Utilization of Fly Ash as a carrier in Biofertilizer and Biopesticide Formulation of Chandrapura Thermal Power Station, India.
Neha Shreya
Junior Research Fellow, Department of Environmental Science and Engineering, Indian School of Mines, Dhanbad-826004, Jharkhand, India.
E-Mail id: [email protected]
Dr. Biswajit Paul
Associate Professor, Department of Environmental Science and Engineering , Indian School of Mines, Dhanbad-826004, Jharkhand, India.
E-Mail id: [email protected]
ABSTRACT
Bio-formulations with the use of fly ash as a carrier have been found as a very effective, inexpensive, low maintenance and environmental friendly technique. The biofertilizer and biopesticides produced using fly ash as carrier should be considered as a valuable resource for use by various agriculture industries and farmers as India is basically an agriculture based country. Using fly ash as carrier in bioformulations could mitigate the environmental crisis of fly ash disposal to some extent by increasing its utilization. This paper assesses the performance of fly ash generated from Chandrapura Thermal Power Station (CTPS) area of Bokaro District by examining the physico-chemical properties. The fly ash of this region has a potential for growth of leguminous plant in various trials. In our experiment fly ash alone and in combination with lignite and talc was tested to explore its possible use as carrier for Azotobacter chroococcum (nitrogen fixing bacteria) and Trichoderma viride (opportunistic avirulent plant symbionts) formulations. The objective of the present study was to develop fly-ash based bio-formulation of selected microbes and to evaluate shelf-life of these bio-formulations through cfu count and statistical analysis. The result showed the decline in cfu count in all the three carrier combination in both the Azotobacter chroococcum and Trichoderma viride and the shelf life of both the formulations was found to be better at 28±2 0 C when compared to room temperature. Fly-ash based bio-fertilizer and bio-pesticide formulations showed slightly better results compared to routine talc or lignite formulations. This paper includes an outline of social and environmental reasons for solid waste utilization, and explores the idea of use of hazardous waste as an environmental friendly biofertilizers and biopesticides.
Key words: Bioformulations, fly ash, talc, charcoal, Azotobacter chroococcum and Trichoderma viride, ANOVA test.
1. Introduction
methods. Physically Fly Ash occurs as very fine particles, having an average diameter of <10mm, low to medium bulk density, high surface area and very light texture. Chemically the composition of Fly Ash varies depending on the quality of coal used and the operating conditions of the Thermal Power Stations. The utilization of Fly ash in India has increased substantially in recent years from 13 million tons to 51 million tons. The future poses challenge to the scientist & technologists, engineers towards sound management of Fly ash. The technical know-how is available in the country. A beginning has been made towards enhanced facilitation for adoption/implementation of better management practices/appropriate technologies towards safe utilization of Fly ash. In spite of an all-out effort, the Fly ash utilization in the country is still very low (<15%).There are several issues responsible for low utilization of Fly-ash, including lack of awareness, confidence and regulation ,non availability of dry ash and most important of all, easy availability of land with topsoil at cheap rates. The study area ‘CTPS’ is situated at Chandrapura, Bokaro district, Jharkhand state, India at 23°45’N and 86°07’E. The power station has a production capacity of 780Mw electricity with a coal consumption of nearly 8000 tons/day. The total amount of ash generated /annum is approximately 15 lakh tons .The emitted fly-ash spread over a large area around the power plant and affects the villagers residing in that locality. As a consequence, a large amount of Fly ash is generated in thermal power plants, and is disposed off in unmanaged landfills, lagoons and ponds. Fly ash disposal in an unscientific way affects the local ecosystems due to the heavy metal pollution through erosion and leachate generation (Sikka and Kansal, 1995). Dumped Fly ash contaminates surface and groundwater, soils and vegetation by mobilization of its hazardous metals. . The other potential adverse effects of fly-ash are leaching of potentially toxic substances from fly-ash into soils and groundwater, changes in plant elemental composition, increased cycling of these toxic elements through food chain and respiratory problems to human beings. Apart from that it causes some chronic occupational diseases like Silicosis (chronic cough and pain in the chest) and Pneumoconiosis or black lung disease (lung problems).
Thus the justification of current study was utilization of solid waste material fly ash as a carrier material in bioformulations of bio-fertilizer and bio-pesticides in an economical manner to mitigate the environmental crisis of fly ash disposal by increasing its utilization as well as improvement of various qualities of soil than chemical fertilizers. Thus it serves as social and environmental reasons for solid and hazardous waste utilization in an environmental friendly manner.
1.1 BOI-FERTILIZERS AND BIO-PESTICIDES:
Intensive agriculture is inevitable in India because of the limited arable land and the high population. Hence, agricultural researches and technologies aimed to increase high quality yields until the 1980s. Since mid-1990s, to ensure sustainable agriculture and sound environment and also aimed to reduce 40%of chemical fertilizers and pesticides produced in India, bio-fertilizers and bio-pesticides have become ideal substitutes for environment-friendly agriculture in India. (Muraleedharan H. , 2010). Biofertilizer “are defined as a substance which contains living micro-organisms which colonizes the rhizosphere or interior of the plant and promotes growth by increasing the supply and availability of primary nutrients and/or growth stimulus to the target crop, when applied to seed, plant surfaces and soil. The purposes of the bio-fertilizers are soil fertilization and conditioning, and increment in plant growth and health. The most commonly used microbial agents are Bacillus spp. Rhizobium spp., Pseudomonas spp., Bradyrhizobium sp., and Azospirillum sp. Basically “Biopesticides" are naturally occurring substances (biochemical pesticides) that control pests, microorganisms that control pests (microbial pesticides), and pesticidal substances produced by plants containing added genetic material, plant-incorporated protectants. The microbial agents include Paenibacillus polymyxa, Bacillus subtilis, Streptomyces goshikiensis, and Bacillus thuringiensis subsp. : Trichoderma viride & Trichoderma harzianum
1.1.1 Azotobater chroococcum
Mode of action
It produces and releases in soil root zone, biologically active metabolites like Iodole acetic acid (IAA), Gibberellins (GA), Vitamin-B and cytokines. It partially solubilises tricalcium phosphate and calcium phosphate thus increasing uptake of phosphorus in plants. Antibiotics, HON, lytic enzymes produced by Azotobacter are directly involved in the reduction of plant pathogens.
1.1.2 Trichoderma viride:
It’s a the potential antagonistic fungus which prevents the crops from diseases viz. Root rots, Wilts, brown rot, damping off, Charcoal rot and other soil born diseases in crops. It is suitable for: Sugarcane, Pulses, Oilseeds, Cotton, Vegetables, Banana and Coconut etc.
Mode of action:
The fungus secrets cellulase and chitinase enzymes which react with cell wall of the disease causative pathogenic fungi and bacteria and dissolve the same.Trichoderma utilizes the protoplasm as a source of food and multiply its spores .By this method the spores of the pathogenic fungi are destroyed. It destroys the fungal pathogens by secreting toxic substances as Glyotoxin, Viridin and Trichodermin.
To reduce input of chemical fertilizers and pesticides yet sustain the productivity of the agro-ecosystem, bio-fertilizers and bio-pesticides have become ideal substitutes for environment-friendly agriculture in India. At present, a total of 138 companies produce hundreds of commercial products, and 23 bio-pesticides are now registered in the country. Among the registered bio-pesticides, 12 are fungicides and the others are insecticides.
2 Aims and Objectives
The objective of this study was the development of bio-formulation of selected Bio- fertilizer and Bio-pesticide in fly-ash and shelf-life studies of fly-ash based bio-fertilizers and bio-pesticides at two different temperatures. Along with this the isolation and identification of microbial strain for bio-formulations and laboratory analysis of physico- chemical properties of fly-ash (to be used) in reference to toxic elements and heavy metals was done.
3 Materials and Methods
3.1 Fly Ash
Huge amounts of fly-ash are generated as solid waste material from thermal power stations. This fly ash gets deposited on soil over a large area in thermal power stations. The fly ash samples are collected and analyzed for toxic elements and compared with unpolluted soil samples having same geochemical properties. The current study was to find out the impacts of fly ash around Chandrapura thermal power station of Damodar Valley Corporation, Jharkhand and to study their potential as a carrier in bio-pesticide and bio-fertilizer formulation. Sampling was done over a period of 2 days from the Chandrapura thermal power station.
3.2 Study
December-January. It is as low as 70 to 90. The monsoon season start in the early June and continue till September. The experiences 80% to 85% annual rain fall. Relative humidity of the area has maximum of 87% and minimum of 55%. In view of deposition of such a huge amount of waste fly ash over a large are, it was felt to study its characteristics and to make the most of it in any beneficial manner like use it as a carrier material in bio-pesticide and bio-fertilizer formulation or in other agricultural and commercial purposes.
3.3 Laboratory Analyses
Various physical and chemical tests on fly-ash were performed in the laboratory like bulk density, moisture content, water holding capacity , cation exchange capacity and E.C using standard laboratory protocols . Along with this pH was determined by by pH in soil to water ratio of 1:2.5 ,organic carbon by rapid dichromate oxidation technique, determination of Mn, Cu and Zn by HNO 3 and HClO 4 Digestion method, sulfate estimation by Turbidimetric method , Estimation of Ca & Mg by EDTA titrimetry method ,determination of N by Kjeldahl method, exchangeable potassium by centrifugation and decantation procedure determination of available phosphorus by bray’s method and heavy metal
detection by DTPA extractable method (Maiti S.K. , 2003).
Fig.1. Map of Bokaro district indicating Chandrapura Thermal Power Station (Gond et al, 2009) 3.4 Isolation of Azotobacter Choroococcum and Trichoderma Viride
Azotobacter choroococcum and Trichoderma viride were isolated from Rhizosphere by Soil Dilution and Plate Count Method (Aneja, K.R., 2002).
3.5 Identification and Purification of Cultures
Isolated cultures thus obtained were identified on the basis of colony characteristics; microscopic examinations and biochemical analysis by using standard manual papers were consulted. These identified strains were obtained on their respective medium in slants or in petriplates at 40C with periodic sub-culturing at fortnightly interval.
3.6 Multiplication of Microbial Strains
3.7 Development of Bio-Formulation in Fly as: In Carrier
The fine neutralized fly ash were packaged in autoclavable propylene polybags and sterilized in an autoclave for 4 hours continuously at 1210C or 15 psi.
3.8 Blending
Broth inoculums of effective strains were blended with fine neutralized and sterilized fly ash @ 10-15% in bio-pesticides and @ 20-25% in bio-fertilizers.
3.9 Packaging and Storage
Fig.2: Packaging of bio-fertilizers and bio-pesticides
The formulations were packed in low density pouch and storage at 2 different temperatures 28±0C and at normal room temperature.
3.10 Shelf-Life Studies
Shelf-life off selected bio-pesticides and bio-fertilizers were tested following serial dilutions pour plate technique and Colony forming unit per gram will be estimated at regular intervals (15 days interval).
Fig.3: Testing shelf-life of T. viride and A. chroococcum 3.11 Statistical Analysis
4. Results and Discussion
4.1 Table-1 Physico- chemical and available heavy metal analysis of fly ash and
1.10 P (mg/kg) 0.004±
From the calculation of all physico –chemical tests of fly-ash it has been deduced that the fly-ash holds a potential for growing some leguminous plants on fly-ash of CTPS and hence an effective utilization of this fly-ash.
4.3 Identification of Azotobacter chroococcum:
The gram stain and biochemical test results were analyzed upon completion in order to identify distinguishing characteristics of each unknown bacterial isolates. The test isolates were found positive when further examined for their biochemical properties for Iodole, Methyl red and Vogus proskauer (MRVP) test, citrate test, catalase test and Oxidase tests .Based on the colony characters, cell shape, presence of cyst, capsule, gram reaction, and utilization of different carbohydrates tested and biochemical tests, the isolates were confirmed as A. chroococcum isolates.
4.4 Identification of Trichoderma viride:
The test isolates were found positive when further examined for their biochemical properties for carbohydrate fermentation ,carbohydrate assimilation, API 20C system, Uni-Yeast Tek system ,urea hydrolysis and nitrate utilization tests .Based on the their reaction, colour appearance, turbidity, ability to ferment sugars, urea hydrolysis and utilization of different carbohydrates and nitrates tested , the isolates were confirmed as T. viride isolates.
4.5 cfu count
factors) was observed at 30 days of incubation whereas the least cfu count (02 at 107 dilution factor) was noticed after120 days of incubation.
Graph –1. Showing comparative study of bio-efficacy of fly-ash vs charcoal as formulation for bio-fertilizer at room temperature
The cfu count of the Azotobacter chroococcum formulation at 28±20C was seen to be slightly increases from 0 days to 30 days. As the data obtained from graph-2 the maximum cfu count (48 at 107 dilution factors) was observed at 30 days of incubation whereas the least cfu count (04 at 107 dilution factor) was noticed after120 days of incubation.
Graph – 2 showing comparative study of bio-efficacy of fly-ash vs charcoal as formulation for bio-fertilizer at 28±20C
The cfu count of the formulation of Trichoderma viride at room temperature was found to be slightly increases from 0 days to 30 days. As the data obtained from the graph -3, the maximum cfu count (34 at 107 dilution factor) was observed at 30 days of incubation whereas the least cfu count (02 at 107 dilution factor) was noticed after120 days of incubation.
The cfu count of the formulation of Trichoderma viride at 28±20C was found to be slightly increases from 0 days to 30 days. As the data obtained from graph -4, the maximum cfu count (33 at 107 dilution factor) was observed at 30 days of incubation whereas the least cfu count (04 at 107 dilution factor) was noticed after120 days of incubation.
Graph – 4 showing comparative study of bio-efficacy of fly-ash vs talc as formulation for bio-pesticide at 28±20C
5. STATISTICAL ANALYSIS BY TWO WAY ANALYSIS OF VARIANCE (ANOVA)
The statistical analysis by comparative 2 Way ANOVA Analysis of fly-ash vs charcoal at room temperature and at 28±2 0 C and fly-ash vs talc at room temperature and at 28±2 0 C
Table -2. TWO Way ANOVA Analysis of fly-ash vs charcoal at room temperature and at 28±2 0 C and fly-ash vs talc at
room temperature and at 28±2 0 C
Rows 3254.111 8 406.7639 480.1148 6.5E-10 3.438101
Columns 6.722222 1 6.722222 7.934426 0.022605 5.317655
Error 6.777778 8 0.847222
Total 3267.611 17
By comparing the values obtained and by analyzing the graph 1 and 2 and 3 and 4 respectively we can conclude that the bio-efficacy of fly-ash is much better at 28±2 0 C for both bio-fertilizer and bio-pesticide than at room temperature. The statistical analysis of data indicates that the shelf life of Azotobacter chroococcum and Trichoderma viride was found to be significant in all treatments over time. Significant decline in cfu/g might be due to decreased viability of propagules and loss of moisture content during passage of time. The population of Azotobacter chroococcum and Trichoderma viride were retained in all the cfu/g formulations, through a significant first increase and then decline in cfu/ g was noticed up to 120 days from zero day stage at 28±2 0 C and up to 120 days at room temperature. Similar results on shelf-life (stored at 28±2 0 C) up to one-year were reported by (Jayaraj et al 1993).
From table-2 the statistical analysis by comparative 2 Way ANOVA Analysis of fly-ash vs charcoal at room temperature and at 28±2 0 C and fly-ash vs talc at room temperature and at 28±20C reveals that the critical value obtained is (F critical=5.317655) shows greater value when compared with the tabular value. By this analysis we can conclude that fly-ash is a better carrier for bio-fertilizer and bio-pesticide formulation than charcoal and talc.
Since the value of fly-ash ( sample) obtained is much nearer to the values of charcoal(control) taken in graph 1 and 2 the value of fly-ash (sample) and talc (control) in graph 3 and 4 respectively obtained is much nearer to the values of charcoal which has been used as a carrier material in formulations of many bio-fertilizers (Kumar and Gupta, 2010) ) and (Kumar et al .2010) and (Narayanasamy, 1994) and talc alone and in combination with fly-ash and other materials have been used as a
carrier material in formulations of many bio- pesticides (Kumar and Gupta, 2008), (Karunanidhi et al, 2011) and (Mythukumar , 2009) also fly-ash being cheaper than charcoal and talc can be used as a carrier in fertilizer and bio-pesticide formulation. Thus from the present study Azotobacter chroococcum and Trichoderma viride formulations showed a better result than lignite and talc respectively and thus can be used as an environmental friendly carrier and a source for another green revolution.
Fly ash holds a potential to improve the physical health of the soil. It can serve as a soil modifier and also enhance the water retaining capacity and fertility of the soil. It improves the plants water and nutrient uptake; helps in development of roots and soil binding, stores carbohydrates and oils for use when needed, protects the soil from soil borne diseases and detoxifies contaminated soil. Use of Fly ash in agriculture can increase the yield of cereals, oil seeds, pulses, cotton and sugarcane by 10-15%, vegetables by about 20-25% and root vegetables by 30-40%. Waste lands, degraded lands, saline alkaline soils, eroded soils etc., can be successfully reclaimed by fly ash.
Fly ash alone and in combination with lignite and talc was tested to explore its possible use as carrier for Azotobacter chroococcum and Trichoderma viride formulations. Fly-ash being generated in large quantity in thermal power stations is
generally recognized as waste and an environmental hazard. However, it has been reported to promote crop growth in various trials conducted under National Fly-ash Utilization Programme. The objective of the present study was to develop fly-ash based bio-formulation of selected microbes and to evaluate shelf-life of these formulations. Bio formulations of Azotobacter chroococcum and Trichoderma viride were obtained by mixing both cultures culture with fly-ash and
lignite/talc, respectively. A general decline in cfu count was noticed in all the three carrier combination in both the Azotobacter chroococcum and Trichoderma viride. Fly-ash based bio-fertilizer and bio-pesticide formulations showed
slightly better results compared to routine talc or lignite formulations. The same process can be used for the bioformulations of different microbial species like Bauveria bassiana , Rhizobium Leguminosarum etc.using fly ash as a carrier material.
Acknowledgement
Authors are grateful to Dr. Vipin Kumar, Assistant Professor, Dept. of Environmental science & Engg., Indian School of Mines , Dhanbad for his help and valuable suggestions leading to this research and our findings.
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