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At least 19 recordsLinked to original sources

Testing of selected pharmacological agents for capturing waterfowl [Annual Progress Report]

The response of game-farm mallards (Frost strain) to seven pharmacological immobilizing agents was evaluated in Phase I of a planned four-phase study. A limited amount of testing was also done with wild mallards. Single dosages were administered to determine the mean effective dose (ED50) and mean lethal dose (LD50), The therapeutic index, or safety factor (LD50/ED50), and palatability were also established. Optimum dosage rates of compounds administered orally on baits were not considered in this phase of the study. Compounds were-administered by intubation and calculated in terms of mg/kg. All except one compound produced narcosis within 5 minutes at the effective dose rate.Immobilization periods for the seven compounds ranged from 7-24 minutes, and recovery periods from 1.0-6.5 hours. Such wide variations in actions of the compounds can be attributed to a compound's rate of absorption, the ease with which it passes the blood-brain barrier, its solubility in tissues, and its rate of metabolism in the liver and kidneys. Length of both the immobilization and recovery periods were extended when dosages were increased. There was no delayed mortality among survivors with any of the seven compounds at either the ED50 or LD50. Females were generally more sensitive to the anesthetizing agents than males. The ED50 for wild mallards was substantially higher than that for the experimental game-farm birds for the two compounds on which this was tested.Tribromoethanol (Avertin of Winthrop Laboratories) satisfied all test criteria an Phase I and will be subjected to more intensive investigation in ensuing tests. Thiopental sodium (Pentothal of Amdal Company) and pentobarbital sodium (Nembutal of Abbott Laboratories) were judged to be marginal. Although their therapeutic indexes were good (5.00), recovery periods were prolonged and toxic convulsions occurred at medium to high dose rates as the LD50 was approached.Alpha-chloralose (Fisher Scientific) proved least promising of the seven compounds, mainly because of its unacceptable therapeutic index (2.25) and because it possesses prolonged induction and recovery periods. Two new experimental drugs, methoxymol and metomidate (Pitman-Moore), appeared effective and safe when administered by intubation but produced a taste aversion when added to bait. Rejection because of taste was also a problem with secobarbital (Seconal of Elanco Products), and its therapeutic index of 2.75 was unacceptable. Monitoring of heart and respiratory rates, and body temperature by telemetry showed promise as a technique for determining physiological response to drug action.

Report

Embryotoxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD): Theembryonic vasculature is a physiological target for TCDD-induced DNA damage and apoptotic cell death in medaka (Orizias latipes)

Vertebrate embryos are extremely sensitive to environmental contaminants known as planar halogenated hydrocarbons (PHHs). The physiological targets that mediate PHH-induced embryotoxicity are not known. We have characterized embryotoxicity in medaka ( Orizias latipes ) caused by 2,3,7,8-tetrachlorodibenzo- p -dioxin (TCDD), the prototypic PHH. DNA degradation in cells of the embryonic vasculature and loss of functional integrity of the medial yolk vein were demonstrated in TCDD-exposed embryos. Pharmacological intervention with piperonyl butoxide inhibited TCDD-induced DNA degradation, restored the functional integrity of the medial yolk vein, and protected against the embryotoxicity of TCDD. Treatment of TCDD-exposed embryos with the antioxidant N -acetylcysteine also provided significant protection against the embryotoxicity of TCDD. These results demonstrate that DNA damage and consequent cell death in the embryonic vasculature are key physiological mediators of TCDD-induced embryotoxicity.

Toxicology and Applied Pharmacology

Toxicokinetics of PAHs in Hexagenia

The accumulation kinetics of two waterborne polycyclic aromatic hydrocarbons (PAHs), benzo[a]pyrene (BAP) and phenanthrene (PHE), were studied in the mayfly nymph ( Hexagenia limbata ). The uptake clearance decreased while the bioconcentration of BAP increased with an increase in weight of the H. limbata nymph. The relationship between uptake clearance and bioconcentration for PHE was variable, and bioconcentration was greater for the heavier animals. Two kinetic models were used to evaluate the effect of nymph weight on disposition of PAHs: (a) the amount-uptake clearance model, similar to models most frequently used in environmental toxicology; and (b) a clearance-volume model, similar to models used in clinical pharmacology. The two models gave similar predictive results but were different in a few cases. These differences in common parameter estimation probably resulted from methodologies used and high data variability rather than the models themselves, since they are mathematically equal. Some of the parameters are unique to each of the models and are defined and described. The clearance of oxygen from water is inversely and linearly related to the weight of the mayfly nymphs, but oxygen clearances were always much less than the uptake clearances of the PAHs. The high PAH uptake clearance compared to oxygen clearance implies a greater surface area or efficiency for PAH accumulation from water.

Environmental Toxicology and Chemistry

Coral reef diseases in the Atlantic-Caribbean

Coral reefs are the jewels of the tropical oceans. They boast the highest diversity of all marine ecosystems, aid in the development and protection of other important, productive coastal marine communities, and have provided millions of people with food, building materials, protection from storms, recreation and social stability over thousands of years, and more recently, income, active pharmacological compounds and other benefits. These communities have been deteriorating rapidly in recent times. The continuous emergence of coral reef diseases and increase in bleaching events caused in part by high water temperatures among other factors underscore the need for intensive assessments of their ecological status and causes and their impact on coral reefs.

Book chapter

A regional assessment of chemicals of concern in surface waters of four Midwestern United States national parks

Anthropogenic chemicals and their potential for adverse biological effects raise concern for aquatic ecosystem health in protected areas. During 2013–15, surface waters of four Midwestern United States national parks were sampled and analyzed for wastewater indicators, pharmaceuticals, personal care products, and pesticides. More chemicals and higher concentrations were detected at the two parks with greater urban influences (Mississippi National River and Recreation Area and Indiana Dunes National Lakeshore) than at the two more remote parks (Apostle Islands National Lakeshore and Isle Royale National Park). Atrazine (10–15 ng/L) and N,N-diethyl-meta-toluamide (16–120 ng/L) were the only chemicals detected in inland lakes of a remote island national park (Isle Royale National Park). Bisphenol A and organophosphate flame retardants were commonly detected at the other sampled parks. Gabapentin and simazine had the highest observed concentrations (> 1000 ng/L) in three and two samples, respectively. At the two parks with urban influences, metolachlor and simazine concentrations were similar to those reported for other major urban rivers in the United States. Environmental concentrations of detected chemicals were often orders of magnitude less than standards or reference values with three exceptions: (1) hydrochlorothiazide exceeded a human health-based screening value in seven samples, (2) estrone exceeded a predicted critical environmental concentration for fish pharmacological effects in one sample, and (3) simazine was approaching the 4000 ng/L Maximum Contaminant Level in one sample even though this concentration is not expected to reflect peak pesticide use. Although few environmental concentrations were approaching or exceeded standards or reference values, concentrations were often in ranges reported to elicit effects in aquatic biota. Data from this study will assist in establishing a baseline for chemicals of concern in Midwestern national parks and highlight the need to better understand the sources, pathways, and potential adverse effects to aquatic systems in national parks.

Indiana, Michigan, Minnesota, Wisconsin

Leveraging invasive mussel contaminant survey data for stepwise prioritization of chemicals of potential concern in the Great Lakes basin

Historical and ongoing anthropogenic activities coupled with advancements in analytical techniques have led to the detection of large numbers of contaminants in the Laurentian Great Lakes. Consequently, identifying and prioritizing chemicals likely to cause ecological harm represents a challenge for natural resource managers. Previous prioritization efforts have focused on contaminants in sediment, water, and passive samplers, which may not be representative of compounds that bioaccumulate in aquatic organisms. Consequently, this study adopted a stepwise method to prioritize chemicals of potential concern detected in dreissenid mussels from samples collected across the Great Lakes from 2009–2018. The stepwise method considered environmental fate, detection frequency, and exceedance of toxicity quotients based on ecotoxicological effect concentrations. Overall, 153 compounds out of 267 analyzed were detected in dreissenid mussels, 47 of which had water quality effect concentrations, 56 had apical effect concentrations (Tier 1 ECOTOX or apical screening), 17 had nonapical effect concentrations (Tier 2 ECOTOX, Cytotoxic Burst, and ToxCast) and 33 had estimated effect concentrations (quantitative structure-activity relationship, estimated screening, and pharmacological potency). Of the compounds with water quality effect concentrations, nine were designated as high priority, including the herbicide atrazine and five polycyclic aromatic hydrocarbons that were previously identified as potentially hazardous within other matrices. Similar contaminants were identified as high priority in a related study of native unionid mussels in the Great Lakes. A total of 27 compounds were low priority, suggesting that these contaminants do not warrant further action based on this dataset. Overall, these findings will facilitate the development of management strategies to mitigate the effects of contaminants on aquatic organisms within the Great Lakes.

Great Lakes basin

Functional characterization and osmoregulatory role of the Na+/K+/2Cl--cotransporter (NKCC1) in the gill of sea lamprey (Petromyzon marinus), a basal vertebrate

The present study provides molecular and functional characterization of Na+/K+/2Cl- cotransporter (nkcc1/NKCC1) in the gills of sea lamprey, the most basal extant vertebrate with an osmoregulatory strategy. We report the full-length peptide sequence for the lamprey NKCC1, which we show to group strongly with and occupy a basal position among other vertebrate NKCC1 sequences. Lamprey nkcc1 mRNA were present in many tissues but was 5-fold higher in the gill than any other tissue. NKCC1 protein was only detected in the gill. Gill mRNA and protein abundances of NKCC1 and Na+/K+-ATPase (NKA) were significantly upregulated (20- to 200-fold) in late metamorphosis in freshwater, coinciding with the development of salinity tolerance, and were upregulated an additional 2-fold after acclimation to seawater. Immunohistochemistry revealed that NKCC1 in the gill is found in filamental ionocytes that develop during metamorphosis. Lamprey treated with bumetanide, a widely used pharmacological inhibitor of NKCC1, exhibited higher plasma Cl- and osmolality and reduced muscle water content after 24 h in seawater, but had no effect in FW. This work provides the first functional characterization of NKCC1 as having a functional role mechanism for branchial salt secretion in lampreys, providing evidence that this mode of Cl- secretion has been present among vertebrates for ~550 million years.

American Journal of Physiology - Regulatory, Integ

A Cftr-independent, Ano1-rich seawater-adaptive ionocyte in sea lamprey gills

All ionoregulating marine fishes examined to date utilize seawater-type ionocytes expressing the apical Cl- channel, cystic fibrosis transmembrane conductance regulator (Cftr) to secrete Cl − . We performed transcriptomic, molecular, and functional studies to identify Cl − transporters in the seawater-type ionocytes of sea lamprey ( Petromyzon marinus ). Gill cftr expression was minimal or undetectable in larvae and post-metamorphic juveniles. We identified other Cl − transporters highly expressed in the gills and/or upregulated following metamorphosis and further investigated two candidates that stood out in our analysis, a Ca 2+ -activated Cl − channel, anoctamin 1 ( ano1 ), and the Clc chloride channel family member 2 ( clcn2 ). Of these, ano1 was expressed 10-100 times more than clcn2 in the gills; moreover, ano1 was upregulated during seawater acclimation, while clcn2 was not. Using an antibody raised against sea lamprey Ano1, we did not detect Ano1 in the gills of larvae, found elevated levels in juveniles and observed a 4-fold increase in juveniles after seawater acclimation. Ano1 was localized to seawater-type branchial ionocytes but, surprisingly, was localized to the basolateral membrane. In vivo pharmacological inhibition experiments demonstrated that a DIDS-sensitive mechanism was critical to the maintenance of osmoregulatory homeostasis in seawater- but not freshwater-acclimated sea lamprey. Taken together, our results provide evidence of a Cftr-independent mechanism for branchial Cl − secretion in sea lamprey that leverages Ano1-expressing ionocytes. Once further characterized, the Cftr-independent, Ano1-rich ionocytes of sea lamprey could reveal novel strategies for branchial Cl − secretion, whether by Ano1 or some other Cl − transporter, not previously known in ionoregulating marine organisms.

Journal of Experimental Biology

High doses of corticosteroid suppress resistance to Ichthyophonus in starry flounder

Application of pharmacological doses of the corticosteroid dexamethasone phosphate to starry flounder Platichthys stellatus resulted in a predisposition to clinical ichthyophoniasis and a progression from latent Ichthyophonus infections to patent, histologically identifiable infections. Among Ichthyophonus -challenged starry flounder, the prevalences of clinical infections and histologically identifiable infections were significantly greater in two groups that received dexamethasone (100% and 31%, respectively) than in the respective control groups (8% and 0%). Proliferation of Ichthyophonus infections in corticosteroid-treated groups may have resulted from suppression of the cellular immune response that typically follows corticosteroid application; however, further studies are needed to determine whether these effects occur at lower, physiological concentrations of corticosteroids.

Journal of Aquatic Animal Health

In vivo/in vitro comparison of pharmacokinetics and pharmacodynamics of 3,3',4,4'-tetrachlorobiphenyl (PCB77)

The rat hepatoma cell line, H4IIE, serves as a useful tool to assess potential biological effects such as induction of cytochrome P4501A1 expression. The objectives of this study were twofold: to investigate the kinetic time course and dosimetry of PCB77 in rat hepatoma cells dosed with PCB77 and in liver of rats given ip doses of PCB77, and to compare in vitro and in vivo P4501A1 enzyme induction responses. For the 4-day time–course study, H4IIE cells were exposed with two doses of [ 14 C]PCB77 (0.9 and 3 μg/plate) and harvested at 15 and 30 min, 1, 2, 4, 8, and 12 hr, and 1, 2, 3, and 4 days. PCB77-derived radioactivity was detected in the cells as early as 15 min postdosing. For the dose–response study, the cells were dosed with various concentrations of PCB77 (0.00316–5.37 μg/plate) and harvested on Day 3 since ethoxyresorufin O -deethylase (EROD) activity in vitro reached its maximum on the third day postdosing. Time–course and dose–response studies revealed that only 1–3% of the total delivered dose was found in the cells, with the remainder in the media and adhering to the culture plates. For the dose–response study in vivo, male Fischer rats were dosed with a single ip injection of various concentrations of PCB77 (0.1–50 mg/kg body wt) and euthanized on Day 3. PCB77-derived radioactivity and EROD induction in vivo were measured. When EROD activity and PCB77-derived radioactivity in the rat hepatoma cells and in the rat liver were compared on an equivalent weight basis, there was a significant correlation ( r 2 = 0.985) between them. Prior to this study, no information on quantitative dosimetry and EROD activities of PCB77 has been reported to validate the in vitro assay with in vivo data.

Toxicology and Applied Pharmacology

Chronic oral DDT toxicity in juvenile coho and chinook salmon

Technical and p,p′ -DDT was incorporated into test diets and fed to juvenile chinook and coho salmon for periods as long as 95 days. Pure p,p′ -DDT was slightly more toxic to young salmon than was the technical DDT mixture. Chinook salmon appeared to be 2–3 times more sensitive to a given concentration of DDT in the diet than were coho salmon. The size of the fish greatly influenced toxicity, smaller younger fish being more susceptible to a given diet than larger older fish. The dose of DDT accumulated within the median survival time ranged from 27–73 mg/kg for chinook salmon and from 56–72 mg/kg for coho salmon. The extrapolated 90-dose LD50 ( Hayes, 1967 ) for young chinook and coho salmon were 0.0275 and 0.064 mg/kg/day, respectively. Liver size decreased on prolonged feeding with DDT, and carcass lipid content was increased. A severe surface ulceration of the nose region appeared in coho salmon fed DDT over long periods. In addition, an interesting localized degeneration of the distal convoluted tubule was observed in the kidney of coho salmon receiving DDT.

Toxicology and Applied Pharmacology

Toxicity of DDT to Japanese quail as influenced by body weight, breeding condition, and sex

Controlled experiments were utilized to simulate the stresses on wild birds of breeding condition and of weight loss due to migration. Light conditions in the laboratory were manipulated to produce Japanese quail ( Coturnix coturnix japonica ) in breeding condition and not in breeding condition. Within each of these groups, some birds were partially starved before dosage and some were fully fed. Birds were then fed dietary levels of 0, 700, 922, 1214, or 1600 ppm dry weight of p,p′ -DDT for a period of 20 days or until death. Birds partially starved before dosage were more susceptible to DDT intoxication than nonstarved ones, and birds not in breeding condition were slightly more so than birds in breeding condition. Similarly, males died earlier than females, and the birds of the lighter weight strain used in the second half of the study died earlier than the birds of the heavier strain used in the first half. The heavier birds of each sex not only survived longer than lighter individuals receiving the same treatments, but they also lost a greater proportion of their weight before death. During the early portion of the dosage period, females in breeding condition were less sensitive to DDT than were females not in breeding condition and males. After 10 days on dosage, however, the cumulative mortality of females in breeding condition rapidly approached that of males and of females not in breeding condition. Food restriction prior to dosage, strains of quail, breeding conditions, and sexes resulted in weight differences and a corresponding accentuation or delay of the effects of the different levels of DDT.

Toxicology and Applied Pharmacology

Comparative acute oral toxicity of pesticides to six species of birds

Acute oral LD50 values were determined for 16 common pesticides on mallard ducks ( Anas platyrhynchos ), ring-necked pheasants ( Phasianus colchicus ), chukar partridges ( Alectoris graeca ), coturnix quail ( Coturnix coturnix japonica ), common pigeons ( Columba livia ), and house sparrows ( Passer domesticus ). Analyses of the data revealed that (1) the average sensitivity of any one species to the 16 pesticides did not differ statistically from that of any other species, and (2) at the same time, the species varied widely in their sensitivity to any given compound. The average range of the LD50 values for single chemicals in the 6 species was nearly 10-fold. With differences of this magnitude, it is recommended that extrapolation of toxicity data from one species to another be avoided.

Toxicology and Applied Pharmacology

The acute oral toxicity of 369 pesticidal, pharmaceutical and other chemicals to wild birds

The acute po toxicity of 369 chemicals was determined for 1 or more species of wild birds, always including either red-winged blackbirds or starlings. Of these, 180 chemicals were toxic to 1 or more species at 100 mg/kg or less. Statistical comparison of redwing, starling and rat data indicated that redwings were more sensitive to chemicals than starlings, and both were more sensitive than rats.

Toxicology and Applied Pharmacology

Plasma enzyme activities in coturnix quail fed graded doses of DDE, polychlorinated biphenyl, malathion, and mercuric chloride

Male Coturnix quail (Coturnix coturnix japonica) were fed diets for 12 weeks containing graded levels of DDE, polychlorinated biphenyl (Aroclor 1254), malathion, and mercuric chloride. Birds were bled prior to exposure and at 2, 4 and 12 weeks, and the plasma used to measure the activities of creatine kinase, aspartate aminotransferase, cholinesterase, fructose-diphosphate aldolase, and lactate dehydrogenase. Abnormal activity of certain plasma enzymes was noted in birds after 2 and 4 weeks, but these changes were not proportional to dose or exposure time. At 12 weeks increases in each of the activities of plasma enzymes of birds fed organochlorines, and decreases in cholinesterase activity of birds fed malathion or mercuric chloride, were proportional to the log dose of the respective agents. In addition, the pattern of enzyme responses in the 4 experimental groups had changed, and was illustrative of the specific type of substance that had been fed. The data suggest that qualitative and quantitative identification of environmental contaminants in birds, and perhaps a variety of wild animals, may be possible by utilization of multiple plasma enzyme assays. Residue analyses after 12 weeks of feeding showed that DDE accumulated in carcasses and livers at concentrations up to 4-fold higher than those in the diets. In contrast residues of Aroclor 1254 attained in carcasses were identical to, and in livers one-half of, the concentration in the feed. Mercury did not accumulate as much in the tissues; residues attained were one-twentieth or less of those in the feed.

Toxicology and Applied Pharmacology

Responsiveness of 6 to 14 generations of birds to dietary dieldrin toxicity

The lethal dietary toxicity of dieldrin was estimated repeatedly during 8 years of testing young bobwhites (Colinus virginianus), Japanese quail (Coturnix c. japonica), ring-necked pheasants (Phasianus colchicus), and mallards (Anas platyrhynchos). Toxicities, quantified as 8-day LC50 values (5 days on toxic diet, followed by 3 days of untreated feed), were estimated at least 18 times per species. Average dietary LC50 values (ppm) and their 95% confidence limits for 14-day-old quail and 10-day-old pheasants and mallards were: bobwhite, 38 (34?42); Japanese quail, 61 (58?64); ring-necked pheasant, 56 (53?59); and mallard, 179 (163?195). No time-related changes were detected in LC50 values for any of the species.

Toxicology and Applied Pharmacology

Embryotoxic effects of crude oil in mallard ducks and chicks

Recent studies in this laboratory have revealed that surface applications of microliter amounts of some crude and fuel oils that coat less than 10% of the egg surface reduce hatching considerably in different avian species. Applications of paraffin compounds that coat equal areas of the egg surface do not reduce hatching suggesting that toxicity is due to causes other than asphyxia. In the present study, 1–10 μl of South Louisiana crude oil, an API reference oil, were applied to the surface of fertile mallard ( Anas platyrhynchos ) and chicken ( Gallus gallus ) eggs. Early embryolethality was greater in mallard embryos than in chick embryos, but later embryolethality that coincided with the time of rapid outgrowth of the chorioallantoic membrane was more prevalent in chick embryos. The overall incidence of embryolethality was similar in both species. Retardation of growth as reflected by embryonic body weight, crown-rump length, beak length, and general appearance was more pronounced in chick than mallard embryos. Teratogenic defects were more frequent in chick embryos, and incomplete or abnormal ossification of the skull was the most common. External application of equivalent amounts of a mixture of paraffin compounds present in crude oil had virtually no embryotoxic effects in either species, suggesting that other components including aromatic hydrocarbons and organometallics may cause the embryotoxicity.

Toxicology and Applied Pharmacology