Teratogenic Responses Research Articles

Sex-Related Embryotoxicity of Pulp Mill Effluent Extracts in Medaka (Oryzias latipes) Female Leucophore-free FLFII Strain.

The aim of the present study was to evaluate the effects of exposure to Chilean pulp mill effluent extracts on developing postfertilized medaka embryos before and after sex definition relative to sex steroids (testosterone and 17beta-estradiol) and a wood phytoestrogen (beta-sitosterol). Our study included 2 waterborne semichronic exposure experiments, using a 24-h post fertilization (hpf) unknown-sex FLFII (female leucophore free) group and a second 72-hpf FLFII phenotypic sex-identified group (male autofluorescence leucophore) strain of medaka embryos. Chronic exposure of both FLFII strain embryo groups showed similar delay in time to hatch and decreased hatchability. Teratogenic responses such as vertebral malformation (fusion, incomplete formation, and lack of vertebral formation process) and pericardial edema were observed in both experiments, with a high percentage related to FLFII fluorescent leucophore-identified males. In addition, high mortality associated with severe malformations was observed in male and female embryos exposed to testosterone. Our research has demonstrated that exposure to Chilean mill effluent extracts caused severe male medaka embryotoxicity (in postfertilized embryos) before and after sex definition and, irrespective of the experimental group and effluent treatment, suggests partial removal following secondary treatment. Furthermore, differences in the severity and type of teratogenic effects with previous experiments (d-rR medaka strain), are associated with the unique phenotypes of this medaka mutant strain. Environ Toxicol Chem 2021;40:2297-2305. © 2021 SETAC.

Teratogenic responses of zebrafish embryos to decabromodiphenyl ether (BDE-209) in the presence of nano-SiO2 particles

This study investigated the influence of nano-SiO2 particles (nSiO2) on the teratogenic responses of zebrafish embryos to decabromodiphenyl ether (BDE-209). Zebrafish embryos were exposed to BDE-209 in the absence and presence of nSiO2 for 96 h post fertilization (hpf). Results showed that formation of nSiO2-BDE-209 associates promoted both extracellular and intracellular uptake of BDE-209 by zebrafish embryos, thereby increasing the bioconcentration of BDE-209 on the chorion surface and the embryos. Results also showed embryos delay hatching temporarily when co-exposure to BDE-209 and nSiO2 at 60 hpf. Furthermore, there was heartbeat decline (28.3 beats/10s) and increase in irregular heartbeat (45.8%) in zebrafish larvae at 96 hpf, compared to the sole exposure to BDE-209 (32.7 beats/10s and 0%). Malformation in terms of spinal curvature (SC), pericardial edema (PE) and yolk sac edema (YSE) were observed on zebrafish larvae at 33.9, 23.4, and 18%, respectively. Overall, abnormal development of zebrafish was apparent when co-exposure to BDE-209 and nSiO2. All relevant evidence considered, nSiO2 could facilitate the transport of BDE-209 towards zebrafish embryos and negatively impact the development of zebrafish.

Global gene expression analysis reveals pathway differences between teratogenic and non-teratogenic exposure concentrations of bisphenol A and 17β-estradiol in embryonic zebrafish

Transient developmental exposure to 0.1μM bisphenol A (BPA) results in larval zebrafish hyperactivity and learning impairments in the adult, while exposure to 80μM BPA results in teratogenic responses, including craniofacial abnormalities and edema. The mode of action underlying these effects is unclear. We used global gene expression analysis to identify candidate genes and signaling pathways that mediate BPA's developmental toxicity in zebrafish. Exposure concentrations were selected and anchored to the positive control, 17β-estradiol (E2), based on previously determined behavioral or teratogenic phenotypes. Functional analysis of differentially expressed genes revealed distinct expression profiles at 24h post fertilization for 0.1μM versus 80μM BPA and 0.1μM versus 15μM E2 exposure, identification of prothrombin activation as a top canonical pathway impacted by both 0.1μM BPA and 0.1μM E2 exposure, and suppressed expression of several genes involved in nervous system development and function following 0.1μM BPA exposure.

Comparative embryotoxicity of pulp mill extracts in rainbow trout ( Oncorhynchus mykiss), American flagfish ( Jordanella floridae) and Japanese medaka ( Oryzias latipes)

This study evaluated the effects of Chilean pulp mill effluent extracts (untreated, primary and secondary treated pulp mill effluents), along with steroid standards (testosterone and 17β-estradiol) and a wood extractive standard (beta-sitosterol) on developing post-fertilized fish embryos. Our study included a cold freshwater species, rainbow trout ( Oncorhynchus mykiss), and two warm freshwater species American flagfish ( Jordanella floridae) and Japanese medaka ( Oryzias latipes). Embryotoxicity results included delay in time to hatch and decreased hatchability but no significant egg and larvae mortality was observed in the pulp mill extract exposed embryos. By contrast, significant early hatching and increased hatchability were observed in beta-sitosterol exposed embryos, along with high mortality of testosterone exposed embryos across species. Teratogenic responses were observed in medaka embryos in all treatments. Abnormalities were detected starting at development stages 19–20 (2–4 somite stages) and included optical deformities (micro-opthalmia, 1 or 2 eyes) and lack of development of brains and hearts. Additionally, phenotypic sex identification of surviving offspring found female-biased sex-ratios in all treatments except testosterone across species. Overall, our study indicated that Chilean pulp and paper mill extractives caused embryotoxicity (post-fertilized embryos) across species and irrespective of the effluent treatment. The effects were mainly associated with delayed time to hatch, decreased hatchability, and species-specific teratogenesis.

Acute lethal and teratogenic effects of tributyltin chloride and copper chloride on mahi mahi (Coryphaena hippurus) eggs and larvae

Acute and chronic bioassays were used to evaluate the lethal and sublethal effects of copper chloride and tributyltin chloride on mahi mahi (Coryphaena hippurus) embryos, a pelagic life stage often found in the surface microlayer where anthropogenic contaminants can accumulate. Acute bioassay testing determined the median lethal concentration (LC50) for the test organism after 48 h of exposure. Chronic toxicity tests were used to determine the measurement of sublethal parameters, such as developmental abnormalities after 72-h exposures to the toxicants. Embryos were collected 4 h postfertilization and subsequently exposed to 1, 5, 25, 50, and 100 microg/L of copper chloride and 3, 10, 20, 30, and 50 microg/L of tributyltin chloride. Analysis of hatch rate percentage determined that the mean 48-h LC50 of copper chloride and tributyltin chloride was 32.8 and 16.7 microg/L, respectively, based on the pooled data of four experiments with four replicates for each metal. Consistent abnormalities, such as yolk sac swelling, spinal deformities, and decreased hatch rates, were observed for each metal. Teratogenic responses to copper chloride and tributyltin chloride demonstrate the need to investigate further the impacts of pollution in the open oceans to a species indigenous to, and commercially important to, the Florida (USA) coasts. This information could then lead to the future development of a surface microlayer bioassay using mahi mahi embryos.

Effects of formulations of the fungicide, pentachloronitrobenzene on early life stage development of the Japanese medaka (Oryzias latipes)

Quintozene is a fungicide containing the active ingredient, pentachloronitrobenzene (PCNB) that is used to control “snow mold” on golf courses in temperate regions of North America. In this study, quintozene and a formulation of quintozene widely used on golf courses, FFIITM were tested for toxicity to early life stages of the Japanese medaka, Oryzias latipes. For medaka exposed in static non-renewal assays to quintozene for 17d from the fertilized egg stage to yolk resorption at the fry stage, the LC50 for mortality was a nominal concentration of 707μgl−1 and the effective concentration for 50% hatch (i.e. EC50) was a nominal concentration of 71μgl−1. Eggs and fry showed developmental abnormalities, including ocular malformations and retarded development of the brain, notochord, organs and body segmentation, which were interpreted as teratogenic responses to exposure to PCNB. For medaka exposed to quintozene, the LOECs for abnormalities of the eye and all other developmental abnormalities were 750 and 100μgl−1, respectively. In medaka exposed to the FFIITM formulation, similar patterns of mortalities, reduced hatching success and developmental abnormalities were observed, but at higher test concentrations that were consistent with the proportion of quintozene in the formulation. For medaka exposed to the formulation, the LOECs for abnormalities of the eye and all other developmental abnormalities were 10000 and 1000μgl−1, respectively. Overall, these data indicate that studies should be conducted to assess the risk of exposure of early life stages of fish to quintozene in watersheds impacted by golf courses.

Comparative developmental toxicity of 2,3,7,8-tetrachlorodibenzo- p-dioxin in the hamster, rat and guinea pig

2,3,7,8-Tetrachlorodibenzo- p-dioxin (TCDD) is a persistent environmental contaminant capable of causing a wide variety of adverse health effects including teratogenesis and altered development. The objective of this study was to compare the developmental toxicity of TCDD in the hamster, rat and guinea pig, which in mature animals exhibit a relatively low, medium and high sensitivity to TCDD, respectively. A single oral dose of TCDD was administered to pregnant rats (0, 1.5, 3.0, 6.0 or 18.0 μg/kg) on gestation day 10, pregnant hamsters (0, 1.5, 3.0, 6.0 or 18.0 μg/kg) on gestation day 9 and pregnant guinea pigs (0, 0.15 or 1.5 μg/kg) on gestation day 14 with fetal analysis on gestation day 20, 15 and 56, respectively. The developmental toxicity of TCDD in the three species included increased fetal mortality, alterations to fetal body weight, body length, organ weight and significant changes to the fetal white blood cell differential counts. Additionally, teratogenic responses were observed in the hamster and rat consisting of cleft palate, kidney congestion, hydronephrosis and intestinal hemorrhaging. Furthermore, the results from this study demonstrate that despite the up to 5000-fold interspecies variability to the acute lethal potency of TCDD observed in mature guinea pigs, rats and hamsters, the developing fetus is uniquely vulnerable to gestational TCDD exposure and displays approximately a 10-fold variability in fetal lethal potency in these species. Together, these results will assist efforts to reduce the uncertainty in the risk assessment for TCDD in sensitive populations, such as the developing embryo and fetus.

Defining critical periods for itraconazole-induced cleft palate, limb defects and axial skeletal malformations in the mouse

The main aim of the study was to identify the critical periods of susceptibility for itraconazole-mediated teratogenesis in the mouse. Pregnant ICR (CD-1) mice received a single oral administration of itraconazole at 50 mg/kg b.w., 150 mg/kg b.w. or 250 mg/kg b.w. on gestation day 8, 9, 10, 11 or 12. Control animals were administered with vehicle on gestation days 8–12. The gestational outcome was evaluated near term, on gestation day 18. Treatment-related morphological findings, as they can be evaluated by external, visceral and skeletal examination, mainly included cleft palate, limb defects and axial skeletal malformations. Cleft palate and limb defects resulted after single exposures between gestation days 9 and 11, with a tendency toward maximal response on gestation day 10. Significant incidences of axial skeletal defects were seen exclusively on gestation days 8 and 9. Exposure on gestation day 12 did not yield significant teratogenic responses. Cleft palate was the most sensitive teratogenic response, being the only developmental lesion associated to exposure to itraconazole at 150 mg/kg b.w.

Assessing contamination levels of Laguna Lake sediments (Philippines) using a contact assay with zebrafish ( Danio rerio) embryos

The present study investigated the suitability of a sediment contact assay using zebrafish ( Danio rerio) embryos to evaluate the degree of lake sediment contamination. As endpoints, developmental parameters (mortality, abnormality, heart rate, and hatching rate) as well as stress protein responses (hsp 70 levels) in the developing embryos were recorded during a 96-h exposure. Fertilized zebrafish eggs were exposed to both the whole as well as organic extract concentrations prepared from collected sediments from 5 sites along Laguna Lake, Philippines. Compared to whole sediment exposure, more severe embryotoxic and teratogenic responses were elicited in embryos exposed to organic extracts. However, since whole sediment-exposed embryos also revealed significant developmental defects, this exposure phase served as the more realistic exposure scenario in our study. Weak to strong upregulation of hsp 70 levels was also registered among embryos exposed to both whole sediments and organic extracts. The observed embryotoxic and proteotoxic responses by zebrafish embryos to Laguna Lake sediment exposures were discussed in relation to the analyzed contaminants in the sediments (heavy metals, PAHs (perylene), solvent). Overall, the present study points out that the sediment contact assay with zebrafish embryos offers a practicable and highly sensitive bioassay for the general assessment of sediment toxicity.

Teratogenic effects of retinoic acid are modulated in mice lacking expression of epidermal growth factor and transforming growth factor-?

Epidermal growth factor (EGF) and transforming growth factor-alpha (TGFalpha) regulate cell proliferation and differentiation in the embryo. The induction of cleft palate (CP) by all trans-retinoic acid (RA) was associated with altered expression of TGFalpha, EGF receptor, and binding of EGF. This study uses knockout (KO) mice to examine the roles of EGF and TGFalpha in teratogenic responses of embryos exposed to RA. Pregnant wild-type (WT) mice of mixed genetic background, EGF KO, C57BL/6J, and TGFalpha KO mice were given a single oral dose of RA (100 mg/kg, 10 ml/kg) or corn oil on GD 10 at 12 PM, GD 11 at 12 PM or 4 PM, or GD 12 at 8 AM or 12 PM (plug day = GD 0). GD 18 fetuses were examined for external, visceral, and skeletal effects. After exposure to RA on GD 12, the incidence of CP in EGF KO was significantly reduced relative to WT. In TGFalpha KO fetuses, RA exposure on GD 10 increased the incidence of CP versus C57BL/6J. The incidence of skeletal defects in the limbs, vertebrae, sternebrae, and ribs were also affected by lack of expression of EGF or TGFalpha with region-specific amelioration or exacerbation of the effects of RA. In TGFalpha KO fetuses, incidences of forelimb long bone and digit defects increased relative to C57BL/6J. In EGF KO fetuses, relative to WT, the incidence of hindlimb oligodactyly was increased. In EGF KO, but not WT, RA produced short, bent radius, humerus, and ulna. Both TGFalpha and EGF KO mice had increased incidences of dilation of the renal pelvis and this was reduced by RA. RA exposure produced skeletal and visceral defects in all genotypes; however, EGF or TGFalpha KO influenced the incidence and severity of defects. This study supports a role for EGF and TGFalpha in the response to RA.

Exposure‐disease continuum for 2‐chloro‐2′‐deoxyadenosine (2‐CdA), a prototype teratogen: Induction of lumbar hernia in the rat and species comparison for the teratogenic responses

The purine analog 2-chloro-2'-deoxyadenosine (2-CdA) caused ocular and limb defects in the mouse and rabbit. The current study examined the teratogenic potential of this drug in the rat and compared the adverse developmental outcomes with the other species. Timed-pregnant Sprague-Dawley rats were given a single intraperitoneal injection of various doses of 2-CdA ranging from 5-60 mg/kg, at gestational day (GD) 9.5 and GD 14. 2-CdA concentrations in maternal serum and embryos were measured by HPLC and termed fetuses were prepared for teratological examination. Full-litter resorption was seen in dams receiving 50 mg/kg of 2-CdA at GD 9.5, whereas post-implantation loss was significantly increased and fetal weights significantly reduced at 40 mg/kg. Gross examination of the surviving fetuses revealed microphthalmia, a shortened body trunk and lumbar hernia, manifested by a soft mass protrusion at the lumbar region on one or both sides of the spine. Incidence of these defects increased in a dose-dependent fashion. Histological examination indicated that the hernia was associated with hypoplasia of the body wall, poorly developed skeletal muscle bundles surrounding the vertebral column in the lumbar region, and an absence of the lateral muscle groups that allowed protrusion of the abdominal viscera. The lumbar hernia was generally accompanied by spina bifida, deformed ribs and a wide spectrum of soft tissue-abnormalities that included kidney, genitourinary and heart defects. At GD 14, exposure to 2-CdA at 60 mg/kg produced oligodactyly in one of six litters. 2-CdA produced similar ocular defects in the rat and mouse, although the incidence was much lower in the former species. In contrast, the drug-induced lumbar hernia was only seen in the rat. These apparent disparities were not readily explained by species differences in pharmacokinetic parameters. the similarities between the teratological features of 2-CdA-induced lumbar hernia in the rat and the clinical description of lumbocostovertebral syndrome, however, may provide a key to unlock the etiology of this rare birth defect in humans.

Vitamin A prevents inner ear defects in mice with congenital homeobox gene deficiency.

For the past 75 years, vitamin A and its biologically active metabolites, the retinoids, have been the object of intense study in biology and medicine. A large body of evidence demonstrates that these nutrients are essential for normal development and survival of vertebrate embryos, including mammals. In fact, it has been known since the mid-1930s that vitamin A deficiency during pregnancy results in death of the fetus and congenital abnormalities. Similarly, excess dietary intake of vitamin A can also cause teratogenic responses. Among the main targets of both deficiency and excess retinoid-induced teratogenesis are the heart, limbs, craniofacial structures, central nervous system, and the inner ear. Specific malformations are induced in a stage- and dose-dependent manner. Thus, these studies indicate that precise levels and timing of action of vitamin A metabolites are required for normal patterning of embryonic structures. In addition, the discovery of the nuclear receptors for retinoic acid (RA) and other vitamin A derivatives provided a molecular basis to explain how distinct doses of these compounds elicit cell-specific responses via the direct transcriptional activation of a panel of target genes.

Significance of excursions of intake above the acceptable daily intake: effect of time and dose in developmental toxicology.

Three major factors to be considered in assessing the possible effects on developmental toxicology of excursions above the acceptable daily intake (ADI) are discussed. If maternal toxicity occurs at lower doses than developmental toxicity, then there may be adequate protection for the fetus if the mother is protected. In other cases, the first adverse developmental effects are usually small and reversible changes in fetal weight and it is unlikely that brief excursions, for a few days, above the ADI, would induce changes in final birth weight. The importance of excursions above the ADI on teratogenic responses would likely depend on the mechanism of teratogenesis and whether the agent acted primarily as a result of a high peak plasma level, a Cmax effect, or depended more on the total body exposure, an area under the curve (AUC) effect. This type of information is usually not available from current safety study designs. Finally, the temporal equivalence factors must be taken into account. There are 10- to 100-fold differences in time span for pre- and postnatal development up to sexual maturity, in rodents compared with humans. This can be contrasted with the relatively small pharmacokinetic differences, perhaps of up to 2- or 3-fold in plasma levels of chemicals with the same administered dose in the two species. Thus, single episodes of high chemical exposure have greater opportunity to produce permanent effects on development in rodents compared with humans. The limited evidence from acute poisonings in pregnant women tends to support this hypothesis and it seems unlikely that occasional excursions by women above the ADI would result in developmental toxic effects.

Developmental effects of urban storm water in Medaka (Oryzias latipes) and inland silverside (Menidia beryllina).

Stormwater runoff in a coastal urban area (San Diego County, CA) produced significant toxicity to early life stages of medaka (Oryzias latipes) and Menidia (M. beryllina). Exposure of embryos to lower concentrations (5 to 25%) increased the incidence of abnormal swim bladder inflation and other teratogenic responses, whereas higher concentrations resulted in mortality or failure to hatch. Comparisons of EC50s for mortality and failure to hatch with concentrations of individual chemical pollutants (including Cd, Cr, Cu, Pb, Ni, and Zn) revealed low correlations; however, the correlation with total metals was high (-0.84) and corresponded with sample exceedences of Water Quality Criteria (WQC) for Cd, Cu, Pb, and Zn. This strong association between developmental toxicity and toxic metal content of storm water compared favorably with developmental anomalies reported in other studies. Analytical chemistry data for pesticides that may have been in these samples were limited to selected pesticides found usually below detection limits. Greater toxicity of the watershed effluents sampled was generally associated with more developed land surface and less open space. Both medaka and Menidia were found to be useful for studying effects of stormwater on embryonic and early larval development. http://link.springer-ny. com/link/service/journals/00244/bibs/37n2p227.html

Preliminary observations on responses of embryonic and larval Pacific herring, Clupea pallasi, to neutral fraction biodegradation products of weathered Alaska North Slope oil.

Weathered Alaska North Slope crude oil (ANS 521) was subjected to biodegradation in vigorously stirred incubations for 14 days at 15 +/- 1 degrees C in 20/1000 salinity sterilized seawater, amended with nutrients and inoculated with a hydrocarbon-degrading microorganism (EI2V) isolated from an oil-contaminated beach in Prince William Sound, Alaska. A total of 13.7 mg/L water-soluble neutral fraction (WSF) was recovered from the incubation of weathered ANS 521. Toxicity/ teratogenicity tests were conducted with WSF recovered from the biodegradation system using embryonic and larval Pacific herring, Clupea pallasi. Exposures were begun at 4, 48, and 96 h postfertilization of herring eggs. Exposure concentrations were 1, 10, and 100% of the original concentration of WSF recovered from incubations (redissolved in 20/1000 salinity sterile seawater at 15 +/- 1 degrees C). Sterile 20/1000 salinity seawater without the addition of redissolved neutral fraction was used as a control. Significant (p < or = 0.05) embryo mortality or teratogenic responses were observed at WSF concentrations of 10 and 100%. On days 5 through 8 of embryogenesis, counts of heart contraction rates were significantly lower (p < or = 0.05) at the 100% WSF concentration for embryos exposed beginning at 4 and 48 h postfertilization. Grow-out of larvae from selected exposures was conducted. High mortality was noted in larvae exposed to the 10% WSF concentration beginning at 4 and 48 h postfertilization. Most of these larvae died 5 to 8 days after hatching when they elicited vertebral displacements at a time concurrent with the onset of feeding behavior.

Coteratogenic Effects of Caffeine

Coteratogenicity studies have been carried out using various physical and chemical agents along with caffeine. For ionizing radiation in mice, enhancement of teratogenic responses (cleft palate, limb malformations) was noted with single systemic bolus doses of 50 to 200 mg/kg. Studies in rats with ethanol or nicotine reveal only an additive effect with caffeine. There are mixed results with chemical carcinogens and caffeine with some studies showing enhancement and others showing that caffeine inhibits the teratogenic effect of some carcinogens. The time of treatment, at the time of carcinogen exposure for the inhibition and later in the gestation period for the enhancement, appears to be the critical factor. For a variety of pharmaceutical agents (acetazolamide, mitomycin C, hydroxyurea, 5-fluorouracil), caffeine was shown to enhance the teratogenic effect of the agent. With 5-azacytidine in rats, caffeine suppressed limb malformations. Administration of inhibitors of β-adrenergic function reduces the teratogenic effect of caffeine in mice. The interpretation of the experimental studies in terms of human hazard is complicated by the general use of high-dose bolus exposures which are not typical of human exposures, and the use of test systems that are not readily applicable to humans. The studies in human populations show clearly that caffeine itself has no link to negative birth outcome, and in the few instances where it has been examined there appears to be no interaction between coffee consumption and either alcohol consumption or smoking on pregnancy outcome.

Detoxification of pentachlorophenol and creosote contaminated groundwater by physical extraction: Chemical and biological assessment

Chemical analyses revealed that polycyclic aromatic hydrocarbons (PAHs) and other organic compounds were present in a perennial freshwater stream that flowed through the abandoned American Creosote Works and into Pensacola Bay, Florida. Moreover, groundwater pumped from a well depth of 21 m at a location adjacent to the site was heavily contaminated with PAHs and other organics. A study was conducted to determine the efficacy of ultrafiltration for removal of organics from groundwater at this U.S. Environmental Protection Agency, Super Fund site. Ultrafiltration reduced the concentration of total identified organics from 210.0 mg/L in groundwater to 1.5 mg/L in the post-filtration permeate. Tests for toxicity/teratogenicity in embryonic inland silversides, Menidia beryllina; and Microtox 15 min EC50's were conducted with: 1) streamwater, 2) untreated groundwater, 3) feedwater used in the ultrafiltration system and 4) permeate water that passed through the ultrafiltration system. A concentration of 100% streamwater caused significant (alpha less than or equal to 0.05) teratogenic responses in fish embryos and larvae; the Microtox EC50 was 3.7% streamwater. Groundwater and feedwater caused significant embryo toxic or teratogenic responses at concentrations of 100, 10, and 1%; the Microtox EC50's were 0.85 and 0.48%, respectively. In contrast, only 100% permeate water caused significant increases in terata, compared to the control response; at 10 and 1% concentrations greater than 90% of hatched larvae appeared normal. The Microtox EC50 was 30% permeate water.

Teratogenicity of three polychlorinated dibenzofurans in [formula omitted] mice

Polychlorinated dibenzofurans (PCDFs) are widespread environmental contaminants which have been detected in human tissues and implicated in several poisoning incidents. Their toxic effects are similar to those observed with other related halogenated aromatic hydrocarbons such as TCDD. The teratogenic effects of three PCDFs, 1,2,3,7,8,-pentachlorodibenzofuran (1-PeCDF), 2,3,4,7,8-pentachlorodibenzofuran (4-PeCDF), and 1,2,3,4,7,8-hexachlorodibenzofuran (HCDF), were assessed in C57BL 6N mice. Pregnant mice were exposed on Gestation Days 10–13 to 10 ml corn oil/kg containing PCDFs. The dams were killed on Gestation Day 18 and maternal and fetal toxicity were assessed. All three compounds were highly teratogenic, with very steep and parallel dose-response curves for the two diagnostic indicators of dioxin-like teratogenicity, hydronephrosis, and cleft palate. 4-PeCDF was the most teratogenic with an ED50 of 36 μg/kg for cleft palate and 7 μg/kg for hydronephrosis. 4-PeCDF was approximately 4 times as potent as 1-PeCDF and 10 times as potent as HCDF. The teratogenic responses occurred at a dose below that where any obvious maternal or fetal toxicity was detected. Thus, these three compounds cause teratogenic responses similar to those seen with TCDD but are only 1 10 to 1 100 as potent.

Dose-response relationship of cadmium embryotoxicity in cultured mouse embryos

Mouse embryos were exposed in vitro to 1.2 to 2.2 μM cadmium, and effects on embryotoxicity were examined after 39 h of culture. Teratogenic responses similar to in vivo were obtained at 1.2 to 2.2 μM with concomitant reduction in embryonic protein, while embryo deaths were increased from 13.8 to 93.3% at 2.0 to 2.2 μM. The response data of both teratogenicity and growth parameters, including embryonic protein, head lenght, crown-rump lenght, somite number, and protein and diameter of yolk sac, were acceptably fitted to a cadmium is a critical parameter in the manifestation of teratogenic potential, (b) as an estimation of interference in the growth of embryos, embryonic protein is one of the most sensitive endpoints while somite number is an insensitive criterion, and (c) a linear log-probit regression is applicable to the analyses of embryotoxicity data, including growth parameters in whole-embryo culture systems.

Metallothionein and mercury tolerance in the killifish, fundulus heteroclitus

Abstract Synthesis of metallothionein (MT) as a possible mechanism by which Fundulus heteroclitus adapts to heavy metal pollution has been investigated in relation to responses to methylmercuric chloride (meHg) and HgCl 2 (Hg 2+ ) at different life stages. Clutches of embryos from unpolluted sites exhibit teratogenic responses to a standard dose of meHg or of Hg 2+ which range from little or no effect (tolerant) to severely affected or almost complete inhibition of development (susceptible). At the time of hatching, untreated embryos from tolerant clutches had twice as much of a protein which co-migrates in SDS-PAGE gels with Fundulus MT as do those of susceptible clutches. This difference could be a mechanism for conveying tolerance to Hg 2+ and also to meHg if demethylation occurs. However, eggs at the time of deposition had virtually no MT, suggesting that the protein is synthesized too late to provide tolerance in the early stages of development which are critical in the etiology of malformations. Treatment of embryos with non-teratogenic doses of either meHg or Hg 2+ did not increase the amount of MT-like protein. This may indicate that the thionein gene is already fully active, and would also explain observations that Hg 2+ pretreatment of embryos does not enhance tolerance to Hg 2+ in the larval stage. MT, isolated from livers of Hg 2+ - or Cu 2+ -dosed or control adults, is associated with Cu and Zn but not with Hg, although it can bind Hg 2+ in vitro. Fundulus , like other fish, methylates much (ca. 75 %) of its Hg burden, and meHg is considered by some not to react with MT. Fundulus liver has its Hg burden distributed between homogenate sediment (25 %), the cytosol (60 %) and the remainder in lipid. The liver responds to Hg 2+ treatment by increasing lipid content. This may allow sequestration of some lipid-soluble meHg. Of the cytosolic Hg, most is associated with larger proteins and some (ca. 25 %) is dialyzable. These data suggest that metallothionein is not a significant factor in Hg tolerance in Fundulus adults, and has a questionable role in embryonic and larval tolerance to Hg 2+ and none to the more toxic meHg.