eChapters

18.1 Estrus Detection
18.1.1 Estrus Detection on the Basis of External Signs

The following signs are suggestive estrous period in cattle and buffaloes

1.    Partial inappetance due to excitement

2.    Drop in milk yield

3.    Restless ness of varying degree

4.    Frequent micturition with raised tail and crouching the back and lumbar region

5.    Marked bellowing

6.    Swollen vulvar lips- Due to swelling the wrinkles on vulvar lips disappear. The vulvar lips appear turgid and stand prominently.

7.    Congestion of vulvar lips 

8.    A clear, shiny mucus discharge seen at the vulvar lips extending down to perineal region and smearing the hind quarters. The discharge is stingy, having good consistency and no odour.

The bacterial load in the semen samples is estimated by standard plate count method. The bacterial contaminants of semen have been a major concern for most of the frozen semen production laboratories as it adversely affects semen quality and hence subsequent fertility. There exists a significant negative correlation of bacterial load and sperm livability, motility and in vitro fertility tests. Macrophages and polymorphonuclear granulocytes, which form the first line of defense against these microorganisms, produce reactive oxygen species (ROS) to kill these microorganisms. ROS, however, is also released outside these cells and may react with molecules and cells such as spermatozoa in their vicinity. The cellular antioxidants, present mostly in the cytoplasm, are scanty and inadequate to counteract this ROS as the sperm cell cytoplasm is very small (20 µm3) and mostly distributed in the midpiece. Therefore, semen samples with minimum bacterial load are expect to give better results on insemination.

The following tests are discussed under this section:
    16.1    Estimation of ascorbic acid concentration
    16.2    Effect of cold shock
    16.3    Mitochondrial activity
    16.4    Capacitation of spermatozoa and calcium influx 
    16.5    Morphometric analysis
    16.6    Flow cytometry

16.1 Estimation of Ascorbic Acid Concentration

Ascorbic acid concentration in semen is beneficial for its antioxidant action. It protects the spermatozoa against oxidative damage of free radical produced in the semen.

Materials Required

    1.    Test tubes with stand
    2.    Burrette with stand
    3.    Centrifuge tube with stand

Semen metabolic activity can be assessed with the help of the following tests:
    
15.1    Incubation test
    
15.2    Methylene Blue Reduction Test
    
15.3    Resazurine Reduction Test
    
15.4    Millovonoo’s Resistance Test (‘r’ test)
    
15.5    Fructolysis Index
    
15.6    Oxygen utilization test
    
15.7     Glutamic oxaloacetic transaminase activity

15.1 Incubation Test

The longevity and viability of spermatozoa during storage is a good index of fertilizing ability of sperm. In this test assessment of motility is carried out over a period time to obtain an idea about the length of time over which acceptable level of motility is maintained in the semen sample. Therefore, this test serves as a mean of determining storage probability within a fairly short period of time.

Only few single sperm viability parameters show a significant correlation with fertility of semen samples. Therefore, need to explore functional in vitro fertility tests having capability to disclose the ability of spermatozoa to undergo complicated processes such as capacitation, binding of zona pellucida, acrosome reaction, fertilization of Oocyte and induction of embryo development invitro have been designed and explored for their correlation with fertility achieved after artificial insemination.

In vitro fertility tests used in evaluation of semen includes: 

14.1 Estimation of membrane integrity
        
    14.1.1 Morphological integrity
        
    14.1.2 Functional integrity
        
    14.1.2.1 Hypoosmotic swelling test
        
    14.1.2.2 Acrosome integrity

14.2     Cervical Mucus Penetration Test

13.1 The Normal Sperm Morphology

A mature spermatozoa is an elongated cell consisting of a flattened head containing the nucleus, a neck and a tail containing the apparatus necessary for cell motility. The sperm cell consists of the following parts covered by the plasmalemma or plasma membrane: 
    
13.1.1    Head
    
13.1.2    Neck
    
13.1.3    Tail / flagellum – The sperm tail consist of     
    
a.    Mid piece
    
b.     Main piece / Principal piece
    
c.     End piece

Sperm Plasma Membrane

The sperm plasma membrane is three layered membrane. It lies in close contact with the underlying cytoplasm and attached at three places i.e. over anterior outline of the sperm head, around the base of the head, in circle around the distal end of the midpiece (Jensen’s ring). This attachment causes creation of three compartments with probably no communication with each other. The sperm plasma membrane is invisible under normal light or phase contrast microscopy but can be visualized after addition of substances with light refractory index e.g. bovine plasma albumin or synthetic polysaccharides. It is very easily permeable to water but there is low permeability for inorganic cations (Na, K, Ca etc.) and even lower permeability for anions (NO3, Cl). Exposure of sperm cells to hypertonic media will cause the water to go from sperm to the surrounding media leading to shrinkage of sperm cell whereas exposure of sperm cells to hypotonic media will induce influx of water causing swelling of the sperm cell. The more water that goes into the tail the more it changes its shape and becomes rounded to give room for increasing amount of water. As the length of the tail decreases as it becomes round, the tail fibrils have less space and tail bends and coils within the membrane vesicle. Tail fibrils are not damaged initially and will continue to contract till the critical volume of bull sperm is 3.8-3.9 times of its volume in an isotonic solution. The swelling and change in shape of the spermatozoa are momentary processes and can be corrected if hypotonicity of the media is corrected before the sperm cells reach the critical volume for rupture.

Sperm membrane is responsible for maintenance of the intracellular biochemical environment. Damaged plasma membrane is responsible for the leakage of electrolytes and organic substances such as enzymes and may also lead to damage of sperm cells by foreign substances from the surrounding media.

The following tests are used for microscopic evaluation of semen: 
    
12.1    Sperm motility
12.2     Per cent Live sperm
12.3     Sperm concentration

12.1 Sperm Motility

The following two methods of estimation of sperm motility are followed in routine practice:
 
12.1.1    Mass activity
12.1.2    Progressive sperm motility
12.1.1 Mass Activity

Mass activity can be defined as ‘en-mass activity of the spermatozoa in the semen, immediately after collection’. Mass activity is generally assessed by vigour of swirls and waves motion in undiluted semen under 20 X magnification and expressed on 0 to 5 scale.

Gross evaluation is generally done to get a rough idea of the quality of semen and to assess the semen producing capacity of the bull. This also gives a fairly good idea about the reproductive health and any pathological conditions of the semen donor. The following are recorded under gross evaluation of semen:

    11.1    Color and appearance of semen
    11.2     Semen Volume
    11.3    pH of semen
    11.4     Semen consistency/ Semen density
    11.5     Presence of foreign matter
    11.6     Gross motility

While collecting and processing semen the following points needs to be considered:

1.    Good quality Artificial Vagina (AV) should be used for semen collection. Strict hygiene should be maintained while collecting semen.

2.    Highly scientific and flawless collection technique should be used for semen collection.   

3.    Neat semen should be maintained at close to body temperature.

4.    Semen should not come in contact with any contaminant such as lubricant, dust, infection etc. in the collection yard or in any stage of semen processing.

5.    Direct exposure of semen to sunlight or cool white fluorescent light should be avoided as it may cause chromosomal damage.

6.    Enclose all cool white light sources e.g. laminar flow hood, cold room etc. in gold or orange sleeves that absorb ultraviolet and low wavelength visible light (< 475 to 500 nm).

Early research in field of semen storage and delivery systems was conducted in USA. The straw technique for the storage of frozen semen was evolved by work of French, Danish and German workers whereas the pellet system of storage of semen was devised by Japanese scientists. The different packing systems of frozen semen are:

    9.1    Ampoule method
    9.2    Pellet method
    9.3    Straw method

9.1 Ampoule Method
In this method the semen is placed in a glass ampoule and is subsequently sealed and frozen in liquid nitrogen. The ampoule is thawed just before insemination and is then cut open. The semen is withdrawn into glass catheter and used for AI. Since the ampoule is sealed there is no risk of contamination during storage. The relevant details can be marked on the ampoule.

Deep freezing or cryopreservation of semen refers to freezing and storage of semen at –79ºC using dry ice and alcohol or at -196ºC using LN2 as refrigerant. Cryopreservation of semen was first attempted by Parkes in 1945. But cryopreservation of semen received a boost by discovery of glycerol a cryoprotectant by Polge in 1949.  Bull semen was first successfully frozen by Stewart in 1951. Cooling rate is a vital factor determining the post thaw survival of the sperm cells. Cell death occurs in rapid freezing due to formation of intracellular and possibly extracellular ice. In contrast slow cooling rates cause cell death due to diffusion of intracellular fluid into the extender, which then become increasingly hypertonic as the extender’s water freezes causing intracellular osmotic damage as the cell dehydrates. Hence, the cooling rate needs to be balanced in a way that it is neither too fast to cause cell death due to cold shock nor too slow to result the same due to osmotic shock. Ultra rapid freezing gives excellent morphological preservation but with poor retention of functions on thawing due to large number of minute crystal formation.

The Tris citric acid egg yolk glycerol extender is the most frequently used extender in the cryopreservation of cattle and buffalo semen. Preparation of the extender plays a very important role in determining the quality of the processed semen. Therefore, care must be taken to ensure that the extender is prepared aseptically and it contains all the ingredients in appropriate concentration. Extender should always be prepared under a laminar air flow. It is also important to check that the pH and osmolarity of the semen extender for a particular species corresponds to the prescribed limits for that particular species. The Tris citric acid egg yolk glycerol contains the following ingredients:

Semen dilution/ extension is done to meet the following objectives

1.    To preserve the fertilizing potential of semen for a long period of time

2.    To increase the number of services per ejaculate. Ideally approximately 500 cows/ buffaloes can be inseminated from diluted/ extended semen of  a single ejaculate.

3.    Extenders provide the spermatozoa with a medium in which they are able to remain viable and fertile for a prolonged time period.

In order to be an ideal extender, it should be able to provide conducive environment to prolong sperm survival and modify the sperm structure to prepare them for freezing through the following properties:

1.    It should be isotonic with blood and should maintain this during preservation.

For the diagnosis of clinical reproductive problems it is essential to collect blood, penile swab, Prepucial swab, prepucial washing, semen, smegma etc from breeding bulls. These samples are dispatched to the nearest laboratory for clinical diagnosis. 

5.1 Collection of Blood Serum

The bull is properly restrained and jugular vein area is cleaned and sterilized with 70% alcohol. An sterile hypodermic needle is used to collect 5-10 ml blood aseptically in a sterile test tube. This test tube is then kept in a slanting position. Serum is separated after the formation of blood clot and dispatched to the laboratory.

5.2 Collection of Seminal Plasma

Semen sample is collected as per the standard procedure. Carbolic acid or 1 % sodium acid is added to preserve seminal plasma at the rate of 2 drops in 1 ml ejaculate in the centrifuge tube. It is then centrifuged at 3000 rpm for 20 minutes. The supernatant is collected and stored in deep freeze till further analysis.

Semen collection is the most important component of any frozen semen production programme as a high quality end product is not possible unless the raw material is of excellent quality. Therefore, all care should be taken to collect semen aseptically with the aim to harvest the best quality neat semen for processing. The person collecting semen should understand bull’s behaviour and should be friendly with him. Professional bull management results in increasing sperm output and enhancing productive life of the bulls. One must know potentials and limitations of the reproductive capacity of bulls. A highly professional and dedicated team of bull handlers and semen collectors is required for this purpose. The persons involved in bull management should be familiar with bull’s sexual behavior and male reproductive physiology. Preferably a veterinarian should collect semen.

3.1 Selection of Breeding Bulls

A sire is called half the herd and the breeding bulls to be used for semen production should be carefully selected. Selection of superior sire for breeding purpose is very important for the success any livestock breeding programme. Therefore, breeding sire must be carefully selected and maintained in scientific way so that its breeding life is prolonged and maximum number of frozen semen doses can be produced from it. 
The selection criteria for future bulls should be meticulously designed keeping into account all the factors/parameters which affects the breeding programme. The selection of breeding bulls is based on three broad parameters-

3.1.1 Pedigree Examination
3.1.2 Gross Physical Examination
3.1.3 Laboratory Examination

Success of AI programme depends on fertility of bulls used for semen production. If infertile bulls are selected in semen station for production of semen doses, large population of cows inseminated with such doses is affected. Therefore, the breeding bulls or bulls to be used for frozen semen production should be selected carefully keeping in view genetic potential, disease condition, reproductive health, libido and semen production potential of the bull. The emphasis should not only be on elimination of bulls with questionable or unsatisfactory potential, but also on induction of bulls with good semen production potential into the semen station. 

The following aspects should be taken into consideration while performing a breeding soundness examination of bulls:

Knowledge of male reproductive system and its physiological functions is essential for understanding male reproductive behavior and for effective reproductive management of breeding bulls. Therefore, personnel involved in management of breeding bulls, semen collection, handling and artificial insemination must have sound knowledge of male reproductive anatomy and physiology of breeding bulls. This section provides a recapitulation of the basic physiology and anatomy of male genitalia.
 

Functions of Male Genital System

    (a)    Primary function of the male genital system is to monitor reproduction by providing cutaneous and glandular system of male gonads

    (b)    Secondary function of male genital organs is to provide glandular, ductular, copulatory and auxillary system associated with male reproduction.

Appendix A: Diagrammatic Presentation of Processes Involved in Obtaining an Indian Patent

6.1    Copyrights
    
6.2    Trademarks
    
6.3    Geographical Indications
    
6.4    Trade Secrets

5.1    TRIPS
    
5.2    WTO
    
5.3    WIPO
    
5.4    Paris Convention
    
5.5    Berne Convention
    
5.6    Budapest Treaty
    
5.7    Madrid Agreement Marks (MAM)
    
5.8    The Patent Law Treaty (PLT)

4.1    Provisional and Complete Specifications
    
4.2    Publication
    
4.3    Request of Examination
    
4.4    PCT Application: Gateway to Foreign Patents

3.1    Competitive and Collaborative Research
    
3.2    The Strategy to Protecting Plants : Plant Variety Protection Act
    
3.3    IPR Issues in Biotechnological Patents Haldi, Neem and Basmati Rice
    
3.4    Patent Litigation

2.1    Specification Drafting
    
2.2    Methods and Precautions in Drafting Patent Specification
    
2.3    Disclosure Requirements
    
2.4    Patent Claims
    
2.5    Filing of A Patent Application

1.1    General Introduction, History, Ownership of Tangible and Intellectual Property
    
1.2    Patentable Subject Matter
    
1.3    Basic Requirements of Patentability
    
1.4    Novelty, Public Domain, Obviousness and Non Obviousness
    
1.5    Inventive Step (Non-Obviousness) and Industrial Application

ABSTRACT
It is impossible to envision a world without energy provisions and has become inevitable and an integral part of a nation’s prosperity. It has unparallel contributions towards the socioeconomic empowerment, industrial growth, and technological advancements, as also in mitigating greenhouse gas (GHG) emissions. Judicious utilisation of energy sources without infringing upon the sustainable energy demand is the need of the hour. Taking India’s huge bovine strength into account, biogas technology has the potential to transform the standard of living of rural millions. Effectively implementing the Govt. of India’s National Biogas and Organic Manure Programme (NBOMP) could save precocious demand on electricity, fertiliser and firewood while promoting self-sustainable organic farming. In this chapter, the cost and benefit analyses of biogas technology (involving biogas plants of different capacities) are discussed along with the long-term approach at the grassroots level to promote digested slurry for organic farming. The commercialisation and carbon crediting of biodigester technology is also narrated. It is necessary to chalk out a revenue model to commercialise slurry at the grassroots level to economically empower the beneficiaries, from family-type to industrial-scale biogas plants. Small-scale biofertiliser hubs at district level may be organised to collect dry slurry from the beneficiaries and process them further to make it market-ready through entities operating at the national level developing manure for agricultural practices, as also feeding materials for animal husbandry practices.

ABSTRACT
The ‘steel-drum type’ biogas plant installation programme in Punjab was initiated in the early seventies, whose construction works were entrusted on the Department of Agriculture. Because of their high cost and high failure rates then, only the affluent farmers could afford to install. In the meantime, a cheaper Chinese model, also known as Janta type (dome-type replacing the drum-type) plant, became popular but was still out of the reach of ordinary farmers. With dual objectives of cost reduction and efficient biogas production to make it affordable to small farmers, a coordinated research was initiated. Detailed extension surveys were carried out in Ludhiana district which inferred that the farmers did not install the gas-holding steel drum though they purchased it. A significantly low cost Katcha-Pucca plant model of 15m3/d capacity was developed and installed in Sangrur district. Similar digester was tried for Janta model in which the inlet, outlet and dome (gas holder) were masonry works but the digester was a Katcha pit, the dome of the plant being modified to a semi-circular dome. More than 80 such plants are functional in a single village, and more than 40–50% of the plants installed in Punjab and Haryana are these PAU designs. The chapter deals in greater details on the design aspects and the extensive field works carried out to achieve these successes, along with some relevant facts and figures. The write-up is written as a commentary, rather than a typical chapter, for the benefit and interest to the readers.

ABSTRACT
Nisargruna biogas technology has been developed in BARC for processing different types of biodegradable waste. This technology is based on biphasic separation of aerobic and anaerobic stages. Introduction of aerobic predigester has increased the efficiency of the process in terms of enhancing the quality and quantity of methane and also reducing the retention time of the process. Process parameters of the plant were studied and found that the modified design has proved very effective in faster digestion of complex waste. The technology has been commercialised and has spread across different parts of country for processing variety of wastes including kitchen waste, municipal waste, agriculture waste, textile waste and poultry waste. Microbial population from different stages of the plant was also studied and found their ability to convert waste to biogas, containing mainly methane.

Keywords: biogas, biphasic digestion, green energy, methane, microbial fermentation, Nisargruna, waste management

ABSTRACT
The global energy demand is increasing day by day. Now-a-days, the supply of global energy is highly dependent on fossil fuels (viz., crude oil, lignite, hard coal, natural gas). But, on one hand the source of fossil fuel is decreasing at a rapid rate, and on the other hand the burning of fossil fuel is also degrading the environment due to huge Greenhouse Gas (GHG) emission (FAO/CMS, 2011). Hence, there is an urgent need of an alternate clean energy source that can sustain the future development undeterred while still protecting the environment. Biomass is considered as a sustainable source of clean energy that is useful in steam, fuel and chemicals production. It can be digested in controlled environment to produce biogas. The biogas is a clean, non-polluting and smoke-free fuel (Weiland, 2010) which can be used for applications that include cooking, space cooling/refrigeration, heating, electricity generation and gaseous fuel (CNG) for vehicles (Weiland, 2003). This paper analyses possible alternatives to process and utilise biogas for energy at industrial scale as a petroleum fuels replacement.

ABSTRACT
The rapid industrialisation and speedy progress towards urbanisation across the world have created the huge demand for energy moreover creating enormous quantity of wastes. The latter is in charge for polluting the environment in an exponential manner, resulting out of the harmful and poisonous compounds released by them. In recent years, there has been a pattern change from ‘waste-to-wealth’, considering the value of high organic fraction accessible in the wastes. The best-accomplished methods are that of anaerobic digestion; contribute to the production of methane (CH₄). Similar biotransformation has partial net energy harvests. Recently, fermentation industries steering their interest towards the production of value added products, viz., biohydrogen (H₂), biopolymers, ethanol, vitamins, acetic acid, enzymes, butanediols, etc. from carbon rich organic waste like cellulosic substrates have been envisioned to flourish in a multi-step process, such as the ‘Biorefinery’. A contemporary development in fermentation technology holds the key to overcome the constraints for such processes that may lead to a sustainable economy. One related technology is electrofermentation that has attracted significant attention recently owing to its capacity to boost the metabolic activities of microbes by means of extracellular electron transfer.

ABSTRACT
Biomethanation is the process of generating energy from anaerobic digestion (AD) of organic refuse. This helps reduce dependency on fossil fuels while reducing CO₂ emission. Organically-rich kitchen refuse (KR) generated daily from the households all over the world could be a good candidate to biogasify, though the quantity and quality of biogas depends on the KR proximate. To enhance biogasification, KR is subjected to physical, chemical and/or biological pretreatment. Physical pretreatments include chopping, grinding, ultrasonication and steam explosion, whereas chemical pretreatments could be by an alkali or a base. Biological treatment involves digestion of the organic matters by individual microbe or its consortium, although it takes place naturally. For biological treatment, standardising the substrate and inoculum ratio is essential. Candidate strains must be mutually compatible to formulate a microbial consortium. Bacillius seems to be a potential candidate as it is a facultative anaerobe exhibiting multiple essential enzyme activities (viz., amylase, cellulase and lipase). Maintaining an optimal pH of 7.0 and temperature of 37°C before and during anaerobic digestion is suggested. Other important factors in digestion include organic loading rate, retention time, C/N ratio, volatile fatty acids (VFA) concentration, etc. VFA content is vital as its accumulation during acidogenesis adversely affects the AD by dropping pH values to unfavourable levels. Acetogens convert VFA to acetate, CO₂ and H₂ in acetogenesis, finally converting it to CH4 in methanogenesis. For effective methanation,

ABSTRACT
The annual accumulating organic wastes generated through milling, brewing and various other agro-industrial and agri-processing activities in India amount to about 500 MMT. Most of these contain three major structural biopolymers, viz., cellulose, hemicellulose and lignin, a large proportion of which is carbohydrate and phenolic in nature. Massive accumulations of this kind of recalcitrant biomass not only deteriorate the environment, but also generate huge amount of potentially useful materials. Biological transformation has globally become increasingly popular among researchers and academicians in treating agricultural, industrial, organic and toxic wastes these days. About 60% by weight of the original pineapple fruit in forms of peeled skin, core, crown end, etc. resulting from the pineapple processing remains unutilised being discharged as waste, causing disposal and pollution owes. Of the 10% dry matter content of such waste, about 96% is organic and 4% is inorganic. Being rich in cellulose and hemicellulose, it can be a valuable resource for numerous value-added products by bioconverting it to valuable products, including bioenergy. Biomethane is one such energy resource that can be produced by valorizing pineapple wastes by employing various bioconversion technologies. This chapter focuses on the bioconverstion technologies to produce biomethane from pineapple wastes.

ABSTRACT
There is a quantum global growth in both number and size of dairy industries in recent times. These industries discharge thousands of liters of wastewater characterised by a heavy chemical oxygen demand (COD), biological oxygen demand (BOD), nutrients, inorganic salts, besides detergents and sanitisers each day (Kushwaha et al., 2011). High carbohydrates, fats and proteins contents in such waste streams originating from milk pose challenge to conventional municipal sewage treatment systems (Demirel et al., 2005). The easily biodegradable dairy effluent leads to precipitation of casein which is toxic and has a strong odour. Without adequate treatment, these effluents could pose serious threats to the receiving water bodies and disturb the ecosystem (Satpathy et al., 2017). For such heavily loaded organic effluents, biogasification through anaerobic digestion seems a promising strategy. This cost-effective energy generation process is also effective in environmental management compared to its other eco-friendly renewable source counterparts, such as, hydro, solar and wind (Satpathy, 2017). The Upflow Anaerobic Sludge Blanket (UASB) reactors are the most popular anaerobic systems to treat the fluctuating organic loads in dairy effluents. Anaerobic filter (AF), Upflow Anaerobic Filter (UPAF), Fixed-bed anaerobic reactor, Anaerobic Hybrid Reactor (AHR) etc. are few other systems that are demonstrably competent removing COD upto 90% and generating biogas. Successful anaerobic systems reported yielded (with upto about 85% CH₄ content) 0.32-0.34m3 CH₄/kg COD removed. Biomethanation process and reactors designs are constantly being optimised for effective treatment of the effluents while maximising the biogas production. The energy-rich gas thus obtained could further be utilised for heat and electricity at the domestic level, and also as a vehicular fuel. Residual digestate can also be used as organic manures (Angelidaki et al., 2007).

ABSTRACT
Rice straw is one of the most abundant lignocellulosic agricultural wastes in India and can be used as a major resource for biogas production. The structure of rice straw is complex and is made up mainly of cellulose, hemicelluloses and lignin. The lignin physically shields the cellulose and hemicelluloses, the main components which can be converted to energy, and hinders the access of microbial enzymes to these carbohydrate moieties. To overcome this making the straw a potential resource of biogas production, effective pretreatment is necessary that can efficiently hydrolyse the straw and make it accessible to cellulolytic enzymes and methanogens. Various pretreatment methods which are employed currently in the market are discussed here. For a process to be economically feasible it is necessary that the pretreatment be efficient as well as cost effective. To gain some insights into this aspect, the techno-commercial feasibility of Biomethanation of rice straw has been discussed.

ABSTRACT
Recent challenges for fossil fuels, associated environmental issues with disposal of organic garbage, and the rising costs for energy has encouraged researchers to search for alternate energy sources. Massive uncontrolled biogas from landfills, sludge and organic material degradation under anoxic conditions is an environmental management issue. Methane emission to atmosphere has a tremendous green house effect, but this can also be a valuable renewable energy resource. Biogas from kitchen wastes through anaerobic digestion by methanogens is such an alternate technology. Kitchen wastes are considered one of the best raw materials for biogasification. Biogas is mainly composed of 50-74% methane (CH₄), 25-28% carbon-di-oxide (CO₂), other volatile compounds including hydrogen sulphide (H₂S), water vapour and other trace gas compounds. The impurities in it, like Hydrogen sulphide (H₂S) which is corrosive, downgrade its quality. Purified biogas of higher calorific value attracts attention as a substitute fuel for additional applications. Its purification using various suitable scalable and economically-viable scrubbing technologies to enhance its quality and performance as an alternate fuel is an important step in this direction. Thus, a technology to generate and purify biogas from kitchen waste has significant prospects in the energy sector, and also as a promising enterprise to employ the youth.

ABSTRACT
Increased environmental pollution and high demand over supply for conventional fossil fuel sources have led to the exploration of other renewable energy sources. Out of the potential alternatives, methane has come up with a promising unconventional fuel source. Several technologies have been developed to establish methanation, including biomethanation, chemical methanation and gas-to-liquid technology. Several factors like process parameters, catalysts and reactors have been studied for process optimisation and making methane production commercially viable. Although methane as a greenhouse gas is a contributor to global warming, its combustion produces relatively ‘environment friendly’ gas CO₂, and H₂O. The released CO₂ is fixed into biomass by plant through photosynthesis process. Only rarely does methane combustion produces CO when combusted in an oxygen-limited condition. Thus, methane production and combustion are carbon neutral process. This chapter focuses on the application of methane as a source of alternate energy. Various technologies used, recent advances and challenges faced in methanation process are discussed.

ABSTRACT
Biomethanation comprises of four interdependent stages in series, hydrolysis, acidogenesis, acetogenesis and methanogenesis. Methanogenesis, the final stage of biomethanation, is accompanied with the reduction of substrates that include organic acids such as CH₃COOH, and others such as CO₂ and CH₃OH to CH₄ by methanogenic and methanotrophic archaea, usually under anaerobic conditions. Strictly anaerobic methanogenic archaea are found in the ruminant intestine, sewage digester, groundwater and deep soil/water etc. In methanogenesis, H₂ acts as the electron donor while substrates CO₂, CH₃OH and probably CoM-S-S-CoB (for CH₃COOH) act as terminal electron acceptors and reduce to methane, a process known as anaerobic respiration. This conversion is associated with the electrogenic proton motive force along with sodium motive force to drive the ATP synthesis by A0A1 ATP synthase, known as respiratory phosphorylation. The mechanism of conversion of methane from CO₂ by methanogens with cytochromes is same as those without cytochromes except that the reduction of heterodisulphide CoM-S-S-CoB is performed by the membrane-bound VhoACG-HdrDE complex for former while by cytoplasmic MvhADG–HdrABC complex for the later.

ABSTRACT
Methanogenesis is an important biogeochemical process takes place in anaerobic ecological sites. It is the last step of terminal electron accepting process of anaerobic respiration. Methanogens synthesise CH₄ using acetate and CO₂ as precursor molecules. Former types of methanogens are referred as acetoclastic methanogens and the later ones as hydrogenotrophic methanogens. After biogenesis of CH₄, it is emitted to atmosphere and causes global warming. In aerobic ecosystem CH₄ is oxidised by methanotrophs to CO₂ a less global warming molecule. However, in anoxic environment CH4 acts as an electron donor and interacts with various electron acceptors. Methane modulates denitrification, Fe reduction, SO₄ reduction and organic matter decomposition. Manuscript elucidates methanogenesis and highlights the pathways of interaction between methanogenesis and cycling of electron acceptors in various anaerobic ecosystems in relation to global C and N balance.

ABSTRACT
Methanogens are primarily anaerobic Archaebacteria representing the most primitive earthly creatures, alive since about 3.5 billion years. While oxygen inhibit their activity, they adapt well in a variety of ecological niche, viz., the intestinal tracts of insects and animals including ruminants, sewage digesters, groundwater and deep soil/water, etc. They are involved in biomethanation that converts organic polymers, including the complex recalcitrant lignocelluloses, to CH₄ and CO₂. Enhancing the process by optimising the physical, chemical and molecular parameters is an interesting biotechnology. Molecular research is rapidly progressing on the genomics, gene and its regulation, and methanogenesis as it expresses. Identifying novel species expressing methanogenesis and manipulating the genetic make-up including through cloning in recent years will set goals and leads for future investigations. The biomethanation enzyme Methyl-CoM-reductase (Mcr) constitutes about 10% of the methanogen protein. Due to its abundance and significance, elucidating its structure and its synthesis and regulation mechanism has received much attention recently. Mcr-coding genes from methanogens have been cloned and sequenced. An approach at the molecular level to enhance biomethanation seems to lie in metagenomics. The microbiology, biochemistry, the roles of the participating microbes, their molecular biology and applications in methanogenesis are discussed in this chapter.

ABSTRACT
Methanogenesis is a ubiquitous process primarily encountered in anaerobic environments. Methanogens are a group of diverse obligate anaerobic bacteria that produce methane from complex organic matter. Despite their differences in morphological characters and habitat, they show coherence in terms of methane-producing metabolic activity. Being distinctive from all other classical prokaryotes, these are grouped under another kingdom of Archea. Other distinctions of the members are based on their unique cell wall structures and membrane components, as well as differences in their highly conserved gene sequences. Methanogens likely owe their cosmopolitan status to their unique mode of energy metabolism of methane generation. They are active in various ecological niches, from rumens of animals to submarine volcanic vents, sewage digestors and landfills. Methanogenic bacteria are abundant in habitats where high affinity electron acceptors are negligible. Methane fermentation involves bacteria that obtain energy by catalysing anaerobically degradable organic matter to CH₄ and CO₂ by consuming H₂, CO₂, and low-carbon organic acids as their substrates. The gaseous products of methanogenesis are greenhouse gases, thus are major contributors to global warming. Methanogens could be used to produce renewable carbon-neutral energy substitute, biogas. Methane cycling archaea play a major role in global methane production as energy for the future.

A. General
        
B. Turf Grass
        
C. Indoor Plants

A. General
        
B. Bulbous Plants
        
C. Greenhouse Technology
        
D. Plant Breeding
        
E. Plant Physiology
        
F. Plant Tissue Culture and Biotechnology
        
G. Post Harvest Technology

Accent : It is created in the garden to avoid the monotonous view. Mostly unusual objects like tall fountain, tree, statue etc are used to create the effect.

Acre: A unit of measurement of land area equivalent to 4000 sq.met or 0.4047 hectare.

Acrylic: It is a glassy thermoplastic that is incorporated in rigid plastic to enhance wither ability.

Aging: Refers to the processes acquiring maturity with the passage of time or normally an increment of time, which may or may not be accompanied by physiological changes including senescence. Aging thus includes a much ‘wider span’ of physiological processes, which may either “weaken the organism or to be neutral”.

Agmark: It is Agriculture Produce, Grading & Marketing Act, 1937 & voluntary in nature.

Anaerobic respiration: An incomplete oxidation of food to release partial energy takes place in the absence of atmospheric oxygen and occurs in cytoplasm.

Analytical chemistry: Deals with detection and estimation of elements and compounds.

A. General
        
B. Leaves and Floral Terminologies
        
C. Diversity in the Flower
        
D. Diversity in Fruits
        
E. Terminologies Related to Seed

• Definition of Bioinformatics
• History of Bioinformatics 
• Structural Bioinformatics
• New Branch of Bioinformatics 
• Bioinformatics - Application in Agriculture
• Analyzing Protein Sequence
• Pairwise Alignment
• Multiple Alignment
• Blast - A Sequence Alignment Tool
• EMBL
• SWISS–PROT
• Rice Genome Project
• Software for Bio-informatics
• Major Categories of Bioinformatics Software
• Internet Sites For Bioinformatics

• Web Portal
• Types of Portals

• Agriculture Database Concepts and Importance
• The Orientation of Topic Selection for the Construction of the Agricultural Database
• The Principle of Specialization in the Construction of the Agricultural Database
• Top Quality Principle of Building Databases
• Agriculture Database Types
• Agriculture Libraries
• Agriculture Journals
• Technical Reports
• Conference Papers
• E-Books in Agriculture

• Database
• Data Model
• Database Software
• DBMS Facility 
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• Data Modeling
• Components of a Data Mode
• RDBMS (Relational Database Management System)
• Client/Server Model of Computing
• Entity Relationship Diagram (E-R diagram)
• Entity Relationship Diagram Notations - Front-end and Back-end
• Codd’s Rules
• SQL - Structured Query Language
• Data Definition Language (DDL)
• Data Manipulation Language (DML)
• DCL
• Data Control Language
• Transaction Control (TCL)

• A Brief Description of Visual Basic
• Types of Application in VB
• Creating the Project
• Steps in Building a Visual Basic Application
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• The InputBox( ) Function
• Methods
• Procedur
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• Data Access
• Data Access Objects (DAO)
• Microsoft Jet Database Engine 
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• Microsoft ActiveX Data Objects
• Data Control
• Object Linking and Embedding (OLE)

• Introduction
• Features of Genetic Algorithm 
• Steps in Executing the Genetic Algorithm
• Applications of Genetic Algorithm
• Artificial Ant Problem