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.     

Assessment of the environmental profile of PLA,PET and PS clamshell containers using LCA methodology

Life cycle assessments of bio-based polymer resin and products historically have shown favorable results in terms of environmental impacts and energy use compared to petroleum-based products. However, calculation of these impacts always depends on the system and boundary conditions considered during the study. This paper reports a cradle-to-cradle Life Cycle Assessment (LCA) of poly(lactic acid) (PLA) in comparison with poly(ethylene terephthalate) (PET) and poly(styrene) (PS) thermoformed clamshell containers, used for packaging of strawberries with emphasis on different end-of-life scenarios. It considers all the inputs such as fertilizers, pesticides, herbicides and seed corn required for the growing and harvesting of corn used for manufacturing PLA. For PET and PS, the extraction of crude oil and the entire cracking processes from crude oil through styrene and ethylene glycol and terephathalic acid are considered. Global warming, aquatic acidification, aquatic eutrophication, aquatic ecotoxicity, ozone depletion, non-renewable energy and respiratory organics, land occupation and respiratory inorganics were the selected midpoint impact categories. The geographical scope of the study reflects data from Europe, North America and the Middle East. PET showed the highest overall values for all the impact categories, mainly due to the higher weight of the containers. The main impacts to the environment were the resin production and the transportation stage of the resins and containers. This implies that the transportation stage of the package is an important contributor to the environmental impact of the packaging systems, and that it cannot be diminished.     

Sorption-desorption behavior of metsulfuron-methyl and sulfosulfuron in soils

Sorption of metsulfuron-methyl and sulfosulfuron were studied in five Indian soils using batch sorption method. Freundlich adsorption equation described the sorption of herbicides with K(f) (adsorption coefficient) values ranging between 0.21 and 1.88 (metsulfuron-methyl) and 0.37 and 1.17 (sulfosulfuron). Adsorption isotherms were L-type suggesting that the herbicides sorption decreased with increase in the initial concentration of the herbicide in the solution. The K(f) for metsulfuron-methyl showed good positive correlation with silt content (significant at p = 0.01) and strong negative correlation with the soil pH (significant at p = 0.05) while sorption of sulfosulfuron did not correlate with any of the soil parameter. Desorption of herbicides was concentration dependent and, in general, sulfosulfuron showed higher desorption than the metsulfuron-methyl. The study indicates that these herbicides are poorly sorbed in the Indian soil types and there may be a possibility of their leaching to lower soil profiles.     

Changing effluent chemistry affect survival,growth and physiological function of <Emphasis Type="Italic">Acacia nilotica</Emphasis> seedlings in northwestern region of India

Recycling and conservation efforts for water are the need of the day because of the lack of new water sources and the ever-increasing demand for drinking water. Seedlings of Acacia nilotica L. were irrigated with: canal water (T₁, control); municipal effluent (T₂); textile effluent (T₃); steel effluent (T₄); textile + municipal effluent in 1:1 ratio (T₅); steel + municipal effluent in 1:2 ratio (T₆); steel + textile in 1:2 ratio (T₈) and steel + municipal + textile in 1:2:2 ratio (T₇) with views to observe effluents effect on the seedlings and its adaptability and to recommend safe disposal of these effluents. Seedlings in T₆, T₇ and T₈ showed 50% lesser height and collar diameter than those in control. Seedlings in T₂ attained greatest height, collar diameter, numbers of branches and produced 140 g dry biomass seedling⁻¹. Highest concentration of manganese (Mn), iron (Fe), copper (Cu) and zinc (Zn) and lowest concentration of nitrogen (N), phosphorus (P), potassium (K), calcium (Ca) and magnesium (Mg) in the seedlings of T₄, T₆, T₇ and T₈ resulted in nutritional imbalance, mineral toxicity and reduction in photosynthetic (Pn) and transpiration (E) rates and caused seedling mortality. Seedlings of T₃ had highest sodium concentration and low concentration of Ca, Mg and micronutrients resulting in nutritional imbalance, augmented chlorosis and reduced gas exchange and biomass by half as compared to control. Increased growth, Pn and E and biomass in seedlings of T₅ over T₃ and survival period in T₆, T₇ and T₈ seedlings suggested a beneficial effect of effluents mixing. Unscientific disposal should be avoided and toxic concentration of metal ions␣may be reduced for long-term application and harmless disposal of effluents in afforestation and urban development.     

A comparative study on characterization of textile wastewaters (untreated and treated) toxicity by chemical and biological tests

Toxicity of textile wastewaters (untreated and treated) and their ingredient chemicals was quantified in terms of their chemical characteristics, fish (Gambusia affinis) mortality and end point growth responses of duckweed (Lemna aequinoctialis) in short-term bioassays. Other parameters of fish bioassay were erythrocyte morphology and its counts. Despite of a definite correlation between data of biological tests (LC/EC(50) values) with that of chemical tests, biological tests were found to be relatively more sensitive to both wastewaters and ingredient chemicals. Amongst all the examined parameters of test organisms, fish RBCs (morphology and counts) sensitivity to pollutants in the wastewaters was usually maximum and therefore, their study should be included in the routine fish bioassay. Other advantage of biological test such as on Lemna is even detection of eutrophic potential of wastewaters, as noted at their higher dilutions. The ingredient chemicals (major) contributing maximum toxicity to textile dye wastewater were, acids (HCl and H(2)SO(4)), alkali (Na(2)O SiO(2)), salt (NaNO(2)) and heavy metal (Cu), whereas dyes (4) were relatively less toxic.     

Growth performance and biochemical responses of three rice (Oryza sativa L.) cultivars grown in fly-ash amended soil

The disposal of fly-ash (FA) from coal-fired power stations causes significant economic and environmental problems. Use of such contaminated sites for crop production and use of contaminated water for irrigation not only decreases crop productivity but also poses health hazards to humans due to accumulation of toxic metals in edible grains. In the present investigation, three rice cultivars viz., Saryu-52, Sabha-5204, and Pant-4 were grown in garden soil (GS, control) and various amendments (10%, 25%, 50%, 75% and 100%) of FA for a period of 90 days and effect on growth and productivity of plant was evaluated vis-a-vis metal accumulation in the plants. The toxicity of FA at higher concentration (50%) was reflected by the reduction in photosynthetic pigments, protein and growth parameters viz., plant height, root biomass, number of tillers, grain and straw weight. However, at lower concentrations (10-25%), FA enhanced growth of the plants as evident by the increase of studied growth parameters. The cysteine and non-protein thiol (NP-SH) content showed increase in their levels up to 100% FA as compared to control, however, maximum content was found at 25% FA in Saryu-52 and Pant-4 and at 50% FA in Sabha-5204. Accumulation of Fe, Si, Cu, Zn, Mn, Ni, Cd and As was investigated in roots, leaves and seeds of the plants. Fe accumulation was maximum in all the parts of plant followed by Si and both showed more translocation to leaves while Mn, Zn, Cu, Ni and Cd showed lower accumulation and most of the metal was confined to roots in all the three cultivars. As was accumulated only in leaves and was not found to be in detectable levels in roots and seeds. The metal accumulation order in three rice cultivars was Fe > Si > Mn > Zn > Ni > Cu > Cd > As in all the plant parts. The results showed that rice varieties Saryu-52 and Sabha-5204 were more tolerant and could show improved growth and yield in lower FA application doses as compared to Pant-4. Thus, Sabha-5204 and Saryu-52 are found suitable for cultivation in FA amended agricultural soils for better crop yields.     

Modeling and optimization of trihalomethanes formation potential of surface water (a drinking water source) using Box–Behnken design

Purpose  

The present research aims to investigate the individual and interactive effects of chlorine dose/dissolved organic carbon ratio, pH, temperature, bromide concentration, and reaction time on trihalomethanes (THMs) formation in surface water (a drinking water source) during disinfection by chlorination in a prototype laboratory-scale simulation and to develop a model for the prediction and optimization of THMs levels in chlorinated water for their effective control.     

Release kinetics of β-cyfluthrin from its encapsulated formulations in water

Controlled release formulations of β-cyfluthrin, a non-systemic, broad spectrum contact insecticide, have been prepared using laboratory synthesized poly(ethylene glycol) (PEG) based amphiphilic copolymers. Copolymers of polyethylene glycols of different molecular weights and various dimethyl esters, viz. dimethyl isophthalate, which self assemble into nano micellar aggregates in aqueous media, have been synthesized. The kinetics of β-cyfluthrin from developed controlled release (CR) formulations were studied in comparison with that of the commercially available 025 SC. Release from the commercial formulation was faster than with the developed CR formulations. The rate of release of encapsulated β-cyfluthrin from nano micellar aggregates is reduced by increasing the molecular weight of PEG. The diffusion exponent (n value) of β-cyfluthrin in water ranged from 0.427 to 0.622 in the tested formulations. The release was diffusion controlled with a half-release time (t(?)) of 3.92 to 7.9 days in water from different formulations, and the period of optimum availability (POA) of β-cyfluthrin ranged from 1.4 to 20.5 days. The results suggest that the application rate of β-cyfluthrin can be optimized to achieve insect control at the desired level and period.     

Development of controlled release formulations of azadirachtin-A employing poly(ethylene glycol) based amphiphilic copolymers

Controlled release (CR) formulations of azadirachtin-A, a bioactive constituent derived from the seed of Azadirachta indica A. Juss (Meliaceae), have been prepared using commercially available polyvinyl chloride, polyethylene glycol (PEG) and laboratory synthesized poly ethylene glycol–based amphiphilic copolymers. Copolymers of polyethylene glycol and various dimethyl esters, which self assemble into nano micellar aggregates in aqueous media, have been synthesized. The kinetics of azadirachtin-A, release in water from the different formulations was studied. Release from the commercial polyethylene glycol (PEG) formulation was faster than the other CR formulations. The rate of release of encapsulated azadirachtin-A from nano micellar aggregates is reduced by increasing the molecular weight of PEG. The diffusion exponent (n value) of azadirachtin-A, in water ranged from 0.47 to 1.18 in the tested formulations. The release was diffusion controlled with a half release time (t_1/2) of 3.05 to 42.80 days in water from different matrices. The results suggest that depending upon the polymer matrix used, the application rate of azadirachtin-A can be optimized to achieve insect control at the desired level and period.     

Trade-offs between elimination and detoxification in rainbow trout and common bivalve molluscs exposed to metal stressors

The purpose of this paper was to examine trade-offs between elimination and detoxification in rainbow trout and three common bivalve molluscs (clam, oyster, and scallop) exposed to cadmium (Cd), copper (Cu), and zinc (Zn) based on recent reported experimental data. We incorporated metal influx threshold with subcellular partitioning to estimate rate constants of detoxification (k_d) and elimination (k₂). We found that the relationships between k₂ and k_d were negative for rainbow trout and positive for bivalve molluscs. However, the relationships between k_d and % metal in metabolically detoxified pool were found positive for rainbow trout and negative for bivalve molluscs. Our results also indicated that rainbow trout had higher accumulation (∼60-90%) in metabolically active pool when exposed to essential metals of Cu and Zn and had only 10-50% accumulation in response to non-essential metal of Cd. Based on a cluster analysis, this study indicated that similarity of physiological regulations among study species was found between Cd and Zn. Our study suggested that detoxification can be predicted by an elimination-detoxification scheme with the known elimination rate constant. We concluded that quantification of trade-offs between subcellular partitioning and detoxification provides valuable insights into the ecotoxicology of aquatic organisms and enhances our understanding of the subcellular biology of trace metals.