Blood lead levels in indigenous peoples living close to oil extraction areas in the Peruvian Amazon.

BACKGROUND: High blood lead levels (BLLs) have been previously reported in indigenous people living in communities in the northern Peruvian Amazon. Oil extraction activities have been conducted in the area since the 1970s and have been identified as a source of lead exposure. OBJECTIVE: Measure BLL and assess risk factors associated with BLL among indigenous populations from four river basins of the northern Peruvian Amazon. METHODS: Participants from 39 communities were selected using a two-stage stratified random selection strategy and were visited between May and June 2016. Information on risk factors was collected using structured questionnaires and blood samples were taken. Overall, complete information was available from 1047 individuals (309 < 12 years old, 738 ≥ 12 years). BLL was determined using atomic absorption spectrophotometry in a graphite chamber. Weighted linear logistic regression models were used to study the association between socio-demographic variables, self-reported life-style factors, environmental, geographical and occupational exposures and BLLs. RESULTS: Geometric mean (95% CI) BLL was 4.9 (4.5, 5.4) µg/dL in participants <12 years and 5.7 (5.4, 6.0) µg/dL in older participants. There were marked differences in BLL between river basins with the highest levels observed in the Corrientes river basin [8.1 (7.2, 9.1) µg/dL <12 years and 8.8 (8.0, 9.6) µg/dL older participants]. High BLL was associated with older age, being male, living in the Pastaza, Tigre or Corrientes river basins and consumption of fish offal in children and adults. Increased Euclidean distance between residence and oil production facilities was associated with a small reduction in BLL. CONCLUSION: BLLs that pose a health risk were detected in the study population of a non-industrialized and remote area of the Amazon. The highest BLLs were observed in those river basins where relative oil extraction activity and environmental levels of contaminants have been reported to be greatest.

Effects of larval atrazine exposure in mesocosms on Blanchard's cricket frogs (Acris blanchardi) reared through overwintering and to reproductive age.

We exposed Blanchard's cricket frog (Acris blanchardi) tadpoles to atrazine in simulated aquatic communities (outdoor mesocosms) at nominal concentrations of 0, 1, 10, 100, and 200 µg/L and tracked the effects of exposure to spring emergence in the laboratory, as well as to reproductive age in outdoor, terrestrial enclosures. We tested hypotheses that 1) atrazine addition increases the prevalence and intensity of testicular ova (TO) among phenotypic males at metamorphosis and after overwintering, 2) atrazine reduces maturation of ova after overwintering among phenotypic females, and 3) atrazine alters mass, time, and survival to metamorphosis, as well as growth and survival across terrestrial life stages. Atrazine addition increased probability of TO presence at metamorphosis, but only when treatments were pooled and compared to the control, where background atrazine was detected. Atrazine did not influence the intensity of TO among metamorphs. We observed TO among males at spring emergence and at reproductive age regardless of exposure concentration. We found no evidence for effects of exposure on gonadal maturation among females after overwintering. Exposure to 200 µg/L reduced survival to metamorphosis, but atrazine did not affect mass at metamorphosis, time to metamorphosis, or survival or mass after overwintering. We demonstrate that atrazine addition can increase TO prevalence relative to background rates at metamorphosis and that TO are also present among phenotypic males after overwintering. We suggest that this non-model species is sensitive to effects of larval EDC exposures on gonadal development and morphology and that further work with cricket frogs is warranted.

Mapping elemental contamination on Palmyra Atoll National Wildlife Refuge.

Palmyra Atoll, once a WWII U.S. Navy air station, is now a U.S. National Wildlife Refuge with nearly 50km2 of coral reef and 275ha of emergent lands with forests of Pisonia grandis trees and colonies of several bird species. Due to the known elemental and organic contamination from chemicals associated with aviation, power generation and transmission, waste management, and other air station activities, a screening survey to map elemental concentrations was conducted. A map of 1944 Navy facilities was georeferenced and identifiable features were digitized. These data informed a targeted survey of 25 elements in soils and sediment at locations known or suspected to be contaminated, using a hand-held X-ray fluorescence spectrometer. At dozens of locations, concentrations of elements exceeded established soil and marine sediment thresholds for adverse ecological effects. Results were compiled into a publically available geospatial dataset to inform potential remediation and habitat restoration activities.

Oil extraction in the Amazon basin and exposure to metals in indigenous populations.

Most oil extraction areas in the Peruvian Amazon are within indigenous territories. Poor environmental practices have exposed the indigenous population to metals. We conducted a survey in two indigenous Kukama communities to assess body burdens of metals after the occurrence of two major oil spills in 2014. Urine levels above those recommended by the Peruvian Ministry of Health were observed in 50% and 17% of the study population for mercury and cadmium, respectively.

Evaluation of potential mechanisms of atrazine-induced reproductive impairment in fathead minnow (Pimephales promelas) and Japanese medaka (Oryzias latipes).

Atrazine has been implicated in reproductive dysfunction of exposed organisms, and previous studies documented decreased egg production in Japanese medaka (Oryzias latipes) and fathead minnows (Pimephales promelas) during 30-d to 38-d exposures to 0.5 µg/L, 5 µg/L, and 50 µg/L atrazine. The authors evaluated possible mechanisms underlying the reduction in egg production. Gene expression in steroidogenesis pathways and the hypothalamus-pituitary-gonad axis of male and female fish was measured. Atrazine did not significantly induce gonad aromatase (cyp19a1a) expression. An atrazine-induced shift in the number of females in an active reproductive state was observed. Expression of the egg maturation genes vitellogenin 1 (vtg1) and zona pellucida glycoprotein 3.1 (zp3.1) in medaka females was correlated and had a bimodal distribution. In both species, females with low vtg1 or zp3.1 expression also had low expression of steroidogenesis genes in the gonad, estrogen receptor in the liver, and gonadotropins in the brain. In the medaka, the number of females per tank that had high expression of zp3.1 was significantly correlated with egg production per tank. The number of medaka females with low expression of zp3.1 increased significantly with atrazine exposure. Thus, the decline in egg production observed in response to atrazine exposure may be the result of a coordinated downregulation of genes required for reproduction in a subset of females. Environ Toxicol Chem 2016;35:2230-2238. Published 2016 Wiley Periodicals Inc. on behalf of SETAC. This article is a US Government work and, as such, is in the public domain in the United States of America.

Bioaccumulation of Stentorin, the Probable Causative Agent for Discolored ("Purple") Eggs and Ovaries in Blue Catfish (Ictalurus furcatus) from Eufaula Lake, Oklahoma, USA.

Observations of reddish to "purple" discolored eggs in the ovaries of adult female blue catfish (Ictalurus furcatus) from the northern arm of Eufaula Lake, a eutrophic multiuse impoundment in east-central Oklahoma, were first reported in 2006. Blue catfish eggs are normally cream to light yellow. Reports peaked in 2007-2008 and declined through 2009-2010; purple eggs have not been reported between 2010 and 2014. In the laboratory, all tissues and fluids of affected fish were strongly orange-red fluorescent under UV illumination, with the fluorescence most apparent in the lipid-rich ovaries and eggs. The causative agent was isolated chromatographically and confirmed by mass spectrometry as stentorin (1,3,4,6,8,10,11,13-octahydroxy-2,5-diisopropyl-phenanthro[1,10,9,8,o,p,q,r,a]perylene-7,14-dione), the fluorescent, lipophilic pigment associated with the photoreceptor protein of the ciliated protozoan Stentor coeruleus (Heterotrichea; Stentoridae). Larval medaka (Orizias latipes) readily consumed S. coeruleus in the laboratory and were observed to fluoresce in the same manner as the affected blue catfish. Potential deleterious effects of stentorin bioaccumulation remain to be determined, as do the geographic extent and the identities of other fluorescent compounds isolated from catfish eggs and ovaries.

Sensitivity of shovelnose sturgeon (Scaphirhynchus platorynchus) and pallid sturgeon (S. albus) early life stages to 3,3',4,4',5-pentachlorobiphenyl and 2,3,7,8-tetrachlorodibenzo-P-dioxin exposure.

Concern exists that polychlorinated biphenyls (PCBs) may be contributing to the current decline of shovelnose sturgeon (Scaphirhynchus platorynchus) and the US federally endangered pallid sturgeon (Scaphirhynchus albus). Waterborne exposures with newly fertilized eggs were used to assess developmental and morphological effects of 2 of the most potent aryl hydrocarbon receptor (AhR) agonists, 3,3',4,4',5-pentachlorobiphenyl (PCB-126) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), on early life stage shovelnose and pallid sturgeon. No dose-related effects of PCB-126 were observed on percent development or hatch in either species at concentrations as high as 1711 ng/g egg. Effects of TCDD on percent development were not assessed in shovelnose sturgeon. However, percent development was not affected by TCDD in pallid sturgeon, and percent hatch was unaffected by TCDD doses as high as 60 ng/g egg to 81 ng/g egg in either species. Morphological pathologies such as yolk sac edema and craniofacial deformities were typical of AhR agonist exposure and were similar in both species. Calculated PCB-126 50% lethal dose (LD50, 95% fiducial limits) values were 196 ng/g egg (188-203 ng/g) for shovelnose and 159 ng/g egg (122-199 ng/g) for pallid sturgeon. Likewise, calculated TCDD LD50 values were 13 ng/g egg (11-15 ng/g) for shovelnose and 12 ng/g egg (10-14 ng/g) for pallid sturgeon. These LD50 values are among the highest recorded in early life stage fish, suggesting that early life stage Scaphirhynchus sturgeon may be comparatively insensitive to AhR agonists.

Atrazine reduces reproduction in Japanese medaka (Oryzias latipes).

Atrazine is an effective broadleaf herbicide and the second most heavily used herbicide in the United States. Effects along the hypothalamus-pituitary-gonad axis in a number of vertebrate taxa have been demonstrated. Seasonally elevated concentrations of atrazine in surface waters may adversely affect fishes, but only a few studies have examined reproductive effects of this chemical. The present study was designed to evaluate a population endpoint (egg production) in conjunction with histological (reproductive stage, gonad pathology) and biochemical (aromatase activity, sex hormone production) phenotypes associated with atrazine exposure in Japanese medaka. Adult virgin breeding groups of one male and four females were exposed to nominal concentrations of 0, 0.5, 5.0, and 50 µg/L (0, 2.3, 23.2, 231 nM) of atrazine in a flow-through diluter for 14 or 38 days. Total egg production was lower (36-42%) in all atrazine-exposed groups as compared to the controls. The decreases in cumulative egg production of atrazine-treated fish were significant by exposure day 24. Reductions in total egg production in atrazine treatment groups were most attributable to a reduced number of eggs ovulated by females in atrazine-treated tanks. Additionally, males exposed to atrazine had a greater number of abnormal germ cells. There was no effect of atrazine on gonadosomatic index, aromatase protein, or whole body 17 ß-estradiol or testosterone. Our results suggest that atrazine reduces egg production through alteration of final maturation of oocytes. The reduced egg production observed in this study was very similar to our previously reported results for fathead minnow. This study provides further information with which to evaluate atrazine's risk to fish populations.

Gonadal abnormalities in frogs (Lithobates spp.) collected from managed wetlands in an agricultural region of Nebraska, USA.

Nebraska's Rainwater Basin (RWB) provides important wetland habitat for North American migratory birds. Concern exists that pesticide and nutrient runoff from surrounding row-crops enters wetlands degrading water quality and adversely affecting birds and wildlife. Frogs may be especially vulnerable. Plains leopard (Lithobates blairi) metamorphs from RWB wetlands with varying concentrations of pesticides were evaluated for a suite of biomarkers of exposure to endocrine active chemicals. Froglets had ovarian dysgenesis, high rates of testicular oocytes, and female-biased sex ratios however, there was no clear statistical association between pesticide concentrations and biomarkers. Data interpretation was hindered because timing and duration of exposures were unknown and due to an incomplete understanding of L. blairi sexual development. Emphasis is on describing the complex developmental biology of closely-related leopard frogs, how this understanding can explain RWB L. blairi anomalies, and the need for sampling at the appropriate life stage.

Atrazine reduces reproduction in fathead minnow (Pimephales promelas).

Atrazine, the widely used herbicide, has shown to affect the hypothalamus-pituitary-gonad axis in certain vertebrate species, but few studies have examined reproductive effects of this chemical on fish. Our study was designed to evaluate a population endpoint (egg production) in conjunction with histological (e.g., gonad development) and biochemical (e.g., hormone production) phenotypes associated with atrazine exposure in fathead minnows. Adult virgin breeding groups of 1 male and 2 females were exposed to nominal concentrations of 0, 0.5, 5.0, and 50 microg/L of atrazine in a flow-through diluter for 14 or 30 days. Total egg production was lower (19-39%) in all atrazine-exposed groups as compared to the controls. The decreases in cumulative egg production of atrazine treated fish were significant by 17-20 days of exposure. Reductions in egg production in atrazine treatment groups were most attributable to reduced numbers of spawning events with increased atrazine exposure concentrations. Gonad abnormalities were observed in both male and female fish of atrazine-exposed fish. Our results also indicate that atrazine reduces egg production through alteration of final maturation of oocytes. The reproductive effects observed in this study warrant further investigation and evaluation of the potential risks posed by atrazine, particularly feral populations of fish from streams in agricultural areas with high use of this herbicide.

Widespread occurrence of intersex in black basses (Micropterus spp.) from U.S. rivers, 1995-2004.

Intersex occurrence in freshwater fishes was evaluated for nine river basins in the United States. Testicular oocytes (predominantly male testes containing female germ cells) were the most pervasive form of intersex observed, even though similar numbers of male (n=1477) and female (n=1633) fish were examined. Intersex was found in 3% of the fish collected. The intersex condition was observed in four of the 16 species examined (25%) and in fish from 34 of 111 sites (31%). Intersex was not found in multiple species from the same site but was most prevalent in largemouth bass (Micropterus salmoides; 18% of males) and smallmouth bass (M. dolomieu; 33% of males). The percentage of intersex fish per site was 8-91% for largemouth bass and 14-73% for smallmouth bass. The incidence of intersex was greatest in the southeastern United States, with intersex largemouth bass present at all sites in the Apalachicola, Savannah, and Pee Dee River Basins. Total mercury, trans-nonachlor, p,p'-DDE, p,p'-DDD, and total PCBs were the most commonly detected chemical contaminants at all sites, regardless of whether intersex was observed. Although the genotype of the intersex fish was not determined, the microscopic appearance of the gonads, the presence of mature sperm, and the concentrations of sex steroid hormones and vitellogenin indicate the intersex bass were males. Few reproductive endpoints differed significantly among male and intersex bass; plasma vitellogenin concentration in males was not a good indicator of intersex presence. Hierarchical linkages of the intersex condition to reproductive function will require a more quantitative measure of intersex (e.g. severity index) rather than presence or absence of the condition. The baseline incidence of intersex gonadal tissue in black basses and other freshwater fishes is unknown, but intersex prevalence may be related to collection season, age, and endocrine active compounds in the environment. Intersex was not found in largemouth bass older than five years and was most common in 1-3-year-old male largemouth bass. The cause(s) of intersex in these species is also unknown, and it remains to be determined whether the intersex we observed in largemouth and smallmouth bass developed during sex differentiation in early life stages, during exposure to environmental factors during adult life stages, or both.

Toxicity of o,p'-DDE to medaka d-rR strain after a one-time embryonic exposure by in ovo nanoinjection: an early through juvenile life cycle assessment.

The toxicity of o,p'-DDE (1,1-dichloro-2-(p-chlorophenyl)-2-(o-chlorophenyl) ethylene) was evaluated in embryos of medaka (Oryzias latipes) following a one time exposure via nanoinjection. Medaka eggs (early gastrula) were injected with 0.5 nl of triolein (vehicle control) or 0.5 nl of 4 graded doses (0.0005-0.5 ng/egg) of o,p'-DDE in triolein. Embryos were allowed to develop, and fry were reared. Embryonic survival was monitored daily during the first 10 d until hatching and thereafter, on a weekly basis until day 59, at which time the fish were monitored for sexual maturity until day 107. In general, o,p'-DDE caused a dose- and time-dependent mortality. No changes in mortality were observed between the last two time points (day 38 and 59, respectively), and hence a 59 day-LD50 of 346 ng o,p'-DDE/egg was derived from the linear dose-response relationship. Prior to late stage death, only isolated cases of cardiovascular lesions and spinal deformities were observed, but were not dose-dependent. The lowest observable adverse effect level (LOAEL), based on upper 95% CI for regression line=0.0018 mg/kg, and the LOAEL based on exposure doses=0.5 mg/kg. Likewise, the no observable adverse effect level (NOAEL) based on linear extrapolation to 100% survival=0.0000388 mg/kg, while the NOAEL based on exposure doses=0.05 mg/kg. The nanoinjection medaka model has potential in the study of hormonally active compounds in the environment.

In ovo exposure to o,p -DDE affects sexual development but not sexual differentiation in Japanese medaka (Oryzias latipes).

Despite being banned in many countries, dichlorodiphenyltrichloroethane (DDT) and its metabolites dichlorodiphenyldichloroethylene (DDE) and dichlorodiphenyldichloroethane (DDD) continue to be found in fish tissues at concentrations of concern. Like o,p -DDT, o,p -DDE is estrogenic and is believed to exert its effects through binding to the estrogen receptor. The limited toxicologic data for o,p -DDE suggest that it decreases fecundity and fertility of fishes. We conducted an egg injection study using the d-rR strain of medaka and environmentally relevant concentrations of o,p -DDE to examine its effects on sexual differentiation and development. The gonads of exposed fish showed no evidence of sex reversal or intersex. However, other gonad abnormalities occurred in exposed individuals. Females exhibited few vitellogenic oocytes and increased atresia. Male testes appeared morphologically normal but were very small. Gonadosomatic index values for both sexes were lower for exposed fish. Our observations of abnormal female and very small male gonads after in ovo o,p -DDE exposure may be indicative of effects on early endocrine processes important for normal ovarian and testicular development.