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M. S. Sepulveda

Publications and source records attributed to M. S. Sepulveda.

15 recordsLinked to original sources

Experimental West Nile virus infection in Eastern Screech Owls (Megascops asio)

This study evaluated the potential effects of different concentrations of bleached/unbleached kraft mill effluent (B/UKME) on several reproductive endpoints in adult largemouth bass ( Micropterus salmoides ). The kraft mill studied produces a 50/50 mix of bleached/unbleached market pulp with an estimated release of 36 million gal of effluent/day. Bleaching sequences were C 90 d 10 EopHDp and CEHD for softwood (pines) and hardwoods (mainly tupelo, gums, magnolia, and water oaks), respectively. Bass were exposed to different effluent concentrations (0 [controls, exposed to well water], 10, 20, 40, or 80%) for either 28 or 56 days. At the end of each exposure period, fish were euthanized, gonads collected for histological evaluation and determination of gonadosomatic index (GSI), and plasma was analyzed for 17β-estradiol, 11-ketotestosterone, and vitellogenin (VTG). Largemouth bass exposed to B/UKME responded with changes at the biochemical level (decline in sex steroids in both sexes and VTG in females) that were usually translated into tissue/organ-level responses (declines in GSI in both sexes and in ovarian development in females). Although most of these responses occurred after exposing fish to 40% B/UKME concentrations or greater, some were observed after exposures to 20% B/UKME. These threshold concentrations fall within the 60% average yearly concentration of effluent that exists in the stream near the point of discharge (Rice Creek), but are above the <10% effluent concentration present in the St. Johns River. The chemical(s) responsible for such changes as well as their mode(s) of action remain unknown at this time.

Florida↗

Dietary exposure of largemouth bass to OCPs changes expression of genes important for reproduction

Dieldrin and p,p???-DDE are ubiquitous contaminants known to act as endocrine disruptors, causing impaired development and reproduction in fish and wildlife. In order to elucidate the mechanisms by which dieldrin and p,p???-DDE cause endocrine disruption in largemouth bass (Micropterus salmoides), fish were exposed subchronically through the diet to both contaminants. Following 120 days of exposure, p,p???-DDE decreased estradiol in females, but increased 11-ketotestosterone in both sexes. Dieldrin on the other hand, decreased estradiol and 11-ketotestosterone in both sexes. Both pesticides also altered steady state mRNA expression levels of a set of genes chosen to represent three possible mechanisms of endocrine disruption: (1) direct interaction with soluble sex steroid receptors, (2) biosynthesis of endogenous sex hormones, and (3) metabolism of endogenous hormones. p,p???-DDE acted as a weak estrogen, increasing the expression of vitellogenin and estrogen receptor ?? in the liver. p,p???-DDE also altered the expression of genes involved in the synthesis of endogenous hormones as well as their metabolism. Dieldrin, on the other hand, only altered expression of vitellogenin and not estrogen receptor ??. Dieldrin also altered the expression of genes involved in hormone synthesis and metabolism, and it dramatically lowered plasma hormone levels. Both pesticides targeted expression of genes involved in all three modes of action, suggesting that they each have multiple modes of action. ?? 2006 Elsevier B.V. All rights reserved.

Aquatic Toxicology↗

Necropsy findings in American alligator late-stage embryos and hatchlings from northcentral Florida lakes contaminated with organochlorine pesticides

Increased American alligator (Alligator mississippiensis) embryo and neonatal mortality has been reported from several northcentral Florida lakes contaminated with old-use organochlorine pesticides (OCPs). However, a clear relationship among these contaminants and egg viability has not been established, suggesting the involvement of additional factors in these mortalities. Thus, the main objective of this study was to determine the ultimate cause of mortality of American alligator late-stage embryos and hatchlings through the conduction of detailed pathological examinations, and to evaluate better the role of OCPs in these mortalities. Between 2000 and 2001, 236 dead alligators were necropsied at or near hatching (after ???65 days of artificial incubation and up to 1 mo of age posthatch). Dead animals were collected from 18 clutches ranging in viability from 0% to 95%. Total OCP concentrations in yolk ranged from ???100 to 52,000 ??g/kg, wet weight. The most common gross findings were generalized edema (34%) and organ hyperemia (29%), followed by severe emaciation (14%) and gross deformities (3%). Histopathologic examination revealed lesions in 35% of the animals, with over half of the cases being pneumonia, pulmonary edema, and atelectasis. Within and across clutches, dead embryos and hatchlings compared with their live cohorts were significantly smaller and lighter. Although alterations in growth and development were not related to yolk OCPs, there was an increase in prevalence of histologic lesions in clutches with high OCPs. Overall, these results indicate that general growth retardation and respiratory abnormalities were a major contributing factor in observed mortalities and that contaminants may increase the susceptibility of animals to developing certain pathologic conditions. ?? Wildlife Disease Association 2006.

Journal of Wildlife Diseases↗

Predicting maternal body burdens of organochlorine pesticides from eggs and evidence of maternal transfer in Alligator mississippiensis

Few data exist regarding maternal-embryonal transfer of organochlorine pesticides (OCPs) in reptiles. The objective of the present study was to evaluate maternal transfer of OCPs in American alligators (Alligator mississippiensis) from low-, intermediate-, and high-OCP-exposure sites. Overall, total OCP burdens ranged from less than 0.8 ppb in blood to more than 44,000 ppb in abdominal adipose tissue (wet wt concentrations). Lipid-adjusted ratios of maternal adipose burdens (total OCPs) to yolk burdens were close to one (0.94 ?? 0.31:1), suggesting that animals were in steady state and that OCPs in eggs originated from adipose lipids. In contrast, lipid-adjusted muscle and liver OCP burdens were greater than yolk OCP burdens, suggesting that lipids in muscle were not utilized during oogenesis and that nonlipid liver tissue sequesters OCPs. Predictive equations were derived for several tissues and several OCP analytes with r2 values ranging from 0.40 to 0.99 (p < 0.05). We suggest that yolk burdens are predictive of maternal tissue burdens for certain tissues and OCPs and that certain OCPs are maternally transferred in the American alligator. Furthermore, we suggest that future studies should investigate the applicability of these predictive equations for assessing maternal exposure in other crocodilian species.

Environmental Toxicology and Chemistry↗

Umbilical scarring in hatchling American alligators

Umbilical scarring is the presence of excess scar tissue deposited between abdominal dermal layers at the site of yolk sac absorption in hatchling American alligators (Alligator mississippiensis). The presence of this dermal condition plays a key evaluatory role in the overall quality and subsequent value for various commercial leather products. Despite the prevalent nature of this condition, currently the industry has no standardized protocols for its quantification. The objectives of this study were to examine the relationship between hatchling weight and age and incidence of umbilical scarring and to develop a quantifiable and reproducible technique to measure this dermal condition in hatchling American alligators. Thirty eggs from each of nine clutches were incubated in two separate incubators at different facilities and hatchling umbilical scarring was measured at 2 and 10 days of age using digital calipers. Umbilical area was calculated by multiplying umbilical length times umbilical width. There was a significant effect of both age and clutch on umbilical area (overall decline of 64%) by 10 days post-hatch. However, only five of the nine clutches utilized expressed a noticeable decline in the size of this dermal condition (range 67-74%). We had hypothesized that larger hatchlings would have larger umbilical areas and a slower rate of improvement in this condition during the first few days post-hatch. The differences in umbilical area and percent decline with age across clutches, however, were not associated with differences in initial hatchling weights. Within clutches and time periods, hatchling weight had no significant effect on the size and/or rate of decline of this condition. ?? 2004 Published by Elsevier B.V.

Aquaculture↗

Reproductive and biochemical biomarkers in largemouth bass sampled downstream of a pulp and paper mill in Florida

The objective of this study was to evaluate the effects of bleached/unbleached kraft mill effluents (B/UKME) on the reproductive parameters of free-ranging Florida largemouth bass (Micropterus salmoides floridanus). The reproductive parameters measured included gonadosomatic index (GSI), histological evaluation of gonads, and plasma concentrations of vitellogenin (VTG), 17??-estradiol, and 11-ketotestosterone (11-KT). Hepatic ethoxyresorufin-O-deethylase (EROD) activity was measured as a marker of exposure to cytochrome P450-inducing agents in these effluents. Endpoints were compared among adult bass sampled from tributary and mainstream effluent-contaminated and reference sites. Females sampled from the site closest to the mill outfall had a significant five-fold increase in EROD activity compared to bass sampled from reference streams. Although sex hormones were significantly reduced in bass from exposed sites, there were no differences in VTG and GSI across sites. The absence of organism-level responses was probably not related to a lack of sensitivity, as previous studies in our laboratory have shown that bass exposed to these effluents exhibit changes in GSI and in other measures associated with reproductive success. In females, inverse relationships were observed between VTG and GSI and EROD activity. These relationship, however, were not consistent within all of the sites studied. Collectively, our findings indicate that hepatic EROD induction is an effective marker of B/UKME exposure in largemouth bass and that it might be associated with antiestrogenic effects in this species. ?? 2003 Elsevier Inc. All rights reserved.

Ecotoxicology and Environmental Safety↗

Achieving environmentally relevant organochlorine pesticide concentrations in eggs through maternal exposure in Alligator mississippiensis

Alligator mississippiensis eggs from organochlorine pesticide (OCP) contaminated sites in Florida exhibit high rates of embryonic mortality compared to reference sites (p<0.05). The objective of the present study was to use captive adult alligators to test the hypotheses that maternal exposure to OCPs results in increased OCP concentrations in eggs, and that increased exposure is associated with increased embryonic mortality. A total of 24 adult alligators (8 males and 16 females) were housed in eight pens. Eight females in four pens were dosed with a mixture of p,p'-DDE, toxaphene, dieldrin, and chlordane at a rate of 0.2 ? 0.01 mg/kg/day for 274 ? 8 days. Treated females produced eggs containing higher OCP concentrations (12,814 ? 813 ng/g yolk) than controls (38 ? 4 ng/g yolk). Eggs of treated females exhibited decreased viability (13 ? 22%) as compared to controls (45 ? 20%). Results indicated that 0.6% of administered OCPs were maternally transferred to the eggs of American alligators, and that maternal exposure is associated with decreased egg/embryo viability in this species.

Marine Environmental Research↗

Gene expression fingerprints of largemouth bass (Micropterus salmoides) exposed to pulp and paper mill effluents

Effluents from pulp and paper mills that historically have used elemental chlorine in the bleaching process have been implicated in inhibiting reproduction in fish. Compounds with estrogenic and androgenic binding affinities have been found in these effluents, suggesting that the impairment of reproduction is through an endocrine-related mode of action. To date, a great deal of attention has been paid to phytoestrogens and resin acids that are present in mill process streams as a result of pulping trees. Estrogen and estrogen mimics interact directly with the estrogen receptor and have near immediate effects on gene transcription by turning on the expression of a unique set of genes. Using differential display (DD) RT-PCR, we examined changes in gene expression induced by exposure to paper mill effluents. Largemouth bass were exposed to 0, 10, 20, 40, and 80% paper mill effluent concentrations in large flow-through tanks for varied periods of time including 7, 28 or 56 days. Plasma hormone levels in males and females and plasma vitellogenin (Vtg) in females decreased with dose and time. Measurements of changes in gene expression using DD RT-PCR suggest that the gene expression patterns of male fish do not change much with exposure, except for the induction of a few genes including CYP 1A, a protein that is induced through the action of the Ah receptor in response to dioxin and similar polyaromatic hydrocarbons. However, in the case of females, exposure to these effluents resulted in an up-regulation of CYP 1A that was accompanied by a generalized down-regulation of genes normally expressed during the reproductive season. These antiestrogenic changes are in agreement with previous studies in bass exposed to these effluents, and could result in decreased reproductive success in affected populations. ?? 2004 Elsevier B.V. All rights reserved.

Mutation Research - Fundamental and Molecular Mech↗

Physiological changes in largemouth bass exposed to paper mill effluents under laboratory and field conditions

We report here on studies designed to asses the effects of paper mill effluents on non-reproductive functions of free-ranging and captive Florida largemouth bass (Micropterus salmoides floridanus) This was accomplished by conducting an outdoor tank study, in which fish were exposed to well water or to 10%, 20%, 40%, and 80% full strength effluent for 28 or 56 days, and by sampling largemouth bass from sites within the St. Johns River, Florida, upstream and downstream from a paper mill plant. Blood and plasma samples from fish from the tank study and from fish sampled from the ambient sites were analyzed for over 20 variables. We also determined liver and spleen weights and examined them histologically. The most significant finding from the tank study was an increase in the concentration of albumin and hepatosomatic index for bass exposed to ???20% effluents for 56 days. Spleenosomatic index and number of melanomacrophage centers were decreased in bass from effluent-dominated sites (Palatka and Rice Creek), whereas concentrations of calcium, phosphorous, glucose, and creatinine were elevated in fish from these sites, compared to fish from reference streams. Fish from Rice Creek also had fewer red blood cells, and male bass from Palatka had lower concentrations of cholesterol. Plasma concentrations of albumin and hepatic concentrations of glutathione were elevated in males from Palatka, and both females and males from Rice Creek had higher concentrations of globulin. These results indicate a complex pattern of effects of paper mill effluents on several physiological functions. However, despite the myriad of treatment and site-related effects, most physiological parameters fell within normal ranges when compared to reports on largemouth bass and other freshwater species.

Ecotoxicology↗

Organochlorine pesticides and thiamine in eggs of largemouth bass and American alligators and their relationship with early life-stage mortality

Thiamine deficiency has been linked to early mortality syndrome in salmonids in the Great Lakes. This study was conducted to compare thiamine concentrations in American alligators ( Alligator mississippiensis ) and Florida largemouth bass ( Micropterus salmoides floridanus ) eggs from sites with high embryo mortality and high exposure to organochlorine pesticides (OCPs) (Lakes Apopka and Griffin, and Emeralda Marsh, Florida, USA) to those from sites that have historically exhibited low embryo mortality and low OCPs (Lakes Woodruff and Orange, Florida). During June-July 2000, 20 alligator clutches were collected from these sites, artificially incubated, and monitored for embryo mortality. Thiamine and OCPs were measured in one egg/clutch. During February 2002, 10 adult female bass were collected from Emeralda Marsh and Lake Woodruff and mature ovaries analyzed for thiamine and OCP concentrations. Although ovaries from the Emeralda Marsh bass contained almost 1,000-fold more OCPs compared with the reference site, Lake Woodruff, there were no differences in thiamine concentrations between sites (11,710 vs. 11,857 pmol/g). In contrast, alligator eggs from the reference site had five times the amount of thiamine compared with the contaminated sites (3,123 vs. 617 pmol/g). Similarly, clutches with > 55% hatch rates had significantly higher concentrations of thiamine compared with clutches with <54% hatch rates (1,119 vs. 201 pmol/g). These results suggest that thiamine deficiency might be playing an important role in alligator embryo survival but not in reproductive failure and recruitment of largemouth bass. The cause(s) of this thiamine deficiency are unknown but might be related to differences in the nutritional value of prey items across the sites studied and/or to the presence of high concentration of contaminants in eggs.

Florida↗

Mercury in feathers from Chilean birds: Influence of location, feeding strategy, and taxonomic affiliation

This study reports baseline concentrations of mercury (Hg) in feathers from different species of birds sampled at various locations off the Chilean coast (Southeastern Pacific). Hg concentrations were evaluated in relation to geographic location, taxonomic affiliation, and feeding strategies. Between January and March of 1995, we collected mature contour feathers from 116 birds belonging to 22 species, mostly seabirds. Birds were collected from 10 different locations (26??09???S, 70??40???W to 54??56???S, 67??37???W). Feather Hg concentrations ranged from 0.11 to 13 ??gg-1 dry weight. We found differences in feather Hg concentrations across taxonomic groups, with highest concentrations in petrels, shearwaters, and albatrosses (Procellaridae), followed by boobies (Sulidae), gulls, terns, skuas (Laridae) and cormorants (Phalacrocoracidae). Ibises and oystercatchers (Threskiornithidae and Charadriidae) had intermediate values, whereas ducks and geese (Anatidae) contained the least amount of Hg. Oceanic species preying on mesopelagic fish (the Procellariformes albatrosses, petrels, and fulmars) had over twice as much Hg (overall average of 3.9 ??gg-1) when compared to the rest of the species sampled (overall average of 1.5 ??gg-1). We did not find higher Hg concentrations in birds inhabiting the more heavily industrialized and urbanized areas of the country (central and northern regions), but in birds inhabiting the remote Juan Ferna??ndez Archipelago. This is not surprising, since all the Procellariformes (the group with highest Hg values in this study) were collected from these islands. Except for Hg in Kermadec petrels (mean of 12 ??gg-1), the range of Hg values reported here (0.11-7.3 ??gg-1) fell below those known to cause adverse health and reproductive effects in birds. ?? 2002 Elsevier Science Ltd. All rights reserved.

Marine Pollution Bulletin↗

An evaluation of biomarkers of reproductive function and potential contaminant effects in Florida largemouth bass (Micropterus salmoides floridanus) sampled from the St. Johns River

The objective of this study was to describe and compare several reproductive parameters for Florida largemouth bass ( Micropterus salmoides floridanus ) inhabiting the St. Johns River and exposed to different types and/or degrees of contamination. Welaka was selected as the reference site in this study because of its low urban and agricultural development, Palatka is in close proximity to a paper mill plant, the Green Cove site is influenced by marine shipping activities and Julington Creek site receives discharges of domestic wastewater and storm water runoff from recreational boating marinas. For this study, bass were sampled both prior to (September 1996) and during the spawning season (February 1997). In order to characterize chemical exposure, bass livers were analyzed for up to 90 trace organics and 11 trace metal contaminants. Reproductive parameters measured included gonadosomatic index (GSI), histological evaluation of gonads and plasma concentrations of vitellogenin (VTG), 17β-estradiol (E 2 ) and 11-ketotestosterone (11-KT). In general, the sum of organic chemicals was highest in livers from Palatka bass and bass from Green Cove and Julington Creek had higher hepatic concentrations of low molecular polycyclic aromatic hydrocarbons and polychlorinated biphenyls when compared to fish from Welaka. Metals were more variable across sites, with highest mean concentrations found in bass from either Julington Creek (Ag, As, Cr, Cu, Zn) or Welaka (Cd, Hg, Pb, Se, Tn). Female bass from Palatka and Green Cove had lower concentrations of E 2 , VTG and lower GSI in relation to Welaka. Males from Palatka and Green Cove showed comparable declines in 11-KT in relation to males from Julington Creek and GSI were decreased only in Palatka males. These results indicate a geographical trend in reproductive effects, with changes being most pronounced at the site closest to the paper mill (Palatka) and decreasing as the St. Johns River flows downstream. Since reproductive alterations were most evident in bass sampled from the site closest to the paper mill discharge, it is possible that exposure to these effluents might explain at least some of the results reported here. However, the presence of reproductive alterations in fish sampled at a considerable distance from the mill discharge (Green Cove, 40 km) would suggest exposure to chemicals released from sources other than the paper mill plant. It is clear that additional studies are needed to evaluate the potential impact of these reproductive changes in populations of Florida largemouth bass inhabiting the St. Johns River.

Florida↗

Characterization of annual reproductive cycles for pond-reared Florida largemouth bass Micropterus salmoides floridanus

The annual reproductive cycle of hatchery-raised largemouth bass (Florida subspecies Micropterus salmoides floridanus) was characterized over a one-year period. Largemouth bass have a distinct annual reproductive cycle with a spring spawning season (approximately between mid-January and mid-June). Cycle characterization focused on an evaluation of gonadal development and plasma concentrations of several sex steroids and vitellogenin (VTG). Adult largemouth bass (n = 20: 10 females and 10 males) were collected monthly from hatchery ponds for one full calendar year. Plasma samples were analyzed for estradiol-17?? (E2), 11-ketotestosterone (11-KT), testosterone (T), progesterone (P), and VTG. Gonadal tissues were weighed to calculate gonadosomatic index (GSI) and evaluated histologically to characterize reproductive stage. In both sexes, GSI began to increase in November, and peaked in February-March. Increases in gonad weights were correlated with maturation of gonads as evidenced by histological evaluations. Bass exhibited seasonal changes in plasma sex steroids and VTG. In males, 11-KT was the only sex steroid that showed strong seasonality, with highest values in February. In females, although E2 and T concentrations followed a similar annual cycle, with highest and lowest values in February and August, respectively, the strongest pattern was observed with E2. 11-KT concentrations were less variable across months, and values were about half of those observed in males. In females, P peaked two months after E2, with high values still in May and June and decreased thereafter, and VTG began to increase in October, but peaked a month prior to the observed peaked in E2. VTG was also detected in males but at concentrations that were about 1/12 that of females, and no seasonal pattern was evident. This study is the first to fully characterize the seasonal endocrine cycle for largemouth bass. These data will be useful when conducting reproductive evaluations of free-ranging populations of largemouth bass and for assessing potential reproductive effects due to environmental contaminants in this species. ?? 2002 by the American Fisheries Society.

American Fisheries Society Symposium↗