David J. Hoffman

EMBRYONIC MORTALITY AND ABNORMALITIES OF AQUATIC BIRDS: APPARENT IMPACTS OF SELENIUM FROM IRRIGATION DRAINWATER

Abstract Severe reproductive impacts were found in aquatic birds nesting on irrigation drainwater ponds in the San Joaquin Valley of California. Of 347 nests studied to late incubation or to hatching, 40.6% had at least one dead embryo and 19.6% had at least one embryo or chick with an obvious external anomaly. The deformities were often multiple and included missing or abnormal eyes, beaks, wings, legs and feet. Brain, heart, liver and skeletal anomalies were also present. Mean selenium concentrations in plants, invertebrates, and fish from the ponds were 22–175 ppm (dry weight), about 12 to 130 times those found at a nearby control area. Bird eggs (2.2–110 ppm) and livers (19–130 ppm) also contained elevated levels of selenium. Aquatic birds may experience similar problems in other areas where selenium occurs at elevated levels.

Selenium toxicosis in wild aquatic birds

Severe gross and microscopic lesions and other changes were found in adult aquatic birds and in embryos from Kesterson Reservoir (a portion of Kesterson National Wildlife Refuge), Merced County, Calif., during 1984. Adult birds from that area were emaciated, had subacute to extensive chronic hepatic lesions, and had excess fluid and fibrin in the peritoneal cavity. Biochemical changes in their livers included elevated glycogen and non-protein-bound sulfhydryl concentrations and glutathione peroxidase activity but lowered protein, total sulfhydryl, and protein-bound sulfhydryl concentrations. Congenital malformations observed grossly in embryos were often multiple and included anophthalmia, microphthalmia, abnormal beaks, amelia, micromelia, ectrodactyly, and hydrocephaly. Mean concentrations of selenium in livers (94.4 ppm, dry weight) and kidneys (96.6 ppm) of birds collected at the Kesterson ponds were about 10 times those found at a nearby control area (8.3 and 12.2 ppm). We conclude that selenium present in the agricultural drainage water supplied to the Kesterson ponds accumulated in the food chain of aquatic birds to toxic concentrations and caused the lesion and other changes observed.

Metal contamination in wildlife living near two zinc smelters

Abstract Wildlife in an oak forest on Blue Mountain was studied 10 km upwind (Bake Oven Knob site) and 2 km downwind (Palmerton site) of two zinc smelters in eastern Pennsylvania, USA. Previous studies at sites near these smelters had shown changes in populations of soil microflora, lichens, green plants and litter-inhabiting arthropods. The 02 soil litter horizon at Palmerton was heavily contaminated with Pb (2700 mg kg −1 ), Zn (24 000 mg kg −1 ), and Cd (710 mg kg −1 ), and to a lesser extent with Cu (440 mg kg −1 ). Various kinds of invertebrates (earthworms, slugs and millipedes) that feed on soil litter or soil organic matter were rare at, or absent from, the Palmerton site. Those collected at Bake Oven Knob tended to have much higher concentrations of metals than did other invertebrates. Frogs, toads and salamanders were very rare at, or absent from, the Palmerton site, but were present at Bake Oven Knob and at other sites on Blue Mountain farther from the smelters. Metal concentrations (dry wt) in different organisms from Palmerton were compared. Concentrations of Pb were highest in shrews (110 mg kg −1 ), followed by songbirds (56 mg kg −1 ), leaves (21 mg kg −1 ), mice (17 mg kg −1 ), carrion insects (14 mg kg −1 ), berries (4·0 mg kg −1 ), moths (4·3 mg kg −1 ) and fungi (3·7 mg kg −1 ). Concentrations of Cd, in contrast, were highest in carrion insects (25 mg kg −1 ), followed by fungi (9·8 mg kg −1 ), leaves (8·1 mg kg −1 ), shrews (7·3 mg kg −1 ), moths (4·9 mg kg −1 ), mice (2·6 mg kg −1 ), songbirds (2·5 mg kg −1 ) and berries (1·2 mg kg −1 ). Concentrations of Zn and Cu tended to be highest in the same organisms that had the highest concentrations of Cd. Only a small proportion of the metals in the soil became incorporated into plant foliage, and much of the metal contamination detected in the biota probably came from aerial deposition. The mice from both sites seemed to be healthy. Shrews had higher concentrations of metals than did mice, and one shrew showed evidence of Pb poisoning; its red blood cell ALAD activity was greatly reduced, one kidney contained acid-fast intranuclear inclusion bodies, and the other kidney had 280 mg kg −1 Pb (wet wt). The livers from two cuckoos from Palmerton had elevated Pb concentrations (18 and 25 mg kg −1 , wet wt). Most of the songbirds seemed to be healthy.

Embryotoxicity, teratogenicity, and aryl hydrocarbon hydroxylase activity in Forster's terns on Green Bay, Lake Michigan.

Known reproductive problems, including congenital malformations and poor hatching success, exist for the state endangered Forsters tern (Sterna forsteri) in Green Bay, Wisconsin. Twenty Forsters tern eggs were collected from separate nests at a natural colony with documented reproductive problems, situated at Green Bay, Lake Michigan, and an inland colony at Lake Poygan (control) where reproduction was documented as normal. Eggs from the two locations were placed in the same laboratory incubator and candled throughout incubation. Hatching success of Green Bay eggs was 52% of that for controls. Several early embryonic deaths occurred, but most mortality occurred close to the time of hatching. Liver microsomal aryl hydrocarbon hydroxylase activity was elevated approximately threefold in Green Bay hatchlings compared to controls. Green Bay terns that hatched weighed less than controls, had an increased liver to body weight ratio, and had a shorter femur length. Two Green Bay embryos that failed to hatch had anomalies, one with a crossed beak and one with poor ossification of the foot. One Green Bay hatchling had an abnormally ossified ilium. These effects were observed in eggs where there were measureable levels of aryl hydrocarbon hydroxylase inducers including polychlorinated biphenyls and polychlorinated dibenzo-p-dioxins.

Selenium teratogenesis in natural populations of aquatic birds in Central California

The frequency and types of malformations are described that were encountered during the spring of 1983 in a natural population of aquatic birds exposed to agricultural drainwater ponds and food items containing high concentrations of selenium in central California. A total of 347 nests of aquatic birds containing 1,681 eggs was selected for study at Kesterson Reservoir located in the Kesterson National Wildlife Refuge (NWR), Merced County, California. Embryos collected during incubation or from eggs that failed to hatch were examined to determine the age at death and presence of malformations. Embryonic death was generally high; approximately 17–60% of the nests of different species contained at least one dead embryo. The incidence of malformed embryos was also high; approximately 22–65% of the nests where at least two embryos were examined contained abnormal embryos. American coots (Fulica americana) and black-necked stilts (Himantopus mexicanus) experienced the highest incidence of malformed embryos. For all species, the average percentage of eggs containing dead or live abnormal embryos was 16.1 whereas the average percentage containing live abnormal embryos was 10.7. Multiple gross malformations of the eyes, brain, and feet were often present. Brain defects included hydrocephaly and exencephaly. Eye defects included both unilateral and bilateral anophthalmia and microphthalmia. Eye and foot defects with ectrodactyly and swollen joints were the most common in coots. Beak defects also occurred frequently and most often included incomplete development of the lower beak of ducks (Anas spp.) and stilts. Wing and leg defects were most prevalent in stilts and ducks, with ectromelia and amelia most prevalent in stilts. Other malformations occurring at lower frequencies included enlarged hearts with thin ventricular walls, liver hypopiasia, and gastroschisis. Based upon simultaneous examination of a control population of aquatic birds of the same species and published studies, the incidences of embryonic mortality and deformities were 9–30 times greater than expected. The role of the form of selenium responsible for teratogenesis in laboratory studies is discussed.

Embryotoxic and teratogenic effects of selenium in the diet of mallards

Mallards (Anas platyrhynchos) were fed a control diet, diets containing 1, 5, 10, or 25 ppm Se as sodium selenite, or a diet containing 10 ppm Se as seleno-DL-methionine in the first of two experiments. Selenium at 10 ppm as selenomethionine or 25 ppm as sodium selenite caused a 40-44% decrease in the total number of eggs that hatched compared to controls. Selenium at 25 ppm (sodium selenite) resulted in a 19% decrease in mean embryonic weight at 18 d of incubation, accompanied by a 6% decrease in crown-rump length. Ten parts per million Se as selenomethionine was more teratogenic than sodium selenite at 25 ppm. Selenomethionine (10 ppm Se) resulted in an incidence of 13.1% malformations that were often multiple, whereas sodium selenite (10 and 25 ppm Se) resulted in 3.6 and 4.2% malformations. The teratogenicity of selenomethionine was confirmed in a second experiment in which mallards received 1, 2, 4, 8, or 16 ppm Se as selenomethionine, resulting in 0.9, 0.5, 1.4, 6.8, and 67.9% malformations, respectively. These malformations included hydrocephaly, microphthalmia, lower bill defects, and foot defects with ectrodactyly. Both forms of selenium increased the incidence of edema and stunted embryonic growth. Selenomethionine (10 ppm Se) resulted in a significant increase of approximately 40% in plasma glutathione peroxidase activity and a 70% increase in sorbitol dehydrogenase activity (indicative of hepatotoxicity) in hatchlings. Sodium selenite (25 ppm Se) resulted in fourfold elevation in plasma uric acid concentration, indicative of renal alteration. Selenomethionine accumulated much better in eggs than did sodium selenite. These findings indicate that selenomethionine is considerably more teratogenic and generally more embryotoxic than sodium selenite, probably due to higher uptake of selenomethionine.

Association between PCBs and lower embryonic weight in black-crowned night herons in San Francisco Bay

Reproductive problems, including congenital malformations, reduced hatching success, and decreased survival of hatchlings, have been observed in colonial-nesting water birds at the San Francisco Bay National Wildlife Refuge (SFBNWR). Twenty-four black-crowned night heron (Nycticorax nycticorax) eggs were collected from SFBNWR in 1983. Twelve of these were collected from separate nests when late-stage embryos were pipping, and an additional egg was randomly collected from each nest for organochlorine analysis. Overt anomalies and skeletal defects were not apparent. Embryonic weights (with partially absorbed yolk sacs removed) were 15% lower (p less than 0.05) in SFBNWR embryos compared to control embryos from the Patuxent Wildlife Research Center (PWRC). Crown-rump length and femur length were shorter for SFBNWR embryos. The geometric mean polychlorinated biphenyl (PCB) concentration in SFBNWR eggs was 4.1 ppm wet weight, with a range of 0.8-52.0 ppm. A negative correlation (r = -0.61; p less than 0.05) existed between embryonic weight and log-transformed PCB residues in whole eggs collected from the same nest at SFBNWR, suggesting a possible impact of PCBs on embryonic growth. A correlation with embryonic weight did not occur for DDE [1,1-dichloro-2,2-bis(p-chlorophenyl) ethylene] residues. Liver microsomal aryl hydrocarbon hydroxylase activity was neither significantly elevated nor correlated with PCB, DDE, or PCB plus DDE log-transformed residues. It is unknown whether the apparent association between PCBs and lower weight is persistent through hatching.

EFFECTS OF LEAD SHOT INGESTION ON δ-AMINOLEVULINIC ACID DEHYDRATASE ACTIVITY, HEMOGLOBIN CONCENTRATION, AND SERUM CHEMISTRY IN BALD EAGLES

Lead shot ingestion by bald eagles (Haliaeetus leucocephalus) is considered to be widespread and has been implicated in the death of eagles in nature. It was recently demonstrated under experimental conditions that ingestion of as few as 10 lead shot resulted in death within 12 to 20 days. In the present study hematological responses to lead toxicity including red blood cell ALAD activity, hemoglobin concentration and 23 different blood serum chemistries were examined in five captive bald eagles that were unsuitable for rehabilitation and release. Eagles were dosed by force-feeding with 10 lead shot; they were redosed if regurgitation occurred. Red blood cell ALAD activity was inhibited by nearly 80% within 24 hours when mean blood lead concentration had increased to 0.8 parts per million (ppm). By the end of 1 week there was a significant decrease (20-25%) in hematocrit and hemoglobin, and the mean blood lead concentration was over 3 ppm. Within as little as 1-2 weeks after dosing, significant elevations in serum creatinine and serum alanine aminotransferase occurred, as well as a significant decrease in the ratio of serum aspartic aminotransferase to serum alanine aminotransferase. The mean blood lead concentration was over 5 ppm by the end of 2 weeks. These changes in serum chemistry may be indicative of kidney and liver alterations.

Lead concentrations and reproductive success in european starlings Sturnus vulgaris nesting within highway roadside verges

In 1981, we studied lead concentrations and reproductive success in freeliving European starlings Sturnus vulgaris nesting within the verges of two Maryland highways with different traffic volumes, Route 197 (average daily traffic volume [ADT] = 10 800 vehicles) and the Baltimore-Washington Parkway (ADT = 52 500 vehicles) and a nearby control area. Concentrations (mgkg−1 dry weight) of lead in the ingesta (84–94 mg kg−1), carcasses (4·0–9·6 mgkg−1) and feathers (6·8–52 mgkg−1) of Parkway nestlings and adults were 3 to 13 times those found in starlings from the control area, whereas lead concentrations in the ingesta and tissues of starlings from the verge of Route 197 were similar to those of controls. Activity of δ-aminolevulinic acid dehydratase (ALAD) in red blood cells (RBCs) of adult and nestling starling from the Parkway was depressed from 43 to 60% compared to controls. RBC ALAD activity in adults from nests along Route 197 was similar to that of adult starlings from the control area, but that of their young was depressed 17%. Haemoglobin concentrations (−16%) and haematocrits (−10%) in Parkway nestlings were depressed compared with those of nestlings from the other two study areas, whereas those of adults were not affected. Clutch size, number of young hatched and the number of young in nests 1 to 3 days before fledging were similar among sites, as were body weights of adults and prefledging weights of their young. However, brain weights of Parkway nestlings were lower (P<0·05) than those of nestlings from the other study areas. Results suggest that lead within verges of major highways probably does not pose a serious hazard to adult ground-foraging songbirds. However, the effects of lead-induced reductions in haemoglobin concentration, haemotocrit, RBC ALAD activity and brain weight on the postfledging survival of their young are not known.

Embryotoxic effects of benzo [a] pyrene, chrysene, and 7,12‐dimethylbenz [a] anthracene in petroleum hydrocarbon mixtures in mallard ducks

Studies with different avian species have revealed that surface applications of microliter amounts of some crude and fuel oils that coat less than 10% of the egg surface result in considerable reduction in hatching with teratogenicity and stunted growth. Other studies have shown that the embryotoxicity is dependent on the aromatic hydrocarbon content, further suggesting that the toxicity is due to causes other than asphyxia. In the present study the effects of three polycyclic aromatic hydrocarbons identified in petroleum were examined on mallard (Anas platyrhynchos) embryo development. Addition of benzo[a]pyrene (BaP), chrysene, or 7,12-dimethylbenz[a]anthracene (DMBA) to a synthetic petroleum hydrocarbon mixture of known composition and relatively low embryotoxicity resulted in embryotoxicity that was enhanced or equal to that of crude oil when 10 microliter was applied externally to eggs at 72 h of development. The order of ability to enhance embryotoxicity was DMBA greater than BaP greater than chrysene. The temporal pattern of embryonic death was similar to that reported after exposure to crude oil, with additional mortality occurring after outgrowth of the chorioallantois. Retarded growth, as reflected by embryonic body weight, crown-rump length, and bill length, was accompanied by teratogenicity. Abnormal embryos exhibited extreme stunting; eye, brain, and bill defects; and incomplete ossification. Gas chromatographic-mass spectral analysis of externally treated eggs showed the passage of aromatic hydrocarbons including chrysene through the shell and shell membranes to the developing embryos. These findings suggest that the presence of polycyclic aromatic hydrocarbons in petroleum, including BaP, chrysene, and DMBA, significantly enhances the overall embryotoxicity in avian species.