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.

Tissue distribution of metals in white-fronted geese and spot-billed ducks from Korea.

This study presents concentrations of Fe, Zn, Mn, Cu, Pb and Cd in livers, kidneys, muscles and bones of white-fronted geese Anser albifrons (geese) and spot-billed ducks Anas poecilorhyncha (ducks). Iron in livers, kidneys and muscles, Zn in muscles, Mn and Cd in every tissue, Cu in livers, muscles and bones and Pb in bones differed between species, and there were significant differences among tissues in both species. Essential elements such as Fe, Zn, Mn and Cu concentrations were within the background levels. Lead concentrations in livers of 7 of 14 geese and 7 of 19 ducks and in bones of 4 of 19 ducks exceeded background concentrations for waterfowl (5 µg/g dw for the liver, 10 µg/g dw for the bone). Almost all samples of both species had the background Cd concentrations in the liver (33 of 33 geese and ducks) and kidney (14 geese and 18 ducks). Tissue concentrations of Cd were greater in geese than ducks. In contrast, tissue concentrations of Pb in bones were greater in ducks than in geese. These different trends for Cd and Pb reflect a short and/or long term difference in exposure and degree of accumulation of these metals.

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.

Evaluation of metallothionein as a biomarker of single and combined Cd/Cu exposure in Dreissena polymorpha.

The effects of metal mixture (Cd+Cu) versus single-metal exposure on total MT response and bioaccumulation were investigated in the freshwater bivalve Dreissena polymorpha. A two-month exposure period, including two levels of contamination, was chosen for each of the two metals: 5, 10 microg/L for Cu, and 2, 20 microg/L for Cd, with mixtures of, respectively, 5 microg/L Cu+2 microg/L Cd, 5 microg/L Cu+20 microg/L Cd, 10 microg/L Cu+2 microg/L Cd, and 10 microg/L Cu+20 microg/L Cd. Total MT contents were assessed by an Ag-saturation method, and metals contents were determined by atomic absorption spectrometry. Results at the whole-organism level showed a significant and early increase of total MT biosynthesis after exposure to Cd. This increase was significantly correlated with Cd bioaccumulation. By contrast, Cu did not modify total MT response, and mussels limited Cu bioaccumulation. The mixture either did not influence or only weakly influenced metal accumulation and MT response to Cu and Cd after long-term exposure. Our results suggest that the form of MT existing in D. polymorpha was not Cu-inducible. This could limit the use of MT in D. polymorpha as a biomarker of heavy metal pollution in freshwater ecosystems.

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.

Pluma de Aguila Calzada (Hieraaetus pennatus) como unidad biomonitora de la exposición de la ambiental a cadmio y plomo / Feather of Booted Eagle (Hieraaetus pennatus) as biomonitor of environmental exposure to cadmium and lead

La determinación de plomo y cadmio en sangre y tejidos de aves rapaces permite obtener información útil sobre el estado de salud de los individuos, de la especie y del medio ambiente en el que viven. Además su medición sistemática en programas de toxicovigilancia facilita la predicción de riesgos y la toma de decisiones de gestión medioambiental. El presente trabajo tiene como objetivo valorar la utilidad de la pluma de aves rapaces como unidad de monitorización de la exposición a metales pesados (cadmio y plomo) en un ambiente considerado a priori como mínimamente contaminado de la Región de Murcia (Sierras de Lavia y Burete, NO de Murcia). Para ello se han tomado muestras de sangre de pollos y plumas de adultos de águila calzada (Hieraaetus pennatus) durante la época de cría de los años 1999 y 2000. Previo al análisis de metales en plumas fue preciso optimizar una técnica de lavado de las muestras capaz de eliminar los metales depositados en su superficie. Todas las muestras de sangre, excepto dos, tenían niveles de cadmio y plomo inferiores a 0,5 y 5 microg/dl, respectivamente, lo que es compatible con situaciones de contaminación ambiental de fondo. Las concentraciones en plumas fueron muy inferiores a las indicativas de efecto, con medias de 16,5 y 750 microg/kg para cadmio y plomo, respectivamente. Las correlaciones entre las concentraciones de ambos metales en sangre y plumas fueron significativas al nivel p< 0,05 para el cadmio y de p <0,01 para el plomo. En conclusión opinamos que la determinación de cadmio y plomo en pluma de águila calzada es útil en los estudios de biomonitorización y toxicovigilancia. (AU)

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.

Metal toxicosis in horses.

The ubiquity and stability of metals in the environment make them unique as a pollutant or an essential dietary component. Metals are neither created nor destroyed by chemical processes but are redistributed in the environment. In combination with other elements, metal compounds and alloys are essential materials of the contemporary world. Inappropriate use or distribution in the environment leads to adverse health effects on all biologic systems, including horses. Gastrointestinal upset is a common feature of acute toxicosis with metals in general. Among the metals discussed, arsenic and inorganic mercury have a propensity to do severe damage to the gut. Deposition of cadmium on forage is the source most likely to intoxicate horses. This subchronic to chronic problem in horses is manifest as disease of the musculoskeletal system and kidneys. Iron-containing hematinics are widely used in racetrack horses and occasionally result in hepatopathy when excessive doses are administered. Lead continues to be recognized as the most significant environmental metal pollutant. Poisoning is encountered routinely in humans and animals. Of the animal species of veterinary concern, lead-poisoned horses are not a frequent encounter. Lead-intoxicated horses show signs of peripheral neuropathy (laryngeal hemiplegia), intermittent colic, and mild anemia. Acute mercury poisoning sometimes occurs from the common use of mercury-containing blistering agents, with most clinical findings related to acute renal failure. Chronic excessive intake of zinc by horses is uncommon but devastating in rapidly growing foals. The mechanism of chronic zinc toxicosis is coupled to the induced copper deficiency. The condition is a disease of cartilage in the articular and growth physes.

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.