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    Biotechnological Approaches To Conservation And Improvement Of Biodiversity In Rajasthan
    By Ashwani Kumar | November 2nd 2009 05:22 PM | 15 comments | Print | E-mail | Track Comments
    About Ashwani

    Professor Emeritus ,Former Head of the Department of Botany, and Director Life Sciences, University of Rajasthan, Jaipur. 302004, India At present...

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    Arid and semi-arid regions :

    Arid and semi arid lands occupy one third of the earth's surface. Indian arid zone occupies an area of about 0.3 million sq. km. 90 percent of which about 2,70,000 sq. km.) is confined to north west Indian covering most of Western Rajasthan, part of Gujrat and small portions of Punjab and Haryana. India with its vast expanse of wasteland unsuitable for agricultural production (nearly 180 million ha) could be considered for economically viable production of bio-fuels, medical and conservation and improvement of biodiversity work on bio-fuels was initiated around 20 years ago. Several of the plant diversely available in Rajasthan is endemic and of high medicinal value. Some of the plant species on threatened.

    Bio-diesel production :

    A recent World Bank report concluded that "Energy policies will need to be as concerned about the supply and use of bio-fuels as they are about modern fuels. (and) they must support ways to use bio-fuels more efficiently and in sustainable manner Although there is significant volume of bio-diesel already produced in Europe there are remaining risks showing down the further expansion to the target set by the European Commission to reach 5% market share in fuels by the year 2000.

    Global scenario :

    Out of 2,50,000 plant species only 10,000 or so have been exploited. At the global level, according to recent estimates by FAO the annual tropical deforestation rate for the decade 1981 to 1990 was about 15.4 million ha (Mha).


    According to the latest data published in 1994, for the assessment period 1989-1991, the total area under forests in 64.01 Mha accounting for 19.5 percent of India's geographic area.

    At present there is hardly 0.4 percent forest below 25 cm rainfall zone and 1.3 percent above 30 cm rainfall zone. There is rapid depletion of forest products and in order to provide alternative energy sources a change in needed in conventional forestry management.

    The fact that nearly 90 percent of the worlds population will reside in developing countries by about 2050 probably implies that biomass energy will be with us forever unless there are drastic changes in the world energy trading pattern. However if these plants could find additional use in medicines also besides providing the bio fuel, the entire economics of biofuel production shall become viable proposition in different parts of the globe.

    India with its varied climate soil and agro-ecology possesses immense plant diversity, with over 15,000 species of higher plants. Both our Indian civilization as well as our diverse tribal heritage have gone a long way in conserving the wild weedy species, native land races and primitive cultivars 9Fig. 1). The Indian gene center is endowed with rich flora especially with regard to several less known yet economically important plants, ca 160 cultivar species of economic plants, plus 56 species of lesser known cultivated food plants. Further there are ca 320 species of wild and weedy economic types 9Paroda 1979; Arora and Nayar, 1984; Kumar, 1998).

    Utilization of biomass for bioenergy and medicine :

    Use of biomass for energy, medicine and industry allows a significant quantity of hydrocarbons to be consumed without increasing the CO2 content of the atmosphere and thus makes a positive contribution to the Greenhouse effect and to the problems of "global change" as occurs in both industrialized and developing countries. Further the advantages from utilization of biomass include : liquid fuels produced from biomass contain so sulfur, thus avoiding SO2 emissions and also reducing emission of NOx. Improved agronomic practices well managed biomass plantations will also provide a basis for environmental improvement by helping to stabilize certain soils, avoiding desertification which is already occurring rapidly in tropical countries, Modern bioenergy technologies and biofuels are relatively benign from environmental view point and produce very little pollution if burned correctly and completely.

    Natural resource :

    A detailed survey on the weeds on wastelands yielded valuable data about the first colonizers. Total land area of Rajasthan is 3,42,239 sq. km out of which 45.25 percent is characterized as wasteland. Large portions of this land were productive at a given time and due to man made deforestation, cattle pressure, water and wind based soil erosion, improper water management, they have turned out to be wastelands. (Kotia and Kumar, 2001a). A large number of such weeds have the medicinal value. Out of the total weeds around 50 weeds having important medicinal values while other produce related compounds. These regions are rich in bio-diversity and weeds were collected from different regions of the developing wastelands. (Kotia and Kumar, 2001b)



    A detailed survey was carried out in different parts of Rajasthan and some of the medicinally important plants of Rajasthan and listed by Ajanta and Kumar, (2001a). Some of the Medicinal plants found in wild in the forest of Rajasthan include :



    Table 1. List of Medicinal plants of Rajasthan



    Plants species : Local name

    1. Asparagus racemosus satavari

    2. chlorophytum arundinaceum Safed musli

    3. Curculigo orchioides Kali Musali

    4. Solanum surattense Kantkari

    5. Boerhaavia diffusa Santhi,

    6. Hamidesmus indicus anantmool

    7. Sida cordifolia bala

    8. Holarrhena antidysenterica Indrajo

    9. Curcuma aromatica Vanhaldi

    10. Oroxylum indicum Shyonaka

    11. Balanites aegyptiaca hingot

    12. Withania somnifera ashwagandha

    13. Aegle marmelos Bael

    14. Cassia fistula Amaltas

    15. Gymnema sysvestre gudmar

    16. Terminalia arjuna arjuna

    17. Butea monosperma palas

    18. Soymida febrifuga rohan

    19. Woodfordia fruticosa dhavri

    20. Tribulus terrestris gokhru

    21. Pedalium murex badagokhru

    22. Vitex negundo negad

    23. Dyerophytum indicum chitral

    24. Plumbago zeylanicum Chitrak

    25. Plantago ovata Isabgol

    26. Colocynthes vulgaris Indrayan

    27. Adhathoda vasica ardusta

    28. Allangium salvifolium aankol

    29. Caesalpinnia bonducella tas

    30. Jatropha curcas ratanjot

    31. Eclipta alba bhringraj

    32. Aloe barbadensis gwarpatha

    33. Mucuna prutita Konch

    34. Terminalia bellerica Baheda

    35. Tamarindus indica imli

    36. Azadirachta indica neem

    37. Achyranthes aspera aandhijhara

    38. Barleria cacrulea bajrandanti s

    39. Barleria cristata Badradanti p

    40. Barleria prinoitis bajradanti p.

    41. Ocimum americanum bapchii

    42. Centella asiatica brahmiduti

    43. Datura metal Dhatura

    44. convolvulus arvensis haranpadi

    45. Evolvulus alsinoides shankhpushpi

    46. Cassia occidentalis kasaundi

    47. Urginea indica Kolikanda

    48. Andrographis paniculata kalmegh

    49. Helicteres ispara marophali

    50. Tinospora cordifolia nimgiloy.



    In addition to this some of the laticiferous with potential for bioenergy and ayurvedic use include the followings :

    (I) Hydrocarbon yielding plants :

    1. Euphorbia lathyris Linn.

    2. Euphorbia tirucalli. Linn.

    3. Euphorbia antisyphilitica Zucc.

    4. Euphorbia caducifolia Haines.

    5. Euphorbia neriifolia Linn.

    6. Pedilanthus tithymalides Linn.

    7. Calotropis procera (Ait.) R.Br.

    8. Calotropis gigantea (Linn) R.Br.



    (II) High molecular weight hydrocarbon yielding plants :

    1. Parthenium argentatum Linn.



    III) Non edible oil yielding plants :

    Jatropha curcas.

    Simmondsia chinenesis



    IV) Short rotation energy plants :

    Acacia tortilis; Holoptelia integrifolia; Parkinsonia aculeata; Cassia siamea; Al'bizzia lebbek; Acacia nilotica; Tecomella undulata; Prosopis juliflora; Pithocellobium



    A large number of energy yielding desertic plants of India used in Ayurvedic system have great potential as Ayurvedic medicine. Negative environmental effects of current agricultural practices, such as emission of greenhouse gases, nutrient leaching, decreased soil fertility, and erosion, may be reduced when traditional annual food crops are replaced by dedicated perennial energy crops and medicinal plants. As they are able to grow and produce valuable products under dessert conditions they have great potential for covering the global desert areas into green belts leading to environmental improvement on one hand and providing valuable Ayurvedic crude drugs in addition to supplementing the bio-energy resources as renewable fuels. However detailed studies on their pharmacognostical characterization and determination of chemical products obtained from them are lacking. Some of the investigations indicated their potential use in Human immuno deficiency (HIV) diseases 9Hattori et al 19950. Such bio-energy plants have not been explored in depth. Here attempt shall be made to provide brief out look of the Indian scene and highlight some of the work being carried out at our place in Rajasthan along with the possible impact assessment for desertic plants for future research strategies.



    Among the desert plants the value of Aloe vera (L.) was recognized more than 3000 years ago when the Egyptian and Greek civilizations used its extract for skin burns, cut and wounds on the skin surface and found that it had a wonderful healing effects on skin. It is claimed that even 3rd degree burn can be cured and healed by Aloe vera. The chemical compounds like Aloein, resins, mixture of polysaccharides containing pectic acid are present. Modern investigation indicate that extracts of Aloe vera act on the dead epithelial cells of skin, aiding their removal from the surface and stimulating the growth of new cells. Thus Aloe is providing to be a great gift of traditional medicine for protecting the smooth skin of human beings especially when radiation damage of human skin has assumed alarming situation due to stratospheric ozone depletion. Fresh juice of leaves are also used in liver and spleen troubles and also for eye troubles, found useful in X-ray burns, dermatitis, coetaneous and other skin disorders.



    In India, Egypt and Sudan around 70 percent of the rural people use traditional medicine. Similar situation exists in a large number of developing countries. In India and China 60 percent of the people affected with cholera and malaria are treated with herbal medicines. In these countries the market for traditional medicines is US $ 500 million while Western type medicine account for only US $ 300. In Singapore 50 percent and in Australia 60 percent of population uses alternative medicine. Around 17,000 herbal products are registered in these countries. In Belgium 40 percent contemporary and 84 percent home medicines and 74 percent acupuncture medicine is utilized. In France 50 percent of the people take advantage of complementary medicine. In Germany 10,000 to 13,000 alternative medical practitioners are thriving well and 75 percent o[f them utilize complementary medicines. 77 percent of pain clinics utilize acupuncture. In UK 90 percent of complementary medical practitioners utilize osteopathy and acupuncture. In US where in 1990 only 30 percent of the people were utilizing complementary medicines in 1997 it grew to 40 percent.



    Extant of natural habitats and vegetation

    Biodiversity characterised&ecological . Complete list of the plant cover is required. These in need for their conservation to maintain essential ecological processes and life support system. It is important contribution to social or economic development. The objective of this study -

    1. To enumerate the plant species adilleir distribute pattern including medicinal plant and their present status.

    2. of stress resistant plant in through genetic manipulating if possible.

    3. Abiolic factors responsible for their loss in habitat.

    4. Antropogenic impact on aminated + vegetation.

    5. Regeneration of vegetation through conservation strategies



    Biotechnological approaches for conservation and improvement of biodiversity in the arid and semi arid region of Rajasthan state :



    India is one of the 12 mega diversity centres in the world. Loss of biologically jila habitats in the main threat to the diversity in developing tropical countries. It is almaring to note that more than 50% of the national habitats are already destroyes in 45 out of 61 old world tropical countries. In India the route destruction recorded 80%. Excessive exploitation and habitat modification by human beings are readily recognised as major factors causing loss of biodiversity large industrial and commercial activities associated with global economy such as mining and dam construction with the object of immediate gains resulted in the depletion to a large improvement of economically important tree species of the arid and semi-arid regions of Rajasthan.



    Origin of proposal : The diversity and extent of forests in the arid and semi-arid regions of Rajasthan is declining and yet the demand for wood world-wife, is expected to double by the century. The demand of forest resource is exceptional in India where the crisis created by explosion in population, increased agricultural activities and urbanisation have created a situation for rapid deforestation leading to degradation of land and destruction of habitats of several plant species. Plant biotechnology is playing a crucial role in ex-situ conservation of endangered species. In vitro propagation now enables a board range of species and potentially all species of higher plants to be cloned under highly controlled conditions. The natural regeneration of many plant species is limited by poor seed germination and lack of proper method for vegetative propagation.



    The Indian Thar desert and the neighbouring Aravallis are two ecologically fragile systems of Rajasthan which harbour germplasm of several tree species. Some of the tree species which have been under pressure of indiscriminate exploitation include Acacia senegal, Zizyphus xylopyrus, Acacia catechu, Commiphora wightii, Prosopis cineraria, Wrightia tirctoria, Wrightia tomentosa, Emblica officinalis and several others.



    Plan of Work :

    Basic protocol for micropropagation of some of these species has been worked out in our laboratory viz. Acacia senegal, Acacia catechu, Zizyphus sps., Acacia leucophlora. It is proposed to follow the basic protocol in the following phases for further studies :

    Phase 0 : Management of donar plant in field

    Phase 1 : Sterile isolation of explant and establishment of cultures

    Phase 2 : Shoot multiplication and elongation

    Phase 3 : Rooting of in vitro raised shoots

    Phase 4 : Hardening, acclimatization and transfer of plantlets in soil

    Phase 5 : Testing of plantlets under greenhouse / field conditions



    The protocols developed in laboratory must be tested at pilot scale to ascertain their viability. It is important that the plants produced in vitro should be successfully established in soil and their performance evaluated in comparison to conventionally raised plants for cost-benefit analysis and agronomic traits. Hence the protocol developed must be highly responsible and commercially feasible.



    IMPORTANCE&EXPECTED OUTCOME OF THE PROPOSAL

    The present project proposal aims to concentrate on various aspects of scaling-up of protocols established for micropropagation of Acacia senegal, Acacia Ccatechu and Zizyphus sps. The growth performance of plants transferred in greenhouse and fields will be evaluated for their success. The investigation will help to standardize the condition leading to scaling-up production and their evaluation at various stages. It will establish a highly reproducible and commercially feasible protocol keeping in view the cost factor, for rapid multiplication for conservation and improvement of these economically important trees of the arid and semi-arid regions of Rajasthan.



    1. Moder biotechnological tools will be developed and utilized in the study for the Isolation of nature Rhizobial isolates from the tree legunies, VAM fungi, phosphate solubilizing lacteria which are efficient against the extremes of environmental factors. Molecular characterisation of each of the above isolates, wing PCR amplification of 16 Sr. DNA gene techniques will also be carried out Development of biomoculants (Rhizobium, VAM fungi and phosphate solubilizers) their mass multiplication and selection of suitable carriers for prolonged establishment in the soil and providing tolerance to tree legumers against environmental extremes.



    2. Development and utilization of modern bio-technology in relation to VAM fungi and associative and free living N2 fixing bacteria (Azospirillum, Azotobacter, Derkia etc.) for increased production of millets in arid and semi-arid regions of Rajasthan. Under this research plan screening, identification, molecular characterization, selection and next multiplication of efficient VAM fungi and free living associative N2 fixing bacteria will be taken up. Development of carrier based bioinoculants for their establishment in soil, necessary for sustainable production of millets.



    3. Application of plant growth promoting rhizobacteria (PGPRof important tree legumes and millets, their molecular characterization and their role in enhancement of growth, nodulation and N2 fixation.



    Physiological biotechnological rhidisors medicinal

    Details of research profile /plans for next 5 yrs.

    Phytochemistry & Bioactives

    1. To study metabolomics, specifically metabolites profiling, where the activity would be concerned with the investigation of the chemical processes especially non-peptide metabolite molecules occurring in living cells, tissue of plant at a particular physiological state.



    2. The use of plant cell cultures to study natural products biosynthesis & the isolation of norel products with emphasis on bioactivities, high level production by biotransformation, and stimulation of growth and yield of end angered species.

    Comments

    I guess the meaning of your article was that there will not be just one answer to every question. It will be a big disappointment in the western world, where a few buzz words and some hyper activity are supposed to fix everything.

    You have given some of the answers that I need to have for my PhD oral exams about 25 years ago.

    The key points were that the future will have a large variety of sources for energy, and much of it will be co-generated with other products.

    I really wonder what type of public policy can accomplish energy independence.

    It seems like a a free market would never accomplish it, but a regulated market might.

    Ashwani Kumar
    Energy independence using variety of energy sources would also include wood fuel, agricultural waste and municipal waste wind energy solar energy and hydrocarbon from plants. Around 90 percent of energy used comes from solar energy directly or indirectly. Efforts are to be directed for making a Global inventory of facts, procedures and options to save the mankind from total disaster of climate and energy supply depletion. I am only beginning to learn how complex the problem is globally in different agroclimatic zones having varied technological advancements.
    I am encouraged to think more as energy solutions are neither  streight nor simple.
    The products you named are special items of limited supply, with price structures that cannot easily be controlled from Chicago or New York. The economics looks like something that will remain regional and under capitalized, but locally important to the economy. Cash flow and reverse flow of cash has a big impact on the regions where it occurs.

    The quality of life is greatly affected over large areas of the country.

    QOL is something we don't talk about much any more, but in a democratic type of country, the quality of life for a majority of people is important.

    Science should be talking about QOL.

    Ashwani Kumar
    Quality of life is not only metrial but the  environmental parameters determine quality of life also and alas there are vast climatic differences and developmental parameters. Efforts are needed to go near to nature to resume from where we started - among woods among the trees among the nature cure among the pollution free world.
    Amaranth and related plants like Celosia greatly improve the quality of life in many regions of the world, but are ignored or resisted in other regions due to ignorance or inertia.

    In North America the plants are treated as weeds, and disposed of as waste, while food prices rise, and many people have an unhealthy diet.

    Amaranth plants offer a variety of benefits for food and fuel. They yield an enormous increase of small seeds 200-700 kg per hectare that are sometimes used as cereal grain.

    From my own experience with native varieties, the plants grow well on dry land, seeding themselves in dense stands without plowing the ground. They produce many crops per year. Sometimes maturity occurs in as little time as two weeks, with no fertilizer or pesticides. A lot of energy is expended in trying to eradicate them, and some varieties are now immune to herbicides.

    The entire plant is edible as a vegetable, and it can also be dried and used as fuel.

    Education would be very useful in some regions about Amaranth. In other places where knowledge is not lacking, there is need for a community program to overcome objections.

    In farms near my home, none of the farmers would consider or even listen to a suggestion of planting Amaranth, because of the community opinion, although the yield and environmental factors would be vastly superior to traditional crops.

    So when we speak about quality of life, the education and community acceptance are sometimes a missing part.

    Ashwani Kumar
    Community acceptance of biogas in India has not taken place and many people detest the idea of cooking in biogas coming from cow dung and insteast prefer to burn the valuable organic matter thus breaking the carbon cycle and depleting the soils of the organic supply. Educating masses about environment globally is very important and community acceptance can come from spreading knowledge.
    Purified methane is the fuel produced from my most recent biogas project. Everything else went into four different types of fertilizer. That is a higher level of science than we are discussing for low technology communities.

    The customs of India are beyond my abilities to change. So are the customs of my home region. That's a big part of the problem.

    Science must tell what the possibilities are, well enough that local leadership can choose which ones to support.

    Biogas can be made from vegetation leaving out the dung, but then the technology level is moving more into the category of fermentation and biological digestion.

    Right now the biological digestion is becoming economical based on bacterial that live inside of cattle to convert cellulose into glucose as a first step for making ethanol fuel.

    It would be possible to make the second step produce methane from bacterial activity on glucose instead of ethanol from yeast activity. This is the biogas from vegetation, with or without dung.

    Ashwani Kumar
    This is significant comment and could you please workout the costs of such installations for a village level of a village of 100 or 200 houses as India has dentralized village system
    Cost it first consideration for the rural poor of India and if this process is cost effective It will be great breakthrough in technology but from lab to land transformation of technology is need of the hour.
    To answer your question, the bacterial technology is new and still under development, but already installed in one full sized factory, and as an auxiliary unit in a small number of ethanol factories.

    That will change in the next 3 years as the demonstration factories provide reliable data. Within 5 years the technology will be well developed and the economics will be integrated into the larger economy.

    At this time the costing is controlled by technology suppliers to make competition with corn based ethanol, giving about 20% discount from corn dry milling economics. This will change because a great many small companies are beginning to offer alternative technology, in a situation where the cellulose fiber is valued at $118 per metric ton compared to $167 for corn

    This is not a stable long lasting situation because the corn has twice as much nutritional energy as fiber per ton in cattle feeding. So there should be an increase of corn price and a constant fiber price to drive the new technology for bacterial decomposition, as the world economy recovers from recession.

    For your question the equipment for bacterial decomposition is exactly the same as for biogas from dung, except the production rates are slower, causing the equipment to be at least 3 times as large for the operation without dung.

    The interesting aspect is the bacteria are derived from cattle that are local to the community, and somewhat familiar to the residents. so a new species is not creating biohazards.

    In considering the equipment size compared to a cow, the cow is more efficient on time and space, because of the long time of experience, climate control, and supplemental nutrients. It gives some hope that the technology will improve for how to nourish the bacteria and promote efficient growth in ways the cow does.

    With advance in technology it is expected that the bacterial decomposition will specialize in fibers that are not attractive to cattle, such that the competition for food or fuel will not occur. To reach that point the testing will pass through a program of gradual substitution and conditioning of the bacteria.

    You chose the best topic in your question because it is on the leading edge of technology and economics at this moment, and the development is just now reaching the point of commercialization. It occupies much of my time.

    From all of this you might find a few research projects for your students that are appropriate to the regions around Jaipur.

    Ashwani Kumar
    Sanganer the world famous printing industry and hand paper industry has totall polluted the water of famous town of Sanganer Jaipur Behind the colourful bedsheets , skirts , and dress material there lies heaps of coloured water which is highly poisonous to the local people. Paper industry causes immense pollution. Three of my students have been working with isolating bacteria from the roots of the plants which can provide some solutions for bioremediation of waste water with only limited success. Pollution rules are there to please someone but who is that someone No adhearance to clean up unit to be established by each industry are adheared to and industries all over are polluting rivers, cities and nations and people are suffering silently.
    However scientists like you have a role in providing solutions What has happend in other parts of the world to control.
    How green are our cities? The key number in measuring that now is the per capita emission or how big a carbon footprint an individual leaves.

    The developed world pegs lndia's per capita emission at 1.7 tonnes. But a study by a non-profit organization of 40 Indian cities says it is much lower at less than 1 tonne.

    The worst big city in India is Kolkata, at 1.83 tonnes, but it is nothing compared to Washington DC, which has per capita emission of 19.7 tonnes, or China's Shanghai and Canada's Toronto, both over 8 tonnes.

    Both Chennai and Bangalore have per capita emission lower than 1 tonne.

    The Carbon Culprits
    • Washington DC: 19.7 tonnes
    • Shanghai, Toronto: Over 8 tonnes
    • Kolkata: 1.83 tonnes
    • Chennai, Bangalore: Lower than 1 tonne
     
    In cities that top the list, the transport sector and industry seem to be playing a big role in upping carbon emissions.

    Interestingly, key Indian cities like Delhi and Mumbai were not part of the survey because the local administration was not too keen about divulging energy data.
     
    This data could give India the edge at the negotiating table in Copenhagen next month.

    But India's fast-growing cities, like Gurgaon, already high at over 2 tonnes per capita emission have to be extra careful.
     
    "We have already started initiating action that will bring down the per capita emissions in Gurgaon. Solid waste is a big contributor so we have put a plant in place to tackle the situation," says RK Khullar, Gurgaon Commissioner.

    Now the challenge is to keep the emission figures at these levels or even lower it.

    Recently in Agrintex several persons approached us as we were representing in that fair by the following questions
    1. I have tannery and we have several hundred litres of waste from there each day . how to deal with waste.

    2. I have coconut factory and while removing the husk there is lot of dust from the coconut shell which goes waste how can we use this dust for making electricity

    3. I have sugarcane factory how can we make use of bagasse in better way.

    4. What is the suitable size of biodiesel plant and what feed stock level it would require

    Source: NDTV Correspondent, Thursday November 5, 2009, New Delhi

    Any solutions
    For the toxic dye remediation, I believe your associates should look deeper in the ground to find the effective bacteria. In North America we had a terrible problem (self inflicted) with agent orange and related chemicals until it was discovered the problem molecules did not survive long in deep earth, although they were nearly impossible to destroy on the surface.

    With toxic materials the aerobic biogas reactor is usually effective as long as the concentration is controlled to a tolerable level and the nutrients are balanced, enough vitamins and minerals. This is also true of phenolics, anilines, and complex ethers. Sometimes more than one stage is required operating at different temperatures and nutrients.

    For questions 1, 2, and 3 the biogas reactor should find some acceptable operating condition, sometime at the upper temperature limit of the thermophylic microbials. Pretreatment is sometimes economical. Hydrogen peroxide is very popular now because it leaves no residue, and is economical to produce. The greatest challenge from individual factories is to balance the nutrients in the best way to manage the sludge production compared to the fuel gas. A poor diet in a heavily loaded digester will give too much sludge and not enough fuel gas. The age of the sludge is important. as is the measure of how much of it is actually alive compared to the demand of the feed materials.

    Carbohydrates provide the chemical oxygen for a biogas reactor so there is an optimum mix of the feed for the best yield of fuel gas. Sometimes a small community of factories pool their waste into a single shared treatment plant to get a balanced diet for the bacteria. It is helpful to look at the chemicals in the effluent and the liquid part of the sludge to decide if there is too much or too little carbohydrate in the feed.

    In the worst case potassium permanganate is almost always effective to oxidize any organic material, but also the most expensive last resort. This is the test that government agencies require for the chemical oxygen demand measurement.

    For the toxic dye, and interesting laboratory comparison would be to compare the consumption of permanganate in one test, to the consumption of peroxide in another test, and possibly some lesser oxidizing compound like iron oxide in a third test. From these you can get a good graph for the presence or absence of resistance to oxidation at three different oxidation potentials.

    For question 4 in north America the economic size is about 100 million gallons per year of bio diesel, mainly for keeping the labor cost low. I guess in a developing country with a locally owned factory, a good size would be around 15 million gallons per year, and a minimum size would be around 3 million gallons per year.

    Feed material for a biodiesel plant is usually an oil based on glycerol and three branches of fatty acids. The branches go into the bio diesel by trans esterification with a light alcohol. The glycerol goes into biogas or is sold into an unfavorable market that has an excess. Much glycerol is burned for fuel so I guess it could also be bought for making biogas.

    Ashwani Kumar
    There seems to be great future in biofuel research. You are making good headway. Would like to share more informations and exchange of views how to overcome fuelwood shortages in this part of the globe and what could be alternative strategy.
    Wood fuel shortages will get worse with bio-energy. That is one of the largest problems, and the main reason many people are against biofuels. We have the example of England in the 16th century, where foundries for casting ion consumed all of the available wood in a few decades. It caused the mining of coal to begin on a large scale.

    Solar energy in the dryer parts of India represents one of the best chances to economize on wood consumption. Here we are speaking about highly reflective surfaces of low cost materials shaped to collect and focus the sun light to a very hot cooking area like an oven. Then if wood is also used in the same device, the solar heat helps to conserve wood and make the heating more effective.

    A similar efficiency is achieved by constructing the air flow such that the rising smoke from a flame heats the incoming air before it reaches the flame. Then a small fire can be very hot from a lower quality of fuel. In traditional wood burning a large part of the fuel goes out the vent without accomplishing much useful work. So you need more fuel of a higher quality. Preheating the air in a rather inexpensive duct makes a fire nearly indifferent to the quality of fuel and cuts the total amount of fuel by maybe a third or sometimes more than a third.

    With preheated air almost any combustible material can be used as fuel, although the starting method might require a better fuel for a while, then changing to the lower quality fuel when the fire is hot enough.

    You need to be careful with preheated air, because the fire can get too hot and damage the equipment. Also the smoke can deposit combustible materials in the vent duct and cause a dangerous fire there if it isn't cleaned on a schedule.

    Ashwani Kumar
    Recently our Government is making policy for generating electricity from biofuel Unless this is done with utmost care and biofuel is generated in sustainable manner what are the alternatives available for electricity generation in decentralised manner . Can ecologically fragile ecosystem support biofuel use or enough wasteland has vast potential for the production of biofuel without harming the food crop areas will depend on the knowledge of climate soil and plants and their productivity , the area we have been working for last three decades but there are more questions than answers .