Accumulation of Environmental Contaminants in Wood Duck (<Emphasis Type="Italic">Aix sponsa</Emphasis>) Eggs,with Emphasis on Polychlorinated Dibenzo-<Emphasis Type="Italic">p</Emphasis>-Dioxins and Polychlorinated Dibenzofurans

We measured polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), polychlorinated biphenyls (PCBs), organochlorine pesticides, and mercury in wood duck (Aix sponsa) eggs collected near a North Carolina (USA) bleached kraft paper mill. Samples were taken a decade after the mill stopped using molecular chlorine. Using avian toxic equivalency factors, 2,3,7,8-tetrachlorodibenzo-p-dioxin toxicity equivalent (TEQ) concentrations were 1–30 pg/g fresh wet weight in eggs (n = 48) collected near the mill in 2002–2005 and were significantly higher than those from a reference site (<1 pg/g) 25 km away. Geometric mean wood duck egg TEQs (6 pg/g) were one-fifth those measured at this site prior to the cessation of molecular chlorine bleaching. Concentrations of mercury in wood duck eggs from nests of the Roanoke River sites ranged from 0.01 to 0.14 μg/g (geometric mean, 0.04 μg/g) and were significantly higher than those from the reference site, where concentrations did not exceed 0.04 μg/g (geometric mean, 0.02 μg/g). All concentrations were lower than those associated with adverse effects in birds. The congener profiles, lack of contamination in reference site eggs, and decline in contaminant concentrations after process changes at the mill provide strong evidence that mill discharges influenced contamination of local wood duck eggs. Collectively, the results indicate that the wood duck is an effective sentinel of the spatial and temporal extent of PCDD, PCDF, and mercury contamination.     

Polychlorinated Dibenzo-<Emphasis Type="Italic">p</Emphasis>-Dioxins,Dibenzofurans, and Dioxin-Like Polychlorinated Biphenyls in Livers of Birds from Japan

Concentrations of 2,3,7,8-substituted polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and non- and mono-ortho-chlorine-substituted polychlorinated biphenyls (dioxin-like PCBs) were measured in livers of 17 species of birds collected from Japan. Birds were grouped according to their feeding habits as granivores, piscivores, omnivores, and predators for discussions. Livers of granivores contained relatively low concentrations of PCDD/DFs (80–660 pg/g) followed in increasing order by omnivores (2,300–8,000 pg/g), piscivores (61–12,000 pg/g) and predators (480–490,000 pg/g on a fat weight basis). Especially, one species of predatory bird (mountain hawk eagle) contained elevated concentrations of PCDDs, PCDFs, and dioxin-like PCBs, and the measured concentration is one of the highest reported to date. Homolog and congener patterns of PCDDs and PCDFs varied among species; hence, the results suggested that feeding habits, specific elimination, and metabolism influence contamination pattern. Concentrations of dioxin-like PCBs were in the order of granivores (32–83 ng/g) < predators [excluding mountain hawk eagle] (32–2,500 ng/g) < piscivore (61–12,000 ng/g) < omnivores (1,800–67,000 ng/g on a fat weight basis). Mountain hawk eagle contained the highest concentration of dioxin-like PCBs (55,000 ng/g fat weight). 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) toxic equivalents (TEQs) ranged from 53–450,000 pg/g fat weight. 23478-PeCDF, 2378-TCDD/TCDF, and PCB congeners IUPAC 126 and 77 were major contributors to TEQs in birds. To our knowledge, this is the first study of PCDD/DFs and dioxin-like PCBs in livers of several species of Japanese birds. Received: 11 March 2001/Accepted: 5 August 2001     

Concentrations of Polychlorinated Dibenzo-p-Dioxins (PCDDs), Polychlorinated Dibenzofurans (PCDFs), and Dioxin-Like PCBs in Adipose Tissue of Infertile Men

Some epidemiological studies suggested the occurrence of an alteration in the male reproductive function in the past 50 years, particularly a decrease in the sperm count and quality, an increase in the malformations frequency of the reproductive apparatus (cryptorchidism and hypospadias) and of testicular cancers. Especially according the laboratory animals studies, polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs) have been suspected to play a crucial and deleterious role in the alteration of human fertility. For this reason, we measured adipose tissue concentrations of PCDDs/Fs and dioxin-like PCBs in 23 fertile and 22 infertile men living in Ankara, Turkey. Adipose tissue samples were analyzed for PCDD/F and 12 dioxin-like PCB congeners using high-resolution gas chromatography/high-resolution mass spectrometry. For the fertile and infertile groups, the World Health Organization (WHO)_PCDD/F-TEQ concentrations ranged from 3.0 to 15.8 pg/g fat and from 2.8 to 17.2 pg/g fat, respectively (4.4–31.5 and 4.7–22.3 WHO-TEQs/g fat, respectively, including dioxin-like PCBs) (p > 0.05). The mean concentrations of WHO_PCDD/F-TEQ and WHO_PCB-TEQ have been calculated as 7.2 and 12.5 pg/g (on a lipid basis) for the fertile group and 7.0 and 9.4 pg/g for the infertile group, respectively. Concentrations of each of the PCDD/F and dioxin-like PCB congeners were compared in fertile and infertile groups among themselves, and no statistical significance was obtained (p > 0.05), except 2,3,7,8-tetrachlorodibenzofuran (p = 0.0029) and 1,2,3,4,6,7,8,9-octachlorodibenzofuran (p = 0.01).     

Microcontaminants and reproductive impairment of the Forster's tern on Green Bay,Lake Michigan-1983

For the 1983 nesting season, Forster's tern (Sterna forsteri) reproductive success was significantly impaired on organochlorine contaminated Green Bay, Lake Michigan compared to a relatively uncontaminated inland location at Lake Poygan, Wisconsin. Compared with tern eggs from Lake Poygan, eggs from Green Bay had significantly higher median concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), other polychlorinated dibenzo-p-dioxins (PCDDs), total polychlorinated biphenyls (PCBs), total (three congeners) non-ortho, ortho' PCBs, five individual PCB congeners known to induce aryl hydrocarbon hydroxylase (AHH) and several other organochlorine contaminants. Conversions of analytical concentrations of TCDD and PCB congeners based on relative AHH induction potencies allowed for estimation of total 2,3,7,8-TCDD equivalents. Two PCB congeners, 2,3,3,4,4- and 3,3,4,4,5-pentachlorobiphenyl (PeCB) accounted for more than 90% of the median estimated TCDD equivalents at both Green Bay and Lake Poygan. The median estimated TCDD equivalents were almost 11-fold higher in tern eggs from Green Bay than in eggs from Lake Poygan (2175 and 201 pg/g). The hatching success of Green Bay sibling eggs from nests where eggs were collected for contaminant analyses was 75% lower at Green Bay than at Lake Poygan. Hatchability of eggs taken from other nests and artificially incubated was about 50% lower for Green Bay than for Lake Poygan. Among hatchlings from laboratory incubation, those from Green Bay weighed approximately 20% less and had a mean liver weight to body weight ratio 26% greater than those from Lake Poygan. In both field and laboratory, mean minimum incubation periods were significantly longer for eggs from Green Bay compared to Lake Poygan (8.25 and 4.58 days, respectively). Mean minimum incubation time for Green Bay eggs in the field was 4.37 days longer than in the laboratory. Hatchability was greatly improved when Green Bay eggs were incubated by Lake Poygan adults in an egg-exchange experiment, but was sharply decreased in Lake Poygan eggs incubated in Green Bay nests. Nest abandonment and egg disappearance were substantial at Green Bay but nil at Lake Poygan. Thus, not only factors intrinsic to the egg, but also extrinsic factors (parental attentiveness), impaired reproductive outcome at Green Bay. The epidemiological evidence from this study strongly suggested that contaminants were a causal factor. AHH-active PCB congeners (intrinsic effects) and PCBs in general (extrinsic effects) appeared to be the only contaminants at the concentrations measured in eggs, capable of producing the effects that were observed at Green Bay.     

A Survey of Dietary Intake of Polychlorinated Dibenzo-<Emphasis Type="Italic">p</Emphasis>-dioxins,Polychlorinated Dibenzofurans,and Dioxin-like Coplanar Polychlorinated Biphenyls from Food During 2000–2002 in Osaka City,Japan

Studies of the dietary intake of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and dioxin-like coplanar polychlorinated biphenyls (dioxin-like PCBs), collectively referred as dioxins, were conducted using the total diet study (TDS) method with food purchased in Osaka City, Japan during 2000–2002. The daily intake of dioxin-TEQ (toxic equivalent) from food per adult person was estimated respectively as 104.24 pg TEQ/person/day in 2000, 72.73 pg TEQ/person/day in 2001, and 87.28 pg TEQ/person/day in 2002, corresponding to 2.08, 1.45, and 1.74 pg TEQ/kg body weight (bw)/day for an adult weighing 50 kg. The highest contribution ratio to the total intake of dioxin-TEQ was from fish and shellfish (group 10) in each year, accounting for 77–92%. The next highest contributor was meat and eggs (groups 11-A and 11-B). An annual decrease of the intake of dioxin-TEQ was not observed clearly. Otherwise, the dietary intake of non-2,3,7,8-substituted PCDDs/PCDFs were estimated together. The intake of 1,3,6,8-TeCDD came mostly from intake of fish and shellfish (group 10), green vegetable (group 7), and the other vegetables, including mushrooms and seaweed (group 8). In addition, the intake of 1,3,6,8-TeCDF, which reportedly antagonizes 2,3,7,8-TeCDD-mediated aryl hydrocarbon hydroxylase (AHH) and ethoxyresorufin O-deethylase (EROD) induction activities, was an insufficient amount to suppress 2,3,7,8-TeCDD-mediated activities. Furthermore, we discussed different TEQ compositions of PCDDs/PCDFs to dioxin-like PCBs from food intake and in human samples and inferred that the difference was caused by low bioaccumulation properties of 3,3′,4,4′,5-PeCB (PCB 126).     

Blood levels of 2,3,7,8-tetrachlorodibenzo-p-dioxin in chemical workers after chloracne and in comparison groups

Summary Nine production workers with a history of chloracne from exposure to polychlorinated dibenzo-p-dioxins (PCDDs) in 1971–1973 had a median level of 340 pg 2,3,7,8-tetrachlorodibenzo p-dioxin (TCDD) per gram blood lipid (range 98–659 pg/g) in 1990. This was significantly higher than blood levels in 4 reference subjects without chloracne and no known exposure from the same plant (median 18, range 8–28 pg/g) and in 17 external referents (median 16, range < 5–23 pg/g). We therefore consider chloracne to be a reliable indicator of heavy dioxin exposure in a cohort of herbicide production workers and will use its former presence in 159 cases as a substitute for exposure when investigating cancer morbidity and mortality in a multicenter cohort study.     

Maternal exposure to 2,3,7,8-tetrachlorodibenzo-<Emphasis Type="Italic">p</Emphasis>-dioxin and the body burden in offspring of long-evans rats

Objectives   In utero and lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) results in a wide variety of developmental effects in pups at doses much lower than those causing overt toxicity in adult animals. We investigated the relationship between tissue concentrations of TCDD in dams and fetuses and developmental effects on pups. Materials and Methods  Pregnant Long-Evans rats were given TCDD at a single oral dose of 12.5, 50, 200, or 800 ng of TCDD or [³H]-TCDD/kg bw on gestation day (GD) 15. Dams were sacrificed on GD16 and GD21, and the tissue concentrations of TCDD were measured in dams and fetuses. Pups were sacrificed on postnatal day (PND) 49 and PND63 for males and PND70 for females, and the reproductive effects and tissue concentrations of TCDD were determined. Results  The sex ratio (male/female) on GD21 was significantly reduced at 50 ng TCDD/kg and at 12.5 and 50 ng TCDD/kg at birth, but not at other doses. Delayed puberty was observed in males at 200 ng TCDD/kg and in males and females at 800 ng TCDD/kg. Anogenital distance, testis weight, epididymal sperm count, sperm motility, and ejaculated sperm count were not affected. Estrous cyclicity was not different from that of the control in any treatment group. A dose-dependent decrease in weight of seminal vesicle and prostate on PND49 was observed. Prostate weight was significantly decreased at 800 ng TCDD/kg. At this dose, maternal body burden and TCDD concentration in fetuses were 290 pg TCDD/g and 52 pg TCDD/g on GD16, respectively. Reduced prostate weight is a sensitive and commonly observed endpoint so that the body burdens of dams and fetuses at the LOAEL of this endpoint could be served as the basis for establishing TDI for dioxins.     

Levels of PCDD/Fs,Dl-PCBs and HCB in Air,Soils and Sediments from a City with Multiple Thermal-Related Industries in China

The levels of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), dioxin-like polychlorinated biphenyls (dl-PCBs) and hexachlorobenzene (HCB) were measured in various environmental compartments in Tangshan, China, which contains multiple thermal-related industries. The total toxic equivalent concentrations of these pollutants were 138 ± 87.2 fg/m³ in air, 3.43 ± 2.88 pg/g in soils, and 1.42 ± 1.5 pg/g in sediments. The 2,3,7,8-PCDD/Fs profiles in atmospheric samples suggest that thermal-related industries are the most likely potential sources. Of the dl-PCBs, CB-77, CB-105 and CB-118 were the most abundant congeners and CB-126 was the dominant contributor to the TEQs from the dl-PCBs.     

Levels and Profiles of Polychlorinated Dibenzo-p-Dioxins, Polychlorinated Dibenzofurans, and Polychlorinated Biphenyls in Feedstuffs and Milk From Farms in the Vicinity of Incineration Plants in Tuscany, Italy

Levels of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and polychlorobiphenyls (PCBs) were determined in samples of bovine and ovine milk collected in farms located in the province of Grosseto, Tuscany, in the vicinity of two incineration plants as well as in farms located in areas with presumable background levels of contamination. Samples of feedstuffs of local origin used in the investigated farms were also collected and analysed. The cumulative levels of PCDDs, PCDFs, and dioxin-like PCBs (DL-PCBs) in feedstuffs ranged from 0.25 to 0.61 pg WHO-TE/g fresh weight (fw) in the farms under impact from incinerator emissions and from 0.21 to 0.34 pg WHO-TE/g fw in the control area farms. The sums of the six non–dioxin-like indicator PCB (NDL-PCB) congeners were 0.13 to 9.3 ng/g fw and 1.2 to 1.9 ng/g fw, respectively. In milk samples, the levels detected were 0.71 to 2.9 pg WHO-TE/g fat and 0.52 to 0.59 pg WHO-TE/g fat in farms under impact from the incinerators and in the control area farms, respectively. The corresponding sums of the six indicator PCB congeners were 1.4 to 8.2 ng/g fat and 0.90 to 1.6 ng/g fat. In all samples, contamination levels were below the limits set by the European Community (EC; Commission Directive 2006/13/EC and Commission Regulation 1881/2006/EC.) No relevant differences were found between samples collected in potentially exposed areas and control areas for total toxic equivalents and cumulative analytic levels of PCDDs, PCDFs, DL-PCBs, and NDL-PCBs (p > 0.05, Mann-Whitney U test). Observed levels were in agreement with those found in other countries in areas with background levels of exposure. Congener levels and profiles of PCDDs, PCDFs, and PCBs in feedstuffs and milk samples that were compared describe congener-specific transfer behavior.     

Factors Affecting the Toxicity of Methylmercury Injected into Eggs

We developed a standardized protocol for comparing the sensitivities of the embryos of different bird species to methylmercury when methylmercury was injected into their eggs. During the course of developing this protocol, we investigated the effects of various factors on the toxicity of the injected methylmercury. Most of our experiments were done with chicken (Gallus domesticus), mallard (Anas platyrhynchos), and ring-necked pheasant (Phasianus colchicus) eggs, all of which were purchased in large numbers from game farms. A smaller amount of work was done with double-crested cormorant (Phalacrocorax auritus) eggs collected from the wild. Several solvents were tested, and corn oil at a rate of 1 μl/g egg contents was selected for the final standardized protocol because it had minimal toxicity to embryos and because methylmercury dissolved in corn oil yielded a dose–response curve in a range of egg concentrations that was similar to the range that causes reproductive impairment when the mother deposits methylmercury into her own eggs. The embryonic stage at which eggs were injected with corn oil altered mercury toxicity; at early stages, the corn oil itself was toxic. Therefore, in the final protocol we standardized the time of injection to occur when each species reached the morphologic equivalent of a 3-day-old chicken embryo. Although solvents can be injected directly into the albumen of an egg, high embryo mortality can occur in the solvent controls because of the formation of air bubbles in the albumen. Our final protocol used corn oil injections into the air cell, which are easier and safer than albumen injections. Most of the methylmercury, when dissolved in corn oil, injected into the air cell passes through the inner shell membrane and into the egg albumen. Most commercial incubators incubate eggs in trays with the air cell end of the egg pointing upward, but we discovered that mercury-induced mortality was too great when eggs were held in this orientation. In addition, some species of bird eggs require incubation on their sides with the eggs being rolled 180° for them to develop normally. Therefore, we adopted a procedure of incubating the eggs of all species on their sides and rolling them 180° every hour. Little has been published about the conditions of temperature, humidity, and the movements to which eggs of wild birds need to be subjected for them to hatch optimally under artificial incubation. Not unexpectedly, hatching success in an artificial incubator is generally less than what natural incubation by the parents can achieve. However, the survival of control embryos of most wild bird species was good (generally ≥ 80%) up to within 1 or 2 days of hatching when we incubated the eggs at 37.5°C (or 37.6°C for gallinaceous species) at a relative humidity that resulted in an approximate 15% to 16% loss in egg weight by the end of incubation and by incubating the eggs on their sides and rolling them 180°/h. To improve statistical comparisons, we used survival through 90% of incubation as our measurement to compare survival of controls with survival of eggs injected with graded concentrations of mercury.     

Instrumental and Bioanalytical Measures of Persistent Organochlorines in Blue Mussel (Mytilus edulis) from Korean Coastal Waters

Blue mussels (Mytilus edulis) collected from 34 locations along the south and east coast of Korea were analyzed for polychlorinated biphenyls (PCBs) and organochlorine (OC) pesticides. Maximum concentrations of PCBs and total OC pesticides were 98.5 and 20.5 ng/g, wet weight, respectively. Extracts were fractionated by Florisil chromatography and each fraction was screened for dioxin-like activity in vitro, using recombinant rat hepatoma cells (H4IIE-luc). Fraction 2 (F-2), which contained hexachlorocyclohexanes, chlordanes, p,p′-DDD, and p,p′-DDT, generally elicited significant dioxin-like activity compared to control, whereas Fraction 1 (F-1), which contained PCBs, p,p′-DDE, and hexachlorobenzene, did not. The greatest magnitude of dioxin-like response observed was 44% of the maximum response elicited by a 2,000 pM 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) standard. The relatively low magnitudes of dioxin-like response observed for F-1 samples were consistent with the relatively low PCB concentrations. At concentrations equal to the maximum observed in the mussel samples, neither individual OC pesticides nor a mixture of OC pesticides yielded a significant dioxin-like response in the H4IIE-luc assay. Thus, the concentrations of OC pesticides in F-2 did not appear to have accounted for the dioxin-like activity observed. This suggests the presence of unidentified and/or unknown, acid-stable, dioxin-like compounds in F-2. This study suggests that in vitro bioassays are useful in assessing the contamination of mussels collected from coastal marine locations. Received: 28 November 1999/Accepted: 3 May 2000     

Exposure and effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin in tree swallows (Tachycineta bicolor) nesting along the Woonasquatucket River, Rhode Island, USA

Concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in tree swallows (Tachycineta bicolor) nesting along the Woonasquatucket River northwest of Providence (RI, USA) in 2000 and 2001 were some of the highest ever reported in avian tissues. Mean concentrations in eggs ranged from 300 to >1,000 pg/g wet weight at the two most contaminated ponds, Allendale and Lyman. Mean egg concentrations at Greystone, the upstream reference pond, were 12 and 29 pg/g. Positive accumulation rates and concentrations in diet samples from 12-day-old nestlings indicated that the contamination was accumulated locally. Concentrations in diet of between 71 and 219 pg/g wet weight were more than 6 and 18 times higher than concentrations considered safe for birds (10-12 pg/g). Hatching success was negatively associated with concentration of TCDD in eggs. Only about half the eggs hatched at Allendale compared with >77% at Greystone. The national average for hatching success in successful nests is 85%. No other contaminants, such as polychlorinated biphenyls and mercury, were present in any sample at concentrations known to affect avian reproduction. Three bioindicators, half-peak coefficient of geometric variation, ethoxyresorufin-O-dealkylase activity, and brain asymmetry were assessed relative to TCDD contamination.     

Transient increase in micronucleus frequency and DNA effects in the comet assay in two patients after intoxication with 2,3,7,8-tetrachlorodibenzo-<Emphasis Type="Italic">p</Emphasis>-dioxin

Background The genotoxic effects in two patients with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) intoxication were measured; an association with the workplace was suspected.Methods We determined sister chromatid exchange (SCE), micronuclei (MN) and comet assay tail factor in peripheral lymphocytes of the patients and of control persons.Results A determination of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) revealed almost pure TCDD at a concentration of 144,000 pg/g blood lipids in patient 1 and 26,000 pg/g blood lipids in patient 2, the first value being the highest level ever reported in humans. The source of TCDD ingestion remains unclear, since TCDD blood levels in 30 employees from the same workplace were in the normal range (1.2–8.6 pg/g blood lipids, average 3.0 pg/g), with the exception of three employees with moderately increased TCDD levels of 93, 149 and 856 pg/g blood lipids, who, however, did not show any clinical symptoms. Systematic measurements of air and equipment did not identify the source of TCDD. We determined SCE and MN in the peripheral lymphocytes of the patients and of control persons. A first determination, a few months after the manifestation of chloracne, showed normal values (2.4 and 2.5 MN/500 binucleated cells; 6.7±2.2 and 6.0±2.5 SCEs/metaphase) in both patients. Two months later, when TCDD levels were 85,600 pg/g blood lipids and 17,700 pg/g blood lipids, respectively, MN had increased enormously to 16 and 21.8/500 binucleated cells, SCE remained within normal range. One employee had a TCDD level of 856 pg/g blood lipids but did not develop any clinical symptoms or genotoxic effects, nor did three healthy employees with TCDD levels within background range (1.5–2.9 pg/g blood lipids). Within a period of 13 months, MN had returned to a nearly normal range in both patients. SCE had been within normal ranges all the time. We also determined the comet assay tail factor (DNA damage level) in peripheral lymphocytes of the patients. A first determination, when the TCDD level was 85,600 pg/g blood lipids (proband 1), showed an enormously high value of 33.5%. Nine months later, when the TCDD level was at 73,900 pg/g blood lipids, we found only a moderately increased value of 6.9%. Proband 2 did not have a significantly elevated comet assay tail factor, neither at the TCDD level of 17,700 pg/g blood lipids nor at the level of 15,700 pg/g blood lipids.Conclusions This delayed and transient effect seems to indicate some kind of indirect or secondary genotoxic effect of TCDD. However, DNA damaging effects, caused directly or indirectly, could be an indicator for a possible carcinogenic risk of TCDD.     

Effects of 3,3′,4,4′,5-pentachlorobiphenyl (PCB 126) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) injected into the yolks of chicken (Gallus domesticus) eggs prior to incubation

The yolks of White Leghorn chicken (Gallus domesticus) eggs were injected prior to incubation with either 3,3,4,4,5- pentachlorobiphenyl (PCB 126) at doses ranging from 0.1 to 12.8 g/kg egg or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) at doses ranging from 0.04 to 0.64 g/kg egg. Chicks were subjected to necropsy within 24 h of hatching. The brain, bursa, heart, liver, and spleen were removed and weighed. Assessment of the rate of hatching indicated an LD₅₀±S.E. of 2.3±0.19 g/kg egg (7.1±0.58 nmol/kg egg) for PCB 126 and 0.15±0.012 g/kg egg (0.47±0.037 nmol/kg egg) for TCDD. No significant differences in the incidence of developmental abnormalities (structural defects and edema) were observed in TCDD-exposed embryos, while PCB 126 caused significantly more developmental abnormalities at 3.2, 6.4, and 12.8 g/kg egg than the vehicle control. PCB 126 caused lower hatchling weights and greater relative brain, heart, and liver weights when compared to the vehicle control group at a dose of 3.2 g/kg egg which is greater than the LD₅₀. TCDD at 0.08 g/kg egg caused relative bursa weights to be less than those of the vehicle control. A toxic equivalency factor (TEF) of 0.07 was determined for PCB 126 in relation to TCDD based on overt lethality.     

Suppression of Endogenous Antioxidant Enzymes by 2,3,7,8-Tetrachlorodibenzo-<Emphasis Type="Italic">p</Emphasis>-dioxin–Induced Oxidative Stress in Chicken Liver During Development

Domestic chickens (Gallus gallus) are an excellent model in which to evaluate developmental toxicity and oxidative stress because of their high sensitivity to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The goal of this study was to measure the effects of environmentally relevant doses of TCDD on endogenous hepatic antioxidant enzyme activity in hatchling chickens. The vehicle (sunflower oil) or 2, 20, or 200 pg/g TCDD was injected into chicken eggs before incubation. On hatching, livers were harvested and quickly frozen. The changes in activity of antioxidant enzymes, including glutathione peroxidase (GPx), glutathione reductase (GRx), copper zinc superoxide dismutase (SOD), and catalase (CAT) were determined as indicators of oxidative stress. TCDD exposure was associated with a significant suppression of the activities of the protective endogenous enzymes GPx, GRx, and SOD in the liver, even at the lowest dose. CAT activity was also suppressed, but not significantly. The measured decreases were 37% to 63% for GPx, 50% to 58% for GRx, 30% to 40% for SOD, and 16% to 24% for CAT. Noncomplex dose–response relationships were evident in GPx and GRx, whereas SOD and CAT curves were U-shaped. These results demonstrate that a decreased ability to scavenge reactive oxygen species may result from developmental TCDD exposure at very low doses, contributing to oxidative stress and thus to the embryotoxicity of TCDD.     

Organochlorine Contaminants in Double-Crested Cormorants from Green Bay, Wisconsin: II. Effects of an Extract Derived from Cormorant Eggs on the Chicken Embryo

White Leghorn chicken (Gallus domesticus) eggs were injected prior to incubation with one of four concentrations (0.001, 0.01, 0.1, and 1.0 egg-equivalent) of an extract derived from 1,000 double-crested cormorant (Phalacrocorax auritus) eggs collected at Spider Island adjacent to Green Bay in Lake Michigan. One egg-equivalent corresponded to the concentration of contaminants present in an average cormorant egg. This was approximately 322 pg toxic equivalents (TEQs)/g, ww egg with polychlorinatedbiphenyl congener 126 (3,3′,4,4′,5-pentachlorobiphenyl) accounting for over 70% of the TEQs. Injection of 1.0 egg-equivalent resulted in 77% mortality at hatch. The incidence of developmental abnormalities (structural defects or edema) was not affected by injection of the extract. Body weight gain of chicks was reduced in the 1.0 egg-equivalent dose group in the first, second, and third week's post-hatch. Relative brain weights were greater and relative bursa weights were less in the 1.0 egg-equivalent dose group than in the vehicle control at three weeks of age. There were no significant differences in the relative weights of the heart, liver, spleen, testes, or comb among treated and control birds. Received: 7 February 1996/Revised: 2 July 1996     

Sensitivity of Common Tern (Sterna hirundo) Embryo Hepatocyte Cultures to CYP1A Induction and Porphyrin Accumulation by Halogenated Aromatic Hydrocarbons and Common Tern Egg Extracts

Experiments were conducted to compare the sensitivity of primary cultures of common tern (Sterna hirundo) and chicken (Gallus domesticus) embryo hepatocytes to cytochrome P4501A (CYP1A) induction and porphyrin accumulation after exposure to halogenated aromatic hydrocarbons (HAHs) or tern egg extracts. The HAHs tested were 2,3,7,8-tetrachlorodibenzo-p -dioxin (TCDD), 2,3,7,8-tetrachlorodibenzofuran (TCDF) 3,3′,4,4′-tetrachlorobiphenyl (PCB 77; IUPAC nomenclature), 3,3′,4,4′,5-pentachlorobiphenyl (PCB 126), 3,3′,4,4′,5,5′-hexachlorobiphenyl (PCB 169) and Aroclor? 1254 (a commercial mixture of PCBs). Extracts were prepared from common tern eggs collected from three sites in the Great Lakes basin and one reference site on the east coast of Canada. CYP1A induction was assayed as ethoxyresorufin-O-deethylase (EROD) activity and/or immunodetectable CYP1A protein, and total intracellular porphyrin accumulation was measured fluorometrically. Unlike chicken embryo hepatocyte (CEH) cultures, no intracellular porphyrin accumulation was observed after treatment of tern embryo hepatocyte (TEH) cultures with HAHs or egg extracts. TEH cultures were ∼50 to >1600 times less sensitive than CEH cultures to HAH-mediated CYP1A induction. In contrast, TEH cultures were either approximately equally sensitive or only 3.5–15 times less sensitive than CEH cultures to CYP1A induction mediated by tern egg extracts. These data suggest that common tern embryos may be more susceptible to the CYP1A inducing effects mediated by complex mixtures of environmental contaminants than indicated by their response to individual HAHs. Received: 28 May 1996/Revised: 27 July 1996     

Embryotoxic Thresholds of Mercury: Estimates from Individual Mallard Eggs

Eighty pairs of mallards (Anas platyrhynchos) were fed an uncontaminated diet until each female had laid 15 eggs. After each female had laid her 15th egg, the pair was randomly assigned to a control diet or diets containing 5, 10, or 20 μg/g mercury as methylmercury until she had laid a second set of 15 eggs. There were 20 pairs in each group. After the second set of 15 eggs, the pair was returned to an uncontaminated diet, and the female was permitted to lay another 30 eggs. For those pairs fed the mercury diets, the even-numbered eggs were incubated and the odd-numbered eggs were saved for possible mercury analysis. Mercury in the even-numbered eggs was estimated as the average of what was in the neighboring odd-numbered eggs. Neurological signs of methylmercury poisoning were observed in ducklings that hatched from eggs containing as little as 2.3 μg/g estimated mercury on a wet-weight basis, and deformities were seen in embryos from eggs containing about 1 μg/g estimated mercury. Although embryo mortality was seen in eggs estimated to contain as little as 0.74 μg/g mercury, there were considerable differences in the sensitivity of mallard embryos, especially from different parents, with some embryos surviving as much as 30 or more μg/g mercury in the egg. Received: 9 February 2002/Accepted: 12 June 2002     

Embryotoxicity of 3,3′,4,4′-tetrachloroazobenzene and 3,3′,4,4′-tetrachioroazoxybenzene in the chick embryo

The toxicity of 3,3,4,4-tetrachloroazobenzene (TCAB) and 3,3,4,4-tetrachloroazoxybenzene (TCAOB) to chick embryos was examined. TCAB or TCAOB was dissolved in corn oil and injected into the air cell of fertile chicken eggs. The time of injection had a major effect on embryo mortality as eggs injected with TCAB or TCAOB on the fourth day of incubation had a higher incidence of embryo mortality than eggs injected on days 11–13. Both TCAB and TCAOB were more toxic than all other chemicals that have been tested in the chick embryo with the exception of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Comparing the potency of the two compounds, TCAOB was more potent than TCAB in the chick embryo with an estimated LD₅₀ of 12 ng and 44 ng respectively. Rump edema was the major abnormality observed in embryos treated with either TCAB or TCAOB. Other malformations included altered feather pattern and lack of down, hemorrhage, external viscera, reduced body size, failure to withdraw the yolk sac, beak malformation, dilation of blood vessels, and monomicropthalmia. The results of this investigation suggest that both TCAB and TCAOB are teratogenic in the chick embryo.This work was presented in part at the Annual Meeting of the Society of Toxicology, San Diego, CA, March 1–5, 1981.     

Congener-Specific Patterns and Toxic Assessment of Polychlorinated Biphenyls in Resident and Migratory Birds from Southern India and Lake Baikal in Russia

Concentrations of polychlorinated biphenyls (PCBs), including non-, mono-, and di-ortho congeners, were determined in migratory and resident birds collected from India and Lake Baikal in Russia. In the 11 different species examined, total PCBs concentrations were in the range of 11–4500 ng/g (wet wt). IUPAC 105, 118, 138, 153, and 180 were the predominant congeners in almost all the birds. White-cheeked tern collected from India and common tern collected from Lake Baikal showed high PCBs concentrations of 4400 ng/g (wet wt) and 4500 ng/g (wet wt), respectively, and accumulated relatively high ratios of penta-CBs (IUPAC 118, 105, 99). Toxic equivalents (TEQs) of non- and mono-ortho PCB congeners in birds collected from India and Lake Baikal were in the range of 1.5–56 and 2.8–370 pg/g wet wt, respectively. Toxic assessment results led by calculated TEQs of the transfer to eggs from female birds revealed that TEQs in most of migratory and resident birds were comparable to the lowest observable effect level (LOAEL) of chicken which is a highly sensitive species against dioxin-like compounds. Calculated transfer TEQs to eggs of common tern collected from Lake Baikal in autumn, however, exceeded the LOAEL of CYP1A induction in bald eagle embryos and ED₅₀ of that in pheasant embryos, suggesting that embryo toxicity by coplanar PCBs in some avian species breeding in Lake Baikal is possible.