled studies, (14)C labeled RDX and TNT were introduced to determine the mass balance. An activity balance of up to 96% was achieved for both (14)C-RDX and (14)C-TNT. For both contaminants, approximately 21% of the influent activity was mineralized to (14)CO(2). The proportion of the initial activity in the dissolved fraction was different for the two test contaminants. Approximately 30% of the initial (14)C-RDX was recovered as unreacted in the dissolved phase. The balance of the (14)C-RDX was recovered as non-volatile, non-nitroso transformation products. None of the (14)C-RDX was sorbed to the column sand packing. For (14)C-TNT approximately 51% of the initial activity was recovered in the dissolved phase, the majority was unreacted TNT. The balance of the (14)C-TNT was either sorbed to the sand packing (approximately 24%) or dissolved/mineralized as unidentified ring cleavage products ( approximately 4%).     

Hydraulic characterization for steam enhanced remediation conducted in fractured rock

To explore the viability of Steam Enhanced Remediation (SER) in fractured rock a small-scale steam injection and water/vapour extraction pilot study was conducted at the former Loring Air Force Base in northern Maine, USA. A detailed well testing program was undertaken to assist in the design of the injection and extraction well array, and to assess the possibility of off-site heat and contaminant migration. A structurally complex limestone having low matrix porosity and a sparse distribution of fractures underlies the study site. To characterize the groundwater and steam flow pathways, single-well slug tests and more than 100 pulse interference tests were conducted. The results of the well testing indicate that the study site is dominated by steeply dipping bedding plane fractures that are interconnected only between some wells in the injection/extraction array. The SER system was designed to take advantage of interconnected fractures located at depth in the eastern end of the site. An array of 29 wells located in an area of 60 by 40 m was used for steam injection and water/vapour extraction. The migration of heat was monitored in several wells using thermistor arrays having a 1.5 m vertical spacing. Temperature measurements obtained during and after the 3 month steam injection period showed that heat migration generally occurred along those fracture features identified by the pulse interference testing. Based on these results, it is concluded that the pulse interference tests were valuable in assisting with the design of the injection/extraction well geometry and in predicting the migration pathways of the hot water associated with the steam injection. The pulse interference test method should also prove useful in support of any other remedial method dependant on the fracture network for delivery of remedial fluid or extraction of contaminants.     

Incidence of ozone symptoms on vegetation within a National Wildlife Refuge in New Jersey, USA

During 1993-1996 and 2001-2003, we evaluated the percentage of plants (incidence) exhibiting ozone-induced foliar symptoms on vegetation within a National Wildlife Refuge located along the Atlantic Ocean coast of New Jersey, USA. Incidence varied among plant species and years. Bioindicator plants most sensitive to ozone, across all years, included native common milkweed (Asclepias syriaca) and wild grape (Vitis spp.), as well as introduced tree-of-heaven (Ailanthus altissima). Less sensitive bioindicators included Virginia creeper (Parthenocissus quinquefolia) and winged sumac (Rhus coppolina). Black cherry (Prunus serotina) and sassafras (Sassafras albidum) were least sensitive. The greatest incidence of ozone symptoms, across all plant species, occurred in 1996, followed by 2001>1995>1994>1993>2003>2002. A model was developed that showed a statistically significant relationship between incidence of ozone symptoms and the following parameters: plant species, Palmer Drought Severity Index, and the interaction of W126 x N100 measures of ambient ozone.     

Soil characteristics and phosphorus level effect on phosphorus loss in runoff

The loss of phosphorus (P) in runoff from agricultural soils may accelerate eutrophication in lakes and streams as well as degrade surface water quality. Limited soil specific data exist on the relationship between runoff P and soil P. This study investigated the relationship between runoff dissolved reactive phosphorus (DRP) and soil P for three Oklahoma benchmark soils: Richfield (fine, smectitic, mesic Aridic Argiustoll), Dennis (fine, mixed, active, thermic Aquic Argiudoll), and Kirkland (fine, mixed, superactive, thermic Udertic Paleustoll) series. These soils were selected to represent the most important agricultural soils in Oklahoma across three major land resource areas. Surface soil (0-15 cm) was collected from three designated locations, treated with diammonium phosphate (18-46-0) to establish a wide range of water-soluble phosphorus (WSP) (3.15-230 mg kg(-1)) and Mehlich-3 phosphorus (M3P) (27.8-925 mg kg(-1)). Amended soils were allowed to reach a steady state 210 d before simulated rainfall (75 mm h(-1)). Runoff was collected for 30 min from bare soil boxes (1.0 x 0.42 m and 5% slope) and analyzed for DRP and total P. Soil samples collected immediately before rainfall simulation were analyzed for the following: M3P, WSP, ammonium oxalate P saturation index (PSI(ox)), water-soluble phosphorus saturation index (PSI(WSP)), and phosphorus saturation index calculated from M3P and phosphorus sorption maxima (P(sat)). The DRP in runoff was highly related (p < 0.001) to M3P for individual soil series (r2 > 0.92). Highly significant relationships (p < 0.001) were found between runoff DRP and soil WSP for the individual soil series (r2 > 0.88). Highly significant relationships (p < 0.001) existed between DRP and different P saturation indexes. Significant differences (p < 0.05) among the slopes of the regressions for the DRP-M3P, DRP-WSP, DRP-PSI(ox), DRP-PSI(WSP), and DRP-P(sat) relationships indicate that the relationships are soil specific and phosphorus management decisions should consider soil characteristics.     

ESCHERICHIA COLI SURVIVAL IN MANTLED KARST SPRINGS AND STREAMS,NORTHWEST ARKANSAS OZARKS,USA1

Recent studies indicate fecal coliform bacterial concentrations, including Escherichia coli (E. coli), characteristically vary by several orders of magnitude, depending on the hydrology of storm recharge and discharge. E. coli concentrations in spring water increase rapidly during the rising limb of a storm hydrograph, peak prior to or coincident with the peak of the storm pulse, and decline rapidly, well before the recession of the storm hydrograph. This suggests E. coli are associated with resuspension of sediment during the onset of turbulent flow, and indicates viable bacteria reside within the spring and stream sediments. E. coli inoculated chambers were placed in spring and stream environments within the mantled karst of northwest Arkansas to assess long term (> 75 days) E. coli viability. During the 75‐day study, a 4‐log die‐off of E. coli was observed for chambers placed in the Illinois River, and a 5‐log die‐off for chambers placed in Copperhead Spring. Extrapolation of the regression line for each environment indicates E. coli concentration would reach 1 most probable number (MPN)/100 g sediment at Copperhead Spring in about 105 days, and about 135 days in the Illinois River, based on a starting inoculation of 2.5 × 107 MPN E. coli/100 g of sediment. These in situ observations indicate it is possible for E. coli to survive in these environments for at least four months with no fresh external inputs.     

Macroinvertebrate biomonitoring in intermittent coastal plain streams impacted by animal agriculture

Little attention has been given to the ecology of intermittent coastal plain streams in the southeastern United States, and it is not known whether available macroinvertebrate biomonitoring methods reliably detect degradation in these streams. This study compared differences in biomonitoring metrics between reference and agricultural streams, and between the flow period (January-April) and the intermittent flow period (May-December). Percentages of crustaceans, isopods, and Ephemeroptera-Plecoptera-Trichoptera (EPT) were significantly higher at the reference site than the two most impacted sites during the flow period, probably resulting from the abundance of leaf litter and lower temperatures. During this same period, the agriculturally impacted sites had a significantly higher percentage of dipterans--a group that thrives in the silty, nutrient-rich waters. Four metrics (percent Crustacea, Isopoda, Diptera, and EPT) had no overlap between values for the most impacted and the least impacted sites during the flow period, but no metrics were able to detect more discrete differences among sites. Sites were physically and biologically similar during the intermittent period when natural stresses (i.e., stagnant water, high temperatures, low dissolved oxygen) were high, with many metrics, such as percentages of dominant family, burrowers, chironomids, and dipterans becoming similar at all sites. Our findings indicate that development of a better understanding of invertebrate fauna in reference conditions and of the natural variation in intermittent streams is necessary to develop effective biomonitoring programs for these systems.