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Long-term trends and sources of organochlorine contamination in Canadian tundra Peregrine Falcons, Falco peregrinus tundrius.

Levels of eggshell thinning, and organochlorine residues in egg contents, blood plasma of adults and juveniles, tissue samples, and prey species were determined for a population of migratory Peregrine Falcons (Falco peregrinus tundrius) breeding in the Canadian Arctic. Temporal trends were assessed by comparing data collected during 1991-1994, with data from 1982-1986, for the same population. Shells (n=54) from 1991-1994 averaged 15% thinner than eggs produced prior to the introduction of DDT. No improvement in shell thickness was detected between decades. Mean DDE residue levels in eggs showed a decline from 7.6 mg kg (1982-1986) to 4.5 mg kg (1991-1994), but there was no significant change in SigmaPCB residues. Moreover, the proportion of clutches with eggs exceeding critical SigmaPCB, DDE, and dieldrin residue levels (10%) did not change between decades. Relative to Greenland and Alaskan populations, F. p. tundrius at Rankin Inlet show high levels of organochlorine contamination and little reduction in residues over the last decade. These Tundra Peregrines continue to be exposed to organochlorines in Latin America; however, results also link relatively high levels in the study population with waterfowl species that do not leave Canada in winter.

Journal Article↗

Comparative pathology of Prudhoe Bay crude oil and inert shell sealants in chicken embryos.

Pathological changes and body weights were compared in 9-day-old chicken embryos exposed to Prudhoe Bay Crude Oil (PBCO), pharmaceutical mineral oil, or a sealing film on the eggshell. Embryos exposed to 10 or 20 microliters PBCO developed marked ascites or subcutaneous edema, extensive liver necrosis, dilation of the heart, and cellular casts and mineralization in renal tubules. Wet and dry body weights were normal or above normal in embryos exposed to PBCO. No changes were found in embryos exposed to 10 microliters mineral oil; embryos exposed to 20 microliters mineral oil had slight dilation of the heart. Very few embryos survived in eggs that were wrapped in sealing film. All embryos that did survive had greatly reduced body weight and marked dilation of the heart. Four of these embryos had vacuolation of hepatocytes at the surface of the liver, one had superficial liver necrosis, and one embryo had slight ascites. These results indicate that the mechanism of the toxic action of PBCO on the avian embryo involves more than simple physical sealing of the shell.

Animals↗

Oxygen uptake in fowl eggs incubated in air and pure oxygen.

1. The oxygen uptake of small (bantam hen) and large (white leghorn) domestic fowl eggs was measured during incubation in air and pure O2. 2. A 2 hr exposure to pure O2 increased the O2-uptake from the 9th day in the large eggs and from the 15th day in the small eggs. 3. On the last 4 days of the incubation period, both small and large eggs increased their O2-uptake by about 22% when transferred from air to pure O2. 4. When 1 cm2 of the eggshell above the air cell was removed, the O2-uptake of the white leghorn eggs incubated in air increased by 13%.

Aerobiosis↗

Hyperphosphataemia and bone resorption in histamine injected laying hens.

The effects of histamine on bone resorption during eggshell formation, were studied. The study included the effect of histamine on the clearance of 45Ca and 32P from plasma, and on the release from the femur or uptake by the egg shell of 45Ca and 32P injected at the time of the first experiment or 10 hr prior to the experiment (experiment 2). Histamine led to a decrease in the disappearance rates of 45Ca and 32P, without any modification of the femur and egg shell levels. However, the increase in the amount of plasma 32P and the decrease in the 32P specific activity (SA) in bone, both elicited by histamine, argues in favour of the skeletal origin of hyperphosphataemia induced by histamine during egg shell formation.

Animals↗

Histamine-induced hyperphosphatemia and hypocalcemia in the laying hen.

The effects of histamine on plasma inorganic phosphorus (Pi) and total calcium values were studied in laying hens during egg formation and in non-laying hens. Histamine induced hyperphosphatemia and slight hypocalcemia during eggshell calcification, whereas the most pronounced hypocalcemia was observed during the early stages of egg formation. The histamine-induced hyperphosphatemia was completely inhibited by cimetidine but only partly by promethazine. However, the histamine-induced hypocalcemia was totally inhibited by cimetidine and promethazine.

Animals↗

The distribution of calmodulin in the mucosa of the avian oviduct and the effect of p-p'-DDE on some of its metabolic parameters.

1. Since calmodulin or some closely related peptide may activate the Ca2(+)-transporting system in the avian eggshell gland, the calmodulin content in different parts of the oviduct mucosa was determined in egg-laying birds killed at 1600 hr. 2. The highest content was noted in the shell gland mucosa both in egg-laying ducks and hens. The calmodulin content was high even in the isthmus part, where the shell formation begins. 3. Treatment of ducks (Indian runner variety) with DDE (40 ppm for 45 days) did not influence the calmodulin content of the shell gland, however. 4. The content of the protein avidin, the formation of which is stimulated by progesterone, was increased significantly in the oviduct. The carbanhydrase activity did not change significantly. 5. The dry weight of the shell gland was reduced by DDE administration in ducks but not in domestic fowls. 6. These and earlier observations indicate that DDE can act as an partial agonist which is able both to stimulate and to inhibit reactions in the shell gland and other parts of the oviduct. 7. In vivo DDE in the dose used probably acted on higher centres, influencing the activity of the shell gland and probably other parts of the oviduct. 8. A regulation centre which influences several sexual functions is the hypothalamic-hypophyseal region, but the endocrine function of the ovary has also been considered.

Animals↗

EM analysis of Drosophila chorion genes: amplification, transcription termination and RNA splicing.

We have used the electron microscope to examine ultrastructurally several events occurring during the biogenesis of two very abundant chorion (eggshell) mRNA molecules in the follicle cells of Drosophila melanogaster--namely, selective gene amplification, transcription initiation and termination, and RNA processing. We find that the highly transcribed s36 and s38 genes are positioned in the central region of large, multi-forked amplified DNA structures. Transcript morphology is consistent with the known presence of a small intron at the 5' end of each gene. Mature transcripts are associated with spliceosomes, demonstrating that splice site selection occurs co-transcriptionally but that splicing is completed after transcript release from the template. We have also mapped the termination sites for the genes. The two genes exhibit efficient termination very near their poly(A) sites--within a 210 bp region for s36 and a 360 bp region for s38.

Animals↗

Medullary bone matrix formation, mineralization, and remodeling related to the daily egg-laying cycle of Japanese quail: a histological and radiological study.

A model is presented concerning the remodeling of medullary bone during the egg-laying cycle of hens. This model is partly based on earlier results that showed that the active period of medullary bone resorption that produces part of the calcium needed for the eggshell coincides with the period of medullary bone matrix formation. It was hypothesized that mineralization of this matrix takes place during the subsequent inactive period. The present study examines this model. The medullary bone matrix volume, regardless of the degree of mineralization, is quantified in quail hens at four time points during the egg-laying cycle. No significant changes in the medullary bone volume (i.e., matrix and bone) during the egg-laying cycle were found, implying that matrix formation and bone resorption are kept in balance within a time interval of only a few hours. In the radiological part of this study the whole body retention (WBR) of 99mTc(Sn)MDP is used as a measure of the amount of low calcium medullary bone matrix formed during the egg-laying cycle. This use of 99mTc(Sn)MDP is justified by an autoradiographic experiment that showed that 99mTc(Sn)MDP actually labels newly formed medullary bone. Localization of the radioactive label showed a high positive correlation with the localization of fluorescent tetracycline labels in the medullary bone. From 4 to 22 hours after ovulation, the amount of medullary bone matrix increases linearly in quail hens.(ABSTRACT TRUNCATED AT 250 WORDS)

Alkaline Phosphatase↗

Development and validation of a direct homologous quantitative sandwich ELISA for fathead minnow (Pimephales promelas) vitellogenin.

Vitellogenin (Vtg) is an established and sensitive endpoint for analysis of exposure to (anti-)oestrogens and their mimics in fish [Sumpter, J.P., 1995. Feminized responses in fish to environmental estrogens. Toxicol. Lett. 82, 737-742; Arukwe, A., Goksøyr, A., 2003. Eggshell and egg yolk proteins in fish: hepatic proteins for the next generation: oogenetic, population, and evolutionary implications of endocrine disruption. Comp. Hepatol. 2, 4. ]. In some instances, links have been drawn between high level induction of Vtg and adverse health effects in fish [Herman, R.L., Kincaide, H.L., 1988. Pathological effects of orally administered estradiol to rainbow trout. Aquaculture 72, 165-172; Schwaiger, J., Spieser, O.H., Bauer, C., Ferling, H., Mallow, U., Kalbfus, W., Negele, R.D., 2000. Chronic toxicity of nonylphenol and ethinyloestraiol: haematological and histopathological effects in juvenile common carp (Cyprinus carpio). Aquat. Toxicol. 51, 69-78]. The widespread use of Vtg as a biomarker has led to the development of a variety of assays to quantitatively measure Vtg concentrations in tissue samples from fish, and hence a need for a standardization of the performance criteria and validation of such assays [Goksøyr, A., Eidem, J.K., Kristiansen, S.I., Nilsen, B.M., 2003. On the need for a standardized set-up for validation studies of fish vitellogenin assays as an endpoint in endocrine disruptor testing and screening-a proposal. ]. One of the most popular test fish species for assessing chemical effects is the fathead minnow (Pimephales promelas), which is now used widely for studies into endocrine disruption [Panter, G.H., Hutchinson, T.H., Lange, R., Lye, C.M., Sumpter, J.P., Zerulla, M., Tyler, C.R., 2002. Utility of a juvenile fathead minnow screening assay for detecting (anti)estrogenic substances. Environ. Toxicol. Chem. 21, 319-326; Hutchinson, T.H., Yokota, H., Hagino, S., Ozato, K., 2003. Development of fish tests for endocrine disruptors. Pure Appl. Chem. 75, 2343-2353]. This paper describes the development and validation of a new, homologous enzyme-linked immunosorbent assay (ELISA) for quantification of Vtg in this fish species.

Animals↗

The relationship between gut contents and supercooling capacity in hatchling painted turtles (Chrysemys picta).

Painted turtles (Chrysemys picta) typically spend their first winter of life in a shallow, subterranean hibernaculum (the natal nest) where they seemingly withstand exposure to ice and cold by resisting freezing and becoming supercooled. However, turtles ingest soil and fragments of eggshell as they are hatching from their eggs, and the ingestate usually contains efficient nucleating agents that cause water to freeze at high subzero temperatures. Consequently, neonatal painted turtles have only a modest ability to undergo supercooling in the period immediately after hatching. We studied the limit for supercooling (SCP) in hatchlings that were acclimating to different thermal regimes and then related SCPs of the turtles to the amount of particulate matter in their gastrointestinal (GI) tract. Turtles that were transferred directly from 26 degrees C (the incubation temperature) to 2 degrees C did not purge soil from their gut, and SCPs for these animals remained near -4 degrees C for the 60 days of the study. Animals that were held at 26 degrees C for the duration of the experiment usually cleared soil from their GI tract within 24 days, but SCPs for these turtles were only slightly lower after 60 days than they were at the outset of the experiment. Hatchlings that were acclimating slowly to 2 degrees C cleared soil from their gut within 24 days and realized a modest reduction in their SCP. However, the limit of supercooling in the slowly acclimating animals continued to decline even after all particulate material had been removed from their GI tract, thereby indicating that factors intrinsic to the nucleating agents themselves also may have been involved in the acclimation of hatchlings to low temperature. The lowest SCPs for turtles that were acclimating slowly to 2 degrees C were similar to SCPs recorded in an earlier study of animals taken from natural nests in late autumn, so the current findings affirm the importance of seasonally declining temperatures in preparing animals in the field to withstand conditions that they will encounter during winter.

Acclimatization↗

The xenoestrogen 4-nonylphenol modulates hepatic gene expression of pregnane X receptor, aryl hydrocarbon receptor, CYP3A and CYP1A1 in juvenile Atlantic salmon (Salmo salar).

Nonylphenol is a degradation product of alkylphenol polyethoxylates (APEs). APEs represent an important class of non-ionic surfactants widely used in many detergent formulations and plastic products for industrial and domestic use. Nonylphenol interacts with xenobiotic- and drug-metabolizing CYP forms, including members of the CYP3A and CYP1A1 subfamilies in fish. The pregnane X receptor (PXR) and aryl hydrocarbon receptor (AhR) are known regulators of the induction of CYP3A and CYP1A1, respectively. Previously in our laboratory, we have shown that nonylphenol, in addition to inducing plasma egg-yolk and eggshell proteins, also modulates hepatic CYP3A- and CYP1A1-mediated enzyme activities in juvenile Atlantic salmon. Given the potential for indirect actions of nonylphenol on xenobiotic and steroid metabolism, studies were carried out to determine the effects of nonylphenol on the expression of two critical enzyme systems using juvenile salmon and waterborne nonylphenol at environmentally relevant concentrations (5, 15 and 50 microg/L) and ethanol solvent control and sampled at different time intervals (days 0 (control), 3 and 7) post-exposure. Our data show that nonylphenol-induced CYP3A and CYP1A1 mRNA levels correlate with PXR and AhR at day 7 post-exposure to 5 and 15 microg NP/L. For CYP1A1 and AhR, nonylphenol caused a temporal decrease at day 3 post-exposure, thereafter CYP1A1 and AhR mRNA levels were significantly induced at day 7. 50 microg NP/L caused a more pronounced effect on CYP1A1 by decreasing the mRNA levels at days 3 and 7, compared to control. Despite the decreased CYP1A1 mRNA levels at day 7, 50 microg NP/L caused a significant induction of AhR mRNA at the same time period. This study suggests a possible regulation by xenoestrogens of fish hepatic CYP3A and CYP1A1 enzymes via PXR and AhR, respectively, and may have impact on the metabolism of endogenous and exogenous substrates. This is the first study that simultaneously examined CYP3A, CYP1A1, PXR and AhR transcripts following nonylphenol treatment in any aquatic species or earlier diverging vertebrates.

Animals↗

Developmental effects of embryonic exposure to toxaphene in the American alligator (Alligator mississippiensis).

A variety of organochlorine pesticides have been shown to adversely affect embryonic development. A number of abnormalities have been documented in alligators (Alligator mississippiensis) from highly-contaminated Lake Apopka, FL, USA that are similar to the results of experimental studies exposing embryos to pesticides. In the current study, we exposed developing alligator embryos to varying concentrations of toxaphene, a broad-spectrum pesticide found in relatively high concentration in Lake Apopka alligator egg yolk. The toxaphene, dissolved in 50 microl of ethanol, was applied topically to the eggshell just prior to the sex-determining period of development. Shortly after hatching, we examined a number of morphological and physiological endpoints to determine the consequences of sub-lethal embryonic exposure to toxaphene. Our results indicate that toxaphene had little or no effect on the morphological endpoints examined including body mass (BM) and size, liver, thyroid, and gonad development. In addition, toxaphene failed to affect sexual differentiation, or in vitro thyroxin, testosterone (T), and estradiol production. However, male plasma T concentration was higher in animals treated with 10 and 0.01 microg toxaphene/kg (based on mean egg mass) than control males. Because in vitro T production was not different among control groups, we suggest the difference in plasma T could be due to differences in hypothalamic-pituitary stimulation of the gonad or hepatic steroid degradation. This study indicates that technical grade toxaphene, at the applied doses, does not induce the same developmental abnormalities associated with alligators living in Lake Apopka. Future studies should consider the effects of embryonic exposure to a mixture of chemicals, including toxaphene metabolites, on development in alligators to better evaluate the consequences of environmental contamination.

Alligators and Crocodiles↗

P and trace metal contents in biomaterials, soils, sediments and plants in colony of red-footed booby (Sula sula) in the Dongdao Island of South China Sea.

Concentrations of P and trace metals Zn, Cu, Cd, Pb and Hg in the faeces, bones, eggshells and feathers of seabirds and in the plants, soils and sediments with and without seabird influence on Dongdao Island, South China Sea, were determined and analyzed. Among the seabird biomaterials, the levels of P, Zn, Cu and Cd are the highest in the droppings and several times those in other materials; the Hg concentration is the highest in the feathers; and the Pb content is comparable among these biomaterials. These marked differences indicate different intake-bioaccumulation-elimination pathways for different trace metals. The levels of P, Zn, Cu, Cd and Hg in the plant, soil and sediment samples with the influence of seabird droppings are significantly higher than those in the samples without, and they are significantly correlated with each other. Thus, P, Zn, Cu, Cd and Hg are very likely to have a common source-predominantly bird guano-and the faeces of red-footed booby is an important vector for the flux of nutrient phosphorus and trace metals Zn, Cu, Cd and Hg from marine to island ecosystems on Dongdao Island.

Animal Structures↗

The effect of prenatal hypoxia and malnutrition on memory consolidation in the chick.

The contribution of hypoxia and malnutrition to cognitive impairments was investigated in chicks incubated in conditions of reduced gas exchange. Previous research has shown that reducing gas exchange during incubation by wrapping half the eggshell with an impermeable membrane results in impaired cognitive ability in young chicks. The results were interpreted within a three stage sequential model of memory using discriminated bead avoidance learning. Reducing gas exchange for 4 days from day 10 or 14, of the 21-day incubation, inhibits memory formation and consolidation into permanent storage. The nature of the cognitive deficit depended on the timing of the insult. Environmental hypoxia (14% oxygen), induced from days 10 to 14 and from days 14 to 18, replicated the memory deficits found previously when eggs were partially wrapped with a membrane. Oxygen is necessary to break down food and to provide energy to build tissue proteins, and therefore hypoxia (partial wrapping or environmental incubation) may indirectly cause malnutrition. Malnutrition, induced by removing 5%, 7.5% or 10% albumin from the egg prior to incubation, had no significant effect on memory consolidation. Raised corticosterone levels occurred in chicks malnourished by 5% and 7.5%, but brain sparing was only evident in chicks with 7.5% albumin removal. Hatch rates were very low in 10% malnourished chicks. Using the chick as a model of prenatal stress, we have been able to isolate the effects of hypoxia from contributing maternal factors.

Age Factors↗

Assessing breeding potential of peregrine falcons based on chlorinated hydrocarbon concentrations in prey.

Peregrine falcons (Falco peregrinus) now breed successfully in most areas of North America from which they were previously extirpated. The loss during the mid-part of the last century of many of the world's peregrine populations was largely a consequence of impaired reproduction caused by the effects of DDE on eggshell quality and embryo hatchability. Population recovery has been attributed to re-introduction efforts, coupled with regulatory restrictions on the use of organochlorine pesticides. Peregrines have not returned to breed in some areas, such as the Okanagan Valley of British Columbia. That region has been extensively planted in fruit orchards which were treated annually with DDT during the early 1950s to the 1970s. Ongoing contamination of avian species, including potential peregrine prey, inhabiting orchards has been documented. In response to an initiative to release peregrines around the city of Kelowna in the Okanagan Valley, we collected potential peregrine prey species and analyzed whole bodies for chlorinated hydrocarbon residues. We used a simple bioaccumulation model to predict concentrations of DDE in peregrine eggs using concentrations in prey and estimates of dietary makeup as input. Peregrines would be expected to breed successfully only if they fed on a diet primarily of doves. Feeding on as little as 10% of other species such as starlings, robins, gulls and magpies would produce DDE concentrations in peregrine eggs greater than the threshold of 15 mg/kg. We also estimated the critical concentration of DDE in total prey to be about 0.5 mg/kg, one half of the previous most conservative criterion for peregrine prey. Concentrations of dieldrin and PCBs in peregrine prey are less than suggested critical levels.

Animals↗

Analysis of chitin synthase function in a plant parasitic nematode, Meloidogyne artiellia, using RNAi.

A single chitin synthase gene is responsible for chitin production in the eggshells of the plant parasitic nematode Meloidogyne artiellia. In this paper we describe a functional analysis of this gene using RNAi as well as further analysis of two similar genes from the free-living nematode Caenorhabditis elegans. The parasitic life-style of M. artiellia required the development of a novel method for delivery of dsRNA to nematode eggs that may be of utility in other experimental systems. C. elegans chitin synthase genes were silenced by feeding nematodes bacteria expressing appropriate chitin synthase dsRNA from a plasmid vector, while M. artiellia egg masses were soaked in dsRNA solution. The results obtained demonstrated that the synthesis of chitin continues to take place in nematode eggs within the egg sac in the parasitic nematode, and that the removal of this activity affects egg development in both C. elegans and M. artiellia. The method described here provides a new way of investigating gene function in plant parasitic nematodes allowing the validity of potential target genes for novel control methods to be assessed. Furthermore, since intact egg cells within the gelatinous matrix of M. artiellia are permeable to dsRNA, eggs of other nematodes may also be similarly permeable to dsRNA and therefore amenable to use with dsRNAi.

Animals↗

A chronology for late prehistoric Madagascar.

A database has been assembled with 278 age determinations for Madagascar. Materials 14C dated include pretreated sediments and plant macrofossils from cores and excavations throughout the island, and bones, teeth, or eggshells of most of the extinct megafaunal taxa, including the giant lemurs, hippopotami, and ratites. Additional measurements come from uranium-series dates on speleothems and thermoluminescence dating of pottery. Changes documented include late Pleistocene climatic events and, in the late Holocene, the apparently human-caused transformation of the environment. Multiple lines of evidence point to the earliest human presence at ca. 2300 14C yr BP (350 cal yr BC). A decline in megafauna, inferred from a drastic decrease in spores of the coprophilous fungus Sporormiella spp. in sediments at 1720+/-40 14C yr BP (230-410 cal yr AD), is followed by large increases in charcoal particles in sediment cores, beginning in the SW part of the island, and spreading to other coasts and the interior over the next millennium. The record of human occupation is initially sparse, but shows large human populations throughout the island by the beginning of the Second Millennium AD. Dating of the "subfossil" megafauna, including pygmy hippos, elephant birds, giant tortoises, and large lemurs, demonstrates that most if not all the extinct taxa were still present on the island when humans arrived. Many taxa overlapped chronologically with humans for a millennium or more. The extinct lemurs Hadropithecus stenognathus, Pachylemur insignis, Mesopropithecus pithecoides, and Daubentonia robusta, and the elephant birds Aepyornis spp. and Mullerornis spp., were still present near the end of the First Millennium AD. Palaeopropithecus ingens, Megaladapis edwardsi, and Archaeolemur sp. (cf. edwardsi) may have survived until the middle of the Second Millennium A.D. One specimen of Hippopotamus of unknown provenance dates to the period of European colonization.

Animals↗

Amyloid fibrillogenesis of silkmoth chorion protein peptide-analogues via a liquid-crystalline intermediate phase.

Chorion, the major component of silkmoth eggshell, consists of the A and B classes of low-molecular weight structural proteins. Chorion protects the oocyte and the developing embryo from environmental hazards and this is due to the extraordinary physical and chemical properties of its constituent proteins. We have shown previously [FEBS Lett. 479 (2000) 141; 499 (2001) 268] that peptide-analogues of the A and B classes of chorion proteins form amyloid fibrils under a variety of conditions, which led us to propose that silkmoth chorion is a natural, protective amyloid. In this work, we present data showing conclusively that, the first main step of amyloid-like fibrillogenesis of chorion peptides is the formation of nuclei of liquid crystalline nature, which is reminiscent of spider-silk formation. We show that these liquid-crystalline nuclei (spherulites) 'collapse'/deteriorate to form amyloid fibrils in a spectacular manner, important, it seems, for chorion morphogenesis and amyloid fibrillogenesis in general. The molecular 'switch' causing this spectacular transformation is, most probably, a conformational transition to the structure of chorion peptides, from a left-handed parallel beta-helix to an antiparallel beta-pleated sheet. Apparently, these peptides were suitably designed to play this role, after millions of years of molecular evolution.

Amino Acid Sequence↗