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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↗

Acute oral and percutaneous toxicity of pesticides to mallards: Correlations with mammalian toxicity data

Acute oral (po) and 24-hr percutaneous (perc) LD50 values for 21 common pesticides (19 anticholinesterases, of which 18 were organophosphates, and one was a carbamate; one was an organochlorine central nervous system stimulant; and one was an organonitrogen pneumotoxicant) were determined in mallards ( Anas platyrhynchos ). Three of the pesticides tested were more toxic percutaneously than orally. An index to the percutaneous hazard of a pesticide, the dermal toxicity index (DTI = po LD50/perc LD50 × 100), was also calculated for each pesticide. These toxicity values in mallards were compared with toxicity data for rats from the literature. Significant positive correlations were found between log po and log percutaneous LD50 values in mallards ( r = 0.65, p < 0.01), between log po LD50 values in mallards and in rats ( r = 0.71, p < 0.01), and between log DTI values in mallards and in rats ( r = 0.52, p < 0.05). Percutaneous toxicity values were not significantly correlated between mallards and rats ( r = 0.36, p > 0.10). Variations in percutaneous methodologies are discussed with reference to interspecies variation in toxicity values. It is recommended that a mammalian DTI value approaching 30 be used as a guideline for the initiation of percutaneous toxicity studies in birds, when the po LD50 and/or projected percutaneous LD50 are less than expected field exposure levels.

Toxicology and Applied Pharmacology↗

Dopamine and norepinephrine depletion in ring doves fed DDE, dieldrin, and Aroclor 1254

The levels of dopamine and norepinephrine were measured in one-half of the brain of ring doves fed a control diet or a diet containing 2, 20, or 200 ppm DDE; 1, 4, or 16 ppm dieldrin; or 1, 10, or 100 ppm Aroclor 1254. Levels of DDE, dieldrin, or Aroclor 1254 were determined in the other half of each brain. The intermediate and high levels of each chemical caused depletions in both neurotransmitters, and brain residues of each chemical were negatively correlated with levels of neurotransmitters. The highest concentrations of DDE, dieldrin, and Aroclor 1254 depressed averages of dopamine to 42.4, 41.4, and 45.2% of the control level and norepinephrine to 61.6, 62.0, and 56.9% of controls, respectively. Depletions of dopamine and norepinephrine could result in abnormal behavior of contaminated birds in the wild, and the detection of such depletions could become an important tool in assessing contaminant-induced behavioral aberrations in birds.

Toxicology and Applied Pharmacology↗

Embryotoxic and biochemical effects of waste crankcase oil on birds' eggs

Waste crankcase oil (WCO) is a major source of oil pollution in both the aquatic and terrestrial environment and has been implicated in the poisoning of mammals and fish. It is also mutagenic. Since birds' eggs are highly sensitive to external microliter applications of environmentally polluting oils, we examined the developmental effects of external applications of WCO on eggs of the mallard duck (Anas platyrhynchos) and the bobwhite quail (Colinus virginianus). At 48 hr of development, mallard eggs were exposed externally to 2, 5, or 15 :l of WCO or 15 :l of clean crankcase oil (CCO) while bobwhite eggs received proportional doses of 0.5, 1, or 3 :l of WCO and 3 :l of CCO in a similar manner. WCO was highly embryotoxic to both species compared to CCO and resulted in dose-dependent mortality, reduced growth, and abnormal survivors. Application of 15 :l WCO resulted in 84% mortality in mallards and 3 :l WCO resulted in 88% mortality in bobwhites. Abnormal survivors included embryos with subcutaneous edema, incomplete ossification, and eye and brain defects. Red blood cell *-aminolevulinic acid dehydratase (ALAD) activity, liver ALAD activity, and hemoglobin concentration were significantly lower after treatment with WCO in embryos and hatchlings of both species. Plasma uric acid, plasma alanine aminotransferase (ALT), and plasma aspartate aminotransferese (AST) were significantly elevated in WCO-treated mallards after hatching. Biochemical effects, growth retardation, and mortality at proportionally lower dose levels were more pronounced in mallards than in bobwhites. Chemical analysis of the WCO and CCO revealed a considerably higher content of aromatic hydrocarbons in WCO than in CCO. Lead levels were highly elevated in WCO (4600 ppm) compared to CCO (2 ppm).

Toxicology and Applied Pharmacology↗

Subchronic organophosphorus ester-induced delayed neurotoxicity in mallards

Eighteenweek-old mallard hens received 0, 10, 30, 90, or 270 ppm technical grade EPN (phenylphosphonothioic acid O -ethyl- O -4-nitrophenyl ester) in the diet for 90 days. Ataxia was first observed in the 270-ppm group after 16 days, in the 90-ppm group after 20 days, in the 30-ppm group after 38 days; 10 ppm failed to produce ataxia. By the end of 90 days all 6 birds in the 270-ppm group exhibited ataxia or paralysis whereas 5 of 6 birds in the 90-ppm group and 2 of 6 birds in the 30-ppm group were visibly affected. Treatment with 30 ppm or more resulted in a significant reduction in body weight. Brain neurotoxic esterase activity was inhibited by averages of 16, 69, 73, and 74% in the 10-, 30-, 90-, and 270-ppm groups, respectively. Brain acetylcholinesterase, plasma cholinesterase, and plasma alkaline phosphatase were significantly inhibited as well. Distinct histopathological effects were seen in the 30-, 90-, and 270-ppm groups which included demyelination and degeneration of axons of the spinal cord. Additional ducks were exposed in a similar manner to 60-, 270-, or 540-ppm leptophos (phosphonothioic acid O -4-bromo-2,5-dichlorophenyl- O -methylphenyl ester) which resulted in similar behavioral, biochemical, and histopathological alterations. these findings indicate that adult mallards are probably somewhat less sensitive than chickens to subchronic dietary exposure to organophosphorus insecticides that induce delayed neurotoxicity.

Toxicology and Applied Pharmacology↗

Neurotoxic and teratogenic effects of an organophosphorus insecticide (phenyl phosphonothioic acid- O -ethyl- O -[4-nitrophenyl] ester) on mallard development

Phenyl phosphonothioic acid- O -ethyl- O -[4-nitrophenyl] ester (EPN) is one of the 10 most frequently used organophosphorus insecticides and caused delayed neurotoxicity in adult chickens and mallards. Small amounts of organophosphorus insecticides placed on birds' eggs are embryotoxic and teratogenic. For this reason, the effects of topical egg application on EPN were examined on mallard ( Anas platyrhynchos ) embryo development. Mallard eggs were treated topically at 72 hr of incubation with 25 μl of a nontoxic oil vehicle or with EPN in the vehicle at concentrations of approximately 12, 36, or 108 μg/g egg, equivalent to one, three, and nine times the agricultural level of application used to spray crops. Treatment with EPN resulted in 22 to 44% mortality over this dose range by 18 days of development compared with 4 and 5% for untreated and vehicle-treated controls. EPN impaired embryonic growth and was highly teratogenic: 37–42% of the surviving embryos at 18 days were abnormal with cervical and axial scoliosis as well as severe edema. Brain weights were significantly lower in EPN-treated groups at different stages of development including hatchlings. Brain neurotoxic esterase (NTE) activity was inhibited by as much as 91% at 11 days, 81% at 18 days, and 79% in hatchlings. Examination of brain NTE activity during the course of normal development revealed an increase of nearly sixfold from Day 11 through hatching. The most rapid increase occurred between Day 20 and hatching. Brain acetylcholinesterase (AChE) activity was inhibited by as much as 41% at 11 days, 47% at 18 days, and 20% in hatchlings. Plasma cholinesterase and alkaline phosphatase activities were inhibited and plasma aspartate aminotransferase activity was increased at one or more stages of development. Hatchlings from EPN-treated eggs were weaker and slower to right themselves. Histopathological examination did not reveal demyelination and axonopathy of the spinal cord that was characteristic of delayed neurotoxicity in adult birds.

Toxicology and Applied Pharmacology↗

Biological effects of dietary T-2 toxin on rainbow trout, Salmo gairdneri

A 16-wk feeding study was conducted to evaluate the chronic toxicity of graded levels (0, 1.0, 2.5.5, 10 and 15 mg/kg of chemically pure dietary T-2 toxin (4,15-diacetoxy-8-(3-methylbutyryloxy)-12,13-epoxy-Δ 9 -tricothecen-3-ol) in 1-g rainbow trout, Salmo gairdneri , held in 9°C single-passage well water. Levels of T-2 toxin > 2.5 mg/kg depressed growth, efficiency of feed use, hematocrit, blood hemoglobin concentration and feed acceptance, and caused a transitory edema in a dose-dependent manner. Growth of trout fed a semipurified diet containing the toxin was described by the function: Y = 0.265 + 142.075 e (0.029 X 1 − 1.554x 2 3.7 ), where Y = gain as percentage starting weight per wk; X 1 is time in wk and 0 ⩽ X 1 ⩽16; and X 2 is T-2 content of diet in mg/kgand 0⩽ X 2 ⩽15. Exposure of fish to T-2 toxin did not affect activity of intestinal lumen chymoirypsin or trypsin, nitrogen digestibility or metabolizabte energy. Feeding of 15 mg/kg T-2 toxin to adult trout caused hemorrhaging in the intestines and regurgitation of subsequently intubated feed regardless of T-2 loxin content.

Aquatic Toxicology↗

Acute and chronic toxicity studies with monochlorobenzene in rainbow trout

The toxicity of monochlorobenzene (CB) was investigated in rainbow trout following acute intraperitoneal (i.p.) administration and chronic exposure via the water in a continuously flowing system for 15 or 30 days. In the acute study overt toxicity and hepatotoxicity were monitored over a 96-h time period. Variables measured to assess toxicity included weight changes, liver weight to body weight ratios, behavioral changes, alanine aminotransferase activity (GPT), sulfobromophthalein (BSP) retention, total plasma protein concentration and liver histopathology. In the chronic study the same measures of toxicity were followed as well as food consumption and alkaline phosphatase (AP) activity. Upon acute i.p. exposure the toxicant (9.8 mmol/kg) caused behavioral changes in the fish which were consistent with the known anesthetic properties of CB in mammals. Elevations in BSP retention and GPT activity, and histopathology indicated that CB was hepatotoxic in fish. The LC 50 of CB in trout exposed via the water for 96 h was 4.7 mg/l. Chronic exposure of trout to 2 or 3 mg/l CB resulted in similar behavioral changes as seen in the acute study. Liver toxicity was evident from elevations in GPT activity. BSP retention and AP activity appeared to be affected by the nutritional status of the trout as much as by the CB treatment. After 30 days of exposure to 3 mg/l CB, trout appeared to have developed some tolerance to the toxic effects.

Aquatic Toxicology↗

Uptake, biotransformation, and elimination of rotenone by bluegills (Lepomis macrochirus )

Yearling bluegills ( Lepomis macrochirus ) were exposed to sublethal concentrations of [ 14 C]rotenone (5.2 &mu;g/l) for 30 days in a continuous flow exposure system and then transferred to clean, flowing water for an additional 21-day depuration period. Rates of uptake and elimination and profile of the rotenoid metabolites in head, viscera, and carcass components were evaluated by 14 C counting and by high performance liquid chromatography. Total [ 14 C]rotenone derived activity was relatively uniform in all body components within 3 days after initial exposure and remained constant during the ensuing 27 days of exposure. Initial uptake rate coefficients were highest in viscera (K u = 80&middot; h -1 ) and were nearly identical for head (K u = 14 &middot; h) and carcass (K u = 10 &middot; h -1 ). Analyses of tissue extracts by high performance liquid chromatography confirmed the presence of at least six biotransformation products of rotenone. More than 60% of the activity extracted from viscera was present as a single peak which represented a compound that was extremely soluble in water. Rotenone composed only 0.3% of the extractable activity in viscera taken from fish exposed to rotenone for 30 days; however, rotenone accounted for 15.4% of extractable activity in the head and 20.1% in the carcass components. Rotenolone and 6 ' ,7 ' -dihydro-6 ' -,7 ' --dihydroxyrotenolone were tentatively identified as oxidation products in all tissue extracts. Elimination of 14 C activity from all body components was biphasic; both phases followed first-order kinetics. The rate of elimination was nearly equal for all body components during the initial phase but was most rapid from viscera during the second phase of elimination. Bioconcentration factors for the head, viscera, and carcass were 165, 3,550, and 125, respectively, when calculated on the basis of total 14 C activity but only 25.4, 11, and 26 when calculated as the concentration of parent material.

Aquatic Toxicology↗

Effect of injected rotenone on the production and composition of urine from the rainbow trout (Salmo gairdneri)

Renal function was evaluated in adult rainbow trout (Salmo gairdneri) dosed i.a. with rotenone at 225 and 275 &mu;g/kg. The chemical composition of urine samples and urine flow rates collected over a 5-h pretreatment period were compared with hourly urine samples collected over a 5-h posttreatment period. Significant increases in osmolality and in concentrations of sodium, potassium, chloride, glucose, and total protein were observed in the urine of treated fish. Urine solute concentrations reached maximum values within 1 to 3 h after treatment and decreased thereafter, indicating that the effects were reversible. Concentrations of sodium and chloride were highly correlated in 2-h posttreatment urine samples at the low ( r = 0.922) and high ( r = 0.981) rotenone treatments. Urine flow rates were reduced in trout at each dose of rotenone but the decrease in volume of urine voided was not dose-dependent. In a separate study, [ 14 C]polyethylene glycol was used as a filtration marker to determine the effect of rotenone treatment (225 &mu:g/kg) on urine flow rate, glomerular filtration rate, and renal water reabsorption. We showed that posttreatment urine flow rates were reduced partly by reduced glomerular filtration and partly by increased water reabsorption. Transient increases in plasma osmolality and hematocrit also were observed 0.5 h after rotenone treatment.

Aquatic Toxicology↗

Tissue distribution and elimination of rotenone in rainbow trout

The fate of a single i.v. dose (120 &mu;g/kg) of the piscicide [ 14 C]rotenone was evaluated in rainbow trout for periods up to 72 h after dosing. Rotenone was rapidly cleared from the plasma; less than 2% of the dose remained in the plasma compartment after 20 min. The highest concentrations of rotenone residues (% dose/g tissue) were in the hepatobiliary system, bile, intestine, and in heart, lateral line swimming muscle, and posterior kidney; tissues that are highly dependent on oxidative metabolism. Although rotenone activity was present in all cell fractions examined, greater than 40% was associated with the mitochondrial fraction of liver, kidney, and muscle. More than 85% of the activity extracted from these tissues, except the liver, was parent rotenone. Elimination from whole body and major tissue depots conformed to simple first-order kinetics; the estimated half-life from whole body was 68.5 h. Branchial elimination accounted for 5% of the injected dose over a 4-h period, and urinary elimination was less than 2% over a 48-h period. Rotenone was eliminated essentially unchanged across the gills; however, parent rotenone was not found in either urine or bile. More than 80% of the activity in both urine and bile eluted from HPLC chromatographs as a highly polar fraction that was not hydrolyzed by incubation with either &beta;-glucuronidase or sulfatase. The results imply that hepatobiliary excretion is the major route of elimination for rotenone residues in the trout and that metabolism to a more polar form is a prerequisite for elimination in both the bile and the urine

Aquatic Toxicology↗

In situ striped bass (Morone saxatilis) contaminant and water quality studies in the Potomac River

The objectives of this study were to evaluate survival of striped bass ( Morone saxatilis ) prolarvae and yearlings in the Potomac River by using in situ test chambers; correlate survival of both striped bass life stages with the presence of water quality conditions, inorganic contaminants, and organic contaminants and conduct histological examinations of surviving yearling striped bass. Survival of striped bass prolarvae ranged from 4.5–22.5% at three field locations during three 96-h experiments; control survival was ≥81%. Yearling survival ranged from 0–77.5% at three river stations during two 7-d experiments; highest mortality occurred at the upriver station. Control survival was 100%. Poor survival of striped bass prolarvae was likely related to the presence of inorganic contaminants (monomeric aluminum, cadmium, and copper) acting singly or synergistically and sudden decreases in water temperature (< 11°C). Yearling mortality at the upriver station was likely correlated with high pH conditions from a point source discharge and perhaps inorganic contaminants. Histological examinations of yearlings suggested that test organisms exposed to Potomac River water exhibited adverse changes in the kidney.

Maryland, Virginia↗

Disposition of pentachlorophenol in rainbow trout (Salmo gairdneri): Effect of inhibition of metabolism

The accumulation kinetics of pentachlorophenol (PCP) were investigated in rainbow trout ( Salmo gairdneri ) in the absence and presence of 25 mg/1 salicylamide, an inhibitor of PCP metabolism. After exposure to 5 μg/1 PCP over 1–96 h, the amount of PCP in the whole fish, its concentration in water and the total amount of metabolites (water, whole fish and bile) were measured. Equations for these variables, based on a two compartment pharmacokinetic model, were fitted simultaneously to the data using the computer program NONLIN, which uses an iterative nonlinear least squares technique. Salicylamide decreased the metabolic clearance of PCP, which resulted in an increase in the bioconcentration factor (BCF); this increase was partially offset by a salicylamide-induced decrease in the apparent volume of distribution of PCP. A clearance-volume compartment model permitted partitioning of the BCF in terms of the underlying physiologic and biochemical processes (uptake clearance, metabolic clearance and apparent volume of distribution). With this approach the BCF can be categorized as either dependent (e.g., PCP) or independent of uptake and metabolism (elimination) based on the relative sizes of the clearances for uptake and metabolism. Inhibition of PCP metabolism resulted in a loss of its dependence on uptake and metabolism. The BCF estimated as the apparent volume of distribution may be useful for assessment of the risk associated with exposure and bioaccumulation potential, as elimination is generally quite variable among aquatic species.

Aquatic Toxicology↗