Inhibitory effect of cadmium on estrogen signaling in zebrafish brain and protection by zinc.

The present study was conducted to assess the effects of Cd exposure on estrogen signaling in the zebrafish brain, as well as the potential protective role of Zn against Cd-induced toxicity. For this purpose, the effects on transcriptional activation of the estrogen receptors (ERs), aromatase B (Aro-B) protein expression and molecular expression of related genes were examined in vivo using wild-type and transgenic zebrafish embryos. For in vitro studies, an ER-negative glial cell line (U251MG) transfected with different zebrafish ER subtypes (ERα, ERß1 and ERß2) was also used. Embryos were exposed either to estradiol (E2 ), Cd, E2 +Cd or E2 +Cd+Zn for 72 h and cells were exposed to the same treatments for 30 h. Our results show that E2 treatment promoted the transcriptional activation of ERs and increased Aro-B expression, at both the protein and mRNA levels. Although exposure to Cd, does not affect the studied parameters when administered alone, it significantly abolished the E2 -stimulated transcriptional response of the reporter gene for the three ER subtypes in U251-MG cells, and clearly inhibited the E2 induction of Aro-B in radial glial cells of zebrafish embryos. These inhibitory effects were accompanied by a significant downregulation of the expression of esr1, esr2a, esr2b and cyp19a1b genes compared to the E2 -treated group used as a positive control. Zn administration during simultaneous exposure to E2 and Cd strongly stimulated zebrafish ERs transactivation and increased Aro-B protein expression, whereas mRNA levels of the three ERs as well as the cyp19a1b remained unchanged in comparison with Cd-treated embryos. In conclusion, our results clearly demonstrate that Cd acts as a potent anti-estrogen in vivo and in vitro, and that Cd-induced E2 antagonism can be reversed, at the protein level, by Zn supplement. Copyright © 2016 John Wiley & Sons, Ltd.

The role of organic selenium in cadmium toxicity: effects on broiler performance and health status.

This work was part of a project designed to assess whether organic selenium (Se) can protect against the toxic effects of cadmium (Cd). A total of 300 1-day-old, as hatched, broilers were randomly distributed in four dietary treatments with five replicate pens per treatment. In T1 treatment, broilers were fed a diet with 0.3 mg/kg added Se, as Se-yeast, without added Cd; in T2, broilers were fed a diet with 0.3 mg/kg Se and 10 mg/kg Cd; in T3, broilers were fed a diet with 0.3 mg/kg Se and 100 mg/kg of Cd; and in T4 treatment broilers were fed a diet with 3 mg/kg Se and 100 mg/kg Cd. The Cd was added to diets T2, T3 and T4 as CdCl2. On the 4th and 6th week, two broilers per replicate pen were killed in order to obtain whole blood, liver, kidney and breast samples. Body mass, feed conversion ratio and mortality were assessed and haematological analyses were performed. Se and Cd levels in tissues were analysed by inductively coupled plasma mass spectrometry. Broilers supplemented with 0.3 mg/kg Se can tolerate low levels of Cd added to the diets, as there were no significant negative effects on the examined performance parameters, whereas addition of excess Cd led to an impairment of broilers' performance. Mortality of broilers did not differ between the four dietary treatments at any interval point or the whole period. The examined haematological parameters such as haematocrit, total blood protein concentration, and leukocytes types ranged within physiological values, revealing no negative health effects after simultaneous Cd and Se addition. The present study indicated that Se can help against the negative effects of Cd, but cannot counteract all of its negative effects.

Parâmetros qualitativos da carne de cordeiros submetidosaos modelos de produção orgânico e convencional / Qualitative parameters of lamb meat submitted to organic and conventional production models

Este trabalho objetivou avaliar parâmetros qualitativos da carne, análise sensorial e quantificação de resíduo de drogaveterinária e metais pesados provenientes de 48 cordeiros Ile de France submetidos aos modelos de produção orgânicoe convencional, os quais foram abatidos aos 32 kg de peso corporal. A carne dos cordeiros do modelo orgânico tevemaior teor de amarelo que a dos cordeiros do modelo convencional aos 45 minutos após o abate, sendo que os demaisparâmetros L* e a* não foram afetados, já a cor da carne dos cordeiros 24 horas após o abate, não foi influenciada pelostratamentos. Não houve efeito dos tratamentos no pH e na temperatura aos 45 minutos e 24 horas após o abate, nacapacidade de retenção de água e na força de cisalhamento, enquanto as perdas de peso na cocção foram influenciadaspelos tratamentos. Na carne dos cordeiros criados no modelo orgânico, a maciez subjetiva e a aceitação global foraminferiores quando comparadas às do modelo convencional.Os tratamentos não influenciaram os teores de arsênio,cádmio e chumbo da carne. Constatou-se inexistência do princípio ativo ivermectina na carne proveniente dos modelosde produção orgânico e convencional.

This work aimed to evaluate qualitative parameters of meat, sensorial analysis, ivermecin residue and heavy metalsfrom 48 Ile de France lambs submitted to organic and conventional production models which were slaughtered at 32 kgof body weight. Lamb meat from organic model had larger yellowness w to compawithto conventional mo to 45 minutesafter the slaught anbut L * and a* parameters were not affeced;, however, not eady the color of lamb meat 24 hoursafter the slaugter, not influenced by treatments. There was not effect of treatments in pH and temperatureat 45 minutesand 24 hours after the slaughter, in water holding capacity and in shear force, while the cooking losses were influencedby treatments. In the meat of lambs submitted to organic model, the subjective tenderness and the global acceptancewere lower when compared to convencional model. Treatments didn’t influence arsenic, cadmium and lead meat tenor.Inexistence of ivermectin was verified in meat from organic and conventional production models.

Milk trace elements in lactating cows environmentally exposed to higher level of lead and cadmium around different industrial units.

The present investigation was carried out to assess the trace mineral profile of milk from lactating cows reared around different industrial units and to examine the effect of blood and milk concentration of lead and cadmium on copper, cobalt, zinc and iron levels in milk. Respective blood and milk samples were collected from a total of 201 apparently healthy lactating cows above 3 years of age including 52 cows reared in areas supposed to be free from pollution. The highest milk lead (0.85+/-0.11 microg/ml) and cadmium (0.23+/-0.02 microg/ml) levels were recorded in lactating cows reared around lead-zinc smelter and steel manufacturing plant, respectively. Significantly (P<0.05) higher concentration of milk copper, cobalt, zinc and iron compared to control animals was recorded in cows around closed lead cum operational zinc smelter. Analysis of correlation between lead and other trace elements in milk from lactating cows with the blood lead level>0.20 microg/ml (n=79) revealed a significant negative correlations between milk iron and milk lead (r=-0.273, P=0.015). However, such trend was not recorded with blood lead level<0.20 microg/ml (n=122). The milk cobalt concentration was significantly correlated (r=0.365, P<0.001) with cadmium level in milk and the highest milk cadmium (>0.10 to 0.39 microg/ml) group had significantly (P<0.05) increased milk cobalt. It is concluded that increased blood and milk lead or cadmium level as a result of natural exposure of lactating cows to these environmental toxicants significantly influences trace minerals composition of milk and such alterations affect the milk quality and nutritional values.

Trace mineral profile in blood and hair from cattle environmentally exposed to lead and cadmium around different industrial units.

The present investigation was carried out to assess the trace mineral profile in blood and hair from cows environmentally exposed to lead and cadmium and to examine if these toxic heavy metals in blood and hair could affect blood copper, cobalt, zinc and iron concentrations and their accumulation in hair. Respective blood and tail hair samples were collected from adult cows above 3 years, reared in different industrial localities. Samples were also collected from urban areas with small industrial units (n = 55) and areas supposed to be free from pollution. The concomitant exposure of animals to both the heavy metal pollutants was not recorded in either of the industrial or urban locality. Blood lead was significantly (P < 0.01) correlated with blood copper (r = -0.339), cobalt (r = -0.224) and iron (r = -0.497). The increasing blood lead concentrations, irrespective of area of collection of samples, was associated with declining blood copper and iron, and cows with blood lead level above 0.60 mug/ml had significantly (P < 0.05) lower blood copper and iron. The higher blood lead but not cadmium significantly influenced the accumulation of lead (r = 0.323, P < 0.01) and cadmium (r = 0.204, P < 0.01) in hair possibly leading to significantly (P < 0.05) higher accumulation of both lead and cadmium in hair from cattle around lead-zinc smelters and closed lead-cum-operational zinc smelter, where blood cadmium level was comparable with that from unpolluted area. Concentration of zinc (r = 0.237, P < 0.01) and iron (r = 0.183, P < 0.01) but not copper and cobalt in tail hair was significantly influenced by their respective blood concentration. Both the hair lead and cadmium had a significant (P < 0.01) positive correlation with hair copper (r = 0.234, 0.294), zinc (r = 0.489, 0.775), and iron (r = 0.385, 0.643) concentrations. Thus, it is concluded from the present study that the higher blood lead concentrations in cattle irrespective of locality/industrial operations areas affected trace elements profile in blood and hair.

Cytotoxic mechanisms of Zn2+ and Cd2+ involve Na+/H+ exchanger (NHE) activation by ROS.

The signaling mechanism induced by cadmium (Cd) and zinc (Zn) in gill cells of Mytilus galloprovincialis was investigated. Both metals cause an increase in *O2- production, with Cd to be more potent (216 +/- 15%) than Zn (150 +/- 9.5%), in relation to control value (100%). The metals effect was reversed after incubation with the amiloride analogue, EIPA, a selective Na+/H+ exchanger (NHE) inhibitor as well as in the presence of calphostin C, a protein kinase C (PKC) inhibitor. The heavy metals effect on *O2- production was mediated via the interaction of metal ions with alpha1- and beta-adrenergic receptors, as shown after incubation with their respective agonists and antagonists. In addition, both metals caused an increase in intracellular pH (pHi) of gill cells. EIPA together with either metal significantly reduced the effect of each metal treatment on pHi. Incubation of gill cells with the oxidants rotenone, antimycin A and pyruvate caused a significant increase in pHi (delta pHi 0.830, 0.272 and 0.610, respectively), while in the presence of the anti-oxidant N-acetyl cysteine (NAC) a decrease in pHi (delta pHi -0.090) was measured, indicating that change in reactive oxygen species (ROS) production by heavy metals affects NHE activity. When rosiglitazone was incubated together with either heavy metal a decrease in O2- production was observed. Our results show a key role of NHE in the signal transduction pathway induced by Zn and Cd in gill cells, with the involvement of ROS, PKC, adrenergic and PPAR-gamma receptors. In addition, differences between the two metals concerning NHE activation, O2- production and interaction with adrenergic receptors were observed.

Histopathological alterations in the edible snail, Babylonia areolata (spotted babylon), in acute and subchronic cadmium poisoning.

Histopathological alterations in 6- to 8-month-old juvenile spotted babylon, Babylonia areolata, from acute and subchronic cadmium exposure were studied by light microscopy. The 96-h LC(50) value of cadmium for B. areolata was found to be 3.35 mg/L, and the maximum acceptable toxicant concentration (MATC) was 1.6 mg/L. Snails were exposed to 3.35 and 0.08 mg/L (5% of MATC) of cadmium for 96 h and 90 days, respectively. After exposure the gill, the organs of the digestive system (proboscis, esophagus, stomach, digestive gland, and rectum), and the foot were analyzed for cadmium accumulation. The results showed that most digestive organs had a high affinity for cadmium. The main target organ was the stomach, which could accumulate on average 1192.18 microg/g dry weight of cadmium. Cadmium was shown to accumulate to a lesser extent in the digestive gland, gill, rectum, esophagus, proboscis, and foot. Histopathological alterations were observed in the gill and digestive organs (proboscis, esophagus, stomach, and rectum). The study showed that the stomach and gill were the primary target organs of both acute and subchronic exposure. Gill alterations included increased size of mucous vacuoles, reduced length of cilia, dilation and pyknosis of nuclei, thickening of basal lamina, and accumulation of hemocytes. The epithelial lining of the digestive tract showed similar alterations such as increased size of mucous vacuoles, reduced length of cilia, and dilation of nuclei. In addition, fragmentation of the muscle sheath was observed.

Association of metals (mercury, cadmium and zinc) with metallothionein-like proteins in storage organs of stranded dolphins from the Mediterranean sea (Southern Italy).

Selected toxic and essential metals (mercury, Hg; cadmium, Cd; and zinc, Zn) were determined in the liver and in the kidney collected from 13 dolphins (Stenella coeruleoalba and Tursiops truncatus) stranded along the southern coasts of Italy from 1991 to 1999. Liver samples were also analysed for their methyl mercury (MeHg) and selenium (Se) contents. For subcellular fractionation, liver and kidney samples were homogenized in Tris-HCl buffer; after centrifugation, the supernatant (cytosol) was separated from pellets (insoluble fraction), heated at 80 degrees C for 10 min and centrifuged in order to separate the precipitate containing the thermolabile high molecular weight proteins. The cytosol heat-stable fraction, including metallothionein-like proteins (MTLPs), was then purified by gel filtration chromatography on Sephadex G-75 column. The three subcellular fractions collected (insoluble fraction, thermolabile fraction and purified heat-stable fraction) were analysed for their Hg, Cd and Zn contents. The analytical results confirm previous similar studies on toothed whales in showing that: (1) in the liver, as well as in the kidney, Hg was mainly found in the insoluble fraction, therefore, metallothioneins seem to have no role in the Hg detoxification; (2) in the liver, the molar ratio between Se and inorganic Hg was very close to 1; this suggests that the final compound of MeHg detoxification could be HgSe (tiemannite); (3) in almost all the samples, Cd and Zn were detected in the purified heat-stable fraction (including MTLPs). The mechanism of Cd detoxification and Zn homeostasis is also discussed.

Quantifying copper and cadmium impacts on intrinsic rate of population increase in the terrestrial oligochaete Lumbricus rubellus.

Demographic methods can translate toxicant effects on individuals into consequences for populations. To date few such studies have been conducted with longer-lived invertebrates. This is because full life-cycle experiments are difficult with such species. Here we report the effects of copper and cadmium on the key demographic parameter intrinsic rate of population increase (r) in a long-lived invertebrate (an earthworm). The approach used to derive r was based on robust measurement of effects on life-cycle traits in three specifically designed toxicity tests and integration of this data within a demographic model. The three laboratory tests used to generate values for specific life-cycle parameters under copper and cadmium exposure were suited to the task. Significant effects on a range of separate adult and juvenile life-cycle parameters were seen. Integration of parameter values within the demographic equation indicated that for copper, r was reduced only at a concentration that also caused adult mortality. For cadmium, a more graded exposure-dependent effect on fitness was seen, with r reduced at sublethal concentrations. The concentration response patterns for r found for the two metals suggest significantly different consequences for earthworm populations exposed to sublethal levels of copper and cadmium.

Impairment of mysid (Neomysis integer) swimming ability: an environmentally realistic assessment of the impact of cadmium exposure.

Neomysis integer (Crustacea: Mysidacea), a euryhaline member of the hyperbenthos of the upper reaches of European estuaries, has been identified as a suitable animal for assessing the impacts of chemical pollutants on these estuarine regions. In this study, the effect of a 7 day pre-exposure to environmentally relevant concentrations of cadmium (0.5 and 1.0 microg l(-1)) on the swimming behaviour of N. integer was examined using an annular flume. Cadmium speciation at two salinities (1 and 10 per thousand) that dominate these upper estuarine regions was modelled to ensure mysids were exposed to the same concentration of the toxic free-ion at each salinity. There was no significant difference in the swimming behaviour of mysids exposed to the same free-ion cadmium concentration at the two different salinities. At each salinity, exposure to 0.5 microg Cd2+ (aq) l(-1)resulted in fewer mysids moving forward into the current (normal behaviour) at free stream velocities typical of their natural habitat (e.g. 3-9 cm s(-1)) than non cadmium-exposed mysids. At these low current speeds, cadmium-exposed mysids were either able to maintain position or were swept by the current. The same general responses were recorded for mysids exposed to 1.0 microg Cd2+ (aq) l(-1)except that more mysids showed disrupted swimming ability compared with 0.5 microg Cd2+ (aq) l(-1). At higher current speeds (>12 cm s(-1)), current velocity was the dominant factor affecting mysid swimming behaviour and there was no effect of cadmium on mysids maintaining position. Exposure to cadmium also caused significant disruption of the hyperbenthic behaviour of N. integer and more cadmium-exposed individuals were in the water column than control mysids; this result was more variable at 10 per thousand than 1 per thousand. Results indicate that exposure to cadmium concentrations of 0.5 microg Cd2+(aq) l(-1)would result in displacement of N. integer from its optimum region within the estuarine environment. This conclusion would not be achieved from standard LC(50) tests (e.g. 7 day LC50 = 2.95 microg Cd2+ (aq) l(-1)), highlighting the value of behavioural disruption as a sensitive indicator of environmental chemical contamination.

Dynamics of (Cd,Zn)-metallothioneins in gills, liver and kidney of common carp Cyprinus carpio during cadmium exposure.

Cadmium concentrations, (Cd,Zn)-metallothionein (MT) concentrations, MT synthesis and the relative amounts of cadmium bound to (Cd,Zn)-MTs were determined in gills, liver and kidney of common carp Cyprinus carpio exposed to 0, 0.5 microM (0.06 mg.l(-1)), 2.5 microM (0.28 mg.l(-1)) and 7 microM (0.79 mg.l(-1)) Cd for up to 29 days. Cadmium accumulation was in the order kidney > liver > gills. Control levels of hepatic (Cd,Zn)-MT were four times higher compared to those of gills and kidney. No increases in (Cd,Zn)-MT concentrations were observed in liver during the exposure period. In comparison with control carp, (Cd,Zn)-MT concentrations increased up to 4.5 times in kidney and two times in gills. In both these organs, (Cd,Zn)-MT concentrations were linearly related with cadmium tissue levels and with the de novo synthesis of MTs. Hepatic cadmium was almost completely bound to (Cd,Zn)-MT, while percentages of non-MT-bound cadmium were at least 40% in gills and 25% in kidney. This corresponded with a total saturation of (Cd,Zn)-MT by cadmium in kidney and a saturation of approximately 50 and 60% in gills and liver, respectively. The final order of non-MT-bound cadmium was kidney > gills > liver. Our results indicate that cadmium exposure causes toxic effects, which cannot be correlated with the accumulated levels of the metal in tissues. Although cadmium clearly leads to the de novo synthesis of MT and higher (Cd,Zn)-MT concentrations, the role of this protein in the detoxification process is clearly organ-specific and its synthesis does not keep track with cadmium accumulation.

Cadmium induces apoptosis and genotoxicity in rainbow trout hepatocytes through generation of reactive oxygene species.

Cadmium poses a serious environmental threat in aquatic ecosystems but the mechanisms of its toxicity remain unclear. The purpose of this work was first to determine whether cadmium induced apoptosis in trout hepatocytes, second to determine whether or not reactive oxygen species (ROS) were involved in cadmium-induced apoptosis and genotoxicity. Hepatocytes exposed to increasing cadmium concentrations (in the range of 1-10 microM) showed a molecular hallmark of apoptosis which is the fragmentation of the nuclear DNA into oligonucleosomal-length fragments, resulting from an activation of endogenous endonucleases and recognized as a 'DNA ladder' on conventional agarose gel electrophoresis. Exposure of hepatocytes to cadmium led clearly to the DEVD-dependent protease activation, acting upstream from the endonucleases and considered as central mediators of apoptosis. DNA strand breaks in cadmium-treated trout hepatocytes was assessed using the comet assay, a rapid and sensitive single-cell gel electrophoresis technique used to detect DNA primary damage in individual cells. Simultaneous treatment of trout hepatocytes with cadmium and the nitroxide radical TEMPO used as a ROS scavenger, reduced significantly DNA fragmentation, DEVD-related protease activity and DNA strand breaks formation. These results lead to a working hypothesis that cadmium-induced apoptosis and DNA strand breaks in trout hepatocytes are partially triggered by the generation of ROS. Additional studies are required for proposing a mechanistic model of cadmium-induced apoptosis and genotoxicity in trout liver cells, in underlying the balance between DNA damage and cellular defence systems in fish.

The use of microangiography in detecting aberrant vasculature in zebrafish embryos exposed to cadmium.

Embryonic vascular patterns in zebrafish (Danio rerio) could be visualised by confocal microscopy coupled with microinjected fluorescent microbeads. This microangiographic technique was adopted here, for the first time, to study the effects of cadmium on cardiovascular development in zebrafish embryos. Zebrafish embryos were incubated in culture medium containing 100 microM cadmium from 5 h post fertilisation (hpf) to 48 hpf. At 48 hpf, embryos were examined for viability and occurrence of malformations. The 100 microM cadmium caused 32.21 +/- 3.65% mortality and 20.33 +/- 4.04% visible malformations in surviving embryos. In the remaining embryos with no visible signs of malformations, further assessments for less obvious abnormalities were performed. Assessments on craniofacial development were made by digital measurements on areas of brains and eyes. Cardiac development was assessed by immunostaining the heart with the antibody MF20 specific for myosin heavy chain. Body lengths of the embryos were also measured. Embryonic development of brains, eyes, hearts and body lengths of visibly healthy embryos in the cadmium treatment group showed no significant difference from the controls. Embryonic vasculature of these visibly healthy embryos was then studied by microinjecting fluorescent microbeads of diameter 0.02 microm into the circulation. All the cadmium treated embryos showed localised vascular defects in the dorsal aortae, segmental and cranial vessels while none of the control embryos showed any aberrant patterns in the networking of the vasculature. Improved image analyses on the anterior regions revealed that cadmium treated embryos had markedly less complex networks of cranial vessels with fewer vessels perfusing the craniofacial regions. The number of branch points in the vascular network was counted. In untreated embryos, there were 135.6 +/- 51 branches in the vasculature in entire body. In the cadmium treated embryos, there were 64.5+/-31 branches. The difference was significant when assessed with Student's t-test. It appeared that although cadmium did not cause any signs of external malformations in these visibly healthy embryos, nonetheless induced impaired branching and anastomsis of the cranial vessels. This study revealed, for the first time, that vital vascular structures in fish embryos could be affected by exposure to cadmium. This technique allowed visualisation of vascular anomalies in embryos showing no external signs of malformations. The impairment of anatomical features during embryonic development might serve as meaningful health endpoints in ecotoxicological studies and in risk assessment.

Morphological and pathological effects of cadmium ingestion on Pekin ducks exposed to saline.

This study examined the effects of simultaneous exposure to saline and cadmium (Cd) on organ mass and histology of a bird with salt glands, the Pekin duck, Anas platyrhynchos. Three mixed-sex groups, each containing 6 birds, ate duck pellets containing 0, 50, or 300 microg Cd/g, respectively, for 4 1/2 mo and drank 300 mM NaCl. Only females on the high-Cd diet lost body mass. Ingestion of Cd reduced heart mass in females. There was increased mass of Harderian and salt glands in both sexes. Mass of kidneys and liver increased only in males, and the gut mass (also length) increased more in males. Cadmium ingestion also induced (1) inflammation of renal interstitium and degenerative tubular changes, (2) marked degenerative changes in testes, (3) increased heart water content, (4) decreased cytoplasmic volume of liver cells, (5) reduced proportion of basophilic granular cells in chromaffin tissue of the adrenal glands, and (6) in the ileum, increased heterophilia in the lamina propria and, only in females, the apoptosis to mitosis ratio in crypt cells of the epithelium. The ducks' outward appearance gave no indication that ingesting large amounts of cadmium for 4 1/2 mo produced deleterious effects, but the physiological consequences were profound. Both sexes had greatly reduced gonadal mass and the males produced no sperm. The higher dietary level greatly hypertrophied the liver, kidneys, and gut only in males. The cadmium-induced changes in organs, particularly in the gonads, kidneys, and adrenal glands, should greatly impair the health and reproductive capacity of these ducks.

A probabilistic model for deriving soil quality criteria based on secondary poisoning of top predators. II. Calculations for dichlorodiphenyltrichloroethane (DDT) and cadmium.

A simplified food web with three trophic levels is designed: plants and invertebrates at the first, small birds and mammals at the second, and birds and beasts of prey at the third trophic level. Exposure of top predators via separate food chains is analyzed. However, most top predator species are exposed via more than one food chain (food web). Therefore, a species-specific approach is followed too, for which four bird of prey species and two beast of prey species with different food choices are selected: sparrow hawk, kestrel, barn owl, little owl, badger, and weasel. The most critical food chains for secondary poisoning of top predators are soil --> worm/insect --> bird --> bird of prey for dichlorodiphenyltrichloroethane (DDT), and soil --> worm --> bird/mammal --> bird of prey for cadmium (Cd). The risk for the selected top predator species is much lower than the risk based on these critical food chains because the critical food chains constitute a minor part of their food webs. Species feeding on birds (sparrow hawk) and small carnivorous mammals (barn owl) are exposed to DDT and Cd to a much higher extent than species mainly feeding on small herbivorous mammals (kestrel and weasel). It is recommended to include exposure via the pathways soil --> worm/insect --> bird/mammal --> top predator in procedures for derivation of environmental quality objectives for persistent and highly lipophilic compounds.

The effect of concurrent lead and cadmium exposure on the cell-mediated immune response in goats.

Cell-mediated immune response was monitored by cutaneous hypersensitivity reaction (CHR) to 2-4 dinitrochlorobenzene in goats given lead or cadmium alone or in combination. Twenty goats were divided into 4 groups of 5 animals each. Group A served as control whereas Groups B, C and D were given po doses of 50 mg lead acetate/kg body weight, 10 mg cadmium chloride/kg body weight or 50 mg lead acetate/kg body weight + 10 mg cadmium chloride/kg body weight, respectively, for 42 d. Primary sensitization was done on day 27 followed by a challenge dose after 14 d. Elicitation of CHR, as measured by average increase in skin thickness, was suppressed significantly in goats of all dosed groups. Suppression was more in the cadmium-dosed than in the lead or lead + cadmium dosed goats. Histopathology demonstrated reduced intensity of cellular reactions in the cadmium and lead + cadmium-dosed animals.

Complex study of the physiological role of cadmium. III. Cadmium loading trials on broiler chickens.

Cadmium (Cd) loading trials were conducted on a total of 110 (3 x 10 and 4 x 20) broiler chickens prereared for 21 days. The control chickens received no cadmium, while chickens in the six treatment groups were given different doses of Cd as an aqueous solution of CdSO4 administered either into the crop or mixed in the feed. The chickens were kept in a climatized animal house and treated usually for 3-5 weeks (maximum 68 days), with the exception of group Cd-75 chickens which were treated up to 239 days of age. The chickens' health status, body mass and feed consumption were monitored throughout the trial. On days 14-20 and on day 42 of the trial 2 chickens per group, then at the end of trial a total of 25 chickens were killed in anaesthesia. These birds, together with chickens that died or were killed during the trial, were subjected to detailed gross pathological examination. From 11 organs (kidney, liver, spleen, testicle, brain, myocardium, skeletal muscle, lungs, digestive tract, pancreas, tubular bones) of these chickens samples were taken for assay for a total of 16 elements, as well as for light and electron microscopic examination. With the exception of groups Cd-30 and Cd-600, no abnormal clinical signs were observed in the first two weeks of the trial. Chickens of group Cd-30 died before day 8-12 of the trial among signs of complete anorexia, rapid emaciation, huddling and diarrhoea, while chickens of group Cd-600 died before day 28, showing similar clinical signs. The body mass of chickens fed a Cd-supplemented diet either remained constant or decreased substantially, in a degree proportional to the Cd load. The only exception was group Cd-2.5, in which the average body mass of birds at the end of week 8 slightly exceeded that of the controls. Four out of the 10 cockerel chicks fed a diet containing 75 ppm Cd up to 239 days of age died of intercurrent diseases; the remaining six grew well and reached a body mass of 3.8-4.3 kg. Feed conversion efficiency was satisfactory in the control group and in group Cd-2.5 (2.1 and 2.4 kg, respectively) and could not be evaluated in a realistic manner in the other groups. At necropsy, the cockerel chicks of groups Cd-30 and Cd-600 showed severe emaciation, liver and kidney degeneration, myocardial hypertrophy and cardiac dilatation.(ABSTRACT TRUNCATED AT 400 WORDS)