1

ORGANISMS 1. Classification The organisms living on the surface of the earth or in the soil have been considered under four headings i.e., Higher plants, Vertebrates, Mesofauna and Microorganisms.

1.1 Higher plants Higher plants by extending their roots into the soil act as binders, prevent erosion, with grasses being more effective, while growing within cracks in rocks force them apart, and when plants die and roots decay, which form a net work of passages through which water and air can circulate more freely. One of the greatest contributions of the higher plants is through the addition of organic matter or litter to the surface. Tropical plant communities contribute annually as much as 25 tons ha–1, tall grass prairie 5.0 tons ha–1 and pine forest 2.5 tons ha–1. Pine forests may have an accumulation of about 15 cm of organic matter at the surface Prairie grass soils have up to 15 % organic matter incorporated in the mineral soil while the soils of tropical rainforests often contain < 5% organic matter. On the other hand dairy cows produce annually about 3.5–4.5tons ha–1. 1.2 Vertebrates A few mammals, including rabbits, moles and the prairie dog, are active within the soil, burrow deeply causing considerable mixing, and bring the subsoil to the surface such as crotovinas in many Chernozems and the blind mole rat in European soils. Uncontrolled grazing by animals leave the surface bare for erosion. 1.3 Mesofauna In this group are included earthworms, enchytraeid worms, nematodes, mites, springtails, millipedes, some gastropods and many insects, particularly termites and ants. Their distribution is dependent upon the food supply, therefore they are concentrated in the top 2 – 5cm soil; only earthworms can penetrate below 10 – 20cm. They require, in general anaerobic environment at around pH 7.0, but can also live in acid and alkaline soils. Under optimum conditions it has been

2

estimated that the biomass of earthworms is about 800 Kg ha–1 and nematodes about 5 – 20 million / m2.

The mesofauna generally ingest both mineral and organic matter after its decomposition and produce faecal material. While transporting material from one place to another they produce passages in the soil, thus improve drainage and aeration. Earthworms generally transport material to the surface while termites transport material to build their termitaria, and harvester ants denude an area 2 – 6 m around their nest. 1.4 Microorganisms The Roman philosopher Lucretius (about 98–55 B.C.) and the physician Girolamo Fracastoro (1478–1553) suspected the existence of microorganisms, even before they were seen, and suggested that diseases are caused by invisible living creatures. The amateur microscopist Antony van Leeuwenhoek (1632–1723) of Delft, Holland was the first person to accurately observe both the bacteria and protozoa and describe them. Agostino Bassi (1773–1856) first showed a microorganism could cause disease when he demonstrated in 1835 that a silkworm disease was due to a fungus infection. He further suggested that many diseases were due to microbial infections. [Microbiology 2nd edition by Lansing M. Prescott, John P. Harley and Donald A. Klein]

1.4.1 Predominant microorganisms Microorganisms are too small to be seem clearly by the unaided eye. The predominant microorganisms are bacteria, fungi, actinomycetes, algae and viruses.

1.4.1.1 Bacteria

Bacteria are the simplest organisms found in most natural environments. Bacteria have a much simpler morphology and lack a true membrane-delimited nucleus, thus all bacteria are prokaryotic. Bacteria are spherical or rod-shaped and are commonly several micrometers in linear dimension.

3

Bacteria are small and can replicate quickly, simply dividing in two by binary fission. Under optimal conditions a single prokaryotic cell can divide every 20 minutes and thereby give rise to 5 billion cells in < 11 hours. The ability to divide quickly enables to populations of bacteria to adapt rapidly to changes in their environment. In nature bacteria live in an enormous variety of ecological niches, and they show a corresponding richness in their underlying biochemical composition. Two distantly related groups can be recognized: the eubacteria, which are the commonly encountered forms that inhabit soil, water, and large living organisms; and the archaebacteria, which are found in such incommodious environments as bogs, oceans depths, salt brines, and hot acid springs.

They are the smallest and most numerous of the free-living microorganisms in the soil, where the distribution is determined by the presence of food supply, therefore they occur in the greatest numbers in the surface horizons which have a teeming mass of biological activity. They number several million per gram with a live weight of 1000- 6000 Kg ha-1 in the top 15 cm. There are species of bacteria that can utilize virtually any type of organic molecules as food, including sugars, amino acids, fats, hydrocarbons, polypeptides, and polysaccharides. Some are even able to obtain their carbon atom from CO2 and their nitrogen atoms from N2. Despite their relative simplicity, bacteria have existed for longer than any other organisms and still are the most abundant type of cell on earth.

1.4.1.2 Actinomycetes

Actinomycetes are second in abundance to bacteria preferring dry warm grassland and neutral conditions. There are a large number of genera of which the streptomycetes are dominant. They have a characteristic musty odour and produce antibiotics and enzymes that kill bacteria another microorganisms. They are very important as decomposers of organic matter particularly polysaccharides and chitin.

4

1.4.1.3 Algae Algae are early colonizers of newly exposed material in wet situations such as paddy fields and the very widespread shallow pools in the arctic. When in sufficient numbers they help to form a crust at the soil surface thereby preventing soil erosion. They are considered as early initiators of the carbon and nitrogen cycle. 1.4.1.4 Viruses Viruses are generally regarded as parasites in larger animals and plants and nearly every class of microorganisms in the soil is subject to viral attack. Each individual virus has a limited host range, some attack bacteria and others fungi, actinomycetes, algae, protozoa, earthworms, etc. The virus enters through the cell wall of the bacterium and attaches itself by its tail then causes lyses-decomposition of the cell walls.

1.4.2 Classification with respect to sources of C & energy In order to survive and grow, microorganisms require a source of energy for nourishment. Depending on their sources of carbon and energy they are classified into four classes:

1.4.2.1 Photoautotrophs: The organisms contain chlorophyll and utilize light as their energy source and CO2 as their principal source of carbon; they include the blue-green algae some of which can fix atmospheric nitrogen. The higher plants are photoautotrophs.

1.4.2.2 Photoheterotrophs:

This is a very restricted group of organisms that use light as a source of energy and derive much of their carbon from organic compounds. 1.4.2.3 Chemoautotrophs: These organisms derive their energy from the oxidation of inorganic compounds and use CO2 as their principal source of carbon. They include several groups of specialized bacteria, including the all important nitrifying bacteria.

5

1.4.2.4 Chemoheterotrophs: This is by far the largest group of microorganisms that utilizes organic compounds both as a source of energy and carbon. They include protozoa, fungi, actinomycetes and most bacteria and are of immense importance through their participation in humification and ammonification.

[-- Introduction to soil science second edition by E A Fitzpatrick --Molecular Biology of the Cell 3rd edition by Bruce Alberts, Dennis Bray, Julian Lewis, Martin Raff, Keith Roberts, James D. Watson --Microbiology 2nd edition by Lansing M. Prescott, John P. Harley and Donald A. Klein]

2. Factors affecting the nourishment of microorganisms

2.1 Chemical factors To obtain energy and construct new cellular component, organisms must have a supply of raw materials or nutrients. Nutrients are substances used in biosynthesis and energy production, and therefore are required for microbial growth. Analysis of microbial cell composition shows over 95% of cell dry weight is made up of few major elements: C, O, H, N, S, P, K, Ca, Mg, Fe. These are required in larger quantities and are called macronutrients. The first six (C, O, H, N, S, P) are components of carbohydrates, lipids, proteins, and nucleic acids. The remaining four macronutrients exist in the cell as cation and play a variety of role, e.g. K is required for activity by a number of enzymes; Ca among other functions contributes to the heat resistance of bacterial endospores. Mg serves as cofactors for many enzymes and Fe2+ and Fe3+ is a part of cytochromes. All microorganisms require trace elements (needed in smaller amounts) such as Mn, Zn, Co, Mo, Ni, Cu for their growth and these are a part of enzymes and cofactors, aid in the catalysis of reactions and maintenance of protein structure [Microbiology 2nd edition by Lansing M. Prescott, John P. Harley and Donald A. Klein].

Like all living things bacteria require mineral salts such as Ca, Mg, K, Fe, Cu and others. Some bacteria need sugars, vitamins, amino acids. Other bacteria can digest proteins down to amino acids and digest

6

complex carbohydrates such as starches and table sugar down to simple sugars. Some bacteria can make their own amino acids and vitamins from carbohydrates. The blue-green bacteria have chlorophyll and can make their own food from light energy + carbon dioxide. Some other bacteria have red chlorophyll and can use light and carbon dioxide to make the sugars they need. In short all the essential nutrients / elements found in the soil or occurring in nature are required for microbial growth. If under certain condition a specific element is required it can be added to the medium. The numerous pieces of bacteria live in an astonishing variety of place and live on every food you can imagine. Some can eat gasoline and other hydrocarbons. 2.1.1 Source of nutrients The main source of nutrients in the soil is organic matter. Under natural conditions trees, shrubs, grasses and other native plants supply large quantities of organic residues. The other sources are earthworms, centipedes, ants, etc, crop residues, farmyard manure (FYM), poultry manure (PM), green manures, filter cake of sugar industry and various types biofertilizers inclusive of Bokashi. The composition of manure and dry plant tissues is given in table -1. Table –1. Composition of a Manure and Dry Plant Tissues*

Compounds Percentage Carbohydrates:

Sugar and starches Celluloses Hemicelluloses

01 – 05 20 – 50 10 – 28

Proteins Simple water soluble and crude

01 – 15

Fats, Oils, waxes, tannins etc

01 – 08

Lignins 10 – 30

· Principles of soil science third edition by M M Rai.

7

The carbohydrates are complex substances and range from simple sugar to very complex celluloses and are made of C, H and O. The fats and oils are glyceride esters of fatty acids such as butyric, stearic and oleic etc. They are more complex than carbohydrates. They are also made of C, H and O. Protein are most complicated substances and contain C, H, O, N, S, Fe, P and a few other substances in lesser amounts. They are formed by a number of amino acids united to each other by peptide linkage.

The microorganisms derive their energy from the oxidation of carbohydrates, proteins and fats present in tissues of plants as well as in dead microbes. The common simple products which are formed due to the activity of soil microorganisms are (Principal of Soil science by M M Rai):

Carbon compounds: CO2, CO3, HCO3, CH4, C

Nitrogen compounds: NH4, NO2, NO3, N

Sulfur compounds: S, H2S, SO3, SO4, CS2

Phosphorus compounds: H2PO4, HPO4, PO4 Other simple products: O2, H2, H2O, H, OH, K, Ca, Mg, etc

2.2 Physical / Environmental Factors The temperature, food supply, oxygen level and pH of the medium are critical in the successful cultivation / growth of microorganisms. The microorganisms occur in the greatest numbers in the topsoil due to the presence of good food supply, optimal temperature and oxygen level, therefore they exhibit maximum biological activity. Among the organisms in the soil there exists and extremely complex inter-relationship for seldom does a single type of organism exist or operate separately from the others. Some highly contrasting organisms co-exist while others are predators, competitors or parasite. Earthworms and bacteria co-exist.

8

2.2.1 Temperature Most microorganisms grow well at the normal temperatures favored by man, higher plants and animals. However, certain bacteria grow at temperatures (extremely hot and cold) at which few higher organisms can survive. Depending upon their preferred temperature range, bacteria are divided into three group: Psychrophiles (cold-loving microorganisms): They have an optimum growth temperature between 0oC and 15oC and a maximum growth temperature of not more than 20oC. Mesophiles (moderate-temperature-loving bacteria): They are found in water, soil and in higher organisms. Their optimum growth temperature ranges between 25oC and 40oC. The optimum temperature for many pathogenic bacteria is 37oC, thus the mesophiles constitute most of our common pathogenic bacteria and disease microbes. Thermophiles (heat-loving microbes): They are capable of growth at high temperatures with an optimum above 60oC. Most thermophiles cannot grow below 45oC but some grow even above 100oC. 2.2.2 pH Most bacteria grow best in an environment with a narrow pH range near neutrality between pH 6.5 and 7.5. These are classified as:

Acidophiles (acid-loving). They grow at pH values below 4 with some bacteria still active at a pH of 1. Alkalinophiles (base-loving). They prefer pH values of 9-10 and most cannot grow in solutions with a pH at or below neutral. Often during bacterial growth organic acids are released into the medium, which lower its pH and so interfere with or totally inhibit further growth. Although common media ingredients such as peptones and amino acids have a small buffering effect, an external buffer is needed in most bacteriological media to neutralize the acids and maintain the correct pH. Phosphate salts are the most commonly used buffer because they buffer in the growth range of most bacteria, are non-toxic and provide a source of phosphorus, and essential nutrient element.

9

3. Advantages and disadvantages of microorganisms

3.1 Advantages / benefits They exist everywhere i.e. in the air, water and soil, and in the body of human beings and other creatures. Society benefits from microorganisms in many ways. They are necessary for the production of bread, cheese, bear, antibiotics, vaccines, vitamins, enzymes, and another important products. Microorganisms are indispensable components of our ecosystem. They make possible the carbon, oxygen, nitrogen, and sulfur cycles that take place in terrestrial and aquatic system, and are a source of nutrients at the base of all ecological food chains and webs. Their benefits are enormous in the field of agriculture and in bioremediation of polluted resources.

3.1.1 Fixation of atmospheric nitrogen In the field of agriculture atmospheric nitrogen is fixed by Rhizobium (rhizobia) bacteria in the nodules of legumes in the form of complex compounds of nitrogen (glutamic acid); it is about 250 – 300 Kg ha-1 year –1 in case of Alfalfa and 70 – 80 Kg ha-1 year –1 in case of Pea (Fertilizers, a text book, by Ranjan Kumar Basak). Some bacteria are free-living organisms and have the capacity to absorb atmospheric N2 to synthesis organic nitrogenous compounds. These organisms (non-symbiotic nitrogen fixing organisms) are bacteria, algae and fungi and are aerobic, anaerobic, heterotrophic, autotrophic, photosynthetic.

3.1.2 Mineralization of organic nitrogen compounds

Mineralization of nitrogen is the conversion of organic form nitrogen to inorganic / mineral form of nitrogen such as NH4

+, NO2- and NO3

-. It takes place in three steps i.e., 1. Aminization, 2. Ammonification and 3. Nitrification.

i) Aminization: the protein breaks down to yield amines, amino acids, carbon dioxide, energy and other products. This process is brought about by some heterotrophic soil microorganisms. They are mostly bacteria and fungi and possibly actinomycetes.

ii) Ammonification: in this case the amines and amino acids released by aminization process are converted to ammonia,

10

NH3. The conversion is caused by another group of heterotrophic of soil microorganisms such as bacteria, fungi and actinomycetes.

iii) Nitrification: this process is completed in two steps i.e., in the first step nitrite (NO2

-) are form and in the second step NO3

- are formed. These two steps are caused by two enzymes, dehydrogenase and oxidase, secreted obligate autotrophic bacteria such as Nitrosomonas, nitrosococcus, nitrosocystis, nitrosospira and nitrosogloea.

From the above it can be concluded that life within soil exceeds the life above the soil in terms of numbers of living organisms and total metabolic activity (Thomson, LM and Troeh, FR. 1978, soil and soil fertility, McGraw Hill Book Company, NY., 516 P). Soil is a unique environment as it contains a vast array of bacteria, actinomycetes, fungi, algae and protozoa, which are important group of micro flora in soils. They are unicellular, of numerous genera and perform vast variety of specialized functions. The important bacteria belong to symbiotic-N-fixation, no symbiotic-N-fixation, Aminization, ammonification and nitrification, P-solubilizer, S-oxidizers and H2S oxidizers. Some important bacteria from plant nutrition point of view are listed in table –2.

11

Table –2: IMPORTANT BACTERIA FROM PLANT

NUTRITION POINT OF VIEW* Sr. # Name of bacteria Function 1. Rhizobium sp symbiotic N fixation 2. Bradyrhizobium symbiotic N fixation 3. Cyanobacteria (BGA) biological N fixation 4. Azotobacter sp biological N fixation 5. Azospirillum sp biological N fixation 6. Nitrobacter sp nitrification 7. Nitrosomonas sp nitrification 8. Pseudomonas denitrification 9. Bacillus sp denitrification 10. Paracoccus denitrification 11. Thiobacillus denitrification 12. T. thioparus denitrification 13. Bacillus megaterium P solubilizer 14. B. circulans P solubilizer 15. B. subtilis P solubilizer 16. Pseudomonas straita P solubilizer 17. P. rathonis P solubilizer 18. Escherichia freundla P solubilizer 19. Thiobacillus thioxidans oxidizes S 20. T. thioparus oxidizes S 21. T. copraliticus oxidizes S 22. T. ferrooxidans oxidizes S 23. Beggiatoa oxidizes H2S to S 24. Thiothrix oxidizes H2S to S

________________________________________________________ · Dictionary of Soil Fertility: Fertilizers and Integrated Nutrient

Management by HLS Tandon, p 13

12

3.1.3 Decomposers Decomposers (fungi, bacteria and actinomycetes) breakdown the organic material, form humus and unlock the useful nutrients (NPK, S, trace elements etc) and made them available to plants. Phosphate solubilizers (fungi and bacteria) secrete organic acids and help in solubilization of insoluble P-compound in soil. Mycorrhizal fungi have been reported to mobilized P and other nutrients in normal as well as saline soils. Some bacteria live in association with roots of cereals, e.g. rice, sugarcane and grasses, and produce phyto hormones to enhance plant growth.

Some microbes act as biological control of pathogens, due to liberation of antibiotic and other compounds. The bacteria which affect insects are broadly classified as spore-formers and non-spore-formers. The spore formers include milky disease organisms, Bacillus popilliae, B. lentimorbus and facultative groups such as B. sphaericus and B. thuringiensis. The non-spore-formers include serratia, pseudomonas, aerobacter and streptococcus. The spore forming bacteria are promising organisms for microbial control (Biological approaches in soil microorganisms for sustainable crop production by K.R. Dadarwal).

The microbial community structure / composition not only affects biological process, but most of them especially fungi and algae influence soil physical properties and help in aggregate formation and water retention in soil. The number of groups of microorganisms that commonly occur in top 0 to 15cm per ha-furrow slice (HFS) may be for bacteria 1017 - 1018 with fresh biomass 450 – 4500 Kg, for actinomycetes 1016 - 1017 with fresh biomass 450 – 4500 Kg, for fungi 1014 - 1015 with fresh bio mass 112 – 1120 Kg, for algae 1013 - 1014 with fresh biomass 56 –500 Kg and for protozoa 1013 - 1014 with fresh biomass, 17 – 170 Kg. The microflora (bacteria, actinomycetes, fungi and algae (together form 2076 to 20760 kg fresh biomass and 415 –5190 Kg dry biomass (20 – 25 % for fresh biomass per HFS. The microbial number in the top soil (3-8 cm) may be 12 million /g, comprising live biomass of bacteria 100-4000 Kg / ha; while double of this biomass for all the microbes (bacteria, actinomycetes, algae and fungi) forming 0.02 – 0.8% of total soil biomass.

13

(Reference given by A. Sikandar: effect of organic and inorganic fertilizer on the dynamics of soil microorganisms, biomass, composition and activity. Nuclear Institute for Agriculture Biology, Faisalabad, Pakistan) 3.1.4 Biodegradation Trials have been made to achieve biodegradation of petroleum hydrocarbons using indigenous bacteria. The bacteria mixtures of 40 to 50 different organisms utilized have shown to work effectively on petroleum hydrocarbon at many sites. The bacteria used were naturally occurring, non engineered, non pathogenic organisms and have the ability to degrade petroleum hydrocarbon to CO2 and water. The indigenous bacteria metabolize hydrocarbons as a carbon source in the presence of an electron acceptor. There are several possible electron acceptors available for intrinsic biodegradation including dissolved oxygen, nitrates, sulfates and iron. Dissolved oxygen is the most favored electron acceptor, and is used as an indicator of biodegradation that occurs by aerobic processes. Each 1.0 mg /L of dissolved oxygen consumed by microorganisms will destroy approximately 0.32 mg/L of BETX. During aerobic bio degradation, oxygen levels in the groundwater decrease. Most or all of the degradation that takes place in aerobic conditions involves the consumption of oxygen, as anaerobic bacteria generally cannot function in dissolved oxygen concentration > about 0.5 mg/L (Remediation of a High Priority Petroleum Site Using a Combination of Remedial Technology by Scott D. Hartsough; Contaminated Soils Volume 7). 3.1.5 Breakdown of pesticides The predominant means of pesticide breakdown in soils is biochemical processes carried out for the most part by microorganisms and to a smaller extent by plants (pesticides taken up by plants may be broken down by plant enzymes). Fertile soils contain millions of microorganisms per ounce of surface soil. Organic pesticides may serve as sources of nutrients and energy for microorganisms. The rate of microbiological breakdown of pesticides varies with their chemical structure and climatic conditions. In a summary of 12 studies, Tinsley (1979) noted that the 50% decomposition times for certain pesticides increased in the order malathion, 2,4-D, diazinon, atrazine, diuron, DDT. The 50% decomposition time for DDT was

14

240 days in a tropical environment, 3840 days in a temperate environment (Agriculture and groundwater quality, Council for Agricultural Science and Technology, Report # 103).

3.1.6 Phytoremediation Phytoremediation has emerged as an innovative strategy for various environmental applications. Including hydraulic control of shallow groundwater impacted by petroleum hydrocarbon. A phytoremediation barrier (phytobarrier) at an active petroleum retail station in Ohio in Spring 1997 was installed by planting hybrid poplar trees within a trench along the Western site border, downgradient of the source area. After the third growing season tree roots were observed to a depth of 10 feet. Based on ground water elevation data of the third year the hydraulic gradient across the phyto barrier had decreased more than 3 feet as compared to control, this indicated the aerobic degradation of petroleum hydrocarbon (Phytoremediation for Hydraulic Control of Shallow Groundwater Impacted by Petroleum Hydrocarbons by William M. Golla and James J. Reid, ARCADIS Geraghty & Miller, Contaminated Soils Volume 6). 3.2 Disadvantages / harmful effects Of course, microorganisms also had harmed humus and disrupted society since the beginning of recorded history. Microbial diseases undoubtedly played a major role in historical events such as the decline of the Roman Empire and the conquest of the New World. In the year 1347, plague or Black Death stuck Europe with brutal force. By 1351 the plague had killed 1/3 of the population (about 25 million people). Over the next 80 years, the disease stuck again and again, eventually wiping out 75% of the European population. Some historians believe that this disaster changed European culture and prepared the way for the Renaissance. Today, the struggle by microbiologists and others against killers like AIDS, HEPATITIS and malaria continues (Microbiology 2nd edition by Lansing. M. Prescott John P. Harley and Donald A. Klein). The pollution caused by solid city wastes, sewage water and industrial effluent and sludge (solid waste) have played havoc in the developed countries where it is being continuously controlled and the pollution is said to be to a lesser extent and to the largest extent in the developing

15

countries where the problem has multiplied and it is beyond their financial as well as technical resources to control it.

In Pakistan all the drainage system ends into the riverbeds, which are presently polluted enormously. The situation is at an alarming stage because under the Indus Water treaty 1958 the rivers Sutlej, Ravi, Chenab and Jehlum have been almost abandoned. For example the untreated sewage water of Lahore city and adjoining municipalities, villages and abbadies, and untreated industrial effluents inclusive of Leather Industry from the cluster of industries of Kala Shah Kaku, Lahore and Sheikhupura, etc are discharged into the Ravi River at an average flow of > 650 cusecs / day. The flow is expected to be above 2000 cusecs by the year 2025. Present BOD load is 470 tons / day and there is a loss of more than 5000 tons / day of fish catch. The untreated heavily polluted sewage and industrial effluent is not only being used for growing agricultural crops especially vegetables as these bring higher prices in the vicinity of Lahore city, but also animals are forced to drink this very effluent. This means that most of the inhabitants of Lahore city are eating vegetables grown with polluted water and are drinking milk of animals taking this polluted effluent. This is the reason that diseases like Hepatitis, Dysentery, Gastroenteritis, Jaundice, Cancer, etc have increase tremendously. The same is true farm other big cities like Karachi, Hydrabad, Multan, Faisalabad and Sargodha (Environmental Degradation by Eng. Col. Mumtaz Huassain).

16

4. INVENTION OF EM TECHNOLOGY Prof. Dr. Teruo Higa, University of Ryukyus, Okinawa, Japan developed the first batch of Effective Microorganism, which eventually called EM in 1980. It is available in the liquid form. It is produced through a natural process of fermentation and not chemically synthesized or genetically engineered. EM is a combination of various beneficial, naturally occurring microorganisms mostly used for or found in food. EM is a liquid concentrate. It is produced in vats from cultivations of over 80 varieties of microorganisms. The microorganisms are drawn from 10 genera belonging to 5 different families. The most outstanding characteristic of EM is this that it includes both aerobic and anaerobic species coexisting symbiotically in a most beneficially productive manner. EM contains beneficial tiny anabiotic microorganisms form 3 main genera: phototrophic bacteria, lactic acid bacteria and yeast.

Photosynthetic bacteria (Rhodopseudomonas): The photosynthetic or phototrophic bacteria are a group of independent, self-supporting microbes. These bacteria synthesize useful substances from secretions of roots, organic matter and / or harmful gases (hydrogen sulphide), by using sunlight and the heat of soil as source of energy. The useful substances developed by these microbes include amino acids, nucleic acid, bioactive substance and sugars, all of which promote plant growth and development. The metabolites developed by these microorganisms are absorbed directly by the plants and act as substrates for increasing beneficial microbial populations. For example, Vesicular Arbuscular (VA) mycorrhizae in the rhizosphere are increased due to the availability of nitrogenous compounds (amino acid) which are secreted by the phototrophic bacteria. The VA mycorrhizae in turn enhance the solubility of phosphates in soils, thereby supplying unavailable phosphorus to plants. VA mycorrhizae can also coexist with Azotobactor and Rhizobium, thereby increasing the capacity of plants to fix atmospheric nitrogen.

17

Lactic acid bacteria (Lactobacillus): Lactic acid bacteria produce lactic acid from sugars and other carbohydrates, developed by photosynthetic bacteria and yeast. Lactic acid is a strong sterilizing compound and suppresses harmful microorganisms and enhances decomposition of organic matter. Moreover, Lactic acid bacteria promote the fermentation and decomposition of material such as lignin and cellulose, thereby removing undesirable effects of undecomposed organic matter. Lactic acid bacteria have the ability to suppress disease-inducing microorganisms such as Fusarium, which occurring continuous cropping programmes. Under normal circumstances, species such as Fusarium weakens crop plants, thereby exposing them to diseases and increased pest population such as nematodes. The use of lactic acid bacteria reduces nematode populations and controls propagation and spread of Fusarium, thereby inducing a better environment for crop growth.

Yeast (Saccharomyces): Yeasts synthesize ant microbial and other useful substances required for plant growth from amino acid and sugars secreted by photosynthetic bacteria, organic matter and plant roots. The bioactive substances such as hormones and enzymes produced by yeasts promote active cell and root division. These secretions are also useful substrates for Effective Microorganisms such as Lactic acid bacteria and Actinomycetes. EM as “co existence and co prosperity” The difference species of Effective Microorganism (Photosynthetic and lactic acid bacteria and yeast) have their respective function. However, photosynthetic bacteria could be considered the pivot of EM activity. Photosynthetic bacteria support the activities of other microorganisms in EM. However, the photosynthetic bacteria also utilize substances produced by other microbes. This phenomenon is termed “Co existence and Co prosperity”. The enhancement of population of EM in soils by application promotes the development of existing beneficial soil microorganisms. Thus, the micro flora of the soil becomes abundant; thereby the soil develops a well-balanced microbial system. In this process soil specific microbes (especially harmful species) are suppressed, thereby reducing microbial diseases that cause soil borne diseases. In contrast, in these developed soil, the

18

Effective Microorganisms maintain a symbiotic process with the roots of plants within the rhizosphere. Plant roots also secrete substances such as carbohydrates, amino and organic acids and active enzymes. Effective microorganisms use these secretions for growth. During this process, they also secrete and provide amino and nucleic acids, a variety of vitamins and hormones to plants. Further more, EM in the rhizosphere co exist with plants. Therefore, plants grow exceptionally well in soils, which are dominated by Effective Microorganisms. EM is a living entity containing active microbes. Manufacturing of EM requires good quality water free of pollutants or chemicals. EM can be stored in a closed container for a period up to 6 months if kept in a dark cool place (Refrigeration is not required). EM always has a sweet sour smell. One may notice a white film on the surface of EM solution when it is stored. This is yeast and does not cause any harm to the EM. The soils having a high population of disease causing microbes (Fusarium) are called Disease inducing soils. These are generally hard and physical characteristics are not conducive for crop growth. The soils having organisms such as Pencillium, Trichaderma, Aspergillus and Sterptomyces, which produce antibiotics, are called disease suppressive soils. These soils have very good physical characteristics. The soils containing zymogenic organisms such as Lactic acid bacteria and yeast are called zymogenic soils. When raw organic matter with high nitrogen contents is applied, the soil develops an aromatic smell, the population of fermenting fungi such as Aspergillus and Rhizopus increases. These soils have very good physical characteristics with a high water holding capacity. EM is a versatile product that uses microorganisms found in all ecosystems. The principle of EM is the conversion of a degraded ecosystem full of harmful microbes to one that is productive and contains useful microorganisms. This simple principle is the foundation of EM Technology in agriculture and environmental management. [An Earth Saving Revolution: A means to resolve our world’s problems through Effective Microorganisms (EM) by Dr. Teruo Higa]

19

5. Benefits of EM Technology The effective microorganisms secrete beneficial substances such as vitamins, organic acids, chelated minerals and antioxidant when in contact with organic matter. EM is being used successfully in the field of agriculture, fisheries, poultry, animal husbandry and for the preparation of compost and Bokashi. Now its application has spread in controlling the environmental pollution caused by city solid waste, sewage water, industrial solid wastes and effluent. EM application has helped to eliminate obnoxious odor at leather and starch industry, livestock farms and zoo etc. Its application has been extended to health sectors with the invention of EM-X, research is being conducted in various countries to see the effect of EM-X as antioxidant in patients suffering from cancer, aids, diabetes and skin diseases. Today, EM Technology has extended its activities to over 116 countries, where it is being used in the field of agriculture, fisheries, poultry, animal husbandry and environments such as recycling of sewage water, city wastes and kitchen garbage. Of 116 countries EM is being manufactured in 45 countries.

5.1 Agriculture EM Technology has brought a major revolution in a number of seemingly diverse areas. EM Technology is effecting significant and on going changes in agriculture. It has increased not only the yield but also improved the quality of the produce. In Japan the average yield of rice has been increased from 540 kg to 840-9000 kg / 1000 m2. The cucumber plant grown with EM produced 4 – 5 cucumber per node instead of one only. In South America, Brazil has been enthusiastically focusing on EM as a way of breaking free from the destructive slash-and-burn farming methods which have grown up there, and of simultaneously protecting the natural environment of the Amazon Basin. As a result Brazil is currently the world’s largest consumer of EM. Brazil’s monthly production capacity of EM is over 700 tons. USA, Canada, China France, Russia, Germany, Holland, Japan, Australia, Spain, Argentina, Malaysia, Kenya, Syria, Egypt, Pakistan, India, Sri Lanka, Colombia, Vietnam, Taiwan, Korea, Philippines, Denmark, Poland, South Africa, Sudan, Lebanon, Indonesia, Thailand, New Zealand, Peru are producing and using EM in various fields. On the other hand in 71 countries EM is being used

20

but not manufactured, among these are Chili, Cambodia, Cuba, Sweden, Namibia, Angola, Mali, Portugal, Greece, Yugoslavia, Saudi Arabia, Nigeria, Afghanistan, Zimbabwe, UAE, Mexico, Hungary, Finland, Congo, Ghana, Morocco, Turkey, Iran, Qatar, Italy, Austria, Israel, and Venezuela, etc. In Hawaii EM (1: 500) was sprayed on Macadamia Nuts trees at Cordoza farm weekly and EM Bokashi was applied around the base of each tree along with EM extended application. EM application improved the health of the trees. EM treatment increased not only the size of the flowers but also caused the formation of clusters of macadamia nuts on the branch instead of only single nut.

In Waliua, Oahu U.S.A. the Dole Food Corporation applied EM Bokashi around the base of 2 years old grape fruit trees and EM (1: 1000) was sprayed over the entire plants weekly. EM increased the quality and quantity of grape fruits. It also prevented the insect attack.

[--An Earth Saving Revolution: A means to resolve our world’s problems through Effective Microorganisms (EM) by Dr. Teruo Higa, p 14 – 15; --EM World Network Map indicating countries manufacturing and using EM / not producing EM but using EM in various field by EM Research Organization, Okinawa, Japan; --EM Research Organization, Inc., 417 Ehako Place Honolulu, Hawaii 96817, January 3, 2000, Report # 10362].

5.2 Reclamation of saline-sodic soils in Pakistan Saline sodic soils can be reclaimed with the application of EM Technology comprising of composting of FYM + PM with EM, preparation of Bokashi from rice bran with EM, seed treatment with EM, EM irrigations and EM sprays in the 1st year with good yields. The reclamation with EM involves no soil amendments as needed for reclaiming sodic soils. The Na+ of the exchange complex is replace with H, NH4, and Ca released during the decomposition of composted material added to the soil. The microbial activity is increased to such an extent that a saline-sodic soil becomes normal agricultural land. The microorganisms which increased their population with the application of EM with all its products and helped to achieve reclamation are given in table –3.

21

(Mechanisms of effective microorganisms (EM) in removing salt from saline soils by A. Syed, N. Satou & T. Higa: 13th Annual West Coast Conference on Contaminated Soils, Sediments and Water: March 17 – 20, 2003 Mission Valley Marriott San Diego, CA, USA)

Table –3. Beneficial microorganisms developed with EM application in the saline-sodic soil.

(Mechanisms of effective microorganisms (EM) in removing salt from saline soils by A. Syed, N. Satou & T. Higa: 13th Annual West Coast Conference on Contaminated Soils, Sediments and Water: March 17 – 20, 2003 Mission Valley Marriott San Diego, CA, USA)

Sr. No

Salt affected soil (control)

Sr. No

EM treated soil

1 Bacillus-sp 1 Azotobacter-sp 2 Entrobacter-sp 2 Bacillus -sp1 3 E. coli group 3 Bacillus-subtile 4 Fungi 4 Clostridium-treponema 5 Pseudomonas-sp 5 Corymebacerium-sp 6 Streptococcus-sp 6 Furabacterum 7 Seratia-sp 7 Gluconobacter-sp 8 Lactobacillus- cassei 9 Lactobacillus-sake 10 Lactobacillus-sp 11 Lactobacillus-sp1 12 Lactobacillus-sp2 13 Micrococcus-sp 14 Micrococcus-sp1 15 Micrococcus-sp2 16 Pseudomonas- aeruginosa 17 Pseudomonas- fluorescens 18 Pseudomonas- putida 19 Pseudomonas- Q1 20 Pseudomonas- type –1 21 Pseudomonas- type –2 22 Pseudomonas-sp 23 Rhodobacter-capsulatus 24 Rhodoseubodomonas-sp 25 Rhodospirillum-sp 26 Streptococcus-sp 27 Treponema-sp

22

5.3 Animal husbandry A ranch in America experienced a high incidence of deformity among its cattle. Investigations showed that the deformities were related to the underground water supply on the ranch. The cattle were drinking this underground water, which was found to be loaded with heavy minerals. EM was introduced to the wells containing this very groundwater. In a short space of time it was noticed that the birth of deformed young dropped away surprisingly.

[An Earth Saving Revolution: A means to resolve our world’s problems through Effective Microorganisms (EM) by Dr. Teruo Higa, p 168]

5.4 Environmental pollution EM resolves problems of environmental pollutions with two types of microorganisms. Zymogenic EM produces antioxidants and certain synthesizing strains of anaerobic microorganisms love to consume contaminants and pollutants. The photosynthetic bacteria, which play central role in EM, are able to tolerate extremely high temperature, in certain cases temperatures in excess of 700oC. The use of EM has purified the wastewater in the public library in Gushikawa, a city in Okinawa, Japan. The effluent has been treated with EM, which has purified it to such a degree that it can safely be used for drinking purposes. The use of EM has caused a lot of savings. EM Technology is being used to banish odors in bathrooms and showers as well as to remove the contaminated and malodorous build-up inside pipes.

[An Earth Saving Revolution: A means to resolve our world’s problems through Effective Microorganisms (EM) by Dr. Teruo Higa, P 148 - 149]

5.5 Deodorization at a waste treatment plant In Switzerland, the Olfar Technology of Switzerland developed a plant with finer system for handling 5 - 20 tons of waste per hour. It is capable of disposing of discarded and waste items. This recycling operation was capable of dealing with a huge miscellany of items ranging from scrap metals like aluminum, iron and steel as well as plastics and vinyl’s, waste paper and textiles of all kinds to kitchen waste, food scraps and other organic substances. The entire accumulated mishmash of waste was finely pulverized and then

23

separated for recycling by means of a most advanced sorting and processing procedure. There was, however, one serious drawback to all of this: the sorted and processed waste gave off the most pungent and awful stench. Despite the fact that ozone sterilization was being used midway through the process to counteract the smell. The stench itself was bad enough, but the ozone used in the sterilization process was downright dangerous. Ozone is a virulent carcinogen and as such potentially life-threatening. EM was introduced into the system at one of the earliest stages of the operation. EM was sprinkled on to the waste materials right at the start of the pulverization process before entry into the crusher. EM acted as a very powerful deodorant and eradicated the terrible stench completely as if by magic. EM further enabled the Swiss recycling operation to do away with the dangerous process of ozone sterilization and improved the running efficiency of the machinery. [An Earth Saving Revolution: A means to resolve our world’s problems through Effective Microorganisms (EM) by Dr. Teruo Higa, P 120 -121]

5.6 Deodorization from kitchen garbage processing Kani City, central western part of mainland Japan, has been exceptionally successful in its use of EM Technology to process kitchen garbage since 1992.The associated unpleasant smell produced by the saprogenic or putrefactive bacteria has been completely eliminated by the use of EM. [An Earth Saving Revolution: A means to resolve our world’s problems through Effective Microorganisms (EM) by Dr. Teruo Higa, P 124 -125]

5.7 Deodorization at a zoo The use of EM has banished the unpleasant odor coming from animals kept at Honolulu zoo, Oahu U.S.A. Offensive odor is generally caused by admixture of among other things ammonia, hydrogen sulfide, trim ethylamine and methylmercapton. These substances just happened to provide substrate for the microorganisms in EM, who go to it and gobble them up, thereby effectively eradicating them. (EM Research Organization, Inc., 417 Ehako Place Honolulu, Hawaii 96817, January 3, 2000, Report # 10362)

24

5.8 Safe disposal of tannery’s sludge and its conversion into biofertilizer EM experts have completed research trial on effluent and sludge treatment with EM products in collaboration with Pakistan Tanner Association Lahore. The tannery sludge of Eastern Leather Company was changed to a powdery farm material, was named as bio sludge / bio fertilizer because of its richness in macro and micronutrients. It is worth mentioning that Cr was reduced from 50 000 ppm to 312 ppm in the sludge. The sludge of Siddiq Leather Works (SLW) was also treated and it changed into a powdery material. Cr was reduced to nil to 1.054% as per analysis of Soil and Water Laboratory, Agriculture Department, Government of Punjab and Environmental Sciences Laboratory, PTA, Lahore respectively. At ELC premises rice crop was grown with the application of bio sludge and EM irrigation and EM spraying. The results were satisfactory. (Pilot Study on Safe Disposal of Effluent and Sludge through Bioremediation using EM Technology at Siddiq Leather Works by EM / PTA Collaboration, March 2003).

5.9 Safe disposal of petroleum sludge and its conversion into biofertilizer

EM team has carried out research in collaboration with NCPC on the bio remediation of oily sludge of Attock Refinery Ltd in October 2002. The oily sludge was converted to bio sludge with the use of various EM products and finally by mixing with equal quantity of dried soil it was converted to a bio fertilizer. Agricultural trials are under progress. The heavy metals in the sludge were diminished to a certain extent. Treatment of 600 tons of oily sludge at large scale with EM Technology is under consideration by the ARL dignitaries. [Final Report on Attock Refinery Limited, Oily Sludge Bioremediation (anaerobic), phase-1: treatment of sludge using EM Technology by EM & NCPC Collaboration, 6th February 2003].

25

6. Microorganisms playing their role in Bioremediation of Petroleum sludge

The application of biological methods for pollution reduction is referred to as bioremediation. In microbial remediation microorganisms: bacteria, actinomycetes and fungi are the active organisms. The increased microbial activity contributes a unique environment conducive to the breakdown of organic compounds. The microorganisms use the pollutants as a source of nutrient, carbon, nitrogen, phosphorus etc and as a source of energy for their growth.

The effective microorganisms contained in the EM is applied to the sludge along with Bokashi (a Japanese word for organic matter and is prepared from rice bran, sugar-cane molasses, water) and EM powder. This helps to maintain a high microbial population of effective microorganisms in the presence of optimal moisture and temperature. These effective microorganisms detoxify the heavy metals by bacterial transformations through oxidation and reduction reactions. The concentration of heavy metals is reasonably reduced.

[A Case Study with UN Collaboration]

7. Microorganisms playing their role in

Bioremediation of Petroleum sludge The application of biological methods for pollution reduction is referred to as bioremediation. In microbial remediation microorganisms: bacteria, actinomycetes and fungi are the active organisms. The increased microbial activity contributes a unique environment conducive to the breakdown of organic compounds. The microorganisms use the pollutants as a source of nutrient, carbon, nitrogen, phosphorus etc and as a source of energy for their growth.

The effective microorganisms contained in the EM is applied to the sludge along with Bokashi (a Japanese word for organic matter and is prepared from rice bran, sugar-cane molasses, water) and EM powder. This helps to maintain a high microbial population of effective microorganisms in the presence of optimal moisture and temperature. These effective microorganisms detoxify the heavy metals by bacterial transformations through oxidation and reduction reactions. The concentration of heavy metals is reasonably reduced.

26

[A Case Study with UN Collaboration] EXECUTIVE SUMMARY

Disposal of refinery sludge is a difficult problem in the overall waste treatment management program of refineries. Even the most advanced methods give residues that are no longer amenable to cost effective treatment. Petroleum refining industries are troubled by the problem of handling substantial quantity of sludge in one form or the other depending upon the nature of the crude, processing capacity, downstream capacities, design of effluent treatment plant, pollution abatement measures and the efficiency-cum-effectiveness of these plants. Due to regulatory or legislative requirements, public pressure, side effects on humans, and enlightened corporate behavior; there is a growing realization and movement to clean-up such environmental messes. Oil refineries need a well-planned oily sludge management strategy to manage oily sludge and there is a need to have a more cost-effective alternative to traditional physical and chemical methods of remediation. UNIDO NCPC has been asked for the disposal of oily sludge from almost all refineries of Pakistan. Different options were proposed by NCPC for the disposal of oily sludge from the refineries. The option of treatment of oily sludge through clean microbes (bioremediation) was mutually agreed and implemented in a southern refinery. Aerobic bioremediation was utilized which is a slow process and the total time period for full bioremediation is 40-50 weeks. However it takes 8-12 weeks for 60~70% treatment, depending upon TPH value and the process is totally environment friendly and after the treatment through clean microbes the soil can be used for agriculture purpose. This project was initiated in collaboration with National Institute for Biotechnology and Genetic Engineering (NIBGE) Faisalabad. Before progressing with a preferred option NCPC is searching other processes and technologies for sludge treatment to provide an effective solution that may be applied to various process sludges, and also for the restoration of polluted sites, treatment of hydrocarbon

27

pollution caused by accidental spills during production, transportation or storage. In this regard, NCPC participated in CPP workshop held at Serena Hotel recently, where EM Research Organization also took part. EM has vast experience of providing bio-solutions in various fields. EM team was approached by NCPC to join hands in providing cost-effective and simple solution for the bio-degradation of oily sludge at refineries. EM team visited NCPC on October, 2002. In the meeting it was agreed to perform the trial treatment in collaboration with NCPC at ARL. EM team then visited NCPC on October 22, 2002 and the pre-requisites to undertake the trial were discussed The activities indicated to undertake the trial included sampling and analysis of the sludge, technicians to be trained, space, materials such as plastic sheets to maintain anaerobic conditions, drums and buckets etc. Effective microorganisms were supplied by EM team. ARL, which is also ISO-14000 certified, is interested in the treatment of its Oily sludge. Accordingly a trial was designed to effectively address ARL requirements and objectives. The trial project was designed to treat and convert 1.7 M ton of oily sludge into environmental friendly residue (compost) under anaerobic conditions. The residue can be used as a bio-fertilizer for agricultural purpose. A room near old bulkasar gantry was selected to carry out the trial. The sludge was shifted from site (near tank 1) to the room and mixed with the EM solution in proper ratio. EM Experts visited the site every week. NCPC carried out activities in sludge mixing, solution making and sludge condition monitoring, and coordination of testing. NCPC monitored the room temperature and logged it for the six weeks on daily basis. Laboratory test samples were taken for the priority parameters i.e. pH, C, N, H, S, TPH and metals analysis from QCL, PINSTECH, HDIP, NIBGE and NARC, before and after treatment of the sludge to find out the response of the EM technology to sludge treatment. (The metals analysis for the ARL sludge before treatment was completed; while after treatment is awaited).

28

After six weeks the sludge which was odorous greasy and black, transformed into dispersed & brownish shape and the pungent odor diminished and changed into sweet fermentation smell. During the process of sludge bioremediation it was observed that houseflies and mice/rats were attracted by the sludge and found there alive which shows that the sludge has lost its hazardous ingredients or nature. With EM application Ba was reduced up to 85%, Pb, Fe, Zn, Ni up to 50% in the bio-sludge while As, Cr, Cu and Mn showed no significant change in concentration. The treated sludge (bio-fertilizer) was then applied in phase II to agriculture land as bio fertilizer, after mixing with dry soil in proper ratio. Here the effectiveness of sludge treatment would be proved through physical growth and chemical analysis of the crop. N, P, K, and Organic Matter of the treated sludge mixed in 1:1 ratio with soil after treatment was compared to same analysis for field soil, where an agricultural experiment is being undertaken as part of phase II of this program. The entire N, P, K, and Organic Matter of the treated sludge have the properties of bio-fertilizer, and indicate it is rich in macro-nutrients. In fact the doses of the treated sludge will have to be determined on the basis of farmyard manure for agricultural purposes for various crops inclusive of vegetables and gardens. Conclusion: The trial on bio-remediation (anaerobic) of pure oily sludge of ARL, Rawalpindi within its premises using EM Technology was successfully completed with the collaboration of EM/NCPC/ARL within a period of 6 weeks (29th October to 10th December, 2002). The effective microorganisms transformed the undiluted oily sludge from ARL into bio-active-sludge; which may be called bio-sludge.

For heavy metal breakdown the trial data shows that Ba has been reduced by 85% in the EM treated oily sludge as compared to original ARL sludge, and Pb, Fe, Zn and Ni have been reduced by about 50% in the treated bio-sludge. The contents of As, Cr, Cu and Mn showed no change.

With this the first phase of the experiment ended showing:

29

i Effective Refinery Sludge Treatment providing environmentally friendly/safe disposal; has been demonstrated;

ii ISO-14000 compliance; ensured; iii Cost-effectiveness; being evaluated in phase II i.e. application

as bio-fertilizer iv By-product (saleable); oily sludge has been converted which is

easy to handle, transport, store, and applicable in agriculture. To start with may be distributed free of cost and later on commercial basis.

v Reduction in concentration of heavy metals; reduction in concentration of heavy metals has occurred up to 50 to 85%.

Recommendation: The EM treated sludge being a bio-fertilizer can be used in agriculture and a substitute to farm-yard manure and as a soil amendment to improve/reclaim salt affected lands (a major problem in Punjab and Sindh). While applying in agriculture the doses are to be determined by experiments carried out under given climate conditions.

· Test application of the ARL treated sludge as bio-fertilizer and test toxicity of chosen crop and also treated sludge/bio-fertilizer dose optimization.

· Compare treated sludge/bio-fertilizer yield and cost with

farmyard manure and chemical fertilizer. · After successful completion of the trial, this technology can be

applied to the estimated 400 – 600 M tons / year of ARL sludge on large-scale and also expanded to other Refineries.

· Using the experience of this trial and the methodology used,

proposal may be developed for other petroleum sector sludges upstream and downstream.

A. INTRODUCTION

A.1 EM Technology EM stands for Effective Microorganisms. EM is a combination of various beneficial, naturally occurring microorganisms mostly used for or found in foods. It contains beneficial organisms from three main

30

genera: phototrophic bacteria, lactic acid bacteria and yeast. These effective microorganisms secrete beneficial substances such as vitamins, organic acids, chelated minerals and antioxidants when in contact with organic matter. At first, EM was considered an alternative for agricultural chemicals, but its use has now spread to applications in environmental, industrial, and health fields. However, it must be stressed that EM is neither a synthetic chemical nor a medicine. EM technology can bio-treat wastes that are impossible to be treated chemically. The EM tech. de-ionizes all elements present in the sludge.

A.2 EM/NCPC Collaboration EM and National Cleaner Production Center (NCPC) took part in a workshop on cleaner production program (CPP) held at Islamabad during August 2002. There after through mutual discussions and meetings it was decided by NCPC that EM possesses vast experience in bio-remediation (anaerobic) in various fields (industrial solid wastes, sewage effluent and solid wastes) using EM Technology, a technology, without using chemicals but involving the active working and effectiveness of effective microorganisms (EM). NCPC/UNIDO is already working on different options inclusive of aerobic bio-remediation for the disposal of petroleum oily sludge. Aerobic bioremediation of oily sludge was utilized, which was a slow process completed in 40–50 weeks period.

A.3 Sludge Disposal Problem Petroleum refining industries are troubled by the problem of handling substantial quantity of sludge in one form or the other depending upon the nature of the crude, processing capacity, downstream capacities, design of effluent treatment plant, pollution abatement measures and the efficiency-cum-effectiveness of these plants. To fulfill all the legislative and environmental requirements oil refineries need a well-planned oily sludge management strategy to manage the oily sludge with more cost-effective alternative to traditional physical and chemical methods of disposal.

31

B. TRIAL FOR ARL SLUDGE DISPOSAL B.1 Objectives of the trial The Oily Sludge poses various kinds of environmental hazards such as:

· Fire, as oil being lighter floats on water and is liable to catch fire;

· Ground-water pollution due to gradual percolation; · Threat to marine life; · Air pollution due to evaporation.

The objective of the trial project is to provide ARL:

i. Effective Refinery Sludge Treatment providing environmentally friendly/safe disposal

ii. ISO-14000 compliance iii. Cost-effectiveness iv. By-product (saleable)

B.2 Advantages of Bioremediation · An ecologically sound, natural process that destroys organic

contaminants; residues are usually harmless products and byproducts are generally innocuous;

· Instead of merely transferring contaminants from one environment medium to another (e.g. from water to the air or to land) bioremediation completely eliminates the target chemicals, represents a closed solution;

· Bioremediation is far less expensive than other technologies that are often used to clean up hazardous waste cost-effective when compared to other treatment technologies;

· Can be performed on-site, low profile.

B.3 Trial Room The initial plan was to treat 5 Tons of ARL sludge in a closed-door environment. In this regard, the old control room of BB STILLS was selected. The main task was to shift the sludge from sludge pit to room. While shifting, it was decided to reduce the sludge quantity, as it would be easy to transport and handle during the trial, especially while mixing of EM products in the sludge.

32

B.4 Trial Requirements As the trial was conducted in collaboration with four Organizations, ARL/NCPC/ UNIDO/EM; it was, therefore, imperative to summarize the pre requisites and responsibilities to complete the designed trial as per schedule:

i. Provision of required material: Few buckets of 50litre

capacity for mixing EM ingredients, few drums (steel or plastic) 250 or 500 lit capacity to prepare homogeneous solution of EM extended as well as for storage purposes, and big plastic sheets to spread on the floor of the room and to cover the sludge heap for anaerobic bio-remediation. These were arranged by ARL .AM (TS). ii. Apparatus and equipment: beakers for liquid measurements and thermometer for temperature measurement. Arranged by ARL Principal Chemist.

iii. Provision of Transport, room and labor: Transport required shifting oily sludge from the storage lagoon to the trial site, a room and few laborers for mixing EM products with the oily sludge. ARL made the arrangements. iv. Gloves: These were provided by NCPC. v. Sampling and analysis of sludge samples: Samples of oily sludge were taken before the start of the trial and after completing the trial. NCPC took responsibility to get these samples analyzed for NPK, C: N ratio, organic matter and metals, etc. from external agencies/organizations. vi. Room temperature: NCPC Engineers took readings of room temperature. vii. Training of Workers/Technicians: EM team provided necessary training to workers.

33

viii. Monitoring of Sludge temperature and moisture: EM team visited the trial at ARL on weekly basis, took inside sludge temperature measurement and maintained moisture up to 30% ix. EM products: EM team provided all EM Products and transported them from Lahore to ARL (Rawalpindi Islamabad).

C. FIRST WEEK ACTIVITY

C.1 Monitoring during first week Actual data on maximum and minimum room temperature was recorded daily once (table-1). Activity of EM was observed; it was seen that after two days a white coloring of the surface sludge was observed showing the presence of fungus on the surface of heap. Moreover, the hydrocarbon (HC) odor started to diminish significantly.

ARL Safety department also undertook the HC measurement and the reading on the explosive meter was zero, showing no HC vapors inside the room, while at the start it was not bearable to breathe (no actual measurement).

The weather pattern observed over the first week was, with clear and dry sunny days and a bit colder nights. It was observed that after a couple of days brownish discharge probably molasses started to leak towards the external slope of the room. This was due to some loose packing of the sides of the heap. However the flow was trivial and stopped itself it was not dangerous from the safety point of view and the leakage was washed with clean water.

34

Mixing and sampling during Bio-remediation of Petroleum Sludge

Sludge covered with plastic sheet for anaerobic bioremediation

EM products mixing

35

C.2 Analysis of Sludge Samples The sludge samples (before treatment) were sent to PINSTECH and HDIP for the analysis. NCPC coordinated the activity. The description is summarized in table –4.

C.3 Observations During EM Team 1st visit EM team visited to monitor the microbial activity, moisture content and temperature of the heap. The heap was uncovered and the mix was physically checked/examined for the moisture content.

i. According to EM team, the action of the EM process was found speedy as compared to the other sludge’s treated by EM before (e.g. Leather industry sludge).

ii. The moisture content was found to be a little bit less therefore

EM solution already prepared on 29 October 2002 was spread over the whole heap in order to bring the moisture content up to 30 %.

iii. The temp of heap was also observed by inserting a thermometer

at various places into the heap. The temperature ranged between 30~32 C. The temperature range of the heap was quite satisfactory to stimulate the growth and action of the bacteria.

iv. The sludge was examined from all over the heap and at places

from the depth. The smell of the sludge revealed that the activity of the microorganisms is encouraging as there was no bad smell of the sludge.

D. SECOND WEEK ACTIVITY

D.1 Monitoring during second week Actual data on maximum and minimum room temperature was continued to be recorded daily once during the second week (table-1). There was a decrease in ambient room temperature with the start of winter season. Subsequently, when the temperature of heap was monitored, a decrease in temperature was also detected. Moreover, the HC odor was totally diminished. The sludge temperature measured since the start of the trial is given in table-1

36

The weather was cloudy during the second week with a cool breeze during the day and night. It was observed that the brownish discharge from the heap has stopped and everything was ok. D.2 Observations During EMRO Team 2nd visit EM team visited NCPC/ARL on November 12, 2002 to monitor the microbial activity, moisture content and temperature of the heap.

· The heap was uncovered and the mixture was physically

checked/examined for the moisture content. Temperature of the heap was immediately measured after uncovering the heap.

· Additional activity for the second week was to shuffle and mix the upper one-foot of half of the heap. This was done to evenly mix the colonies developed on the surface of the heap, as the penetration through the depth is difficult for microbes.

· Moreover, approx. 2 Sq Ft area of the heap was thoroughly mixed to the depth. This was done to see the difference between the totally mixed portion with the portion, which was mixed till one foot only.

· According to EM team, activity was found a bit slower during the second week. The main reason behind this slow rate of microbial activity could be due to lowering of the temperature by 6°C as compared to the first week inside the heap.

· The moisture content was found enough, as the activity is low during the second week with the decrease in temperature. Therefore, EM Solution was not added.

· The temperature of heap was also observed by inserting a thermometer at various places into the heap. During first week the temperature of the heap was in the range of 30-32°C and the current temperature was in the range of 24-26°C. From EM team point of view temperature range of the heap was still satisfactory to stimulate the growth and action of the bacteria.

· The sludge was examined from all over the heap and at places from the depth. The smell of the sludge revealed that the activity of the microorganisms is encouraging, as there was no bad smell of the sludge. Additionally, houseflies were seen

37

attracted towards the heap after uncovering, which is not the case with the oily sludge in its original condition.

· By mixing it was observed that the original physical condition of the sludge has totally changed. The lump of sludge, which was greasy and cohesive in nature, has transformed in to a brittle and dispersed condition.

E. THIRD WEEK ACTIVITY

E.1 Monitoring During Third Week Actual data on maximum and minimum room temperature was also recorded daily once during the third week. There was a decrease in ambient room temperature with the start of winter season but the constant room temperature range of 19~21°C was observed during the whole week. Subsequently, when the temperature of heap was monitored, the temperature range was same as that of second week.

· The weather was partly sunny during the third week with colder

nights and sunny days.

E.2 Observations During EM Team 3rd Visit EM team visited NCPC/ARL on November 19, 2002 to monitor the microbial activity, moisture content and temperature of the heap.

The heap was uncovered and the mixture was physically checked/examined for the moisture content. The moisture content was found enough for microbes to remain active. Temperature of the heap was immediately measured after uncovering the heap.

During last week EM visit, half of the heap was mixed. This week, the other side of the sludge heap was also mixed/shuffled till upper one-foot of the heap in order to accelerate the microbial activity. This was done to evenly mix the colonies developed on the surface of the heap, as the penetration through the depth is difficult for microbes.

38

i. According to EM team, activity was found faster on the side, which was mixed during last visit.

ii. The moisture content was found enough; therefore EM Solution

was not added. iii. The temperature of heap was also observed by inserting a

thermometer at various places into the heap. The temperature in the mixed side (during last week) was 1~2 C higher than the undisturbed side. This is due to the higher rate of microbial activity in the mixed side. The temperature was in the range of 24-26°C.

iv. The sludge was examined from all over the heap and at places

from the depth. The smell of the sludge revealed that the activity of the microorganisms is encouraging, as there was no bad smell of the sludge but the smell of fermentation has increased.

F. FOURTH WEEK ACTIVITY F.1 Monitoring During Fourth Week Actual data on maximum and minimum room temperature was recorded daily once during the fourth week. Further decrease in ambient room temperature was observed as we reached the fourth week of the trial. Subsequently, when the temperature of heap was monitored, the temperature range was same as that of last week, i.e. it was dropped.

The weather was partly sunny during the fourth week and a bit colder than the third week.

F.2 Observations During EM Team 4th Visit

EM team visited NCPC/ARL on November 26, 2002 to monitor the microbial activity, moisture content and temperature of the heap.

The heap was uncovered and the mixture was physically checked/examined for the moisture content. The moisture content was found enough for microbes to remain active.

39

Temperature of the heap was immediately measured after uncovering the heap. This week no mixing was conducted.

i. According to EM team, activity is slower on the side, which was mixed during last visit.

ii. The moisture content was found enough; therefore EM Solution was not added.

iii. The temperature of heap was also observed by inserting a thermometer at various places into the heap. The temperature was in the range of 23-25°C. The current temperature range is lower than before, which could be due to the lower rate of microbial activity.

iv. The sludge was examined from all over the heap and at places from the depth. The smell of the sludge revealed that the activity of the microorganisms is still encouraging but slow.

G. FIFTH WEEK ACTIVITY

G.1 Monitoring During Fifth Week Actual data on maximum and minimum room temperature was recorded daily once during the fifth week (table-1) . The weather was sunny during the whole fifth week.

G.2 Observations During EM Team 5th Visit

EM team visited NCPC/ARL on December 3, 2002 to monitor the microbial activity, moisture content and temperature of the heap. The heap was uncovered and the mixture was physically checked/examined for the moisture content. The moisture content was found enough for microbes to remain active. Temperature of the heap was immediately measured after uncovering the heap. This week mixing was also conducted.

i. The moisture content was found enough; therefore EM

Solution was not added.

ii. The temperature of heap was also observed by inserting a thermometer at various places into the heap. The

40

temperature was around 22°C. The temperature range was lower than before and low ambient room temperature could be the dominant factor.

Table I: Temperature Monitoring of the Sludge

Under Treatment

*Note: Temperature was monitored at 1200 hrs daily.

S.# Days Date *Max °C

*Min °C

1. Tuesday 29-10-02 2. Wednesday 30-10-02 3. Thursday 31-10-02 26 22 4. Friday 01-11-02 29 23 5. Monday 04-11-02 26 22 6. Tuesday 05-11-02 24 22 7. Wednesday 06-11-02 24 19 8. Thursday 07-11-02 24 19 9. Friday 08-11-02 24 19 10. Monday 11-11-02 23 19 11. Tuesday 12-11-02 21 18 12. Wednesday 13-11-02 21 19 13. Thursday 14-11-02 21 18 14. Friday 15-11-02 20 19 15. Monday 18-11-02 21 19 16. Tuesday 19-11-02 21 19 17. Wednesday 20-11-02 21 18 18. Thursday 21-11-02 20 16 19. Friday 22-11-02 20 16 20. Monday 25-11-02 21 18 21. Tuesday 26-11-02 20 17 22. Wednesday 27-11-02 20 18 23. Thursday 28-11-02 20 19 24. Friday 29-11-02 20 17 25. Monday 02-12-02 21 17 26. Tuesday 10-12-02 22 16

41

Table 2: Average Weekly Temperature of Sludge Treatment Heap

Week Heap Temp

(Celsius) First 30~32 Second 24~26 Third 24~26 Fourth 23~25 Fifth 22 Sixth 23

Table 3: Average room temperature during 6-weeks of sludge treatment heap (Celsius)

Max Reading Max (Avg.) Min Reading Min (Avg.)

29 20.4 16 17.2

iii. The sludge was examined from all over the heap and at places from the depth. The smell of the sludge revealed that the activity of the microorganisms is encouraging but slow. Areas of activity (brown colored spots in sludge) have been observed within the heap. Two samples (brownish & blackish) from the heap have been taken for TPH analysis from QCL, ARL. The results are stated in table-4. (NCPC: ARL analysis results)

iv. Over all bioremediation process was encouraging and was slow expectedly due to winter season.

H. SIXTH WEEK ACTIVITY

H.1 Monitoring During Sixth Week Actual data on maximum and minimum room temperature was recorded daily once during the sixth week

42

H.2 Observations During EM Team 6th Visit EM team visited NCPC/ARL on December 10, 2002 to monitor the microbial activity, moisture content and temperature of the heap.

· The heap was uncovered and the mixture was physically checked/examined for the moisture content.

· The moisture content was found enough for microbes to remain active.

· Temperature of the heap was immediately measured after uncovering the heap.

· This week mixing was also conducted. · The moisture content was found enough; therefore EM Solution

was not added. · The temperature of heap was also observed by inserting a

thermometer at various places into the heap. The temperature was around 23°C.

· The sludge was examined from all over the heap and at places from the depth. It was observed that the sludge is fully fermented, the color is changed into light brown and black mix, the smell is fermenting smell and the sludge is decomposed well so that the brittleness is formed. Two samples (brownish & blackish) from the heap have been taken for TPH analysis from QCL, ARL.

· After observing the completion of bioremediation/composting process, it was concluded that the oily sludge has been converted to a bio-fertilizer. The first phase of the trial was considered accomplished. This sludge can be used as by-product for horticulture and agro purposes. Once the compost is ready it must be used as by-product with in 10-15 days of its preparation.

· First phase of 100% (undiluted) oily sludge bioremediation trial has been successfully completed.

· This week, about 20kg of composted sludge was brought to NCPC for pot experiment. Treated sludge was mixed with soil 1:1. Two pots of treated sludge and 2 pots of control ? were designed for a preliminary test.

43

I. RESULTS AND DISCUSSION During the first week (29th October to 5th November) it was observed that the action of EM was found speedy and huge number of fungus colonies was visible. The temperature of the heap by inserting a thermometer into the heap at 4-6 places was noted. On an average it was 32ºC. The moisture content was maintained up to 30% with EM solution already prepared on 29th October. The temperature of the heap fell to 25ºC in the 2nd week (12.11.2002) and it was 24ºC on 03.12.2002(5th visit). This means that the temperature remained at 32ºC for 7 days after that at 24ºC for 28 days (05.11.2002 to 03.12.2002) and at 23ºC for 7 days (table-2). With even this low temperature range, the 100% pure undiluted oily sludge of ARL was completely changed to a bio-sludge within a period of 6 weeks.

If the temperature remained a little higher in the range of optimum (30 to 40oC), one could expect even earlier anaerobic bio-remediation. The heap temperature dropped to 23oC after 6 weeks, simply due to the beginning of winter season. In spite of this, the EM-micro-organisms remained active and a lot of fungus, white in color, appeared on the surface of the sludge, even the activity in deeper layers was medium to high. The typical odor of the sludge disappeared, it was perceptible to smell the fermented odor. The original sludge was more cohesive in structure. With this trial it has become brittle and breaks into small aggregates.

It is worth mentioning here that the increased activity of microorganisms not only enhanced the number of microorganisms manifold (105 to 107) but also produced heat and energy during the phase of increasing their population while consuming organic matter and certain heavy metals. The is the reason that the inside heap temperature remained on reasonably higher side (table-2) as compared to average minimum and maximum room temperature (table-3).

44

1.1 Reduction in concentration of Heavy Metals

I.1.1 ARL Sludge Treatment Trial The perusal of data of oily sludge given in table-4 and its graphical presentation in Fig-1 indicate that Ba has been reduced by 85% in the EM treated oily sludge as compared to original oily sludge, and Pb, Fe, Zn and Ni have been reduced by about 50% in the bio-sludge. The contents of As, Cr, Cu and Mn showed no change.

I.1.2 Other Successful EM Experiences i. The EM does reduce the contents of heavy metals in EM-

treated sludge. EM reduced Cr from 50,000ppm to 450ppm in Leather industry sludge in Pakistan (PTA trial 2002).

ii. The research carried out by Eric Nielsen in Denmark on

wastewater sludge indicated a reduction in Cd, Cr, Cu, Hg, Ni, Pb, and Zn by more than 50% with EM application (6th International Conference on Kyusei Nature Farming held at University of Pretoria, South Africa October 1999).

iii. EM application reduced Dioxin generation in the garbage

incinerator in the fly ash as well as in the residual ash by 99.9% and 74% respectively (Masato Miyajima et al 6th International Conference, Pretoria, South Africa 1999).

iv. The research carried out by Dr. Syed Ali in Egypt on

sewage/industrial wastewater indicated a reduction in Cd 40%, Pb 90%, Cu 20%, Zn 99.9%, Ni 80% and Cr 98% with EM application (EM Treatment on Sadat City Sewage/Industrial Waste Water report of Dec. 1997, Egypt).

1.2 Preparation of by-product

The EM treated sludge being a bio-fertilizer can be used in agriculture and a substitute to farm yard manure and as a soil amendment to improve/reclaim salt affected lands (a major problem in Punjab and Sindh). The bio-sludge is to mixed with dry soil in 1:1 ratio before use elsewhere. With this it will become a by-product which is easy to handle, transport, store and applicable in agriculture, horticulture, floriculture and

45

house gardening etc. as and when needed if the by-product is well- dried and packed in bags. While applying in agriculture the dozes are to be determined by experiments carried out under given climate conditions. The application of known quantity of by-product will help to further dilute the concentration of heavy metals as it will be mixed with one acre of ploughed layer (0-9 inches) weighing 1176120kg soil as per calculation methodology.

Calculation Methodology The soil possesses a definite weight depending upon its texture, structure and bulk density (gm/cm3). The weight of a ploughed layer (1ft x 1ft x 3/4ft) or (30.48cm x 30.48cm x 22.86cm) with bulk density of 1.2gm/cm3 varies from 25kg to 30kg.

For practical purposes especially for bio-by-product trial let us take 27 kg weight of one square foot soil up to a depth of 9 inches. Calculated further for one acre (220ft x 198ft = 43560ft2) the weight of ploughed layer comes to 1176120kg.

Normally, one-ton (1000kg) farmyard manure is added to one acre to keep organic matter content within satisfactory limits. The weight of farmyard manure per ft2 (1000 divided by 43560) comes to about 23g but it is 0.85g/kg of soil or it is about 23g/ft2 up to a depth of 9 inches. Now our bio-by-product is rich in effective microorganisms and other elements, it is practicable to add 0.5 ton/acre to achieve the same results. To arrive at a reasonable correct decision the trial, on newly created plot by spreading soil material on existing ground, be conducted with at least 4 or more replications. On the formulation, the doses of bio-fertilizer will be 12g, 25g, 50g, and 100g/ft2 area for vegetables.

Similarly, the doses can be formulated for cash crops e.g. wheat, rice, cotton, etc. At least 2 years trial period results will be reasonably decisive.

46

1.3 Fate of Heavy Metal Cr Cr in its hexavalent form is dangerous for health. It is 26ppm in the bio-sludge (table-4). It will become 13ppm in the by-product (bio-sludge+soil 1:1 ratio), in simple words one kg by-product contain 13mg Cr. Normally, one M.ton of farm yard manure is added to ones acre to have good physical conditions and microbial activity. By-product being very rich or high in microbial activity and other NPK nutrients (table-5) the quantity can be reduced to 500kg/acre. This means 6.5kg Cr will be added to one acre (1176120kg soil); this means 181kg soil (6.60ft2) will receive 1mg Cr. In other words 0.149mg Cr will be in one ft2 of land. Now the question arises that how much Cr will be taken up by Wheat, Rice or Vegetables when many plants of wheat/rice per ft2 are growing, again with a number of grains in ear/spike. For reference purposes WHO (Environmental Degradation by Engineer Col. Rtd. Mumtaz Hussain page 73) has allowed 0.05mg/lit of Cr in drinking water for human consumption. At least 5-10 liters/day of drinking water is the requirement. It will add 0.25 to 0.5mg of Cr daily to the human body. While in our case it is 0.149mg Cr per ft2 of land. It means WHO has kept in view the daily absorption of Cr into the blood. On the basis of this simple mathematical calculation the ARL-by-product (ARL oily sludge treated with EM and mixed with soil 1:1 ratio) may safely be declared useful for application in agriculture but in known quantity. It will have to be determined by agricultural experiment with all prevailing practices of soil and water in different areas with heterogene soil.

The special odor of Hydrocarbons vanished within a week and at the end of 2nd week, it was observed by all that houseflies were attracted by the treated sludge. The concentration of Hydrocarbons was tested and it was found nil.

47

Table 4 Analysis of Pure Oily Sludge and *Bio-Oily Sludge

Test Before Treatment After Treatment

PH

HDIP 6.7

HDIP 7.0

Nitrogen Carbon Hydrogen Sulphur

NIBGE 0.83 81.5 9.25 3.61

NARC

Metals Pb Mn Ba Fe Zn Ni As Cr Cu

PINSTECH % ppm 0.020 200 0.022 220 0.257 2570 3.443 34430 0.144 1440 0.009 90 0.0029 29 0.0024 24 0.005 50

PINSTECH ppm 83 207 382 17373 765 52 36 26 42

QCL TPH

65.10

76.2

Table –5. NPK & OM analysis before and after treatment

Tests Original Soil Untreated Sludge Treated Sludge + Soil

N ppm 800 0.336 1700 P ppm 2.5 584 7.48 K ppm 76 0 374 Oragnic Matter %

1.8 65.1 5.1

48

Fig.1: Metal Analysis of Oily Sludge before and after Treatment with

EM

Metal Analysis

0

50

100

150

200

250

PPM

-4,650

350

5,350

10,350

15,350

20,350

25,350

30,350

35,350

40,350

Fe, B

a, Z

n PP

M

Pb Mn Ni As Cr Cu Ba Fe Zn

The above results show that the metals level has generally decreased across the board.

49

While mixing Bokashi on 3rd December (after 35 days) with the sludge, it was quite distinct and clearly perceivable that the sludge after treatment with EM has changed its physical properties. Sludge became brittle instead of cohesive mass and breaks into small aggregates with pleasant fermenting smell. This made the mixing of Bokashi with EM sludge easy. Beside these, colonies of effective microorganisms were clearly visible with human eye everywhere.

After six weeks the sludge which was odorous greasy and black, transformed into dispersed & brownish shape and the pungent odor diminished and changed into sweet fermentation smell. During the process of sludge bioremediation it was observed that houseflies and mice/rats were attracted by the sludge and found there alive which shows that the sludge has lost its hazardous ingredients or nature.

On 10th December 2002 (after 43 days) the trial was declared successful by EM team. This means oily sludge was completely converted to a bio-sludge (bio-fertilizer). This was authenticated by Prof. Dr. Teruo Higa, the founder/inventor of EM Technology, in the presence of NCPC and ARL dignitaries during his visit to this very trial on 14th December 2002 after the symposium at NCPC. This is being tested through application as fertilizer in the field where seasonal crop onions are to be planted.

Symposium On Bio-Remediation Of Industrial Sludge A half-day symposium on Bio-remediation (anaerobic) of industrial sludge using EM Technology was arranged by National Cleaner Production Center (NCPC/UNIDO) in honor of Prof. Dr. Teruo Higa, founder and inventor of EM at NCPC, Rawalpindi inviting dignitaries of ARL, Federal Government, and Ministry of Petroleum.

Environmental Protection Department, and UNIDO representative Dr. Carlos E. Chanduvi-Suarez were briefed about the NCPC role to introduce and promote Cleaner Production Techniques and processes and pollution control at source along with integrated waste management by the Advisor,

50

NCPC, and opening remarks about the industrial waste especially sludge being a menace and threat to environment, Prof. Dr. Teruo Higa was invited to deliver his lecture and experience on industrial solid waste management. Prof. Dr. Teruo Higa presented application of EM Technology to not only the waste management but also for agriculture with proven benefits and success stories all over the world with pictures and slides. The trial study on treatment of EM Technology on ARL oily sludge was completed successfully within 6 weeks by EM team (presented by Dr. Syed Ali) in collaboration with ARL and NCPC/UNIDO. In the end the UNIDO Representative Dr. Carlos thanked Dr. Teruo Higa and while concluding the symposium informed thus UNIDO will promote EM Technology in Pakistan, and UNIDO network worldwide will consider its application internationally as well.

Dr. Syed Ali’s Lecture Prof. Dr. Teruo Higa’s Lecture

at the Symposium at the Symposium

51

Original sludge Mixing EM products

Microbial activity (fungus) Sludge during bioremediation

Sludge after fermentation

52

Rat enjoying EM Treated sludge as food

Fatty rat is hiding at bioremediation site

53

1.4 Mechanism

The mechanism of bioremediation involves EM-microorganisms (non-GM) which belong to 3 main genera:

· Phototropic bacteria; · Lactic acid bacteria; and · Yeast.

EM secrete beneficial substances:

· vitamins; · organic acids; · chelated minerals; and · antioxidants

Phototrohpic bacteria have the characteristics to accept alkaline substances and produce protein. Yeast contains Apo-protein-A&B which also converts such elements into protein and chelates. EM Microorganisms produce NH3, CH4, CO2, etc. during fermentation to enhance the nutrient value of organic matter. Thus EM regenerates the organic matter by eliminating the harmful microorganisms. The acidic nature of EM reduces pH of the alkaline substances. The anti-oxidation nature of EM stops from chemical reactions and bonding.

A compound aluminum ammonium dibutylamine and bacteria like:

· Rhodobacter; · Pseudomonas; · Lactobacillus; · Furabacterum; · Gluconobacter; etc.

have the characteristics to de-ionize the harmful elements and de-toxify them.

54

J. CONCLUSION It can safely be concluded that the pure oily sludge after treatment with EM Technology has transformed into odorless disposable bio-by product having pleasant fermenting smell, easy to handle, not dangerous to workers and safe to transport else where for use. With this the first phase of the trial is completed successfully. Phase II is being planned. As per advice of Prof. Dr. Teruo Higa, the bio-sludge can be mixed well with the soil (1:1 ratio) for its comfortable transport and practical application in agriculture, orchards, household gardening, pot flowering and for reclamation of salt affected lands, a burning problem in Punjab and Sindh.

The main conclusions are:

1. Effective Refinery Sludge Treatment providing

environmentally friendly/safe disposal; this has been demonstrated;

2. ISO-14000 compliance; ensured; 3. Cost-effectiveness; being evaluated in phase II i.e.

application as bio-fertilizer 4. By-products (saleable); oily sludge has been converted

which is easy to handle, transport, store, and applicable in agriculture. To start with may be distributed free of cost and later on commercial basis.

K. RECOMMENDATIONS:

K.1 Application as an Organic Fertilizer: The treated sludge of two tons after full fermentation is to be treated with equal amount of dry soil and after mixing the treated sludge would be ready to apply and use for agricultural purpose as an organic fertilizer. In order to reduce the cost of the sludge treatment, it is planned to have two application trials: (i) comparison of yields and cost with farmyard manure, (ii) comparison of yield/cost with synthetic fertilizer.

55

K.1.1 Comparison with Farm Yard Manure (Original Cost and Dose) The cost of treating sludge here is considered as $44/Mtons. It is planned to use half canal plot divided into two portions, each of 1 / 4 canal area. The first plot would be cultivated with EM treated sludge fertilizer and second with the conventional fertilizer of farmyard manure. (In both the trial and control plots onions have already been planted).

K.1.2 Comparison with Synthetic Fertilizer (Optimizing Cost and dose) It is practicable to add 0.5 ton/acre to achieve the same results. To arrive at a reasonable correct decision the trial, on newly created plot by spreading soil material on existing ground, be conducted with at least 4 or more replications. On the formulation, the doses of bio-fertilizer will be 12g, 25g, 50g, and 100g/ft2 area for vegetables.

Similarly, the doses can be formulated for cash crops e.g. wheat, rice, cotton, etc. or we would use one kanal plot divide into two portions each of half kanal area and similarly one plot would be cultivated with the EM treated sludge of 151. 50 Kg quantity and the other with synthetic fertilizer @ Rs. 3000/acre so that the process of sludge treatment with EM technology can be justified and shown with the break-even concept. In both the cases the comparison would be carried out by Observing / Checking

i. The physical growth of the crop ii. Chemical analysis of the crop iii. Chemical analysis of the soil before and after cultivation for the

following parameters iv. Nitrogen (N), Phosphorus (P), Potassium (K) Carbon Nitrogen

ratio(C: N), Organic Matter.

56

K.2 Full Scale Sludge treatment at ARL It is recommended that based on the success of this trial, this technology may be applied to the estimated 400 – 600 M Tons/year of ARL sludge on large-scale. Immediate feasibility should be prepared.

K.3 Expansion to other Refineries As the trial is the first of its kind in Pakistan so it can be applied on full-scale to other Refineries as well i.e. NRL, PRL, and PARCO.

K.4 Proposal for other Upstream and Downstream Petroleum Sector Using the experience of this trial and the methodology used, proposal may be developed for other petroleum sector sludge upstream and downstream.

59

Phase II- Application of EM-Bio-Sludge in Agriculture

Having completed the Phase -I trial at Attock Refinery Limited (ARL) on “Microorganisms playing their role in Bioremediation of Petroleum sludge using EM Technology” within a period of 6 weeks (29th Oct to 10th Dec 02) in collaboration with UN/NCPC Rawalpindi, it was decided to apply the byproduct biologically rich in effective microorganisms in agriculture in order to reuse the by product (beneficial and practical use of a biologically active resource prepared from a waste resource). Here the effectiveness of EM treated sludge would be proved through physical growth and chemical analysis of the crop. ARL has its own field area within its boundary and is presently being used for growing vegetables for its staff. A Horticulturist (MSc. Agri) is supervising all the agricultural operations for growing vegetables. A ten marlas plot (126 ft x 18 ft) was earmarked in the vegetables area of New Abadi. The area was already being used for growing vegetables meaning thereby that the soil was in good tilth. Chilies were the previous crop. After its harvest the land was ploughed well. It was divided into two equal plots. One plot was kept for control in which farmyard manure (FYM) was added and in the other plot EM-Bio-Sludge, after mixing with equal quantity of soil to eliminate its oily nature. The samples of the original soil and biofertilizer (treated sludge + soil) were taken and got analyzed. The results are presented in figure-2. A comparison of NPK and organic matter of the biofertilizer and original soil shows that the NPK and organic matter values are much higher in the biofertilizer than in the original soil. This means that the biofertilizer has all the properties of a biofertilizer. In fact the dozes of the biofertilizer for vegetables, gardens, floriculture and crops are to be determined through experiments under various climatic and soil conditions. To start with a pot experiment was also undertaken to see the effect of biofertilizer in growing flowers.

60

Of the biofertilizer about 1.2 tons was spread over 5 marlas of land (63 ft x 18 ft) ands mixed well with the upper layer of the soil. EM irrigation was given to accelerate the decomposition of biofertilizer. Each plot was sub divided in ten small plots in order to irrigate with groundwater pumped out with a hand pump fixed with an electric motor to accommodate the small discharge of the hand pump. The onion nursery was transplanted in both the plots during the end of second week of February 2003.

Fig-2: NPK & OM Analysis before and after treatment

NPK & OM Analysis

800

1700

584

7.48

76

374

65.1

5.1

0.336

2.5

0

1.8

0 500 1000 1500 2000

Original Soil

UntreatedSludge

Treated Sludge +Soil

N ppm P ppm K ppm Oragnic Matter %

Oragnic Matter % 1.8 65.1 5.1

K ppm 76 0 374

P ppm 2.5 584 7.48

N ppm 800 0.336 1700

Original Soil Untreated Sludge Treated Sludge + Soil

61

The biofertilizer plot received groundwater irrigation augmented with EM extended solution whereas the control plot got normal groundwater irrigations. On 4th March 2003 the experiment was visited and it was not possible to observe any difference in the growth between the two plots as the crop was too young. Later on the crop was visited on 7th May 2003 along with the honorable guest from U.A.E., Mr. Jabber Al-Mazroui, Chief Executive Officer, Emirates Science and Environmental Safety (ESES) and NCPC / EM experts. At this stage it was observed that EM plot was showing better growth both in the bulb and aerial part (photo).

Preliminary Pot Experiment

62

Experimental Plot

EM biofertilizer plot Controls

Honourable guest Mr. Jabber Al Mazroui,

UAE, in conversation with UN/NCPC expert

APPLICATION OF BIOFERTILIZER IN AGRICULTURE