Assessing the risk to nontarget organisms from Bt corn resistant to corn rootworms (Coleoptera: Chrysomelidae): Tier-I testing with Orius insidiosus (Heteroptera: Anthocoridae)

A 14-d continuous dietary exposure bioassay using nymphs of the insidious flower bug, Orius insidiosus (Say) (Heteroptera: Anthocoridae), was conducted to assess nontarget impacts of genetically modified corn event MON 863 expressing the Cry3Bb1 protein for management of corn rootworms, Diabrotica spp. (Coleoptera: Chrysomelidae). Nymphs of O. insidiosus were continuously fed a bee pollen diet inoculated with a maximum hazard exposure dose (930 microg/g of diet) of the Cry3Bb1 protein for 14 d. The Cry3Bb1 protein at a concentration of 930 microg/g of diet had no adverse effect on the survival and development (to adults) of O. insidiosus nymphs. In contrast, when O. insidiosus nymphs were fed bee pollen diet treated with a hazard dose of the protease inhibitor E64 (53 microg/g of diet) or the stomach poison potassium arsenate (8.9 microg/g of diet), all nymphs died before developing to adults. Furthermore, statistical power analysis indicated that at levels of 80% power and a 5% type I error rate, the study design would have been able to detect a minimum 30% reduction in survival of test nymphs and a 20% reduction in nymphal development to the adults relative to the buffer control groups. Based on the maximum level (93 microg/g) of the Cry3Bb1 protein expressed in MON 863 corn tissues including leaves, roots, and pollen, findings from this study indicate that corn hybrids containing the MON 863 event have a minimum 10 times safety factor for nymphs of O. insidiosus and thus pose minimal risk to this beneficial insect.     

Development and survival of Orius insidiosus (Say) nymphs on encapsulated bee pollen-based diet in a Tier-I toxicity assay

The insidious flower bug, Orius insidiosus (Say) (Heteroptera: Anthocoridae) is an important surrogate species for assessing potential effects of plant-incorporated protectants (PIPs) on nontarget heterotrophic predators. In this study, a continuous dietary exposure system was optimized by assessing the effect of diet composition and age on the survival and development of nymphs of O. insidiosus. Greater than 85% control survival and an acceptable rate of development from nymph hatching to adult was achieved using 5-d-old nymphs at test initiation and a bee pollen-based diet supplemented with 25% Ephestia eggs. There was an unacceptable level of mortality (>40%) and/or a significantly prolonged development time when nymphs were <5 d old at test initiation. When 5-d-old nymphs were fed a bee pollen diet containing 25% Ephestia eggs and 100 μg/g potassium arsenate, time-dependent mortality was observed with a median lethal time (LT50) of 4.4 d and 100% mortality was observed after 10 d of feeding, indicating the effectiveness of the test system to detect adverse effects by dietary exposure. It is recommended that well-defined 5-d-old nymphs and an encapsulated bee pollen-based diet containing 25% ground Ephestia eggs be used in a Tier-I dietary feeding exposure assay for detecting potential effects of PIPs on O. insidiosus nymphs.     

Bioavailability of selenium from meat and broccoli as determined by retention and distribution of 75Se

The concentration of selenium (Se), an essential nutrient, is variable in foods, depending, in part, on how and where foods are produced; some foods accumulate substantial amounts of Se when produced on high-Se soils. The chemical form of Se also differs among foods. Broccoli is a Se-accumulating plant that contains many methylated forms of Se, and Se bioavailability from broccoli has been reported to be low. Red meats such as pork or beef could accumulate Se when the animal is fed high-Se diets, and Se from such meats has been reported to be highly bioavailable for selenoprotein synthesis. In a further attempt to characterize the utilization of Se from broccoli and meats such as pork or beef, we have fed rats diets adequate (0.1 μg Se/g diet) in Se or high in Se (1.5 μg S/g diet), with the Se source being either high-Se broccoli or beef. Rats were then given test meals of broccoli or pork intrinsically labeled with ⁷⁵Se. When dietary Se was nutritionally adequate (0.1 μg/g diet), more ⁷⁵Se from pork than broccoli was retained in tissues; however, there were no significant differences in whole-body retention when dietary Se was high (1.5 μg/g diet). A significantly greater percentage of ⁷⁵Se from broccoli than pork was excreted in the urine and dietary Se did not affect urinary excretion of broccoli ⁷⁵Se, but the amount excreted from pork varied directly with dietary Se intake. Radiolabeled ⁷⁵Se derived from pork effectively labeled selenoproteins in all tissues examined, but ⁷⁵Se from broccoli was undetectable in selenoproteins. These differences in retention and distribution of Se from broccoli or pork are consistent with reported differences in bioavailability of Se from beef and broccoli. They also suggest that there are fewer differences in bioavailability when Se is consumed in supranutritional amounts.     

Toxicity of pesticides used in chrysanthemum crops to eggs and nymphs of Orius insidiosus (Say) (Hemiptera: Anthocoridae)

The objective of this study was to evaluate the toxicity of some pesticides used in chrysanthemum crops to eggs and nymphs of Orius insidiosus (Say). The bioassays were carried out at 25 +/- 2 degrees C, 70 +/- 10% RH and 12h photophase. For the tests with eggs, stems of Bidens pilosa L. previously exposed to predator oviposition were dipped for five seconds in aqueous dilution of the pesticides, using 40 eggs per treatment. The pesticides were applied on nymphs using a Potter's tower. Forty first-instar and twenty second-instar nymphs of O. insidiosus were used per treatment in the tests with nymphs. The adults from eggs and nymphs treated were grouped in couples to study the pesticides effects on its reproductive parameters. None tested pesticide affected the viability of treated eggs. Azoxystrobin, benomyl, imibenconazole, iprodione, metalaxyl + mancozeb and triforine were harmless to first- and second-instar nymphs of O. insidiosus, whereas abamectin, acephate and chlorfenapyr were shown to be harmful to nymphs of both first and second instars. The pre-oviposition period, the daily number of eggs, number of eggs in ten days, and egg viability were not affected by azoxystrobin, benomyl, imibenconazole, iprodione, metalaxyl + mancozeb and triforine. These pesticides, for showing low toxicity to the predator, can be recommended in disease management programs for chrysanthemum crops, in association with O. insidiosus.     

Use of a dietary exposure system for screening of insecticidal compounds for their toxicity to the planthopper Laodelphax striatellus

We developed a dietary exposure assay for screening insecticidal compounds for their toxicity and for assessing the side effects of insecticidal proteins produced by genetically engineered （GE） plants on the planthopper Laodelphax striatellus Fallen. The fitness bioassay confirmed that the diet fulfills the requirements to be used in the dietary exposure system. To validate the efficacy of the dietary exposure system, nymphs of L. striatellus were fed diets treated with different concentrations of an inorganic stomach poison, potassium arsenate （PA）, or a cysteine protease inhibitor, E-64. The results showed that with increasing concentrations of E-64, the larval development time was prolonged, the adult weight was reduced and the survival rate of L. striatellus was decreased. Similarly the survival rates of L. striatellus consistently decreased with increasing PA content in the diet. The data indicate that the dietary exposure assay is able to detect the effects of insecticidal compounds on L. striatellus. Subsequently, this assay was successfully used for assessing the potential toxicity of Cry2Aa. The results showed that L. striatellus larvae were not negatively affected when fed the artificial diet containing purified Cry2Aa at 300 μg/g diet. In the assay, the stability and bioactivity of crystal （Cry） proteins in the food sources were confirmed by enzyme-linked immunosorbent assay and sensitive-insect bioassays. These results show that L. striatellus is not sensitive to Cry2Aa. We conclude that the dietary exposure system is valid and useful for assessing the toxicity of insecticidal compounds produced by GE plants on planthoppers.     

Use of a Pollen-Based Diet to Expose the Ladybird Beetle Propylea japonica to Insecticidal Proteins

A rape seed pollen-based diet was developed and found to be suitable for use in a dietary exposure assay for Propylea japonica. Using the diet, we established and validated a dietary exposure assay by using the protease inhibitor E-64 as positive control. Dose-dependent responses were documented for all observed life-table parameters of P. japonica including survival, pupation and eclosion rates, development time and adult weight. Results suggested that the dietary assay can detect the effects of insecticidal compounds on the survival and development of P. japonica. Using the established dietary assay, we subsequently tested the toxicity of Cry1Ab, Cry1Ac and Cry1F proteins that are expressed by transgenic maize, cotton or rice plants to P. japonica larvae. The diet containing E-64 was included as a positive control. Survival and development of P. japonica larvae were not adversely affected when the diet contained purified Cry1Ab, Cry1Ac, or Cry1F at 500 µg/g diet representing a worst-case exposure scenario. In contrast, P. japonica larvae were adversely affected when the diet contained E-64. The bioactivity and stability of the Cry proteins in the diet and Cry protein uptake by the ladybird larvae were confirmed by bioassay with a Cry-sensitive insect species and by ELISA. The current study describes a suitable experimental system for assessing the potential effects of gut-active insecticidal compounds on ladybird beetle larvae. The experiments with the Cry proteins demonstrate that P. japonica larvae are not sensitive to Cry1Ab, Cry1Ac and Cry1F.     

Evaluation of seven plant species/cultivars for their suitability as banker plants for Orius insidiosus (Say)

Marigold (cv. Lemon Gem), castor bean, ornamental pepper (cv. Black Pearl and Purple Flash), gerbera daisy (cv. Festival), feverfew, and sunflower (cv. Choco Sun) were evaluated for their suitability as banker plants (BP) for Orius insidiosus (Hemiptera: Anthocoridae) in commercial greenhouses. Oviposition, egg hatch, nymphal development to adulthood, and population increase were quantified in laboratory trials. Assessments of oviposition and egg hatch indicated that all plants tested were equally accepted by O. insidiosus. Nymphal development to adulthood and survival tests indicated that gerbera may be a suitable BP as survival was the highest (58.1 %), whereas marigold would not be an acceptable BP as only 10.7 % of nymphs survived to adulthood. Nymphal development time differed by only one day among all plants. In greenhouse cage experiments, Purple Flash pepper supported the greatest population growth over a ten week period. Based on the combined results from all tests, Purple Flash pepper appears to have the greatest potential as a BP species for O. insidiosus.     

Voracity and prey preference of insidious flower bug (hemiptera: anthocoridae) for immature stages of soybean aphid (hemiptera: aphididae) and soybean thrips (thysanoptera: thripidae)

The generalist predator, Orius insidiosus (Say), is an important natural enemy of the soybean aphid, Aphis glycines Matsumura. Soybean thrips, Neohydatothrips variabilis (Beach), serve as an important prey resource for O. insidiosus in soybeans, sustaining the predator's population before the arrival of the soybean aphid. Although generalist predators can forage on a broad range of prey, they may show distinct preferences for particular prey, attacking prey at levels disproportionate to their relative numbers. To assess the preference of O. insidiosus for soybean aphid and soybean thrips, attack rates of nymphal and adult O. insidiosus were measured in the laboratory. For both adults and nymphs, the number of prey attacked increased as more prey were provided. For nymphs, the total number of prey attacked increased as the predator matured. In general, the number of prey attacked by adult predators was relatively constant as the predator aged. Both O. insidiosus nymphs and adults displayed a preference for soybean thrips, by disproportionately attacking soybean thrips over soybean aphid regardless of the relative densities of the two prey. We discuss implications of this preference on O. insidiosus life history characteristics and the potential impact on O. insidiosus-prey dynamics in the field.     

Hepatobiliary and hematological effects of dietary disodium arsenate heptahydrate in juvenile rainbow trout (Oncorhynchus mykiss)

1. The impact of 8–12 weeks exposure to up to 55–60 μg arsenic/g diet as disodium arsenate (DSA) on the hepatobiliary and hematological systems of rainbow trout was assessed.2. Bile and washed gallbladder tissue arsenic residues in exposed fish were 2–4 times higher than in liver. Plasma arsenic concentration was slightly elevated following dietary DSA exposure.3. Chronic exposure to dietary DSA caused a mild to moderate responsive anemia.4. Lipid, but not protein, digestibility after 12 weeks of exposure showed an inverse relationship with severity of gallbladder inflammation in these fish.5. Changes in the hepatobiliary system appear to provide the most sensitive indicators of dietary DSA exposure in rainbow trout.     

Response of hepatic function to hepatic copper deposition in rats fed a diet containing copper

Fischer rats were a fed diet supplied with copper chloride (150–600 ppm) for 60 d from weaning. Serum (glutamic-oxaloacetic transaminase (GOT) and glutamic-pyruvic transaminase (GPT) activities were increased with the increase of Cu concentration in the diet. Biliary excretion of Cu was related to the dietary Cu level. Depositions of hepatic and renal Cu were also related to the dietary Cu level in a dose-dependent manner. In particular, hepatic (155.2±13.3 μg/g) and renal (44.9±4.4 μg/g) Cu concentrations increased abruptly in the Cu-600 ppm group. In the liver, about 60% of Cu was distributed in the soluble fraction (100,000 g supernatant). In the Cu-600 ppm group, 25% of cystosolic Cu was bound to metallothionein (MT). Our results suggest that chronic exposure to Cu appears to have a deleterious effect on the hepatic function, and further, that even in rats with normal biliary Cu excretion, clearance of Cu from the liver may be marginal when dietary Cu is near the 600-ppm level. Although Cu is an essential nutrient, an overload of Cu should be avoided.     

Effect of dietary protein, zinc, and carbon monoxide on fetal zinc concentration in mice

Dietary protein and zinc deficiencies known to be detrimental to the developing fetus are common in pregnant women in developing countries. Everyone in modern society is at risk of exposure to carbon monoxide (CO). This study was conducted to observe the effect of dietary protein, zinc, and exposure to CO on the fetal zinc concentrations by factorial experimentation. Pregnant mice of CD-1 strain were maintained on 17% (control) or 9% (deficient) protein diets mixed with deficient, normal (control), or supplemental zinc throughout gestation. The dams in each dietary group were exposed to air (control) or 500 ppm CO in air in environmental chambers from gestation day 8 to gestational day 18. The dams were sacrificed on d 18 and fetal zinc levels were measured by atomic absorption spectrophotometry. Carbon monoxide levels used in this study had no significant effect on fetal zinc concentration in any treatment group. When both dietary protein and zinc levels were normal, the mean fetal zinc concentrations were higher than all other dietary protein/zinc combinations (15.2±6.0 and 14.2±4.1 μg Zn/g of tissue for 0 and 500 ppm CO levels). However, when dietary protein levels were deficient, supplemental zinc increased the fetal zinc concentrations significantly (12.7±3.8 and 13.1±0.3.6 μg Zn/g of tissue, in 0 and 500 ppm CO groups) as compared to zinc-deficient groups (8.7±3.0 and 10.0±3.3 μg Zn/g of tissue in 0 and 500 ppm CO groups). The results of this study may be relevant to populations that experience both marginal zinc and protein diets during gestation.     

Dietary fat composition modifies the effect of boron on bone characteristics and plasma lipids in rats

Female and male rats weighing about 170 g and 200 g, respectively, were fed diets (approximately 70 microg boron/kg) in a factorial arrangement with supplemental boron at 0 (deficient) and 3 (adequate) mg/kg and canola oil or palm oil at 75 g/kg of diet as variables. After 5 weeks, six females in each treatment were bred. Dams and pups continued on their respective dietary treatments through gestation, lactation and post-weaning. Thirteen weeks after weaning, plasma and bones were collected from 12 male and 12 female offspring in each treatment. Boron supplementation increased femur strength measured by the breaking variable bending moment; tibial calcium and phosphorus concentrations; and plasma alkaline phosphatase. Femur breaking stress was greatest in boron-supplemented rats fed canola oil, and lowest in boron-deprived females fed canola oil; this group also exhibited the lowest femur bending moment. Minerals associated with bone organic matrix, zinc and potassium, were increased by boron supplementation in tibia. Plasma phospholipids were decreased by boron deprivation in females, but not males. Plasma cholesterol was decreased in boron-supplemented males by replacing canola oil with palm oil. The findings suggest that a diet high in omega-3 alpha-linolenic acid promotes femur strength best when the dietary boron is adequate.     

Investigations on the effects of rape oil quality, choline and methionine concentration in diets for laying hens on the trimethylamine content of the eggs, on trimethylamine metabolism and on laying performance

An experiment was conducted to study the effects of graded levels of choline addition (0, 500, 1000 and 4000 mg/kg diet) in laying hen diets prepared either with degummed or refined rape oil on the performance, sensory properties and trimethylamine (TMA) contents of the eggs. Furthermore, the diets containing no supplemented choline or 4000mg choline/kg diet were tested with adequate or inadequate methionine supply (4.2 vs. 2.8 g methionine/kg diet). TMA metabolism and N-balance were measured for the latter diet types, but only with the diets containing refined rape oil. Therefore, a total of 12 and 4 diets were tested in the feeding (n = 60) and balance study (n = 9). Laying performance (23 -75 weeks of age) was not significantly influenced by increasing choline additions with the exception of feed-to-egg mass ratio which decreased significantly linearly (P(linear) = 0.003). However, a significant interaction between choline addition and laying month was detected which was caused by a depression of performance of the unsupplemented control group occurring from the sixth laying month. The most obvious effect of an inadequate methionine supply was a temporary drop in performance between the third and sixth laying months. The mean TMA-concentration in pooled egg yolks [microg/g] increased with dietary choline concentration [mg/kg] in an exponentially related fashion (y = 1.14 + 4E(-10) x x(2.71), r2 = 0.962) and suggested only a minor influence of total dietary choline on TMA content up to approximately 2000mg choline/kg. Individual TMA-concentrations varied greatly from 0.4 - 1.5 microg/g, from 2.2 - 34 microg/g and from 18.4 - 75 microg/g for eggs with a normal, aberrant and heavily aberrant odour, respectively. It is concluded that a total choline concentration of at least approximately 1500 mg/kg is necessary to maintain a maximal laying performance. An inadequate methionine supply cannot be compensated by an increased addition of choline. Neither degummed nor refined rape oil influenced the TMA content of eggs.     

Dietary silicon and arginine affect mineral element composition of rat femur and vertebra

Both arginine and silicon affect collagen formation and bone mineralization. Thus, an experiment was designed to determine if dietary arginine would alter the effect of dietary silicon on bone mineralization and vice versa. Male weanling Sprague-Dawley rats were assigned to groups of 12 in a 2×2 factorially arranged experiment. Supplemented to a ground corn/casein basal diet containing 2.3 μg Si/g and adequate arginine were silicon as sodium metasilicate at 0 or 35 μg/g diet and arginine at 0 or 5 mg/g diet. The rats were fed ad libitum deionized water and their respective diets for 8 wk. Body weight, liver weight/body weight ratio, and plasma silicon were decreased, and plasma alkaline phosphatase activity was increased by silicon deprivation. Silicon deprivation also decreased femoral calcium, copper, potassium, and zinc concentrations, but increased the femoral manganese concentration. Arginine supplementation decreased femoral molybdenum concentration but increased the femoral manganese concentration. Vertebral concentrations of phosphorus, sodium, potassium, copper, manganese, and zinc were decreased by silicon deprivation. Arginine supplementation increased vertebral concentrations of sodium, potassium, manganese, zinc, and iron. The arginine effects were more marked in the silicon-deprived animals, especially in the vertebra. Germanium concentrations of the femur and vertebra were affected by an interaction between silicon and arginine; the concentrations were decreased by silicon deprivation in those animals not fed supplemental arginine. The change in germanium is consistent with a previous finding by us suggesting that this element may be physiologically important, especially as related to bone DNA concentrations. The femoral and vertebral mineral findings support the contention that silicon has a physiological role in bone formation and that arginine intake can affect that role. The U.S. Department of Agriculture, Agricultural Research Service, Northern Plains Area is an equal opportunity/affirmative action employer, and all agency services are available without discrimination. Mention of a trademark or proprietary product does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products that may be suitable.     

Nutritional assessment of somatolactin function in gilthead sea bream (Sparus aurata): concurrent changes in somatotropic axis and pancreatic hormones

The role of somatolactin (SL) in the regulation of energy homeostasis in gilthead sea bream (Sparus aurata) has been analysed. First, a down-regulation of plasma SL levels in response to gross shifts in dietary amino acid profile and the graded replacement of fish meal by plant protein sources (50%, 75% and 100%) has been observed. Thus, the impaired growth performance with changes in dietary amino acid profile and dietary protein source was accompanied by a decrease in plasma SL levels, which also decreased over the course of the post-prandial period irrespective of dietary nitrogen source. Secondly, we examined the effect of SL and growth hormone (GH) administration on voluntary feed intake. A single intraperitoneal injection of recombinant gilthead sea bream SL (0.1 microg/g fish) evoked a short-term inhibition of feed intake, whereas the same dose of GH exerted a marked enhancement of feed intake that still persisted 1 week later. Further, we addressed the effect of arginine (Arg) injection upon SL and related metabolic hormones (GH, insulin-like growth factor-I (IGF-I), insulin and glucagon) in fish fed diets with different nitrogen sources. A consistent effect of Arg injection (6.6 micromol/g fish) on plasma GH and IGF-I levels was not found regardless of dietary treatment. In contrast, the insulinotropic effect of Arg was found irrespective of dietary treatment, although the up-regulation of plasma glucagon and glucose levels was more persistent in fish fed a fish meal based diet (diet FM) than in those fed a plant protein diet with a 75% replacement (diet PP75). In the same way, a persistent and two-fold increase in plasma SL levels was observed in fish fed diet FM, whereas no effect was found in fish fed diet PP75. Taken together, these findings provide additional evidence for a role of SL as a marker of energy status, which may be perceived by fish as a daily and seasonal signal of abundant energy at a precise calendar time.     

Use of an artificial diet system to study the toxicity of gut-active insecticidal compounds on larvae of the green lacewing Chrysoperla sinica

A semi-liquid artificial diet was established and found to be a suitable food source for Chrysoperla sinica larvae, comparable to aphid prey. Using the artificial diet, we established and validated a dietary exposure assay by using the insecticidal potassium arsenate (PA) as positive control. Dose-dependent responses were documented for all observed life-table parameters of C. sinica larvae such as survival rate, pupation rate, larval weight, and larval development time. Thus, the dietary assay can detect the effects of insecticidal compounds on the survival and development of C. sinica larvae. Using the established dietary assay, we subsequently tested the toxicity of Cry1Ab, Cry1Ac, and Cry2Aa proteins (which are produced by transgenic maize, cotton or rice plants) to C. sinica larvae. Artificial diets containing Galanthus nivalis agglutinin (GNA) or PA were included as positive controls. Survival and development of C. sinica larvae were not affected when the artificial diet contained purified Cry1Ab, Cry1Ac, or Cry2Aa at 200 μg/g diet. In contrast, C. sinica larvae were adversely affected when the diet contained PA and GNA. The stability and bioactivity of the Cry proteins in the diet and Cry protein uptake by the lacewing larvae were confirmed by bioassay with a Cry-sensitive insect species and by ELISA. The current study describes a suitable experimental system for assessing the potential effects of gut-active insecticidal compounds on green lacewing larvae. The experiments with the Cry proteins demonstrate that C. sinica larvae are not sensitive to Cry1Ab, Cry1Ac, and Cry2Aa.     

Estrogens and phytoestrogens: brain plasticity of sexually dimorphic brain volumes

Sexually dimorphic brain volumes (sexually dimorphic nucleus of the preoptic area (SDN-POA) and anteroventral periventricular (AVPV) nucleus) are influenced by estrogens. Phytoestrogens, derived from plants (especially soy products), are molecules structurally and functionally similar to estradiol. The purpose of this study was to examine: the consumption of phytoestrogen (using a phytoestrogen-rich (Phyto-600) versus a phytoestrogen-free (Phyto-free)) diets from conception to adulthood (or changing the diets during adulthood) and characterizing (a) circulating plasma phytoestrogen levels, (b) testosterone levels in males, (c) sexually dimorphic brain volumes (i.e. the SDN-POA and AVPV) and (d) the presence of apoptotic cells in these brain structures in Long-Evans rats. Phyto-600 fed animals displayed total serum phytoestrogens levels 37-fold higher compared to Phyto-free values. Circulating testosterone levels were not significantly altered by the diets. Female SDN-POA volumes were not altered by the diets. Whereas, males fed a Phyto-free diet displayed decreased SDN-POA volumes compared to male Phyto-600 values. Females fed the Phyto-600 diet displayed larger AVPV volumes compared to males on the same diet or females on the Phyto-free diet. Males fed the Phyto-free diet had the largest AVPV values compared to Phyto-600 fed males. When the SDN-POA region was examined in lifelong Phyto-free fed males, apoptotic cells were present versus males fed the Phyto-600 diet and in the AVPV region the opposite results were obtained. In summary, consumption of dietary phytoestrogens (estrogen mimics) can alter hormone-sensitive hypothalamic brain volumes in rodents during adulthood.     

Responses of hybrid striped bass to waterborne and dietary copper in freshwater and saltwater

Mechanisms of copper toxicity and consequences of exposure vary due to uptake route and ionoregulatory status. The goal of this research was to develop a model fish system to assess the influence of different Cu exposure routes (waterborne or dietary) on bioavailability, uptake, and effects in hybrid striped bass (Morone chrysops x Morone saxatilis) acclimated to fresh- or saltwater. Initially, hybrid striped bass were exposed to dietary Cu concentrations of 571, 785, and 1013 mug Cu/g, along with a control (approximately 5 microg Cu/g), for 14 days in saltwater. Intestinal and liver Cu accumulated in a dose-dependent manner in fish exposed to increasing levels of dietary Cu. Chronic (42 days) experiments were then conducted to determine sub-lethal effects of aqueous, dietary, and combined aqueous and dietary Cu exposures to both freshwater- and saltwater-acclimated hybrid striped bass. Growth and Cu accumulation in the gill, intestine, and liver were measured. Although no significant effects were observed in fish exposed to waterborne Cu, those exposed through the diet accumulated significant liver and intestinal Cu but showed no significant change in growth. Overall, these results suggest that at the levels tested, exposure to elevated waterborne Cu did not cause significant long-term tissue Cu accumulation, whereas dietary Cu exposure caused significant liver and intestinal Cu accumulation in hybrid striped bass which was comparable in both freshwater and saltwater (15 g/L).     

Dietary selenium supplementation prolongs pentobarbital induced hypnosis

The present studies characterized the influence of dietary selenium (Na2SeO3) on the duration of pentobarbital (PB) induced hypnosis (sleep) in the rat. Rats were fed semipurified diets varying from 0.01 to 2.0 mg Se/kg for up to 4 weeks. Consumption of diets containing 1.0 and 2.0 mg Se/kg significantly prolonged PB induced hypnosis. Hepatic selenium, but not hepatic glutathione peroxidase activity, correlated with the length of PB induced hypnosis. The prolongation of hypnosis caused by diets containing 1.0 mg Se/kg was substantially reduced or eliminated by repeated exposure to PB. Although single exposure to increasing quantities of PB (60-100 mg/kg body weight) led to a progressive increase in sleep duration, the proportional increase caused by supplemental selenium (2.0 vs 0.1 microg Se/g) remained relatively constant (approximately 25%). Increasing maturity was inversely related to the duration of PB induced hypnosis, regardless of dietary selenium provided. Consumption of the 2.0 mg Se/kg diet prolonged PB induced hypnosis to a greater degree in immature than in mature rats (P < 0.05). Consumption of the selenium enriched diet (2 microg Se/g) resulted in an increase in cytochrome 2B, but had no effect on cytochrome 1A compared to controls (0.1 microg Se/g). Pretreatment of rats with P450 enzymes activators (i.e., PB, Aroclor 1254, or 3-methylcholanthrene) shortened the duration of PB induced sleep and masked the effects of dietary selenium. The current studies document that dietary selenium can influence the response to pentobarbital induced hypnosis and likely relates to changes in drug detoxification enzymes.     

Effect of estrogenic activity, and phytoestrogen and organochlorine pesticide contents in an experimental fish diet on reproduction and hepatic vitellogenin production in medaka (Oryzias latipes)

Endocrine-disrupting chemicals (EDCs) are giving rise to serious concerns for humans and wildlife. Phytoestrogens, such as daidzein and genistein in plants, and organochlorine pesticides are suspected EDCs, because their chemical structure is similar to that of natural or synthetic estrogens and they have estrogenic activity in vitro and in vivo. We assessed estrogenic activity and dietary phytoestrogen and organochlorine pesticide contents of various fish diets made in the United Kingdom, and compared them with those features of diets made in Japan that were tested in a previous study. Genistein and daidzein were detected in all of the diets. Using an in vitro bioassay, many of these diets had higher activation of estrogen beta-receptors than estrogen alpha-receptors. Organochlorine pesticides such as hexachlorobenzene, beta-benzene hexachloride (BHC), and gamma-BHC were detected in all fish diets. On the basis of these data, we investigated the effect of differing dietary phytoestrogen content in Japanese fish diets on hepatic vitellogenin production and reproduction (fecundity and fertility) in medaka (Oryzias latipes). Assessment of the effects of a 28-day feeding period on reproduction of paired medaka did not indicate significant differences in the number of eggs produced and fertility among all feeding groups. However, hepatic vitellogenin values were significantly higher for male medaka fed diet C (genistein, 58.5 +/- 0.6 microg/g; daidzein, 37.3 +/- 0.2 microg/g) for 28 days compared with those fed diet A (genistein, < 0.8 microg/g; daidzein, < 0.8 microg/g) or diet B (genistein, 1.4 +/- 0.1 microg/g; daidzein, 2.0 +/- 0.1 microg/g). Our findings indicate that fish diets containing high amounts of phytoestrogens, such as diet C, have the potential to induce hepatic vitellogenin production in male medaka, even if reproductive parameters are unaffected. Therefore, some diets, by affecting vitellogenin production in males, may alter estrogenic activity of in vivo tests designed to determine activity of test compounds added to the diet.