Amelioration potential of Moringa oleifera extracts against sodium arsenate induced embryotoxicity and genotoxicity in mouse (Mus musculus)

Previous studies have suggested that arsenic crosses the placenta and affects the fetus development. The study under consideration aims to show comparative ameliorative effect of Moringa oleifera leaf and flower extracts against sodium arsenate induced fetus toxicity of mice. Pregnant mice (N=44) were kept in lab and divided into eleven group from (A to K) and were orally administered the doses 6 mg/kg, 12 mg/kg for sodium arsenate, 150 mg/kg and 300 mg/kg for Moringa oleifera leaf extracts (MOLE) and 150 mg/kg and 300 mg/kg for Moringa oleifera flower extracts (MOFE) comparing with control. The investigation revealed evident reduction in the fetuses weight, hind limb, fore limb, tail and snout length, crown rump and head circumferences well as malformations in tail, feet, arms, legs, skin and eyes in the negative control group (only administered with sodium arsenate). Co-administration of sodium arsenate with MOLE and MOFE ameliorate the reversed effect of sodium arsenate on the shape, length, body weight and DNA damage of fetus significantly at 95% confidence interval. However, Moringa oleifera leaf extract showed more significant results in comparison to Moringa oleifera flower extract. Hence concluded that Moringa oleifera leaf extract ameliorated the embryo toxic effects of sodium arsenate and can be used against environmental teratogens.

Alkaline refining of crude lead: a method of arsenic removal and the behavior of arsenic in the process

In this work, the alkaline refining of arsenic in crude lead was studied with a mixture of sodium hydroxide and sodium carbonate as alkaline refining agents. Taking the arsenic removal rate as the research object, the effects of reaction temperature, holding time, Na2CO3:NaOH, the dosage of refining agent were investigated. The arsenic removal rate is 79.09% under the optimum experimental conditions as follows: reaction temperature 823 K, holding time 60 min, Na2CO3:NaOH 1:4, refining agent dosage 10%. The oxidation purification mechanism of arsenic was studied by XPS, SEM-EDS, XRD and FT-IR. The results show that arsenic in the crude lead is gradually oxidized by oxygen and lead oxide during arsenic removal process, and the arsenic trioxide is eventually converted into sodium arsenate (Na3AsO4) and lead arsenate (Pb2As2O7) in the slag.

Embryotoxicity and oxidative stress induced by arsenic in albino mice and protective role of Moringa oleifera

Chronic exposure to arsenic causes the abnormal development of embryo during pregnancy. So the study is designed to investigate the antioxidant and tissue protective properties of Moringa oleifera extracts against sodium arsenate induced embryo toxicity. Moringa oleifera extracts (leaf and flower) were prepared using soxhelt apparatus. Fourty four pregnant females were equally divided in 11 experimental groups (A-K). Group A was of control while B and C were sodium arsenate treated groups as group A (0.00), B (6.00, 0.00), C (12.00, 0.00). Group D to G were of sodium arsenate + Moringa oleifera flower extract treated groups with doses D (6.00, 150.00), E (6.00, 300.00), F (12.00, 150.00), G (12.00, 300.00) and groups H to K were sodium arsenate + Moringa oleifera leaf extracts treated groups H (6.00, 150.00), I (6.00, 300.00), J (12.00, 150.00) and K (12.00, 300.00) mg/kg B.W. Significant (p < 0.05) increased in MDA (malanodialdehyde) values 41.75 ± 3.40 and decreased GSH (glutathione) values 7.75 ± 0.95 in sodium arsenate treated groups were observed as compared to control 25 ± 0.81(MDA) and 18.5 ± 0.57 (GSH). Moringa oleifera extracts treated groups especially Moringa oleifera leaf extract at dose of 300mg/kg B.W normalized the MDA (25.25 ± 0.50) and GSH (18.25 ± 1.70) values. Sodium arsenate induced the histopathologiacl changes like malformed heart, meningocoel, spina bifida, anopathalmia, cavitization, degenerated kidney and intestine. Whereas Moringa oleifera leaf extract as ameliorant minimized these histopathological abnormalities. It is concluded that Moringa oleifera leaf extract is mixture of biomolecules as it has antioxidant activity, so can be used as ameliorant against sodium arsenate.

Characterization of Arsenic-Resistant Endophytic Bacteria From Alfalfa and Chickpea Plants

The present investigation was carried out to isolate arsenic (As)-resistant endophytic bacteria from the roots of alfalfa and chickpea plants grown in arsenic-contamination soil, characterize their As tolerance ability, plant growth-promoting characteristics, and their role to induce As resistance by the plant. A total of four root endophytic bacteria were isolated from plants grown in As-contaminated soil (160–260-mg As kg−1 of soil). These isolates were studied for plant growth-promoting (PGP) characteristics through siderophore, phosphate solubilization, nitrogen fixation, protease, and lipase production, and the presence of the arsenate reductase (arsC) gene. Based on 16S rDNA sequence analysis, these isolates belong to the genera Acinetobacter, Pseudomonas, and Rahnella. All isolates were found As tolerant, of which one isolate, Pseudomonas sp. QNC1, showed the highest tolerance up to 350-mM concentration in the LB medium. All isolates exhibited phosphate solubilization activity. Siderophore production activity was shown by only Pseudomonas sp. QNC1, while nitrogen fixation activity was shown by only Rahnella sp. QNC2 isolate. Acinetobacter sp. QNA1, QNA2, and Rahnella sp. QNC2 exhibited lipase production, while only Pseudomonas sp. QNC1 was able to produce protease. The presence of the arsC gene was detected in all isolates. The effect of endophytic bacteria on biomass production of alfalfa and chickpea in five levels of arsenic concentrations (0-, 10-, 50-, 75-, and 100-mg kg−1 soil) was evaluated. The fresh and dry weights of roots of alfalfa and chickpea plants were decreased as the arsenic concentration of the soil was increased. Results indicate that the fresh and dry root weights of alfalfa and chickpea plants were significantly higher in endophytic bacteria-treated plants compared with non-treated plants. Inoculation of chickpea plants with Pseudomonas sp. QNC1 and Rahnella sp. QNC2 induced lower NPR3 gene expression in chickpea roots grown in soil with the final concentration of 100-mg kg−1 sodium arsenate compared with the non-endophyte-treated control. The same results were obtained in Acinetobacter sp. QNA2-treated alfalfa plants grown in the soil plus 50-mg kg−1 sodium arsenate. These results demonstrated that arsenic-resistant endophytic bacteria are potential candidates to enhance plant-growth promotion in As contamination soils. Characterization of bacterial endophytes with plant growth potential can help us apply them to improve plant yield under stress conditions.

Neurobehavioral assessment of Danio rerio intoxicated by sodium arsenate and the use of Arsenicum Album to reverse the condition of anxiety

Background: Various studies have indicated that the zebrafish serves as a useful and easily executable experimental model. Environmental pollution caused by heavy metal waste is very relevant due to the widespread use of heavy metals in industrial and agricultural practices, and the fact that many effluents are released into the environment without any treatment. Sodium arsenate changes behavioral parameters and brain ectonucleotidase activities in zebrafish1. Aims: The objective of this paper is to assess behavioral changes in zebrafish (Danio rerio) induced by sodium arsenate, through an analysis of locomotor activity and anxiety-related parameters, and determine whether ultra-diluted substances are able to mitigate neurobehavioral effects. Materials and methods: Zebrafish were held in aquariums with controlled temperature and pH, until the time of the toxicological experiment. The animals were exposed for 96 hours to the heavy metal sodium arsenate and divided into four groups with eight fish in each: Arsenicum Album 6 cH, Arsenicum Album 30 cH and inert aqueous solution (positive control) groups received drugs made in distilled water, according to the Brazilian Homeopathic Pharmacopoeia; the experiment was blinded. The fourth white control group (negative control) did not receive the sodium arsenate or drugs. All the homeopathic drugs that treated the heavy metal toxicity were initiated 5 days before the end of the experiment (therefore, the animals were already taking the medication one day before exposure). Afterwards, the animals underwent behavioral tests (locomotion and anxiety) to assess neurotoxicity. All the animals went through various steps: assessment of neurotoxicity signs where the following was observed: time and frequency on the surface and erratic movements; general activity test in an Open Field, where it was observed: time in seconds of locomotion and immobility; and light/dark test (anxiety-like behavior), where it was observed: the number of times the animals crossed from the light to the dark compartment, how long they spent on each side, number of attempts to enter each compartment and immobility time on the light side. The data obtained was analyzed statistically by ANOVA, followed by Bartlett's Test and the Tukey Test, with p?0.052. CEUA Protocol No. 07/2016. Results and Discussion: The heavy metal sodium arsenate produced an anxiogenic effect in the animals that were subjected to it without medication (aqueous solution, positive control group). This effect was observed in the light/dark test through increased time (sec) of the animals on the dark side (245.361.82) in relation to the control group (101.162.3) and a consequent decrease in time of the animals on the light side (54.336.3) compared to the control group (198.962.3). There was also a decrease in locomotion (quadrants) on the light side (96.263.7) in relation to the control group (201.889.2). In the Open Field, there was a decrease in the total of quadrants crossed (346.241.33) compared to the control group (520.5131.6), as well as an increase in the erratic movements (3.50.7) in relation to the control group (1.70.9), proving the toxic effect of heavy metals in the animals. The group that was medicated with Arsenicum Album 30 cH showed a decrease in the erratic movements (0.661.0) in relation to the intoxicated animals, resulting values close to the control group. The group that took Arsenicum Album 6 cH reversed all the positive control parameters (aqueous solution) in a statistically significant way, maintaining the values of the dark side time, the light side time and the clear side locomotion (161.663.7; 138.496.3; 121.049.8, respectively) close to the values of the control animals. Arsenicum 6 cH also increased the total number of quadrants in the Open Field (525.0142.3) and reduced the erratic movements (2.51.1), demonstrating anxiolytic ability. Clear side time for the animals treated with Arsenicum Album 6 cH, in relation to the control group, was more similar, and the same occurred in regard to the dark side time. When fish are treated with anxiolytics, they tend to remain more time on the light side, which indicates decreased anxiety (Serra et al. 1999). In relation to locomotion on the light side, there was a greater similarity between the control group and the fish treated with Arsenicum Album 6 cH, as well as in the displacement behavior in the Open Field with respect to the total number of quadrants of the fish displacement, which coincides with data from the literature that reports increased locomotor activity, as anxiety decreases. Locomotion or swimming activity is also used as an anxiety index and suppression of this activity is indicative of anxiety3. In relation to erratic movements, it was observed that this behavior decreased in animals treated with a concentration of 6 cH as well as with those treated with a concentration of 30 cH, compared to the positive control. Conclusion: The animals intoxicated by the heavy metal sodium arsenate showed increased anxiety, which was reversed by Arsenicum Album that in a concentration of 6cH proved to be more effective as a possible anxiolytic. Other studies are already being conducted by our group. Keywords: Homeopathy, High Dilution, Zebrafish, Intoxication. References 1. Baldissarelli, L.A.; Capiotti, K.M.; Bogo, M.R.; Ghisleni, G; Bonan, C.D. Arsenic alters behavioral parameters and brain ectonucleotidases activities in zebrafish (Danio rerio). Comparative Biochemistry and Physiology, Part C 155: 566-572, 2012. 2. Zar, J. H. Biostatistical analysis. 5 ed. Upper Saddle River: Prentice Hall, 2010. 944 p. 3. Blaser R.E., Chadwick L., Mcginnis G.C. Behavioral measures of anxiety in zebrafish (Danio rerio). Behav Brain Res. 2009.

Studies on the effect of sodium arsenate & cadmium chloride on Pithophora oedogonia (Mont.) Wittrock 1877

Cadmium and Arsenic are heavy metals although not common in our environment, its threat in certain places are aggravated due to anthropogenic factors. To know its critical role on plants the investigation was made using Na2HAsO4 and CdCl2 treatment on Pithophora oedogonia (Mont.) Wittrock 1877. The observations were made after 14 days of treatment. The changes were noted. In both cases, the treated cells exhibited gradual disruption of cell wall and cell membrane. The chlorophyll content initially increased and finally decreased due to the notable destruction of chloroplasts in both treated cells. A profuse number of akinetes were observed at 100 ppm and 150 ppm of Na2HAsO4 and CdCl2 treated media. Decrease in protein content was started at 100 ppm in both cases. The lipid content initially decreased at 50 ppm and at 100 ppm lipid profile increased in terms of toleration to the Na2HAsO4 and CdCl2 stress. Pithophora oedogonia (Mont.) Wittrock 1877 exhibited more sensitivity to CdCl2 stress & showing abrupt changes in chlorophyll-a and chlorophyll-b production. The carotenoid production shown more sensitivity in Na2HAsO4 stress. Total phenol production was decreased initially and at 200 ppm CdCl2 stress had shown significant enhancement than the control set but at the 200 ppm of Na2HAsO4 shown inhibitory effect.

The Protective Effects of Nicotine and Bucladesine on Impaired Avoidance Memory Caused by Sodium Arsenate Toxicity in Mice

Background: The toxic effect of sodium arsenate on nervous system has been shown; but the protective effects of several compounds against sodium arsenate are not clear. This study aimed to investigate the protective effects of nicotine and bucladesine, two positive modulators of neuronal function, on sodium arsenate toxicity against avoidance memory impairment. Methods: Male mice (N=154) were assigned to 22 groups (12 experimental and 10 control) of seven animals each and were treated as follows: sodium arsenate (2.5, 5, or 10 mg/kg) for 28 days, nicotine (1 mg/kg) for either 1, 2, or 4 days, bucladesine (600 nM/mouse) for either 1, 2, or 4 days, and nicotine (1 mg/kg)+bucladesine (600 nM/mouse)+sodium arsenate (2.5 mg/kg). The last group was treated with 2.5 mg/kg sodium arsenate first, and then received the combination of nicotine and bucladesine for 1, 2, or 4-days. The corresponding control groups did not receive any drug but either saline, deionized water, or combination of deionized water and DMSO, but went through the same procedure as other animals. All mice were trained 24 h in the step-through passive avoidance task. The avoidance memory retention was assessed at 24, 48, 96, and 168 h after the training period by measuring the time they stayed in a dark chamber. Results: All sodium arsenate doses significantly reduced the time stayed in the dark chamber regardless of the treatment duration (24, 48, 96 & 168 h) after training. Both nicotine and bucladesine, whether used singly or combined for 1, 2, and 4 days significantly enhanced the time latency compared to the controls at all of the experimental timepoints following the training. Conclusion: Nicotine and bucladesine showed synergistic effects and reversed the sodium arsenate-induced avoidance memory deficits in mice.

Arsenic Induced Vascular Endothelial Dysfunction Prevented by Antioxidants in Fetal kidneys of Albino Mice

Background &Objectives: Cardiovascular diseases and hypertension has a significant correlation with chronic arsenic exposure through drinking water. This study was designed to investigate the prevention of sodium arsenate induced vascular disorders by Vitamins C& E in fetal renal blood vessels of albino mice. Materials & Methods: Gravid albino mice of BALB/c strain (twenty four) were randomly divided into 4 groups having 6 animals each. Control group A was inoculated with 0.1ml/kg/day distilled water I/P for 18 days. Animals of groups B, C & D were given a single I/P injection of sodium arsenate 35mg/kg on 8th GD, whereas groups C and D were also injected with Vitamin C, 9 mg/kg/day and vitamin E 15 mg/kg/day by I/P route, beginning from 8th GD and continued for the entire pregnancy period. On 18th day of gestation fetal kidneys were extracted. Histological examination of renal blood vessels was performed for any discernable congestion, endothelial disruption and hyalinization and frequency of changes were expressed as percentages. Results: In group (B) sodium arsenate induction resulted in congestion of blood vessels, hemorrhages in glomerular capillaries and thickening of endothelial walls. The addition of Vitamins C and E in groups C & D respectively had reduced the congestion and endothelial thickening. Mean score of histological changes was statistically significant. Conclusions: The results showed the antioxidant prospective of Vitamins C and E in combating against the vascular lesions induced by sodium arsenate.