Characterization of the effect of Cr(VI) on humoral innate immunity using Drosophila melanogaster.

With the advancement of human race, different anthropogenic activities have heaped the environment with chemicals that can cause alteration in the immune system of exposed organism. As a first line of barrier, the evolutionary conserved innate immunity is crucial for the health of an organism. However, there is paucity of information regarding in vivo assessment of the effect of environmental chemicals on innate immunity. Therefore, we examined the effect of a widely used environmental chemical, Cr(VI), on humoral innate immune response using Drosophila melanogaster. The adverse effect of Cr(VI) on host humoral response was characterized by decreased gene expression of antimicrobial peptides (AMPs) in the exposed organism. Concurrently, a significantly decreased transcription of humoral pathway receptors (Toll and PGRP) and triglyceride level along with inhibition of antioxidant enzyme activities were observed in exposed organism. This in turn weakened the immune response of exposed organism that was manifested by their reduced resistance against bacterial infection. In addition, overexpression of the components of humoral immunity particularly Diptericin benefits Drosophila from Cr(VI)-induced humoral immune-suppressive effect. To our knowledge, this is the first report regarding negative impact of an environmental chemical on humoral innate immune response of Drosophila along with subsequent protection by AMPs, which may provide novel insight into host-chemical interactions. Also, our data validate the utility and sensitivity of Drosophila as a model that could be used for screening the possible risk of environmental chemicals on innate immunity with minimum ethical concern that can be further extrapolated to higher organisms.

Comparative toxicity of carbaryl, carbofuran, cypermethrin and fenvalerate in Metaphire posthuma and Eisenia fetida -a possible mechanism.

To establish the use of Metaphire posthuma as a sensitive test model for ecotoxicological studies, acute toxicity testing of carbaryl, carbofuran, cypermethrin and fenvalerate on Eisenia fetida and Metaphire posthuma were carried out. Two different types of bioassays, contact filter paper toxicity and soil toxicity bioassays were used to determine LC50 values for these insecticides. Among the tested chemicals, carbofuran was the most toxic to both the earthworm species. In paper contact method, 72 h-LC50 values of carbofuran in M. posthuma and E. fetida were found to be 0.08 µg/cm(2) and 1.55 µg/cm(2) respectively while in soil test, 14-d LC50 values were 0.49 mg/kg and 21.15 mg/kg respectively. On comparing the toxicity data of these chemicals for both the earthworm species, M. posthuma was found to be more sensitive than E. fetida. Based on the acute toxicity data, the order of toxicity of insecticides in both the test procedures was carbofuran>cypermethrin>carbaryl>fenvalerate for M. posthuma whereas for E. fetida it was carbofuran>carbaryl>fenvalerate>cypermethrin. Morphological changes also appeared in the organisms exposed to these chemicals which were more pronounced in M. posthuma at lower concentrations than E. fetida in both the test procedures. The results of the present study advocates the use of M. posthuma for ecotoxicity studies, being a more sensitive and reliable model than E. fetida. Based on the data on partial atomic charges, structural features and spectroscopic studies on carbaryl and carbofuran, a possible mechanism of toxicity of carbamate insecticides in earthworm was proposed.

Determination of t,t-muconic acid in urine samples using a molecular imprinted polymer combined with simultaneous ethyl chloroformate derivatization and pre-concentration by dispersive liquid-liquid microextraction.

The present communication describes the preparation and evaluation of a molecularly imprinted polymer (MIP) as a solid-phase extraction (SPE) sorbent and simultaneous ethyl chloroformate (ECF) derivatization and pre-concentration by dispersive liquid-liquid microextraction (DLLME) for the analysis of t,t-muconic acid (t,t-MA) in urine samples using gas chromatography-mass spectrometry. The imprinting polymer was prepared using methacrylic acid as a functional monomer, ethylene glycol dimethacrylate as a cross-linker, 2,2-azobisisobutyronitrile as the initiator and t,t-MA as a template molecule. The imprinted polymer was evaluated for its use as a SPE sorbent by comparing both imprinted and non-imprinted polymers in terms of the recovery of t,t-MA from urine samples. Molecular modelling studies were performed in order to estimate the binding energy and efficiency of the MIP complex formed between the monomer and the t,t-MA. Various factors that can affect the extraction efficiency of MIP, such as the loading, washing and eluting conditions, were optimized; other factors that can affect the derivatization and DLLME pre-concentration were also optimized. MIP in combination with ECF derivatization and DLLME pre-concentration for t,t-MA exhibits good linearity, ranging from 0.125 to 2 µg mL(-1) (R(2) = 0.9971), with limit of detection of 0.037 µg mL(-1) and limit of quantification of 0.109 µg mL(-1). Intra- and inter-day precision was found to be <6%. The proposed method has been proven to be effective and sensitive for the selective pre-concentration and determination of t,t-MA in urine samples of cigarette smokers.

Transcriptomic analysis provides insights on hexavalent chromium induced DNA double strand breaks and their possible repair in midgut cells of Drosophila melanogaster larvae.

Hexavalent chromium [Cr(VI)] is a well known mutagen and carcinogen. Since genomic instability due to generation of double strand breaks (DSBs) is causally linked to carcinogenesis, we tested a hypothesis that Cr(VI) causes in vivo generation of DSBs and elicits DNA damage response. We fed repair proficient Drosophila melanogaster (Oregon R(+)) larvae Cr(VI) (20.0µg/ml) mixed food for 24 and 48h and observed a significant (p<0.05) induction of DSBs in their midgut cells after 48h using neutral Comet assay. Global gene expression profiling in Cr(VI)-exposed Oregon R(+) larvae unveiled mis-regulation of DSBs responsive repair genes both after 24 and 48h. In vivo generation of DSBs in exposed Drosophila was confirmed by an increased pH2Av immunostaining along with the activation of cell cycle regulation genes. Analysis of mis-regulated genes grouped under DSB response by GOEAST indicated the participation of non-homologous end joining (NHEJ) DSB repair pathway. We selected two strains, one mutant (ligIV) and another ku80-RNAi (knockdown of ku80), whose functions are essentially linked to NHEJ-DSB repair pathway. As a proof of principle, we compared the DSBs generation in larvae of these two strains with that of repair proficient Oregon R(+). Along with this, DSBs generation in spn-A and okr [essential genes in homologous recombination repair (HR) pathway] mutants was also tested for the possible involvement of HR-DSB repair. A significantly increased DSBs generation in the exposed ku80-RNAi and ligIV (mutant) larvae because of impaired repair, concomitant with an insignificant DSBs generation in okr and spn-A mutant larvae indicates an active participation of NHEJ repair pathway. The study, first of its kind to our knowledge, while providing evidences for in vivo generation of DSBs in Cr(VI) exposed Drosophila larvae, assumes significance for its relevance to higher organisms due to causal link between DSB generation and Cr(VI)-induced carcinogenesis.

Temporal distribution of fine particulates (PM2.5:PM10), potentially toxic metals, PAHs and Metal-bound carcinogenic risk in the population of Lucknow City, India.

Ubiquitous fine particulates can readily be bound to toxic metals and polycyclic aromatic hydrocarbons and are considered to be a great threat to human health. The purpose of this study was to assess the magnitude of air pollution risks to public health by determining four crucial parameters- inhalable particulates, metals in particulates and PAHs which are associated with PM10 in the air environment of Lucknow, India during 2007-09. The values of PM10 and PM2.5 ranged between 102.3-240.5 and 28.0-196.9 µg/m³ whilst the average PM10 was 1.7 times and PM was 1.5 times higher than their respective NAAQS of 100 and 60 µg/m³ respectively. The estimated relative death rate and hospital admissions for each increase in the PM10 levels of 10 µg/m³ ranged from 1.5-8% and from 3.9-8.0% (as per APHEA2 1990) respectively in persons > 65 yrs. Among the locations; AQ, AQ and AQ (with diversified activities and heavy traffic) recorded higher concentrations of both the particulate fractions than the AQ (residential area with low traffic). The average concentrations of Fe, Pb, Ni, Cu, Cr, Cd in PM10 were 219.4, 40.6, 35.1, 27.3, 22.2 and 16.2 ng/m³ and that in PM2.5 were 54.3, 33.9, 38.5, 29.4, 8.4, and 1.17 ng/m³ respectively Regression analysis revealed that correlation of metals with PM2.5 was stronger than PM. The ratio of metals adsorbed on surface of particles (PM2.5:PM10) reveals that PM2.5 has more affinity for Ni, Cu and Pb and PM10 for Cd, Fe and Cr. Health risk due to carcinogenic metals bound to respirable particulates was predicted by estimating excess cancer risk (ECR). The highest ECR value was estimated for Cr, 266.70 × 10⁻6, which was associated with PM10 and 100.92 × 10⁻6 which was associated with PM2.5, whereas lead has the lowest ECR value. Amongst PAHs, benzo(a)pyrene (51.96 ± 19.71 ng/m) was maximum in PM10 samples. Maximum concentrations of PM10, PM2.5, metals and PAHs were detected during winter, and the lowest was during monsoon. The higher prevalence of diseases among the population may be due to high concentration of particulates coated with toxic metals and PAHs present in air environment.

Studies on urban drinking water quality in a tropical zone.

Anthropogenic activities associated with industrialization, agriculture and urbanization have led to the deterioration in water quality due to various contaminants. To assess the status of urban drinking water quality, samples were collected from the piped supplies as well as groundwater sources from different localities of residential, commercial and industrial areas of Lucknow City in a tropical zone of India during pre-monsoon for estimation of coliform and faecal coliform bacteria, organochlorine pesticides (OCPs) and heavy metals. Bacterial contamination was found to be more in the samples from commercial areas than residential and industrial areas. OCPs like α,γ-hexachlorocyclohexane and 1,1 p,p-DDE {dichloro-2, 2-bis(p-chlorophenyl) ethene)} were found to be present in most of the samples from study area. The total organochlorine pesticide levels were found to be within the European Union limit (0.5 µg/L) in most of the samples. Most of the heavy metals estimated in the samples were also found to be within the permissible limits as prescribed by World Health Organization for drinking water. Thus, these observations show that contamination of drinking water in urban areas may be mainly due to municipal, industrial and agricultural activities along with improper disposal of solid waste. This is an alarm to safety of public health and aquatic environment in tropics.

Application of ethyl chloroformate derivatization for solid-phase microextraction-gas chromatography-mass spectrometric determination of bisphenol-A in water and milk samples.

A simple and rapid analytical method based on in-matrix ethyl chloroformate (ECF) derivatization has been developed for the quantitative determination of bisphenol-A (BPA) in milk and water samples. The samples containing BPA were derivatised with ECF in the presence of pyridine for 20 s at room temperature, and the non-polar derivative thus formed was extracted using polydimethylsiloxane solid-phase microextraction (SPME) fibres with thicknesses of 100 µm followed by analysis using gas chromatography-mass spectrometry. Three alkyl chloroformates (methyl, ethyl and isobutyl chloroformate) were tested for optimum derivatisation yields, and ECF has been found to be optimum for the derivatisation of BPA. Several parameters such as amount of ECF, pyridine and reaction time as well as SPME parameters were studied and optimised in the present work. The limit of detection for BPA in milk and water samples was found to be 0.1 and 0.01 µg L(-1), respectively, with a signal-to-noise ratio of 3:1. The limit of quantitation for BPA in milk and water was found to be 0.38 and 0.052 µg L(-1), respectively, with a signal-to-noise ratio of 10:1. In conclusion, the method developed was found to be rapid, reliable and cost-effective in comparison to silylation and highly suitable for the routine analysis of BPA by various food and environmental laboratories.

Multiple approaches to evaluate the toxicity of the biomass fuel cow dung (kanda) smoke.

Cow dung (Kanda) is a major source of energy in rural and urban population of developing countries and is burnt in traditional open stoves in confined space of kitchen without proper ventilation. In epidemiological studies, biomass fuel smoke has been reported to be responsible for several respiratory disorders in exposed population. In a laboratory experiment, female wistar rats were exposed to kanda smoke for 60 min/day over a period of 12 weeks. Chemical analysis of smoke showed the presence of PAHs. The increase in CYP1A1, GST-ya, GST-yc expression was found in 12 week exposed lung tissues as compared with controls. The exposure to smoke resulted in significant alteration in the BALF cells in the form of clustering of alveolar macrophages and giant cell formation with vacuolated cytoplasm. The macrophages also showed thickness and villi like projections on the cell surface thus reducing their phagocytic activities. Histopathological changes in lung tissue were manifested in the form of damage to bronchiolar epithelium, edema and thickening of alveolar septa and emphysema after 4 and 8 week of exposure. These findings suggest that exposure to kanda smoke increases pulmonary tissue damage and may result in various forms of respiratory infections in the exposed popultion.

Assessment of urban air pollution and it's probable health impact.

The present study deals with the quantitative effect of vehicular emission on ambient air quality during May 2006 in urban area of Lucknow city. In this study SPM, RSPM, SO2, NOx and 7 trace metals associated with RSPM were estimated at 10 representative locations in urban area and one village area for control. Beside this, air quality index (AQI), health effects of different metals and mortality were assessed. The 24 hr average concentration of SPM, RSPM, SO2 and NOx was found to be 382.3, 171.5, 24.3 and 33.8 microg m(-3) respectively in urban area and these concentrations were found to be significantly (p < 0.01) higher by 94.8, 134.8, 107.4 and 129.6% than control site respectively The 24 hr mean of SPM and RSPM at each location of urban area were found to be higher than prescribed limit of National Ambient Air Quality Standard (NAAQS) except SPM for industrial area. The 24 hr mean concentration of metals associated with RSPM was found to be higher than the control site by 52.3, 271.8, 408.9, 75.81, 62.7, 487.54 and 189.5% for Fe, Cu, Pb, Zn, Ni, Mn and Cr respectively. The inter correlation of metals Pb with Mn, Fe and Cr; Zn with Ni and Cr; Ni with Cr; Mn with Fe and Cu with Cr showed significant positive relation either at p < 0.05 or p < 0.01 level. Metals Pb, Mn and Cr (p < 0.01) and Cu (p < 0.05) showed significant positive correlation with RSPM. These results indicate that ambient air quality in the urban area is affected adversely due to emission and accumulation of SPM, RSPM, SO2, NOx and trace metals. These pollutants may pose detrimental effect on human health, as exposure of these are associated with cardiovascular and respiratory diseases, neurological impairments, increased risk of preterm birth and even mortality and morbidity.

Combined effect of ultraviolet-B radiation and cadmium contamination on nutrient uptake and photosynthetic pigments in Brassica campestris L. seedlings.

Environmental and industrial pollution along with increase in ground level UV-B radiation, because of stratospheric ozone depletion, present multiple stresses, which may affect crop photosynthesis and productivity. The present study was undertaken to see interactive effects of heavy metal contamination (Cd(2+)) and UV-B exposure on essential nutrient (Ca(2+), Mg(2+), K(+)) uptake, biomass, and chlorophyll content in mustard (Brassica campestris L.) seedlings. Plants grown in 0.5, 1.0, 2.5, and 5.0 mg L(-1) Cd(2+) supplemented medium were exposed to UV-B for 30 min (0.4 mW cm(-2)) per day. The interactive effect of two stresses measured after 5 and 10 days showed an overall decline in biomass. Under dual stress (5 mg Cd(2+) L(-1)) significant (P < 0.001) decrease in chlorophyll a (43%), chlorophyll b (23%), and carotenoid (53%) was observed. Ca(2+) uptake was reduced by 51% in roots under high doses of Cd(2+) (5 mg L(-1)) and simultaneous exposure to 0.4 mW cm(-2) UV-B for 10 days. Mg(2+) content was reduced by 48% and K(+) by 62% under similar exposure conditions. Decline in nutrient uptake in Brassica campestris L. seedlings was observed both in root and shoot leaf in the initial growth period under controlled lab conditions. Cadmium ion (Cd(2+)) uptake was significantly enhanced by 33% (P < 0.001) in the presence of UV-B. The findings are significant as multiple stress conditions prevalent in the environment play an important role during the early growth period, a period critical for crop yield.

Measurement of pH and calcium ions release from different calcium hydroxide pastes at different intervals of time: Atomic spectrophotometric analysis.

AIM: To evaluate the effect of different vehicles on pH and release of calcium ions from calcium hydroxide (CH) paste from apical third of root canals. METHODS: 40 single rooted extracted human mandibular premolars were instrumented with RevoS files (MicroMega) up to ISO size 40. The teeth were divided into 4 groups on the basis of vehicle as follows: Group I - calcium hydroxide mixed with 2% chlorhexidine; Group II - calcium hydroxide mixed with propylene glycol; Group III - calcium hydroxide mixed with glycerine; and Control - calcium hydroxide mixed with double distilled water. Each group had two subgroups (n = 5) on the basis of the calcium hydroxide delivery. Subgroup A - calcium hydroxide paste placed with spiral filler (Lentulospiral) subgroup B - calcium hydroxide paste placed with flat wire filler (Paste Inject). pH and calcium ion release was evaluated at 24 h, 48 h, 7th day, 15th day and 30th day using a pH meter and atomic absorption spectrophotometer respectively. RESULTS: The calcium release from various groups was highest for CH+glycerine (Group III) followed by CH+chlorhexidine (Group I), CH+propylene glycol (Group II) and CH+double distilled water (control). At all the intervals the differences in calcium ion release among the groups were statistically significant (p < 0.05), except on day 7. Delivery technique did not have a significant effect on calcium ion release. Highest pH values were recorded from CH+glycerine group at day 30 for both the delivery systems, however rise in pH from day 1 to day 30 were non-significant in all groups with both delivery systems. CONCLUSION: Demonstrable changes in calcium ion release occurred from the calcium hydroxide mixed with various vehicles and CH+glycerine group showed the maximum calcium ion release at all intervals and highest pH day 30.

Genotoxicity of industrial solid waste leachates in Drosophila melanogaster.

The potential toxicity of industrial solid wastes is a major environmental concern. The present study evaluated the genotoxicity of industrial waste leachates on the gut cells of Drosophila melanogaster (Oregon R+), using a modified alkaline comet assay. Leachates were prepared from control soil and solid wastes generated by a flashlight battery factory, a pigment plant, and a tannery, using different pHs (7.0, 4.93, and 2.88). Newly emerged first instar Drosophila larvae (22 +/- 2 hr) were transferred to standard Drosophila diet containing 0.05%, 0.1%, 0.5%, 1.0%, and 2.0% of the leachates, and allowed to grow. At 96 +/- 2 hr, the anterior midgut of control and treated larvae was dissected out; single cell suspensions were prepared; and the comet assay was performed on the cells. All the leachates produced significant (P < 0.05), dose-dependent increases in DNA damage, in the gut cells. Leachates prepared at pH 7.0 were significantly less genotoxic than leachates prepared at pH 4.93 or 2.88. A comparison of the comet parameters among the exposed groups indicated that leachates of the pigment plant solid waste produced the least DNA damage, while leachates prepared from the flashlight battery factory solid waste were the most genotoxic. The present study indicates that leachates of solid wastes from flashlight battery factories, pigment plants, and tanneries possess genotoxic activity and that D. melanogaster is a useful in vivo model for assessing the genotoxicity of these potential environmental contaminants.

Comparative biomonitoring of leachates from hazardous solid waste of two industries using Allium test.

Hazardous industrial wastes are inevitable source of environmental pollution. Leachates from these wastes might contaminate the origins of potable water and affect human health. The study was carried out to determine the possible genotoxic effects of leachates from solid waste of a metal and dye industry using the Allium cepa chromosome aberrations assay. The 10% leachates were prepared from solid wastes obtained from both the industries and examined for the presence of heavy metal content and genotoxicity. To simulate the field and laboratory conditions, A. cepa bulbs were exposed through soil and aqueous medium for 48 h to 2.5-10% leachates. The results revealed that both metal waste leachate (MWL) and dye waste leachate (DWL) contained high concentrations of chromium, nickel and iron that significantly induced cytogenetic alterations. Significant inhibition of mitotic index (MI), inductions of chromosomal/mitotic aberrations (CA/MA) and micronuclei (MN) formation were found in all experimental groups exposed to MWL and DWL. The effects observed were concentration dependent and the frequency of aberrations was higher with treatment of MWL than DWL. The MI was severely inhibited at 10% aqueous exposure it was 4.59+/-0.69 (P<0.001) in MWL and almost half to that induced by DWL that was 8.62+/-0.69 (P<0.05). Significant frequency of CA/MA and MN induced by MWL was 14.21 (P<0.001) and 0.33 (P<0.001) whereas CA/MA and MN induced by DWL was 7.81 (P<0.001) and 0.13 (P<0.05) in the aqueous medium. The investigations inferred that abnormalities caused by MWL were higher than DWL both in soil and aqueous media. These toxic responses may have relied on raised heavy metal concentrations of metal-based than dye industrial wastes.

Release of nickel and chromium ions in the saliva of patients with fixed orthodontic appliance: An in-vivo study.

INTRODUCTION: Various components of fixed orthodontic appliances are continuously interacting with saliva and other fluids in the mouth releasing various metal ions including nickel and chromium that can cause damaging effects if their concentration exceeds above the toxic dose. AIM: To determine and compare the level of nickel and chromium in the saliva of patients undergoing fixed orthodontic treatment at different time periods. MATERIALS AND METHODS: The sample of saliva of 13 patients was taken at different time periods that is: Group 1 (before appliance placement), Group II, III, and IV (after 1-week, 1-month, and 3 months of appliance placement respectively). The fixed appliance comprised of brackets, bands, buccal tubes, lingual sheath, transpalatal arch and wires composed of Ni-Ti and stainless steel. The level of ions was determined using graphite furnace atomic absorption spectro-photometry. The data thus obtained were statistically analyzed using SPSS Statistical Analysis Software (Version 15.0). RESULTS: Level of nickel and chromium in saliva was highest in Group II and lowest in Groups I for both the ions. On comparison among different Groups, it was statistically significant for all the groups (<0.001) except between Group III and Group IV. CONCLUSION: The release of nickel and chromium was maximum at 1-week and then the level gradually declined. These values were well below the toxic dose of these ions. The results should be viewed with caution in subjects with Ni hypersensitivity.

Effect of manganese and copper interaction on behavior and biogenic amines in rats fed a 10% casein diet.

Manganese chloride (1 mg manganese/ml drinking water) and copper sulphate (250 mg copper/kg diet) were administered daily for 30 days to growing rats maintained on a 10% casein diet. Manganese in combination with copper produced impairment in the learning ability and memory of rats in either dietary group, with greater alterations in those receiving a 10% casein diet. Hyperactivation due to the combined effects of the metal ions was identical in both dietary groups. The behavioral aberrations were associated with a marked accumulation of Cu in the brain of rats receiving the 10% casein diet. Combined exposure of Mn and Cu also produced a greater elevation in the levels of dopamine (DA) and norepinephrine (NE) and a depression of 5-hydroxytryptamine (5-HT) in the brain of animals fed with a 10% casein diet compared to the animals receiving the 21% casein diet. Thus the animals receiving the 10% casein diet appear to be more vulnerable to the neurotoxic effects of a combined exposure to manganese and copper. However rats maintained on the 21% casein diet and receiving both manganese and copper showed almost identical changes to those observed after manganese administration alone.

Chromium induced teratogenicity in female rat.

Exposure to chromium (VI) (250, 500 and 750 ppm as potassium dichromate) via drinking water pregestationally in rats revealed embryo- and fetotoxic effects in the form of a significant reduction in the number of implantations and number of fetuses. An increase in the number of resorptions, pre-implantation and post-implantation loss in chromium (VI)-treated mothers was also observed. No significant visceral abnormality was found. A significant increase in sub-dermal hemorrhagic patches on thoracic and abdominal areas was found. Skeletal abnormality in the form of reduced ossification in parietal, interparietal and caudal bones was found in the fetuses of chromium (VI)-treated mothers. Chromium levels in blood, placenta and fetuses were found to be significantly increased in the 500 ppm and 750 ppm dosed groups. The duration of estrus cycle was significantly altered after chromium (VI) exposure. This study suggests that chromium exposure in rat causes a lower degree of toxicity than in mice as observed in our earlier studies.

Ovarian dysfunction in mice following chromium (VI) exposure.

Chromium (VI) was given through drinking water in two sets of adult Swiss albino female mice in three doses; 250 ppm, 500 ppm and 750 ppm for 20 days in set 1 and 0.05 ppm, 0.5 ppm and 5.0 ppm in set II for 90 days. At the termination of the treatment, the animals of both the sets were euthanized for histopathology, follicle counting, counting of the superovulated ova, duration of estrus cycle and for ultrastructural studies. Ovaries of the highest dose group (750 ppm) showed large numbers of atretic follicles and congestion in stromal tissue compared to the rest of the treated groups. Also, there was a dose-dependent reduction in the number of follicles at different stages of their maturation. The number of ova recovered from superovulated chromium (VI)-treated animals showed significant decreases in the 500 and 750 ppm dosed groups compared to lower dosed (250 ppm) and control groups. The duration of estrus cycle increased in highest dosed (750 ppm) group. A dose-dependent increase in blood chromium level was also seen in treated mice. Ultrastructural observations revealed disintegrated cell membranes of two layered follicular cells and altered villiform mitochondria in thecal cells of 5 ppm dosed group. From the study it was concluded that ovarian physiology and rate of ovulation might be altered if females are exposed to sufficiently high chromium through oral route.

Embryo and fetotoxicity of hexavalent chromium: a long-term study.

Ingestion of chromium(VI) (250, 500 or 750 ppm as potassium dichromate, K2Cr2O7) through drinking water by female rats for 3 months prior to gestation was toxic to embryo and fetus. There was a significant reduction in number of implantations and number of fetuses and an increase in number of resorptions and pre-implantation and post-implantation losses. No significant visceral abnormality was found. The increase in the number of subdermal hemorrhagic patches on the thorax and abdomen was significant. Skeletal abnormality in the form of reduced ossification in parietal, interparietal and caudal bones was observed in fetuses. Chromium levels in the blood of mothers, placenta and fetuses showed a significant increase. Duration of the estrous cycle was also increased significantly. The study revealed that long-term chromium exposure in rats did not cause embryo and fetotoxicity in a duration-dependent manner compared to short-term treatment as observed earlier. A possible explanation could be that, in the 90-day study, the female rats did not mate for three estrous cycles, thus giving time for clearance of a sizable amount of chromium from their bodies.

Embryotoxicity of orally administered chromium in mice: exposure during the period of organogenesis.

Administration of chromium (VI)(250, 500 and 750 ppm as potassium dichromate) via drinking water during organogenesis (days 6-14 of gestation) in mice revealed embryo- and fetotoxic effects. Reduced fetal weight, retarded fetal development, number of fetuses (live and dead) per mother and high incidences of dead fetuses and resorptions in treated mothers in the highest dosed group were evident. No significant gross structural abnormalities were observed in any of the fetuses of chromium (VI)-treated mothers. Significant incidences of reduced ossification were found in the highest dosed group. Chromium levels were increased in a dose-dependent manner in maternal blood, placenta and fetuses. The present study suggests a risk to the developing embryo if the mother is exposed to a sufficiently high concentration of chromium (VI) through drinking water during the period of organogenesis.

Serum ceruloplasmin in manganese-treated rats.

Rats were exposed to manganese (Mn) (6 mg/kg) daily for 30 days. The content of ceruloplasmin decreased significantly by 19.5% while copper (Cu) content was reduced by 27.8% in the serum of Mn-treated animals. These alterations were associated with a marked increase in the Cu content of liver and brain. The significant decrease in serum Cu and ceruloplasmin may serve as useful parameters for early diagnosis of Mn poisoning.