Spore inhibition-based enzyme substrate assay for monitoring of aflatoxin M1 in milk

A spore germination-based concept and its transformation into a field level prototype for monitoring aflatoxin M₁ (AFM₁) in milk was developed. Initially, 15 strains of Bacillus spp. procured from different culture collection were screened for AFM₁ sensitivity using spot assay and marker strain showing inhibition at 0.5 ppb was selected based upon maximum zone of inhibition. The selected strain B. megaterium 2949 was further screened for different enzymes activities and subsequently its spores were produced to an extent of 73.13% ± 3.197% in newly developed sporulation medium containing beef extract (0.0075% ± 0.0004%), yeast extract (0.015% ± 0.001%), peptone (0.0375% ± 0.0016%), and sodium chloride (0.0375% ± 0.0018%). A spore germination-based concept/ assay was optimized by immobilizing spores in eppendorf with pretreated milk (80°C/15 min) containing germinant and chromogenic substrate followed by incubation at 37°C. The appearance of sky blue color within real time of 45 min indicated spores germination and release of specific marker enzyme such as acetyl esterase and its specific action on chromogenic substrate which demonstrates absence of AFM₁ in milk. However, if there was no color change, presence of AFM₁ at 0.5 ppb MRL was denoted by Codex. The developed concept on AFM₁ detection was validated and a correlation of 0.97 was established with AOAC approved Charm 6602 and ELISA at Codex MRL with minimal false positive and negative results. The cost effective test has potential application in dairy farms, manufacturing, and R&D units for routine monitoring of AFM₁ in milk.     

Trihalomethanes formation in water treatment plants and distribution lines: a monitoring and modeling scheme

Monitoring and modeling of the concentrations of trihalomethanes (THMs) within four water treatment plants (WTPs) and distribution lines in Fayoum City, Egypt, were studied. Sampling sites for raw and treated waters were determined by global positioning system and the Arc geographic information system software was used for mapping. THMs were monitored using 25 sampling points located at the plant exit and through the distribution lines up to the plant extremity. Results indicated that the THMs concentration varied significantly but it was very rarely higher than the allowed maximum contaminant level of 100 μg L^?1. However, at the dead zones in the distribution lines, the THMs increased by about 160% compared to in-plant level. Furthermore, the level of THMs in hot months was about 1.3 times higher than in cold ones. The influence of some independent variables such as temperature, pH, ammonia concentration, total organic carbon and color were investigated by Pearson model to find their correlation with the rate of THMs formation in WTPs. The resulted R values ranged from 0.81 to 0.98. Successful application of the model to a selected region on the distribution lines has resulted in a correlation coefficient of 0.98.     

Biochemical properties of beryllium potentially relevant to its carcinogenicity†

The carcinogenicity of beryllium to several animal species is well established and evidence exists which strongly suggests that this is the case in human exposure. In this review several biochemical properties of the metallocarcinogen are considered including, the causation of cell transformation, and infidelity of DNA synthesis, inhibition of cell division and enzyme induction, and interference with regulatory mechanisms controlling gene expression. These effects are discussed in relation to beryllium chemistry, cellular accumulation mechanisms and distribution to subcellular organdies and molecular targets. It is suggested that the ultimate location and interactions of the metal ion in cell nuclei and its selective inhibition of certain protein phosphorylation reactions in particular are the biochemical effects potentially most relevant to induction of beryllium carcinogenesis.     

Voltammetric studies of damages in DNA by mutagenic and carcinogenic alkylating chemicals

Voltammetry provides new insights into the effects exerted in vitro by methylation of native DNA. Applying single sweep voltammetry at a stationary mercury electrode the successive steps of the destabilization of alkylated DNA are investigated. The methylation of the nucleic acids is manifested by a specific electrochemical response, due to the 7‐methylguanine, a major product of the methylation of DNA. Short time effects of the methylation include the labilization of 4 to 5 base pairs per methylated guanine base. Furthermore, uncommon protonation properties of the base 7‐methylguanine‐cytosine have been detected. Long term effects of the methylation are ultimately spontaneous hydrolytic strand breaks induced by the prior depurination connected with the release of the 7‐methylguanine from the methylated DNA. A half time t_1/2 of 102 h for the depurination at 37°C has been determined. The depurination and the subsequent strand breaks alter the hydrodynamic properties of the damaged DNA, an effect which can be sensitively followed with voltammetry via the resulting changes in the diffusion coefficient of the damaged biopolymer.     

Catalytic decomposition of hypochlorite ion in petrochemical plant waste water

During the incineration of the chlorinated hydrocarbons originating as the side products in vinyl chloride plant, hypochlorite ion appears as a pollutant in plant waste water. To remove it to the permitted level of 2 mg/l the process of the catalytic oxidation by cobalt (II, III) oxide, followed by sodium sulphite treatment has been chosen. It was shown that the above choice gives satisfactory results. The influence of some parameters such as retention time, effluent pH and temperature versus removal efficiency have been determined.     

Effects of the level of dietary protein on the toxicity of hexachlorocyclohexane in rats

Male albino rats were fed for 28 days from weaning on diets containing 5% (group 1), 10% (group 2) and 21% (group 3, normal protein) protein as casein. At the end of dietary period, HCH was administered daily for 30 days to investigate the interaction between protein deficiency and pesticide toxicity. The results indicated that rats fed a lower protein diet and HCH had a higher mortality, lower rate of growth, increased liver weight and deposition of the pesticide in blood and tissues in larger amounts. Blood pressure (systolic and diastolic) was significantly increased and the heart rate showed tachycardia in low protein exposed animals. A significant increase of total lipids, cholesterol, triglycirides, free fatty acids in serum and tissues of animals exposed to low protein was observed. A close correlation existed between lipid accumulation and storage of HCH in tissues and dietary protein seemed to play an important role in detoxification.     

Concentrations of Hchs and Ddts in the Tissues of River Dolphins,Platanista Gangetica,From the River Ganges,India

Abstract

Concentrations of HCH and DDT organochlorine insecticide residues were measured in blubber, muscle and oil samples from three specimens of river dolphins, Platanista gangetica, from the River Ganges, India. Concentrations of HCH and DDT ranged from 94 to 289 ng g^?1 and from 1324 to 9388 ng g^?1 on wet wt. basis respectively. Comparisons are made with other aquatic mammals and other studies on river dolphins. P. gangetica appears to exhibit similar patterns of accumulation with age and with ß-HCH and p-p′-DDE being accumulated to higher levels than other HCH isomers and parent DDT and its other metabolites, respectively. These organochlorines may pose a health risk to river dolphin populations that are already showing evidence of environmental stress. Further studies are recommended.     

Distribution and speciation of heavy metals in Apulian coastal sediments

In the framework of the Interreg II Project Italy–Greece, distribution and speciation of trace metals in surface coastal sediments of Apulian regions were studied. A sequential extraction technique was used to determine the chemical association of heavy metals (Zn, Pb, Ni, Fe, Cr, Mn, Cu and Cd) with major sedimentary phases (acid soluble phases, easily reducible fraction and oxidizable matter). The relationship between the metals' content and the grain-size composition of the sediments was also considered: generally the elements resulted associated with the fine fraction, even if in some cases (Cu in the oxidizable phase) the opposite behaviour was observed. The studied area is not particularly polluted, except for some defined zones, as for example Brindisi's harbour. Speciation analysis revealed that the major part of the heavy metals are not easily mobilizable and bioavailable. Even in this case, there are some exceptions to the general behaviour: Cu and Pb in several samples are largely present in the oxidizable phase, which even if not immediately available can become a source of associated elements when redox conditions change.     

Indigo carmine removal by charcoal from rice bran as an agricultural by-product

On the utilization of agricultural by-product, charcoal from rice bran was produced. The adsorption characteristics of indigo carmine onto charcoals from rice bran were investigated by the batch method. The yield and surface area of charcoal were decreased and increased with increase of carbonization temperature, respectively. The removal ratio of indigo carmine was high with the charcoal carbonized at higher temperature. In the relationship between the amount of indigo carmine adsorbed and the square root of elapsed time, a good linearity was recognized. The kinetic constant of adsorption removal for indigo carmine was rapid in the charcoal carbonized at higher temperature. The adsorption characteristics for indigo carmine removal by charcoal from rice bran were dominated by the value of their surface area.