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Jill A. Jenkins

Publications and source records attributed to Jill A. Jenkins.

At least 37 records · Page 2Linked to original sources

Photographic images captured while sampling for bald eagles near the Davis Pond freshwater diversion structure in Barataria Bay, Louisiana (2009-10)

The implementation of freshwater diversions in large-scale coastal restoration schemes presents several scientific and management considerations. Large-scale environmental restructuring necessitates aquatic biomonitoring, and during such field studies, photographs that document animals and habitat may be captured. Among the biomonitoring studies performed in conjunction with the Davis Pond freshwater diversion structure south of New Orleans, Louisiana, only postdiversion study images are readily available, and these are presented here.

Data Series

Occurrence and distribution of Asian carps in Louisiana

In the 1970s, commercial fishers reported sightings of grass carp Ctenopharyngodon idella in large rivers and associated backwaters of Louisiana; the first specimen in Louisiana Department of Wildlife and Fisheries' fishery independent sampling was recorded in 1976. Beginning in the early 1980s, commercial fishers noted increasing populations of bighead carp Hypophthalmichthys nobilis and silver carp H. molitrix (together, the bigheaded carps). Populations of bigheaded carps appear to be increasing at a much slower rate than in the Midwest, possibly due to limited suitability of and access to backwater habitat for juvenile fish. In 2002, harvester reports of sporadic captures of "different-looking" grass carp indicated the possible presence of black carp Mylopharyngodon piceus . Because both normal diploid and triploid (in which triploidy has been induced to cause sterility) black carp have been stocked in the Mississippi basin, determination of the ploidy (number of chromosome sets) of these fishes is important. Since 2002, postmortem ploidy determinations using cells from eyeballs removed from six wild black carp captured in Louisiana showed each to be a normal diploid, indicative of breeding capability and potential reproducing populations. Although reported commercial landings of grass and bigheaded carps have been as high as 34,830 kg/year, limited market demand in past years resulted in many captures being discarded. A protocol for obtaining samples for easily determining ploidy is reported here. Accurate data on Asian carp distributions and their reproductive potential provides information to fisheries researchers that will be constructive in documenting the spread of these invasive species and in the assessment of risk to habitats.

Louisiana

Review of concepts useful for maintaining quality of male reproductive field samples for laboratory study

Investigations into cellular and molecular characteristics of male gametes obtained from fish in natural ecosystems require careful sample handling and shipping in order to minimize artifacts. Maintaining sample integrity engenders confident assessments of ecosystem health, whereby animal condition is often reflected by gamete biomarkers - indicators that respond in measurable ways to changes. A number of our investigations have addressed the hypothesis that biomarkers from fish along a pollution gradient are reflective of site location. Species biology and the selected biological endpoints direct choice of parameters such as: temperature, buffer osmolality, time in transit, fixation, cryoprotectants, protease inhibition, and antibiotic inclusion in extender. This paper will highlight case studies, and outline parameters and thoughts on approaches for use by field and laboratory researchers.

Conference Paper

Derivation of cat embryonic stem-like cells from in vitro-produced blastocysts on homologous and heterologous feeder cells

The domestic cat is a focal mammalian species that is used as a model for developing assisted reproductive technologies for preserving endangered cats and for studying human diseases. The generation of stable characterized cat embryonic stem cells (ESC) lines to use as donor nuclei may help to improve the efficiency of interspecies somatic cell nuclear transfer for preserving endangered cats and allow the creation of knockout cell lines to generate knockout cats for studying function of specific genes related to human diseases. It will also enable the possibility of producing gametes in vitro from ESC of endangered cats. In the present study, we report the generation of cat embryonic stem-like (cESL) cells from blastocysts derived entirely in vitro. We generated 32 cESL cell lines from 331 in vitro derived blastocysts from which inner cell masses were isolated by immunosurgery or by a mechanical method. Inhibition of cat dermal fibroblast (CDF) proliferation after exposure to mitomycin-C was both dose and time dependent, where doses of 30 to 40 microg/mL for 5 h were most efficient. These dosages were higher than that required to inhibit cell proliferation of mouse fetal fibroblasts (MFF; 10 microg/mL for 2.5 h). Mitomycin-C did not significantly increase necrosis of cells from either species, and had an anti-proliferative effect at concentrations below cytotoxicity. A clear species-specific relationship between feeder layers and derivation of cESL cell lines was observed, where higher numbers of cESL cell lines were generated on homologous cat feeder layers (n = 26) than from those derived on heterologous mouse feeder layers (n = 6). Three cESL cell lines generated from immunosurgery and cultured on CDF maintained self-renewal and were morphologically undifferentiated for nine and twelve passages (69-102 days). These lines showed a tightly packed dome shaped morphology, exhibited alkaline phosphatase activity and immuno-expression of the pluripotent marker OCT-4 and surface marker SSEA-1. Primary colonies at P0 to P3 and cat blastocysts expressed transcription factors OCT-4, NANOG and SOX-2 and the proto-oncogene C-MYC. However, expression was at levels significantly lower than in vitro produced blastocysts. During culture, cESL colonies spontaneously differentiated into fibroblasts, cardiomyocytes, and embryoid bodies. Development of techniques to prevent differentiation of cESL cells will be essential for maintaining defined cell lines

Theriogenology

Effects of wastewater discharges on endocrine and reproductive function of Western Mosquitofish (Gambusia spp.) and implications for the threatened Santa Ana Sucker (Catostomus santaanae)

The Santa Ana River (SAR) in southern California is impacted by effluents from wastewater treatment plants (WWTP), which are sources of organic wastewater compounds (OWCs) and urban runoff. The Santa Ana River is one of only three river basins supporting native populations of the federally listed Santa Ana sucker ( Catostomus santaanae ) at the time the fish was included on the list 2000. In 2004 and 2005, a U.S. Geological Survey and U.S. Fish and Wildlife Service study was undertaken to determine if the threatened Santa Ana sucker was potentially exposed to OWCs and endocrine disrupting compounds (EDCs) in the SAR by using the western mosquitofish ( Gambusia affinis ) as a surrogate fish model. Four Santa Ana River sites were chosen along a gradient of proximity to WWTP effluents: (1) a point source of tertiary treated wastewater effluent (TTWE), (2) Rialto Drain (just below a WWTP), (3) Prado Dam (11 kilometers [km] below WWTPs), and (4) Sunnyslope Creek (no WWTP but having urban runoff influence). A reference site having no WWTPs or urban runoff, Thousand Palms, was also sampled. Chemical analyses of passive sampler extracts results showed that 15 OWCs and EDCs were detected in water from the Santa Ana River sites. Many of these compounds contributed to activity from an estrogenic in-vitro assay that showed a significant potential for impacting endocrine and reproductive systems compared to the 25 organochlorine compounds detected in aquatic biota. The site showing compounds having highest influence on sex steroid hormone activities was the point source for TTWE. Sex steroid hormone levels, secondary sex characteristics, organosomatic indices, and sperm quality parameters indicated impairment of endocrine and reproductive function of male western mosquitofish in the Santa Ana River. Exposure to EDCs and consequent impairment in mosquitofish followed the gradient of proximity to WWTP effluents, where the most significant effects were found at TTWE point source and Rialto Drain, followed by Prado Dam and Sunnyslope Creek. Each of these sites is suitable habitat for the Santa Ana sucker, especially Sunnyslope Creek and Rialto Drain where juveniles reside. Various OWCs and EDCs were detected at each Santa Ana River site, although one specific compound or group of compounds could not be singled out as a causative factor. Di (2-ethylhexyl) phthalate was strongly negatively correlated with testosterone in male mosquitofish. One group of potent environmental estrogens that likely contributed to endocrine and reproductive impairment are the natural and synthetic estrogen hormones, especially ethinyl estradiol; however, this compound was not targeted in these investigations. The multiple lines of evidence for impaired reproductive and endocrine function in western mosquitofish due to OWCs and EDCs from the Santa Ana River can be used to identify potential problems for the Santa Ana sucker inhabiting the same and nearby sites.

California

Davis Pond freshwater prediversion biomonitoring study: freshwater fisheries and eagles

In January 2001, the construction of the Davis Pond freshwater diversion structure was completed by the U.S. Army Corps of Engineers. The diversion of freshwater from the Mississippi River is intended to mitigate saltwater intrusion from the Gulf of Mexico and to lessen the concomitant loss of wetland areas. In addition to the freshwater inflow, Barataria Bay basin would receive nutrients, increased flows of sediments, and water-borne and sediment-bound compounds. The purpose of this biomonitoring study was, therefore, to serve as a baseline for prediversion concentrations of selected contaminants in bald eagle ( Haliaeetus leucocephalus ) nestlings (hereafter referred to as eaglets), representative freshwater fish, and bivalves. Samples were collected from January through June 2001. Two similarly designed postdiversion studies, as described in the biological monitoring program, are planned. Active bald eagle nests targeted for sampling eaglet blood (n = 6) were generally located southwest and south of the diversion structure. The designated sites for aquatic animal sampling were at Lake Salvador, at Lake Cataouatche, at Bayou Couba, and along the Mississippi River. Aquatic animals representative of eagle prey were collected. Fish were from three different trophic levels and have varying feeding strategies and life histories. These included herbivorous striped mullet ( Mugil cephalus ), omnivorous blue catfish ( Ictalurus furcatus ), and carnivorous largemouth bass ( Micropterus salmoides ). Three individuals per species were collected at each of the four sampling sites. Freshwater Atlantic rangia clams ( Rangia cuneata ) were collected at the downstream marsh sites, and zebra mussels ( Dreissena spp.) were collected on the Mississippi River. The U.S. Geological Survey (USGS) Biomonitoring of Environmental Status and Trends (BEST) protocols served as guides for fish sampling and health assessments. Fish are useful for monitoring aquatic ecosystems because they accumulate pesticides and other contaminants. Biomarker data on individual fish, generated at the USGS National Wetlands Research Center (Lafayette, La.), included percent white blood cells in whole blood, spleen weight to body weight ratio, liver weight to body weight ratio, condition factor, splenic macrophage aggregates, and liver microsomal 7-ethoxyresorufin-o-deethylase (EROD) activity. Fish age was estimated by comparing total lengths with values from the same species in the Southeast United States as determined from the literature. Contaminant analyses were coordinated by the U.S. Fish and Wildlife Service (USFWS) Analytical Control Facility (Laurel, Md.), where residues of organochlorine (OC) pesticides, total polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), aliphatic hydrocarbons (AHs), and trace elements were determined. The organic contaminant data were generated at the Mississippi State University Chemical Lab (Mississippi State, Miss.), and the inorganic contaminant data were generated by the Texas A&M University Geochemical and Environmental Research Group (College Station, Tex.). Statistical tests were performed to assess relationships among contaminants, fish age, fish species, and collection sites. Trends in interspecific differences among fish in concentrations of contaminants were noted. Striped mullet (hereafter mullet) frequently displayed the highest chemical concentrations. Levels of contaminants were generally higher in samples obtained from the Mississippi River than in those collected from the diversion area and were higher in mussels and clams (hereafter bivalves) than in fish. Because the Mississippi River sampling site for mullet and largemouth bass was downriver of the structure and south of New Orleans and the catfish site was upriver, the downriver data may not be directly reflective of the results from the receiving waters at the Davis Pond structure. Compared to the Caernarvon freshwater prediversion study in 1990 that assessed possible influx of contaminants with the freshwater diversion, contaminant levels in fishes and bivalves in this study were generally lower, yet three nontoxic inorganic elements in Davis Pond fish samples exhibited ranges of concentrations that were more than two times higher than did those from Caernarvon. Levels in bivalves were different between diversions but about equal in the numbers of trace elements showing high levels per location. Contaminant values were compared to those listed in various literature and agency sources, both regional and national, including the National Contaminant Biomonitoring Program (NCBP), in which the 85th percentile and above represents what is considered to be an elevated contaminant concentration and cause for concern. Generally, bivalves were at the high end of their ranges for both organic and inorganic contaminants. In this study, OCs were detectable in 67 percent of fish from the Mississippi River site, ranging from 0.15 to 1.09 μg/g wet weight (ww) or fresh weight (fw), and in 11 percent of the fish from the marsh sites, ranging from 0.06 to 0.612 μg/g ww. Bivalves from the Mississippi River had OC levels of 0.096 μg/g ww, whereas none were detectable in bivalves at the marsh sites. In this study, p,p ’-dichlorodiphenyldichloroethylene ( p,p’ -DDE) (a biodegradation product of DDT [dichlorodiphenyl trichloroethane]) and total PCBs were the most frequently detected OCs and were primarily from the Mississippi River. For total OC content, using adjusted least squares means, some significant interactions were noted between fish species and sites. PAHs were detected in aquatic animals at all sites (range of 0.017–17.534 μg/g ww), as were AHs (range of 0.423–4.549 μg/g ww); the highest levels of PAHs and AHs were found in bivalves from the Mississippi River. When analysis of variance (α = 0.05) was performed with data from aquatic animals, there were only two significant relationships between PAHs, AHs, and OCs between species, site, and age or the interaction among these variables. There was an interaction between fish species and n-decane (an AH) in that mullet and largemouth bass had significantly higher levels than did catfish ( P = 0.0175). When general linear means were used to investigate associations of inorganic contaminants among fish species, site, and age or any interactions among these variables, no significant results were noted for arsenic, cadmium, lead, beryllium, boron, molybdenum, or nickel. The range of mercury in fish in this study was 0.04–0.14 μg/g ww (0.14– 0.48 μg/g dry weight [dw]), with the most elevated levels detected in predatory largemouth bass at the sampling point farthest downstream from the structure and within the marsh area. Mercury was positively correlated with fish age ( P = 0.0152), where levels were estimated to increase 0.0253 parts per million (ppm) dw per year. In the Mississippi River, catfish showed significantly higher levels of mercury than did mullet or largemouth bass ( P = 0.00167). Among fish species, mullet displayed the highest levels in fish of aluminum, barium, manganese, and iron, all considered to have low toxicity in hydrologic systems. An interaction between fish and site was seen with aluminum ( P = 0.0031), where concentrations in mullet were significantly higher in the Mississippi River than at the other sites, as was also seen with barium ( P = 0.0009), chromium ( P = <0.0001), manganese ( P = 0.0004), strontium ( P = 0.0074), vanadium ( P = 0.0156), and zinc ( P = 0.0059). For iron ( P = 0.0.0001), mullet and largemouth bass at both the Mississippi River and Lake Salvador showed higher levels than did catfish, and these two species showed higher levels at two of the four sites. An interaction between fish and site was also seen with chromium ( P = <0.0001) in that concentrations in mullet were significantly higher in the Mississippi River than at the other sites, as was also seen with strontium ( P = 0.0074), vanadium ( P = 0.0156), and zinc ( P = 0.0059), metals for which deleterious effects have been demonstrated in other ecosystems. The NCBP program lists the 85th percentile for zinc at 34.2 μg/g fw (117.9 μg/g dw). In the Davis Pond prediversion biomonitoring study (hereafter the current study), one fish (MUL31RIVER, fish ID 8) showed values higher than that (125.4 μg/g dw or 37.54 μg/g ww), and the Mississippi River bivalve sample (MUSSRIVER) had a value of 140 μg/g dw (41.2 μg/g ww). In the current study, approximately 86 percent of the fish had measurable selenium levels, yet none reached the 85th percentile. The 85th percentile for selenium from the NCBP was 0.73 μg/g ww. Significantly higher levels of selenium were seen in mullet than in largemouth bass and catfish ( P = 0.0023). The NCBP 85th percentile for lead is 0.22 μg/g ww (0.76 μg/g dw). In the current study, the range of concentrations of lead was as much as 18.3 ppm dw (MUL31RIVER, fish ID 8), with the three most elevated values (range of 3.46–5.31 μg/g ww) coming from mullet from the Mississippi River. Biomarker data are measurable and directly reflect the condition of the animal, and measuring more than one biomarker in an individual increases confidence in health assessments. In the current study, biomarkers included macrophage aggregates (MAs), liver (hepatosomatic index [HSI]) and spleen (splenosomatic index [SSI]) weight to body weight ratios, percent white blood cells (WBCs) in whole blood, and condition factor. Few significant differences were noted with any of the biomarkers between sites, and there were no relationships between species and sites. For improved use of biomarker assessments, an increase in fish sample size would be useful for postdiversion sampling, as would comparisons of fish of the same sex and reproductive condition. During the current study, success for eagle nests in the diversion area and reference sites was similar as determined by numbers of nestlings fledged. When temperatures were below average during winter 2000, nests in both regions similarly failed. At each nest, the primary evidence of food items was small mammals. Eaglets (n = 6) generally appeared healthy, and whole blood concentrations of organic contaminants exceeded detection limits with three incidences of p,p’ -DDE (0.002–0.006 μg/L ww) and one incidence of oxychlordane (0.002 μg/L ww). The levels of p,p’ -DDE were well below those that have been inversely correlated with productivity and success rates of nesting bald eagles on a regional scale. The low values found in the whole blood samples for OC pesticides and PCBs were even lower when corrected for plasma volume. Aluminum values were 3.66 and 5.75 μg/L in two samples, zinc ranged from 5.21 to 6.77 μg/L ww in six samples, and silicon ranged from 1.7 to 4.6 μg/L in four samples. Selenium was detectable in each bird with the range at 0.332–0.566 μg/L ww, and strontium ranged from 0.0581 to 0.0975 μg/L ww. Mercury was detectable in blood samples from each bird and ranged from 0.0254 to 0.0845 μg/L ww, whereas lead was detectable in four samples and ranged from 0.0042 to 0.0136 μg/L ww. Although no detectable levels of total PCBs were found (also correlated with decreased reproductive productivity), 70 percent of the aquatic animals from the Mississippi River contained total PCBs (range 0.13–0.79 μg/L), whereas only about 7 percent of the aquatic animals sampled from the marsh area contained PCBs. Suggestions for postdiversion sampling include lowering the analytical detection limit for some metals, sampling aquatic animals over the course of a single season, obtaining a higher sample number of mature fish of one species (for example, blue catfish) within a range of total lengths for biomarker analyses, obtaining otoliths for estimating fish ages, assessing dioxins in eaglet blood, examining triazines in water, and obtaining all Mississippi River fish samples as close to the Davis Pond structure intake as possible. Because contaminants found in blood of eaglets reflect their prey species and because of the contaminant levels found in fish in the current study, eaglets may not be consuming primarily these species; therefore, obtaining juvenile nutria ( Myocastor coypus ) or turtle species for contaminant analyses might be considered, as well as collecting greater blood volume and using plasma to measure OCs and PCBs. Data obtained postdiversion will be compared with prediversion data to monitor changes.

Louisiana

Organic Chemical Concentrations and Reproductive Biomarkers in Common Carp (Cyprinus carpio) Collected from Two Areas in Lake Mead, Nevada, May 1999-May 2000

The U.S. Geological Survey, in cooperation with the U.S. Fish and Wildlife Service, National Park Service, Bureau of Reclamation, and Nevada Department of Wildlife, collected and assessed data to determine the general health and reproductive status of common carp (Cyprinus carpio) at two study areas in Lake Mead, Nevada, during May 1999-May 2000. These data will form the basis of interpretations and provide a comparison for continuing studies on the health of the ecosystem in Lake Mead. One study area, Las Vegas Bay, is in the western part of Lake Mead. Las Vegas Bay receives inflows from Las Vegas Wash, which is predominantly tertiary-treated wastewater effluent, and to a lesser extent stormwater runoff from Las Vegas, Henderson, and other nearby communities, and from ground water underlying Las Vegas Valley. The other study area, Overton Arm, is in the northern extent of Lake Mead. Overton Arm receives inflow from the Virgin and Muddy Rivers, which historically are not influenced by wastewater effluent. Both sexes of common carp were collected bimonthly for 12 months using boat-mounted electrofishing gear (a direct electric current is used to temporarily immobilize fish for capture) to determine their health and reproductive status and any relation between these factors and environmental contaminants. This report presents fish tissue chemistry, organic chemical compound concentrations, and biomarker data for 83 male common carp collected from Las Vegas Bay, similar organic chemistry results for 15 male common carp, and similar biomarker measures for 80 male common carp collected from Overton Arm. Tissue chemistry results also are presented for 16 female common carp and biomarker measures for 79 female common carp collected from Las Vegas Bay, and tissue chemistry results for 15 female common carp and biomarker measures for 81 female common carp collected from Overton Arm. Thirty-three organic chemical compounds plus total concentrations for four groups of compounds (chlordanes, polychlorinated biphenyls [PCBs], brominated diphenyl ethers [BDEs], and triclosans) were analyzed from extracts of whole-body tissue using gas chromatography/mass spectrometry in male common carp from Las Vegas Bay during May 1999 through May 2000. All 33 compounds were detected in at least one sample of whole-body tissue from male common carp collected in Las Vegas Bay. In Overton Arm, 37 organic compounds plus total concentrations of three groups of compounds (PCBs, BDEs, and triclosans) were analyzed in male common carp where 20 (54 percent) of the compounds were detected. Sixteen of the 33 compounds detected in male common carp from Las Vegas Bay and 10 compounds detected in males from Overton Arm have the potential to disrupt the endocrine system in fish in Lake Mead. During May and June 1999, the mean concentration of all organic compounds detected in male common carp was 670 micrograms per kilogram from Las Vegas Bay and 109 micrograms per kilogram from Overton Arm. Twenty-seven organic compounds plus total PCBs were analyzed from extracts of whole-body tissue in female common carp collected in Las Vegas Bay and Overton Arm during May 1999. Twenty-four (86 percent) of these compounds were detected in at least one sample of whole-body tissue from female common carp collected from Las Vegas Bay while 10 (36 percent) chemical compounds were detected in female common carp from Overton Arm during that same period. Median concentrations of all chemical compounds were higher in female common carp from Las Vegas Bay compared to those collected from Overton Arm except Dacthal (DCPA), which was similar between sites. Biomarker measures obtained for male and female common carp include gonadosomatic index (percentage of gonad weight to total body weight), plasma vitellogenin (a phospholipid protein normally produced by female common carp and other oviparous fish), and condition factor [body weight/(fork length)3]. Biomarker measures for male c

Data Series

Sperm cryopreservation in fish and shellfish

Initial success in sperm cryopreservation came at about the same time for aquatic species and livestock. However, in the 50-plus years since then cryopreserved sperm of livestock has grown into a billion-dollar global industry, while despite work in some 200 species with well over 200 published reports, cryopreservation of aquatic species sperm remains essentially a research activity with little commercial application. Most research has focused on large-bodied culture and sport fishes, such as salmonids, carps, and catfishes, and mollusks such as commercially important oyster and abalone species. However, only a handful of studies have addressed sperm cryopreservation in small fishes, such as zebrafish, and in endangered species. Overall, this work has yielded techniques that are being applied with varying levels of success around the world. Barriers to expanded application include a diverse and widely distributed literature base, technical problems, small sperm volumes, variable results, a general lack of access to the technology, and most importantly, the lack of standardization in practices and reporting. The benefits of cryopreservation include at least five levels of improvements for existing industries and for creation of new industries. First, cryopreservation can be used to improve existing hatchery operations by providing sperm on demand and simplifying the timing of induced spawning. Second, frozen sperm can enhance efficient use of facilities and create new opportunities in the hatchery by eliminating the need to maintain live males, potentially freeing resources for use with females and larvae. Third, valuable genetic lineages such as endangered species, research models, or improved farmed strains can be protected by storage of frozen sperm. Fourth, cryopreservation opens the door for rapid genetic improvement. Frozen sperm can be used in breeding programs to create improved lines and shape the genetic resources available for aquaculture. Finally, cryopreserved sperm of aquatic species will at some point become an entirely new industry itself. A successful industry will require integrated practices for sample collection, refrigerated storage, freezing, thawing, rules for use and disposal, transfer agreements, and database development. Indeed the development of this new industry is currently constrained by factors including the technical requirements for scaling-up to commercial operations during the transition from research, and the absence of uniform quality control practices, industry standards, marketing and price structures, and appropriate biosecurity safeguards.

Society for Reproduction and Fertility Supplement

Cell proliferation detected with flow cytometric cell cycle analysis and immunohistochemical detection of Proliferating Cell Nuclear Antigen (PCNA) from somatic tissues of Eastern Oysters, Crassostrea virginica

Two novel biomarker of response assays were developed and compared for use with bivalves. Bivalve mollusks are often used as bioindicators to monitor contaminant body burdens and are employed globally in pollution monitoring and as sentinels of environmental quality. The prevalence of proliferating cells in tissues of the eastern oyster, Crassostrea virginica , (n = 10) was investigated by using immunohistochemical staining of proliferating cell nuclear antigen (PCNA) and by flow cytometric analysis of DNA in cell cycle phases. The PCNA protein is highly conserved and central to DNA synthesis in dividing cells. This protein was detected by using a commercial antibody in the histological staining of gills, mantle, digestive gland, stomach, and labial palps. Cells obtained from these same organs and from the hearts of the oysters were used for flow cytometry (FCM). The proportions of nuclei in the S plus G2/M and G2/M fractions were compared with percentages of PCNA-positive cells scored through microscopy. Proliferation responses generated by microscopy and FCM (S plus G2/M) were equal for digestive gland, gills, stomach, and mantle. The proliferation response in labial palps was significantly higher by PCNA microscopy than by DNA FCM (S plus G2/M). For PCNA microscopy results, a significantly higher proliferation response was noted both for labial palps as compared to gills and mantle and for stomach and digestive gland as compared to gills (P < 0.005). For DNA FCM results, the percentages of proliferating cells in G2M fractions were significantly higher for labial palps and heart when compared to gills, mantle, and digestive gland. By FCM (S plus G2M), the percentages of proliferating cells followed a similar trend, although no significant differences were found between organs. The choice of a relatively highly proliferative tissue, such as labial palps as determined by this study, has utility for developing a cell line, for examining proliferation as a biomarker due to stressor impacts in feral oysters, and for detecting impacts of compounds on proliferative mechanisms.

Environmental Bioindicators

Bioindicators from Mosquitofish (Gambusia affinis) Sampled from the Imperial Valley in Southern California

The Sonny Bono Salton Sea National Wildlife Refuge (SSNWR) is located 64 km north of the Mexican border at the southern end of the Salton Sea in California's Imperial Valley. Freshwater ponds and managed habitats at the SSNWR, Calipatria, Calif. are supplied with Colorado River water that carries compounds from upstream sources. Components include municipal and industrial discharges, agricultural drainage, and sewage plant inputs. Aquatic animals in these ecosystems are continuously exposed to multiple constituents, several of which have been demonstrated to be associated with hormonal disturbances. We investigated possible endocrine impacts to fish in the Imperial Valley, Calif., by addressing the null hypothesis that aquatic species in impacted sites did not exhibit evidence of endocrine disruption as compared with those from nonimpacted sites. The results presented are intended to provide managers with science-based information and interpretations about the condition of the animals in their ecosystems for the minimization of potential adverse effects to trust fish and wildlife resources and for the maximization of available water resources.

Open-File Report

Viability of male gametes in common carp ( Cyprinus carpio ) along the Lower Colorado River from the Cibola National Wildlife Refuge (NWR), Havasu NWR, and Lake Mohave of Lake Mead National Recreation Area

To contribute to an investigation on possible endocrine impacts in three sites along the lower Colorado River in Arizona, especially in male fishes, this study addressed the null hypothesis that aquatic species in southern sites did not exhibit evidence of endocrine disruption as compared with those in nonimpacted sites. The results presented are intended to provide managers with science-based information and interpretations about the reproductive condition of biota in their habitat along the lower Colorado River to minimize any potential adverse effects to trust fish and wildlife resources and to identify water resources of acceptable quality. In particular, these data can inform decision making about wastewater discharges into the Colorado River that directly supplies water to Arizona refuges located along the river. These data are integral to the USFWS proposal entitled 'AZ - Endocrine Disruption in Razorback Sucker and Common Carp on National Wildlife Refuges along the Lower Colorado River' that was proposed to assess evidence of endocrine disruption in carp and razorback suckers downstream of Hoover Dam.

Open-File Report

Pallid sturgeon in the Lower Mississippi Region: Hematology and genome information

This project (Project 1448-43270-2M-002) has been coordinated through the Natchitoches National Fish Hatchery (NNFH) and the U.S. Geological Survey’s National Wetlands Research Center (NWRC). From November 2001 to April 2002, over 280 sturgeon of the genus Scaphirhynchus (including pallid sturgeon, shovelnose, and their hybrids) were sampled from the outflow channel of the Old River Control Structure Complex (ORCC) in Concordia Parish, La. In the overall project, several datasets were collected (see Appendix), including species identification by using microsatellites and morphometric characters, food habits, physical anomalies, information on blood cells, and pathologic evidence of iridovirus – the first indication in the lower Mississippi population of pallid sturgeon. In this study, data on blood cells were obtained from the sturgeon collected monthly from approximately 20 different animals at each sampling time. This report presents preliminary information on differential blood cell identifications in sturgeon, data on comparative genomic DNA content and DNA degradation, and summaries and interpretations of data collected in light of available scientific literature addressing blood parameters of fish and sturgeon, in particular. Results obtained from collection and examination of blood and body fluids are often essential in establishing the health of fish (Blaxhall, 1972; Fange, 1992). Blood cells and sperm cells can be obtained nondestructively from fishes, even from small specimens that weigh less than 100 g (Stoskopf, 1992a). For flow cytometry assays, whereby cells are analyzed individually in a fluid stream, less than 1 :L of blood is needed. Examinations of blood by microscopy and flow cytometry were performed at NWRC in assisting in the efforts directed at recovery of the pallid sturgeon population in the Lower Mississippi River Basin.

Open-File Report