Evaluation of interference to conventional and real-time PCR for detection and quantification of fungi in dust

Advances in polymerase chain reaction (PCR) have permitted accurate, rapid and quantitative identification of microorganisms in pure cultures regardless of viability or culturability. In this study, a simple sample processing method was investigated for rapid identification and quantification of fungal spores from dust samples using both conventional and real-time PCR. The proposed method was evaluated for susceptibility to interference from environmental dust samples. Stachybotrys chartarum and Aspergillus fumigatus were used as test organisms. The sensitivity of detection in pure culture was 0.1 spore DNA equivalents per PCR reaction corresponding to 20 spores ml(-1) in the sample. However, 1 spore DNA equivalent per PCR reaction corresponding to 200 spores ml(-1) in the sample was the lowest amount of spores tested without interference in dust samples spiked with spores of either fungal species. The extent of inhibition was calculated using conventional and real-time PCR reactions containing fungal spores, specific primers, specific probes (for real-time PCR) and various amounts of dust. The results indicate that the extent of inhibition by dust on PCR varies with the type and amount of dust, and number of spores. No interference in the analysis of spiked samples was detected from 0.2 mg ml(-1) of four real-life dust samples at p-value >0.05 using 2 x 10(4) spores for conventional PCR and 2 x 10(5) spores for real-time PCR. However, samples containing >0.2 mg ml(-1) real-life dust compromised the PCR assay. These results suggest the potential usefulness of a simple sample processing method in conjunction with PCR for monitoring the fungal content of aerosols collected from indoor environments.     

Biodegradation of atrazine by Rhodococcus sp. BCH2 to N-isopropylammelide with subsequent assessment of toxicity of biodegraded metabolites

Atrazine is a persistent organic pollutant in the environment which affects not only terrestrial and aquatic biota but also human health. Since its removal from the environment is needed, atrazine biodegradation is achieved in the present study using the bacterium Rhodococcus sp. BCH2 isolated from soil, long-term treated with atrazine. The bacterium was capable of degrading about 75 % atrazine in liquid medium having pH 7 under aerobic and dark condition within 7 days. The degradation ability of the bacterium at various temperatures (20–60 °C), pH (range 3–11), carbon (glucose, fructose, sucrose, starch, lactose, and maltose), and nitrogen (ammonium molybdate, sodium nitrate, potassium nitrate, and urea) sources were studied for triumph optimum atrazine degradation. The results indicate that atrazine degradation at higher concentrations (100 ppm) was pH and temperature dependent. However, glucose and potassium nitrate were optimum carbon and nitrogen source, respectively. Atrazine biodegradation analysis was carried out by using high-performance thin-layer chromatography (HPTLC), Fourier transform infrared spectroscopy (FTIR), and liquid chromatography quadrupole time-of-flight (LC/Q-TOF-MS) techniques. LC/Q-TOF-MS analysis revealed formation of various intermediate metabolites including hydroxyatrazine, N-isopropylammelide, deisopropylhydroxyatrazine, deethylatrazine, deisopropylatrazine, and deisopropyldeethylatrazine which was helpful to propose biochemical degradation pathway of atrazine. Furthermore, the toxicological studies of atrazine and its biodegraded metabolites were executed on earthworm Eisenia foetida as a model organism with respect to enzymatic (SOD and Catalase) antioxidant defense mechanism and lipid peroxidation studies. These results suggest innocuous degradation of atrazine by Rhodococcus sp. BCH2 in nontoxic form. Therefore the Rhodococcus sp.BCH2 could prove a valuable source for the eco-friendly biodegradation of atrazine pesticide.     

Development of a purpose built landfill system for the control of methane emissions from municipal solid waste

In the present paper, a new system of purpose built landfill (PBLF) has been proposed for the control of methane emissions from municipal solid waste (MSW), by considering all favourable conditions for improved methane generation in tropical climates. Based on certain theoretical considerations multivariate functional models (MFMs) are developed to estimate methane mitigation and energy generating potential of the proposed system. Comparison was made between the existing waste management system and proposed PBLF system. It has been found that the proposed methodology not only controlled methane emissions to the atmosphere but also could yield considerable energy in terms of landfill gas (LFG). Economic feasibility of the proposed system has been tested by comparing unit cost of waste disposal in conventional as well as PBLF systems. In a case study of MSW management in Mumbai (INDIA), it was found that the unit cost of waste disposal with PBLF system is seven times lesser than that of the conventional waste management system. The proposed system showed promising energy generation potential with production of methane worth of Rs. 244 millions/y ($5.2 million/y). Thus, the new waste management methodology could give an adaptable solution for the conflict between development, environmental degradation and natural resources depletion.     

Green energy generation through PEHF – a blueprint of alternate energy harvesting

In this work, an attempt has been made to harvest green energy from piezoelectric material using fluid flow in a conduit. Piezoelectric Energy Harvesting using Fluid Flow (PEHF) experimental model has been designed and the outputs obtained are compared with results obtained from simulations using ANSYS (computational fluid dynamics) and also with the mathematical modeling. The PEHF model has been utilized to analyze the effect of flow rate of water with reference to energy extracted. The full wave bridge rectifier and voltage doubler circuits have been used to obtain the direct current (DC) from the PEHF model. It is observed that the output obtained using experiments holds good in agreement with the results retrieved through simulations and mathematical results. The increase in flow rate of fluid leads to initially increase and then decrease in output of PEHF model as the maximum energy generated when flow rates (external force) matches with the frequency of excitation of the systems, i.e., at its resonance. The maximum energy output is generated at its resonance frequencies. It is observed that the full wave bridge rectifier circuit gives greater output as compared to a voltage doubler circuit.     

Arsenic-induced health crisis in peri-urban Moyna and Ardebok villages, West Bengal, India: an exposure assessment study

Drinking of arsenic (As)-contaminated groundwater has adverse effects on health of millions of people worldwide. This study aimed to determine the degree of severity of As exposure from drinking water in peri-urban Moyna and Ardebok villages, West Bengal, India. Arsenic concentrations in hair, nail and urine samp les of the individuals were determined. Arsenical dermatosis, keratosis and melanosis were investigated through medical evaluation. We have evaluated the association between As exposure from drinking water, and keratosis and melanosis outcomes. The results showed that 82.7?% of the sampled tube wells contain As concentrations above 10?μg/L, while 57.7?% contain As concentrations above 50?μg/L. The hair, nail and urine As concentrations were positively correlated with As concentrations in drinking water. In our study population, we observed a strong association between As concentrations ranging 51-99?μg/L and keratosis and melanosis outcomes, although the probability decreases at higher concentration ranges perhaps due to switching away from the use of As-contaminated tube wells for drinking and cooking purposes. High As concentrations in hair, nail and urine were observed to be associated with the age of the study population. The level of As concentrations in hair, nail and urine samples of the study population indicated the degree of severity of As exposure in the study region.     

Studies on oxidative stress induced nerve conduction deficits in cigarette smokers

An important role of oxidative stress for the development of vascular and neurological complications has encouraged us to undertake a study to assess the oxidative stress induced nerve conduction deficits among cigarette smokers. Eighteen regular male cigarette smokers and twenty nine male non-smokers were diagnosed for clinical neuro-physiological tests viz., motor and sensory nerve conduction velocity (MNCV and SNCV) and redox status. Significant depletion of reduced glutathione (GSH) level (p < 0.05) and significant increase in malondialdehyde (MDA) level (p < 0.01) was found in smokers compared to non-smokers. Motor and sensory nerve conduction velocity showed no significant difference among smokers compared to non-smokers. The present study shows that smoking can induce oxidative stress among smokers but could not exacerbate to nerve conduction deficits.     

Strong variability in bacterioplankton abundance and production in central and western Bay of Bengal

With large influx of freshwater that decreases sea-surface salinities, weak wind forcing of <10 m s⁻¹ and almost always warm (>28°C) sea-surface temperature that stratifies and shallows the mixed layer leading to low or no nutrient injections into the surface, primary production in Bay of Bengal is reportedly low. As a consequence, the Bay of Bengal is considered as a region of low biological productivity. Along with many biological parameters, bacterioplankton abundance and production were measured in the Bay of Bengal during post monsoon (September–October 2002) along an open ocean transect, in the central Bay (CB, 88°E) and the other transect in the western Bay (WB). The latter representing the coastal influenced shelf/slope waters. Bacterioplankton abundances (<2 × 10⁹cells l⁻¹) were similar to those reported from the HNLC equatorial Pacific and the highly productive northern Arabian Sea. Yet, the thymidine uptake rates along CB (average of 1.46 pM h⁻¹) and WB (average of 1.40 pM h⁻¹) were less than those from the northwestern Indian Ocean. These abundances and uptake rates were higher than those in the oligotrophic northwestern Sargasso Sea (<7 × 10⁸ cells l⁻¹; av 1.0 pM h⁻¹). Concentrations of chlorophyll a (chl a), primary production rates and total organic carbon (TOC) were also measured for a comparison of heterotrophic and autotrophic production. In the WB, bacterioplankton carbon biomass equaled ∼ 95% of chl a carbon than just 31% in the CB. Average bacterial:primary production (BP:PP) ratios accounted for 29% in the CB and 31% in the WB. This is mainly due to lower primary productivity (PP) in the WB (281 mg C m⁻² d⁻¹) than in the CB (306 mg C m⁻² day⁻¹). This study indicates that bacteria–phytoplankton relationship differs in the open (CB) and coastal waters (WB). Higher abundance and contrastingly low bacterial production (BP) in WB may be because of the riverine bacteria, brought in through discharges, becoming dormant and unable to reproduce in salinities of 28 or more psu. Heterotrophic bacteria appear to utilize in situ DOC rather rapidly and their carbon demand is ∼50% of daily primary production. It is also apparent that allochthonous organic matter, in particular in the western Bay, is important for meeting their carbon demand.     

Gender response and safe carrying load limit for sugar industry workers

This study investigates the safe carrying load limit in relation to gender of workers in the sugar industry. A total of 632 workers (male and female) were interviewed following a physiological and psycho-physiological methodology. All of these workers were in the age groups of 21–30, 31–40 and ≥41 years. In this study, for the physiological methodology, both field and laboratory data were collected, which included heart rate, heart rate variability, energy expenditure and maximum rate of oxygen consumption. Information regarding hemoglobin content, weight, age, lean body weight, etc., was recorded to know the physical health of the workers. In the psycho-physiological evaluation, load-carrying limits were determined by the psychophysical rating of the load by workers from the selected age groups. By considering the physiological stresses and psycho-physiological evaluation, the safe carrying load limit was determined in head and back mode for male and female workers as per their age groups.     

Utilization of anaerobically treated distillery spent wash for production of cellulases under solid-state fermentation

Pollution caused by distillery spent wash on one hand has stimulated the need to develop new technologies to treat the waste and on the other, forced us to reevaluate the efficient utilization of its nutritive potential for production of various high value compounds. In this study, anaerobically treated distillery spent wash was used for the production of cellulases by Aspergillus ellipticus under solid-state fermentation using wheat straw as a substrate. The interactions between distillery effluent concentration, initial pH, moisture content and inoculum size were investigated and modeled using response surface methodology (RSM) involving Box–Behnken design (BBD). Under optimized conditions, filter paper activity, β-glucosidase and endo-β-1,4-glucanase activities were found to be 13.38, 26.68 and 130.92 U/g of substrate respectively. Characterization of endo-β-1,4-glucanase and β-glucosidase was done after partial purification by ammonium sulfate fractionation followed by desalting. The partially purified endo-β-1,4-glucanase and β-glucosidase showed maximum activity at 60 °C. Saccharification studies performed with different lignocellulosic substrates showed that wheat bran was most susceptible to enzymatic hydrolysis. The study suggests that anaerobically treated distillery spent wash can be used as a viable nutrient source for cellulase production under solid-state fermentation by A. ellipticus.     

Metallic components of traffic-induced urban aerosol, their spatial variation, and source apportionment

This study proposes a practical method to estimate elemental composition and distribution in order to attribute source and quantify impacts of aerosol particles at an urban region in Kolkata, India. Twelve-hour total particulates were collected in winter (2005–2006) and analyzed by energy-dispersive X-ray fluorescence technique to determine multi-elemental composition, especially trace metals. The aerosols consist of various elements including K, Ca, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Se, and Pb which exhibit significant concentration at various sites (p?<?0.05). The concentration of different metallic elements were found in the order of Zn ?> ?Pb ?> ?Ni ?> ?Cu ?> ?Cr ?>? Co. Statistical multivariate analysis and correlation matrix analyses were performed for factor identification and consequent source apportionment. Contour profiles demonstrate spatial variation of elemental compositions indicating possible source contribution along with meteorological influences. Spatial differences were clearly most significant for Zn, Ni, Pb, and Cu reflecting the importance of anthropogenic inputs, primarily from automobile sources.