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Variation of cisco egg size among Laurentian Great Lakes populations

Many fish species display inter-population and inter-individual egg size variation. Intra-specific differences in egg size seemingly reflect both energetic experiences of individual spawning fish and long-term population responses to differing ecosystems. Optimal egg size theory implies that selection influences a population’s mean egg size in response to its early-life environment, given the well-established trade-off between egg size and fecundity. Currently, there is strong interest in rehabilitation of Laurentian Great Lakes cisco, Coregonus artedi, which is characterized by inter-population variation of morphological and behavioral traits. However, the extent of cisco egg size variation is under-described. In fall 2018 and 2019, we collected egg samples by stripping ripe females at seven total locations in four Great Lakes. We measured unfertilized egg diameters using imaging software and compared mean egg diameters among locations with and without including maternal total length as a covariate. Lake Michigan females produced the largest eggs overall but were excluded from analyses using the total length covariate because of their significantly larger body sizes. Maternal length had a positive effect on egg size, and when accounting for this effect, females in Lake Huron produced the largest eggs followed by Lake Ontario and Lake Superior. We also found that egg size varied among locations within Lake Superior. These findings aligned with observations of morphological and behavioral differences among populations and suggest that cisco phenotypic variation at a fine spatial scale extends to reproductive biology. Consideration of cisco reproductive traits, such as egg size, may inform restoration strategies, including supplemental stocking.

Journal of Great Lakes Research↗

Field and laboratory validation of new sampling gear to quantify coregonine egg deposition and larval emergence across spawning habitat gradients

The influence of habitat and environmental conditions on Great Lakes coregonine reproduction is not well described, in part, because we lack sampling gears for early life stages that are effective across habitats. We designed new egg and larval emergence traps to quantify coregonine reproductive success across variable depths and substrates and tested them in laboratory and field settings. In the laboratory, our new metal ring egg traps had greater egg retention (94–100%) and faster post-catch processing (5–7 min) relative to a commonly employed fiber mat trap (30–67% and 30–60 min). In Lake Ontario’s Chaumont Bay, egg densities for lake whitefish Coregonus clupeaformis (0–5,832 eggs m −2 ) and cisco Coregonus artedi (0–426,501 eggs m −2 ) measured with metal ring traps (n = 112) varied across habitats but were greatest between 2–5 m on rock and dreissenid mussel substrates. Emergence traps used an inverted cone, fine mesh, and a clear collection chamber to capture positively phototactic emerging larvae. In the laboratory, traps captured 69–80% of emerged larvae. In Chaumont Bay, emergence traps deployed for 21 days after ice out caught only cisco larvae. Emergence rates varied across habitats (0–118 larvae m −2 day −1 , n = 85) but were highest on dreissenid mussel reef substrate. Our samplers improved processing efficiency and facilitated large sample sizes to quantify variability in egg deposition densities and emergence rates across habitats. These methods can advance coregonine conservation by determining how anthropogenic changes to habitat and environmental conditions influence incubation success.

Journal of Great Lakes Research↗

On the importance of ichthyoplankton monitoring for invasive grass carp control in the Laurentian Great Lakes

Grass carp ( Ctenopharyngodon idella ) is an invasive herbivore observed in small numbers in the Laurentian Great Lakes since the 1980 s with records from all lakes except Lake Superior. Identification of diploid, age-1 + grass carp from the Sandusky River, a Lake Erie tributary, in 2012, prompted targeted efforts to evaluate the status of grass carp reproduction in the Lake Erie Basin. In 2014, researchers from the U.S. Geological Survey and University of Toledo began surveys to assess grass carp spawning in the Great Lakes, with the first fertilized eggs discovered in 2015 in the Sandusky River. Eight Great Lakes tributaries in the United States have been surveyed for evidence of grass carp spawning, identifying three confirmed spawning tributaries (Sandusky, Maumee, and Huron Rivers in Ohio). Initial work identified specific flow and temperature conditions that likely stimulate spawning. Egg and larval drift modeling estimated spawning locations based on egg development and flow characteristics, which helped removal crews locate and target spawning fish in the Sandusky River and later, the Maumee River. The research synthesized herein demonstrates the value of ichthyoplankton research to inform control of emerging invasive species and could inform management of other invasive carps with similar reproductive life histories, should they enter the Great Lakes.

Michigan, Ohio↗

A post-restoration evaluation of fish spawning on Coreyon Reef in Saginaw Bay, Lake Huron

Habitat diversity is important to fisheries for promoting resiliency to interannual environmental variation. Historical land use change resulting in increased sedimentation has impaired fish rock reef habitats in Saginaw Bay, Lake Huron. Pursuant to the recovery of Saginaw Bay’s walleye population, and to benefit other species, came the decision to restore the offshore habitat Coreyon Reef, a 0.8 ha rock reef complex. Following a pre-restoration assessment in 2014–2016 and construction in 2019, we conducted a comprehensive assessment from fall 2020 to spring 2022 to document the restored habitat’s viability for fish spawning. This assessment featured two objectives: (1) to evaluate reproductive usage by lake whitefish and walleye, and (2) to evaluate their corresponding egg incubation environment. We sampled directly on Coreyon Reef and at nearby unrestored control sites for comparison, with gears standardized to the pre-restoration assessment. We confirmed spawning of lake whitefish and walleye on the restored reef less than 2 years post-construction. Relative to the pre-restoration assessment, peak egg deposition increased substantially for walleye and marginally for lake whitefish. However, concurrent reproductive utilization at nearby unrestored sites was consistent or even greater than directly on the restored site. Dissolved oxygen concentration was sufficient for embryo development, but invasive round goby were abundant and consumed eggs. The fish assemblage at our sampling sites was consistent before and after restoration. We predict eggs deposited on the restored site have improved survival relative to unrestored sites, as the new reef structure may protect incubating embryos from predation and sedimentation.

Michigan↗

Defining oyster resource zones across coastal Louisiana for restoration and aquaculture

Eastern oysters ( Crassostrea virginica ) are a critical ecological and commercial resource in the northern Gulf of Mexico facing changing environmental conditions from river management and climate change. In Louisiana, USA, development of restored reefs, and off-bottom aquaculture would benefit from the identification of locations supportive of sustainable oyster populations ( i.e., metapopulations) and high consistent production. This study defines four oyster resource zones across coastal Louisiana based on environmental conditions known to affect oyster survival, growth, and reproduction. Daily data from 2015 to 2019 were interpolated to generate salinity and temperature profiles across Louisiana's estuaries, which were then used to classify zones based on monthly and annual salinity mean and variance. Zones were classified as supportive of (1) broodstock sanctuary reefs (i.e., support reproductive populations), (2) productive reefs during dry (salty) years, (3) productive reefs during wet (fresh) years, and (4) off-bottom aquaculture development. Of the 38,000 km 2 investigated, over 11,000 km 2 of potential oyster zone area was identified across the Louisiana coast. The Broodstock Sanctuary Zone was the smallest (∼540 km 2 ), as salinity variance limited this zone in many areas, as it is driven largely by riverine inputs across many estuaries. Located up-estuary (Dry Restoration Zone) and down-estuary (Wet Restoration Zone) of the Broodstock Sanctuary Zone, Dry and Wet Restoration Zone areas covered ∼2400 km 2 and ∼3900 km 2 , respectively. Mapped reefs in Louisiana currently exist largely within the Dry Restoration zones, suggesting a potential strategy to focus reef development in Wet Restoration zones to ensure reef network sustainability through years with high precipitation and river inflow. The off-bottom Aquaculture Zone was the largest (∼6400 km 2 ) zone identified, with much of this area located more down-estuary and off-shore. Accounting for variable water quality conditions enables the development of a network of reefs resilient to environmental variability, and more stable areas for consistent off-bottom aquaculture production. Spatial planning and identification of oyster resource zones reduces focus on individual reef success and supports management of oyster metapopulation outcomes, while identifying zones supportive of off-bottom aquaculture.

Louisiana↗

Environmental contaminants and biomarker responses in fish from the Rio Grande and its U.S. tributaries: Spatial and temporal trends

We collected, examined, and analyzed 368 fish of seven species from 10 sites on rivers of the Rio Grande Basin (RGB) during late 1997 and early 1998 to document temporal and geographic trends in the concentrations of accumulative contaminants and to assess contaminant effects on the fish. Sites were located on the mainstem of the Rio Grande and on the Arroyo Colorado and Pecos River in Texas (TX), New Mexico (NM), and Colorado. Common carp (Cyprinus carpio) and largemouth bass (Micropterus salmoides) were the targeted species. Fish were examined in the field for internal and external visible gross lesions, selected organs were weighed to compute ponderal and organosomatic indices, and samples of tissues and fluids were obtained and preserved for analysis of fish health and reproductive biomarkers. Whole fish from each station were composited by species and gender and analyzed for organochlorine chemical residues and elemental contaminants using instrumental methods, and for 2,3,7,8-tetrachloro dibenzo-p-dioxin-like activity (TCDD-EQ) using the H4IIE rat hepatoma cell bioassay. Overall, fish from lower RGB stations contained greater concentrations of organochlorine pesticide residues and appeared to be less healthy than those from sites in the central and upper parts of the basin, as indicated by a general gradient of residue concentrations and biomarker responses. A minimal number of altered biomarkers and few or no elevated contaminant concentrations were noted in fish from the upper RGB. The exception was elevated concentrations [up to 0.46 ??g/g wet-weight (ww)] of total mercury (Hg) in predatory species from the Rio Grande at Elephant Butte Reservoir, NM, a condition documented in previous studies. Arsenic (As) and selenium (Se) concentrations were greatest in fish from sites in the central RGB; Se concentrations in fish from the Pecos River at Red Bluff Lake, TX and from the Rio Grande at Langtry, TX and Amistad International Reservoir, TX exceeded published fish and wildlife toxicity thresholds. In the lower RGB, residues of p,p???-DDT metabolites (???1.69 ??g/g ww), chlordane-related compounds (???0.21 ??g/g ww), dieldrin (???0.0.05 ??g/g ww), and toxaphene (???2.4 ??g/g ww) were detected in fish from most sites; maximum concentrations were in channel catfish (Ictalurus punctatus) from the Arroyo Colorado at Harlingen, TX. Concentrations of one or more residues exceeded toxicity thresholds for fish and wildlife in fish from this site and from the Rio Grande at Mission, TX and Brownsville, TX; however, concentrations were lower than those reported by previous studies. In addition, the proportional concentrations of p,p???-DDT at all sites were low, indicating weathered DDT rather than the influx of new material. Concentrations of total PCBs (<0.05 ??g/g ww) and TCDD-EQ (???6 pg/g ww) were comparatively low in all samples. Hepatic ethoxyresorufin O-deethylase (EROD) activity in some fish was elevated relative to reference rates at most sites, but was generally lower than previously reported activity in fish from heavily contaminated locations. The comparatively low PCB and TCDD-EQ concentrations together with elevated EROD activity may reflect exposure to polycyclic aromatic hydrocarbons. Reproductive biomarkers were consistent with chronic contaminant exposure at lower RGB sites; comparatively large percentages of intersex male largemouth bass, relatively low gonadosomatic indices, and elevated plasma vitellogenin concentrations in male fish were noted at three of the four stations. Large percentages of atretic eggs were also observed in the ovaries of female common carp from the Rio Grande at Brownsville, TX. Although many of the conditions noted may have other causes in addition to contaminant exposure, the biomarker results for the lower RGB sites are consistent with subtle responses of fish to contaminants, an interpretation supported by the chemical data of this and other investigations.

Science of the Total Environment↗

Environmental contaminants and biomarker responses in fish from the Columbia River and its tributaries: spatial and temporal trends

Fish were collected from 16 sites on rivers in the Columbia River Basin (CRB) from September 1997 to April 1998 to document temporal and spatial trends in the concentrations of accumulative contaminants and to assess contaminant effects on the fish. Sites were located on the mainstem of the Columbia River and on the Snake, Willamette, Yakima, Salmon, and Flathead Rivers. Common carp (Cyprinus carpio), black bass (Micropterus sp.), and largescale sucker (Catostomus macrocheilus) were the targeted species. Fish were field-examined for external and internal lesions, selected organs were weighed to compute somatic indices, and tissue and fluid samples were preserved for fish health and reproductive biomarker analyses. Composite samples of whole fish, grouped by species and gender, from each site were analyzed for organochlorine and elemental contaminants using instrumental methods and for 2,3,7,8-tetrachloro dibenzo-p-dioxin-like activity (TCDD-EQ) using the H4IIE rat hepatoma cell bioassay. Overall, pesticide concentrations were greatest in fish from lower CRB sites and elemental concentrations were greatest in fish from upper CRB sites. These patterns reflected land uses. Lead (Pb) concentrations in fish from the Columbia River at Northport and Grand Coulee, Washington (WA) exceeded fish and wildlife toxicity thresholds (> 0.4 ??g/g). Selenium (Se) concentrations in fish from the Salmon River at Riggins, Idaho (ID), the Columbia River at Vernita Bridge, WA, and the Yakima River at Granger, WA exceeded toxicity thresholds for piscivorous wildlife (> 0.6 ??g/g). Mercury (Hg) concentrations in fish were elevated throughout the basin but were greatest (> 0.4 ??g/g) in predatory fish from the Salmon River at Riggins, ID, the Yakima River at Granger, WA, and the Columbia River at Warrendale, Oregon (OR). Residues of p,p???-DDE were greatest (> 0.8 ??g/g) in fish from agricultural areas of the Snake, Yakima, and Columbia River basins but were not detected in upper CRB fish. Other organochlorine pesticides did not exceed toxicity thresholds in fish or were detected infrequently. Total polychlorinated biphenyls (PCBs; > 0.11 ??g/g) and TCDD-EQs (> 5 pg/g) exceeded wildlife guidelines in fish from the middle and lower CRB, and ethoxyresorufin O-deethylase (EROD) activity was also elevated at many of the same sites. Temporal trend analysis indicated decreasing or stable concentrations of Pb, Se, Hg, p,p???-DDE, and PCBs at most sites where historical data were available. Altered biomarkers were noted in fish throughout the CRB. Fish from some stations had responded to chronic contaminant exposure as indicated by fish health and reproductive biomarker results. Although most fish from some sites had grossly visible external or internal lesions, histopathological analysis determined these to be inflammatory responses associated with helminth or myxosporidian parasites. Many largescale sucker from the Columbia River at Northport and Grand Coulee, WA had external lesions and enlarged spleens, which were likely associated with infections. Intersex male smallmouth bass (Micropterus dolomieu) were found in the Snake River at Lewiston, ID and the Columbia River at Warrendale, OR. Male bass, carp, and largescale sucker containing low concentrations of vitellogenin were common in the CRB, and comparatively high concentrations (> 0.3 mg/mL) were measured in male fish from the Flathead River at Creston, Montana, the Snake River at Ice Harbor Dam, WA, and the Columbia River at Vernita Bridge, WA and Warrendale, OR. Results from our study and other investigations indicate that continued monitoring in the CRB is warranted to identify consistently degraded sites and those with emerging problems. ?? 2005 Elsevier B.V. All rights reserved.

Columbia River Basin↗

Effects of biologically-active chemical mixtures on fish in a wastewater-impacted urban stream

Stream flow in urban aquatic ecosystems often is maintained by water-reclamation plant (WRP) effluents that contain mixtures of natural and anthropogenic chemicals that persist through the treatment processes. In effluent-impactedstreams, aquatic organisms such as fish are continuously exposed to biologically-activechemicals throughout their life cycles. The North Shore Channel of the Chicago River (Chicago, Illinois) is part of an urban ecosystem in which > 80% of the annual flow consists of effluent from the North Side WRP. In this study, multiple samplings of the effluent and stream water were conducted and fish (largemouth bass and carp) were collected on 2 occasions from the North Shore Channel. Fish also were collected once from the Outer Chicago Harbor in Lake Michigan, a reference site not impacted by WRP discharges. Over 100 organic chemicals with differing behaviors and biological effects were measured, and 23 compounds were detected in all of the water samples analyzed. The most frequently detected and highest concentration (> 100 &mu;g/L) compounds were ethylenediaminetetraacetic acid and 4-nonylphenolmono-to-tetraethoxycarboxylic acids. Other biologically-activechemicals including bisphenol A, 4-nonylphenol, 4-nonylphenolmono-to-tetraethoxylates, 4- tert -octylphenol, and 4- tert -octylphenolmono-to-tetraethoxylates were detected at lower concentrations (< 5 &mu;g/L). The biogenic steroidal hormones 17&beta;-estradiol, estrone, testosterone, 4-androstene-3,17-dione, and cis -androsterone were detected at even lower concentrations (< 0.005 &mu;g/L). There were slight differences in concentrations between the North Side WRP effluent and the North Shore Channel, indicating minimal in-stream attenuation. Fish populations are continuously exposed to mixtures of biologically-activechemicals because of the relative persistency of the chemicals with respect to stream hydraulic residence time, and the lack of a fresh water source for dilution. The majority of male fish exhibited vitellogenin induction, a physiological response consistent with exposure to estrogenic compounds. Tissue-level signs of reproductive disruption, such as ovatestis, were not observed.

Illinois↗

Earthworm bioassays and seedling emergence for monitoring toxicity, aging and bioaccumulation of anthropogenic waste indicator compounds in biosolids-amended soil

Land application of biosolids (treated sewage sludge) can be an important route for introducing xenobiotic compounds into terrestrial environments. There is a paucity of available information on the effects of biosolids amendment on terrestrial organisms. In this study, the influence of biosolids and biosolids aging on earthworm (Eisenia fetida) reproduction and survival and lettuce (Lactuca sativa) seedling emergence was investigated. Earthworms were exposed to soils amended with varying quantities of biosolids (0, 1, 2, 3, or 4% dry mass). To investigate the influence of biosolids aging, the biosolids used in the study were aged for differing lengths of time (2 or 8 weeks) prior to exposure. All of the adult earthworms survived in the biosolids–amended soils at all concentrations that were aged for 2 weeks; however, only 20% of the adults survived in the soil amended with the highest concentration of biosolids and aged for 8 weeks. Reproduction as measured by mean number of juveniles and unhatched cocoons produced per treatment correlated inversely with biosolids concentration, although the effects were generally more pronounced in the 8-week aged biosolids–soil samples. Latent seedling emergence and reduced seedling fitness correlated inversely with biosolids concentration, but these effects were tempered in the 8-week aged versus the 2-week aged soil–biosolids mixtures. Anthropogenic waste indicator compounds (AWIs) were measured in the biosolids, biosolids–soil mixtures, and earthworm samples. Where possible, bioaccumulation factors (BAFs) were calculated or estimated. A wide variety of AWIs were detected in the biosolids (51 AWIs) and earthworm samples (≤ 19 AWI). The earthworms exposed to the 8-week aged biosolids–soil mixtures tended to accumulate greater quantities of AWIs compared to the 2-week aged mixture, suggesting that the bioavailability of some AWIs was enhanced with aging. The BAFs for a given AWI varied with treatment. Notably large BAFs were determined for some AWIs. For example, the maximum BAF determined for para-cresol, methyl salicylate, bisphenol-A, and cholesterol was greater than 100 in some treatments.

Science of the Total Environment↗

Current state of knowledge on biological effects from contaminants on arctic wildlife and fish

Since the last Arctic Monitoring and Assessment Programme (AMAP) effort to review biological effects of the exposure to organohalogen compounds (OHCs) in Arctic biota, there has been a considerable number of new Arctic effect studies. Here, we provide an update on the state of the knowledge of OHC, and also include mercury, exposure and/or associated effects in key Arctic marine and terrestrial mammal and bird species as well as in fish by reviewing the literature published since the last AMAP assessment in 2010. We aimed at updating the knowledge of how single but also combined health effects are or can be associated to the exposure to single compounds or mixtures of OHCs. We also focussed on assessing both potential individual as well as population health impacts using population-specific exposure data post 2000. We have identified quantifiable effects on vitamin metabolism, immune functioning, thyroid and steroid hormone balances, oxidative stress, tissue pathology, and reproduction. As with the previous assessment, a wealth of documentation is available for biological effects in marine mammals and seabirds, and sentinel species such as the sledge dog and Arctic fox, but information for terrestrial vertebrates and fish remain scarce. While hormones and vitamins are thoroughly studied, oxidative stress, immunotoxic and reproductive effects need further investigation. Depending on the species and population, some OHCs and mercury tissue contaminant burdens post 2000 were observed to be high enough to exceed putative risk threshold levels that have been previously estimated for non-target species or populations outside the Arctic. In this assessment, we made use of risk quotient calculations to summarize the cumulative effects of different OHC classes and mercury for which critical body burdens can be estimated for wildlife across the Arctic. As our ultimate goal is to better predict or estimate the effects of OHCs and mercury in Arctic wildlife at the individual, population and ecosystem level, there remain numerous knowledge gaps on the biological effects of exposure in Arctic biota. These knowledge gaps include the establishment of concentration thresholds for individual compounds as well as for realistic cocktail mixtures that in fact indicate biologically relevant, and not statistically determined, health effects for specific species and subpopulations. Finally, we provide future perspectives on understanding Arctic wildlife health using new in vivo, in vitro, and in silico techniques, and provide case studies on multiple stressors to show that future assessments would benefit from significant efforts to integrate human health, wildlife ecology and retrospective and forecasting aspects into assessing the biological effects of OHC and mercury exposure in Arctic wildlife and fish.

Science of the Total Environment↗

PCB exposure is associated with reduction of endosymbionts in riparian spider microbiomes

Microbial communities, including endosymbionts, play diverse and critical roles in host biology and reproduction, but contaminant exposure may cause an imbalance in the microbiome composition with subsequent impacts on host health. Here, we examined whether there was a significant alteration of the microbiome community within two taxa of riparian spiders (Tetragnathidae and Araneidae) from a site with historical polychlorinated biphenyl (PCB) contamination in southern Ontario, Canada. Riparian spiders specialize in the predation of adult aquatic insects and, as such, their contaminant levels closely track those of nearby aquatic ecosystems. DNA from whole spiders from sites with either low or high PCB contamination was extracted, and spider microbiota profiled by partial 16S rRNA gene amplicon sequencing. The most prevalent shift in microbial communities we observed was a large reduction in endosymbionts in spiders at the high PCB site. The abundance of endosymbionts at the high PCB site was 63 % and 98 % lower for tetragnathids and araneids, respectively, than at the low PCB site. Overall, this has potential implications for spider reproductive success and food webs, as riparian spiders are critical gatekeepers of energy and material fluxes at the land-water interface.

Science of the Total Environment↗

Multi-omic responses of fish exposed to complex chemical mixtures in the Shenandoah River watershed

To evaluate relationships between different anthropogenic impacts, contaminant occurrence, and fish health, we conducted in situ fish exposures across the Shenandoah River watershed at five sites with different land use. Exposure water was analyzed for over 500 chemical constituents, and organismal, metabolomic, and transcriptomic endpoints were measured in fathead minnows. Adverse reproductive outcomes were observed in fish exposed in the upper watershed at both wastewater treatment plant (WWTP) effluent- and agriculture-impacted sites, including decreased gonadosomatic index and altered secondary sex characteristics. This was accompanied with increased mortality at the site most impacted by agricultural activities. Molecular biomarkers of estrogen exposure were unchanged and consistent with low or non-detectable concentrations of common estrogens, indicating that alternative mechanisms were involved in organismal adverse outcomes. Hepatic metabolomic and transcriptomic profiles were altered in a site-specific manner, consistent with variation in land use and contaminant profiles. Integrated biomarker response data were useful for evaluating mechanistic linkages between contaminants and adverse outcomes, suggesting that reproductive endocrine disruption, altered lipid processes, and immunosuppression may have been involved in these organismal impacts. This study demonstrated linkages between human-impact, contaminant occurrence, and exposure effects in the Shenandoah River watershed and showed increased risk of adverse outcomes in fathead minnows exposed to complex mixtures at sites impacted by municipal wastewater discharges and agricultural practices.

Virginia, West Virginia↗

Flow cytometric method for measuring chromatin fragmentation in fixed sperm from yellow perch ( Perca flavescens )

Declining harvests of yellow perch, Perca flavescens , in urbanized watersheds of Chesapeake Bay have prompted investigations of their reproductive fitness. The purpose of this study was to establish a flow cytometric technique for DNA analysis of fixed samples sent from the field to provide reliable gamete quality measurements. Similar to the sperm chromatin structure assay, measures were made on the susceptibility of nuclear DNA to acid-induced denaturation, but used fixed rather than live or thawed cells. Nuclei were best exposed to the acid treatment for 1 minute at 37 &deg;C followed by the addition of cold (4 &deg;C) propidium iodide staining solution before flow cytometry. The rationale for protocol development is presented graphically through cytograms. Field results collected in 2008 and 2009 revealed DNA fragmentation up to 14.5%. In 2008, DNA fragmentation from the more urbanized watersheds was significantly greater than from reference sites (P = 0.026) and in 2009, higher percentages of haploid testicular cells were noted from the less urbanized watersheds (P = 0.032) indicating better reproductive condition at sites with less urbanization. For both years, total and progressive live sperm motilities by computer-assisted sperm motion analysis ranged from 19.1% to 76.5%, being significantly higher at the less urbanized sites (P < 0.05). This flow cytometric method takes advantage of the propensity of fragmented DNA to be denatured under standard conditions, or 1 minute at 37 &deg;C with 10% buffered formalin&ndash;fixed cells. The study of fixed sperm makes possible the restrospective investigation of germplasm fragmentation, spermatogenic ploidy patterns, and chromatin compaction levels from samples translocated over distance and time. The protocol provides an approach that can be modified for other species across taxa.

Theriogenology↗

Environmental enrichment affects adrenocortical stress responses in the endangered black-footed ferret

Potential stressors of wildlife living in captivity, such as artificial living conditions and frequent human contact, may lead to a higher occurrence of disease and reduced reproductive function. One successful method used by wildlife managers to improve general well-being is the provision of environmental enrichment, which is the practice of providing animals under managed care with environmental stimuli. The black-footed ferret ( Mustela nigripes ) is a highly-endangered carnivore species that was rescued from extinction by removal of the last remaining individuals from the wild to begin an ex situ breeding program. Our goal was to examine the effect of environmental enrichment on adrenocortical activity in ferrets by monitoring fecal glucocorticoid metabolites (FGM). Results demonstrated that enrichment lowered FGM in juvenile male ferrets, while increasing it in adult females; enrichment had no effect on FGM in juvenile females and adult males. These results correspond with our findings that juvenile males interacted more with the enrichment items than did adult females. However, we did not detect an impact of FGM on the incidence of disease or on the ability of ferrets to become reproductive during the following breeding season. We conclude that an environmental enrichment program could benefit captive juvenile male ferrets by reducing adrenocortical activity.

General and Comparative Endocrinology↗

California gull chicks raised near colony edges have elevated stress levels

Coloniality in nesting birds represents an important life history strategy for maximizing reproductive success. Birds nesting near the edge of colonies tend to have lower reproductive success than individuals nesting near colony centers, and offspring of edge-nesting parents may be impaired relative to those of central-nesting parents. We used fecal corticosterone metabolites in California gull chicks (Larus californicus) to examine whether colony size or location within the colony influenced a chick's physiological condition. We found that chicks being raised near colony edges had higher fecal corticosterone metabolite concentrations than chicks raised near colony centers, but that colony size (ranging from 150 to 11,554 nests) had no influence on fecal corticosterone levels. Fecal corticosterone metabolite concentrations also increased with chick age. Our results suggest that similarly aged California gull chicks raised near colony edges may be more physiologically stressed, as indicated by corticosterone metabolites, than chicks raised near colony centers.

General and Comparative Endocrinology↗

Low estradiol production of non-laying whooping cranes (Grus americana) is associated with the failure of small follicles to enter follicular hierarchy

For endangered species managed ex situ , production of offspring is a key factor to ensure healthy and self-sustaining populations. However, current breeding goals for the whooping crane ( Grus americana ) are impeded by poor reproduction. Our study sought to better understand mechanisms regulating ovarian function in ex situ managed whooping cranes and the regulatory function of the hypothalamic-pituitary-gonadal (HPG) axis in relation to follicle formation and egg laying. To characterize hormonal regulation of follicular development and ovulation, we collected weekly blood samples from six female whooping cranes during two breeding seasons, for a total of 11 reproductive cycles. The plasma samples were assessed for follicle stimulating hormone, luteinizing hormone, estradiol, and progesterone and the yolk precursors vitellogenin and very low-density lipoprotein. Ultrasonographic examination of the ovary was conducted at the time of blood collection. Preovulatory follicles (>12 mm) were present in laying cycles (n = 6) but absent in non-laying cycles (n = 5). The patterns of plasma hormone and yolk precursor concentrations corresponded to the stage of follicle development. Specifically, gonadotropin and yolk precursors concentrations increased as follicles transitioned from the non-yolky to yolky stage but did not increase further as the follicle advanced to preovulatory and ovulatory stages. Estrogen and progesterone concentrations increased as follicle size increased and reached peak concentrations (P < 0.05) when follicles developed to ovulatory and preovulatory stages, respectively. While overall mean circulating gonadotropin, progesterone, and yolk precursor concentrations did not differ for laying versus non-laying cycles, mean plasma estradiol in laying cycles was significantly higher than that in non-laying cycles. In summary, the findings suggested that disruption of mechanisms regulating follicle recruitment is likely responsible for the oviposition failure of the captive female whooping crane.

General and Comparative Endocrinology↗

Decreased glutathione S-transferase expression and activity and altered sex steroids in Lake Apopka brown bullheads (Ameriurus nebulosus)

A number of freshwater lakes and reclaimed agricultural sites in Central Florida have been the receiving waters for agrochemical and municipal runoff. One of these sites, Lake Apopka, is also a eutrophic system that has been the focus of several case studies reporting altered reproductive activity linked to bioaccumulation of persistent organochlorine chemicals in aquatic species. The present study was initiated to determine if brown bullheads (Ameriurus nebulosus) from the north marsh of Lake Apopka (Lake Apopka Marsh) exhibit an altered capacity to detoxify environmental chemicals through hepatic glutathione S-transferase (GST)-mediated conjugation as compared with bullheads from a nearby reference site (Lake Woodruff). We also compared plasma sex hormone concentrations (testosterone, 17-?? estradiol, and 11 keto-testosterone) in bullheads from the two sites. Female bullheads from Lake Apopka had 40% lower initial rate GST conjugative activity toward 1-chloro-2,4-dinitrobenzene (CDNB), 50% lower activity towards p-nitrobutyl chloride (NBC), 33% lower activity toward ethacrynic acid (ECA), and 43% lower activity toward ??5-androstene-3,17-dione (??5-ADI), as compared with female bullheads from Lake Woodruff. Enzyme kinetic analyses demonstrated that female bullheads from Lake Apopka had lower GST-catalyzed CDNB clearance than did female Lake Woodruff bullheads. Western blotting studies of bullhead liver cytosolic proteins demonstrated that the reduced GST catalytic activities in female Lake Apopka bullheads were accompanied by lower expression of hepatic GST protein. No site differences were observed with respect to GST activities or GST protein expression in male bullheads. Female Lake Apopka bullheads also had elevated concentrations of plasma androgens (testosterone and 11-ketotestosterone) as compared with females from Lake Woodruff. In contrast, male Lake Apopka bullheads had elevated levels of plasma estrogen but similar levels of androgens as compared with male bullheads from Lake Woodruff. Collectively, our studies indicate the presence of reduced GST protein expression, reduced GST conjugative capacity and altered sex steroid homeostasis in female bullheads from a contaminated field site in Central Florida. The implications of these physiological alterations in terms of pollutant biotransformation and reproduction are discussed. ?? 2001 Elsevier Science B.V. All rights reserved.

Aquatic Toxicology↗

Role of selenium toxicity and oxidative stress in aquatic birds

Adverse effects of selenium (Se) in wild aquatic birds have been documented as a consequence of pollution of the aquatic environment by subsurface agricultural drainwater and other sources. These effects include mortality, impaired reproduction with teratogenesis, reduced growth, histopathological lesions and alterations in hepatic glutathione metabolism. A review is provided, relating adverse biological effects of Se in aquatic birds to altered glutathione metabolism and oxidative stress. Laboratory studies, mainly with an organic form of Se, selenomethionine, have revealed oxidative stress in different stages of the mallard (Anas platyrhynchos) life cycle. As dietary and tissue concentrations of Se increase, increases in plasma and hepatic GSH peroxidase activities occur, followed by dose-dependent increases in the ratio of hepatic oxidized to reduced glutathione (GSSG:GSH) and ultimately hepatic lipid peroxidation measured as an increase in thiobarbituric acid reactive substances (TBARS). One or more of these oxidative effects were associated with teratogenesis (4.6 ppm wet weight Se in eggs), reduced growth in ducklings (15 ppm Se in liver), diminished immune function (5 ppm Se in liver) and histopathological lesions (29 ppm Se in liver) in adults. Manifestations of Serelated effects on glutathione metabolism were also apparent in field studies in seven species of aquatic birds. Reduced growth and possibly immune function but increased liver:body weight and hepatic GSSG:GSH ratios were apparent in American avocet (Recurvirostra americana) hatchlings from eggs containing 9 ppm Se. In blacknecked stilts (Himantopus mexicanus), which contained somewhat lower Se concentrations, a decrease in hepatic GSH was apparent with few other effects. In adult American coots (Fulica americana), signs of Se toxicosis included emaciation, abnormal feather loss and histopathological lesions. Mean liver concentrations of 28 ppm Se (ww) in the coots were associated with elevated hepatic GSH peroxidase, depletion of hepatic protein bound thiols and total thiols, but a small increase in GSH. Diving ducks in the San Francisco Bay area exhibited a positive correlation between hepatic Se concentration and GSH peroxidase activity (r=0.63, P<0.05), but a negative correlation between hepatic Se and GSH concentration (r=0.740, P<0.05). In willets (Catoptrophorus semipalmatus) from the San Diego area, positive correlations occurred between hepatic Se concentration and GSSG (r=0.70, P<0.001), GSSG:GSH ratio, and TBARS. In emperor geese (Chen canagica) from western Alaska, blood levels of up to 9.4 ppm occurred and were associated with increased plasma GSH peroxidase activity (r=0.62, P<0.001), but with decreased plasma GSSG reductase activity. When evaluating Se toxicity, interactive nutritional factors, including other elements and dietary protein, should also be taken into consideration. Further studies are needed to examine the relationship between different forms of environmentally occurring selenium, arsenic and mercury on reproduction, hepatotoxicity and immune function of aquatic birds. Further selenium nutritional interaction studies may also help to illucidate the mechanism of selenium induced teratogenesis, by optimizing GSH and other antioxidant defense mechanisms in a manner that would stabilize or raise the cell's threshold for susceptibility to toxic attack from excess selenium. It is concluded that Se-related manifestations of oxidative stress may serve as useful bioindicators of Se exposure and toxicity in wild aquatic birds.

Aquatic Toxicology↗