eChapters

Experiment - 1
 
Aim: Micropropagation of Tobacco plant by leaf disc culture

Principle: The totipotency of the plant cells and tissues form the basis for in vitro cloning i.e., generation or multiplication of genetically identical plants in in vitro culture. The ability to propagate new plants from a cells or tissues of parent plant has many interesting possibilities.

Micropropagation /Clonal propagation

Micropropagation or clonal propagation is the growing of plants from meristematic tissue or somatic cells of superior plants on nutrient suitable media under controlled aseptic physical conditions. Sexually propagated plants (through generation of seeds) demonstrate a high amount of heterogeneity since their seed progenies are not true to type whereas asexual reproduction (by multiplication of vegetative parts) gives rise to genetically identical copies of parent plant. Thus, it permits perpetuation of the parental characters of the cultivars among the plants resulting from micropropagation. Micropropagation proves useful for propagation of: 

Modes of Culture 

Callus is a mass of undifferentiated plant tissues formed from plant cells or tissue cuttings when grown in culture. Tissues and cells of plant cultured in a liquid medium aerated by agitation grow as suspension of single cells and cell clumps. For growth, the cells require to divide, whereas, the cells breaking up from explant are mature, nondividing. Hence, the differentiated tissue undergoes modifications to become meristematic. This phenomenon of a mature cell reverting back to meristematic state to form undifferentiated callus tissue is called dedifferentiation.

Explant

The tissue obtained from plant to be cultured is known as Explant. It may include the parts of shoots, leaves, stems, roots, flowers and callus.

Maintenance of Aseptic Environment

All culture vessels, media and instruments used in handling tissues as well as the explants must be sterilized. The importance is to keep the air surface and floor free of dust. All operations are carried out in laminar air-flow, a sterile cabinet.In general, the methods of elimination of these sources of infection can be grouped under different categories of sterilization procedures: 

Sterilization Techniques

Sterilization: All the materials, e.g., vessels, instruments, medium, explants, etc., used in culture work must be freed from microbes. This can be achieved by one of the following approaches:

One of the most essential factors governing the growth and morphogenesis of plant tissues in culture is the composition of the culture medium. The basic nutrient requirements of cultured plant cells are very similar to those of whole plants. The basic nutritional requirements of cultured plant cells as well as plants are very similar. However, the nutritional composition varies according to the cells, tissues, organs and protoplasts and also with respect to particular plant species.  The appropriate composition of the medium largely determines the success of the culture. A wide variety of salt mixtures  have been reported in various media. A nutrient medium is defined by its mineral salt composition, carbon source, vitamins, plant growth regulators (PGR) and other organic supplements. When referring to a particular medium, the intention is to identify only the salt composition unless otherwise specified. Any number and concentration of amino acids, vitamins, growth regulators and organic supplements can be added in an infinite variety of compositions to a given salt composition in order to achieve the desired product.

Common rules for fine laboratory techniques call for:

    ·    A laboratory should have a record and a complete up to-date record of all the equipments and their operating manual.
    ·    A laboratory should have a record and detailed up to-date record of all the chemicals including the name of manufacturer with their     grade and arranged alphabetically in specific area
    ·    Strong acid and bases in safe area.
    ·    Chloroform, alcohol which are volatile or toxic must be stored in fume hood.

A tissue culture laboratory can be placed in any geographical area. The internal controlled environmental conditions permit isolation with a minimum of external influence. To decide where to build the tissue culture laboratory, the following factors must be taken into consideration:

Year    Contributor    Description
1756    Duharmel du Monceau H.L.    Discovered callus formation in decorticated elm tree. 
1839    Schwann, T.H.    Proposed the view that each living cell of a multicellular organism would be capable of developing independently if provided with proper external conditions.
1853    Trecul, A.    Performed experiment on callus formation by decorticated trees such as Robinia, Pawlonia and Ulmus.

Practical have a significant role in the study of any theoretical subject. Whatever a student learns in theory, he verifies it by actual doing with his own hands.  Thus, these reduce any doubts and misbelieve from his mind. These verifications by actually doing the things are performed in the laboratories.  A laboratory is a place with the basic and essential equipments and adequate facilities for students and teachers to perform and observe experiments.  Plants will continue to provide novel products as well as chemical models for new drugs in the coming centuries.  The arrival of chemical analysis and the characterization of molecular structures have helped in precisely identifying these plants and correlating them with their activity under controlled experimentation.

Collection of faecal samples: To diagnose gastro-intestinal parasites of ruminants or other animals, the parasites or their eggs/larvae must be recovered from the digestive tract of animal or human or from the faecal/stool material.
 
The faecal samples for parasitological examination should be collected from the rectum of animals. If rectum samples cannot be obtained, fresh faecal samples may be collected from the pastures. Several samples should be collected and immediately dispatched to the diagnostic laboratory in suitable containers such as:

Small ruminants is a composite term used for sheep and goats. Throughout the developing countries, small ruminants make a very valuable contribution, especially to the poor in the rural areas. These contributions range from precious animal proteins (meat and milk) to fibre and skins, draught power in the highlands, food security and stable households. By and large, the importance and extent of the contributions are inadequately understood as a consequence of which these valuable genetic resources continue to be generally neglected. Research and development investments to improve the relatively low level of contribution do not match their potential importance, the contribution to nutritional security and livelihoods of the poor in rural areas. Species like goat and sheep are of short-generation interval, have a high prolificacy rate and require less land, investment and operational expenses and are better suited to the resource endowment of the poor.

Feeds and feeding of dairy animals are arguably the most important part of the dairy farm management. And it is not only what we feed that is important but how we manage our feeding is equally as critical. What we feed today not only determines the animal’s milk production but also influences their entire lactation cycle that follows. Dairy animal feeding with balanced ration influences health status, reproductive, productive performance and better economic return in organic environment, all of which are critical to a profitable and sustainable dairy business as well as welfare of the dairy farmers. Feed costs currently account for around 45 percent of total milk price on average, with concentrates representing around 35 percent and forages around 10 percent.

The livestock play a significant role in generating gainful employment in rural areas, particularly among the landless and marginal farmers. The Jammu region of state of Jammu and Kashmir possesses a huge livestock population which is maintained under different management systems. The region has a vast diversity in agro-climatic conditions (comprising low altitude, middle altitude and high altitude), and animal husbandry and pasture management practices. Despite having huge livestock, the production potentials are very low. There are various factors related with low production and among them parasitic diseases rank very high.

Small ruminant sector, after agriculture, is the second largest means for economic sustainability and livelihood in India. It contributed Rs. 32000 crore to the gross domestic product of India in 2012 -13 despite having a potential to contribute over Rs. 150,000 crore but there are certain lacunae in system which act as its growth inhibitor. Small ruminants are the main source for rural economies due to participation of women (70% of total) and weaker sections of the rural society. Thus there is urgent need for formulating policies to support the development of the small ruminant sector.
 
Sheep and goat husbandry is a tool for poverty alleviation of the rural masses because of wide adaptability with low investments, high fertility and fecundity, low feed and management costs, higher feed conversion efficiency, quick pay off and low risk in comparison to large ruminants. The above traits of small ruminants need to be exploited for economic upliftment of small farmers.

Livestock sector has emerged as one of the key components of agricultural growth in developing countries in last few decades. This sector plays a significant role in the economic progress of India as it contributes 26% to the agricultural GDP and provides employment to around 18 million people. Over 70% of the rural households in India depends on livestock farming for supplementary income and is considered as an important instrument in poverty alleviation. It provides the major extra contribution to agriculture through manure, fuel fertilizer, apart from meat and milk that grant a regular supplementary income and employment not only to producers but also to huge size of the population engaged in secondary and tertiary business related to livestock. The economy of Jammu and Kashmir is also agriculture dependent and is the main occupation of majority of the rural people who earn their livelihood from it. Census figures 2011 signify that about 73% of the population of J&K lives in rural areas and are associated with agriculture and allied sectors including livestock rearing as main occupation.

Introduction

Mankind has been utilizing different animal species from the dawn of civilization for a variety of purposes viz., production of milk, meat, wool, egg and leather. Apart from these, various animal species are also used for draught power, companionship, entertainment, research experimentation, sports, security etc. Livestock sector plays a crucial role in rural economy and livelihood. As per report of the working group on animal husbandry and dairying, 11th five year plan: 2007-12, the livestock sector employs eight percent of the countries labour force, including many small and marginal farmers, women and landless agricultural workers. Milk production alone involves more than 30 million small producers, each raising one or two cows or buffaloes. The organic fertilizer produced by the sector is an important input to crop production and dung from livestock is widely used as fuel in rural areas. Livestock also serves as an insurance substitute, especially for poor rural households; it can easily be sold during time of distress.

Helminths include the nematodes (roundworms), trematodes (flukes) and the cestodes (tapeworms). The adult worms live in intestine and discharge the eggs or larvae that comes out with the faeces. Therefore, diagnosis is based on the detection of eggs or larvae in faecal samples.

Overview of Nematode infections in humans and animals

The phylum Nematoda, (roundworms), is the second largest phylum in the animal kingdom, encompassing up to 500,000 species. Members of Nematoda are elongated, with bilaterally symmetric bodies that contain an intestinal system and a large body cavity. According to a report approximately 60 species of roundworms parasitize humans. Intestinal roundworm infections constitute the largest group of helminthic diseases in humans. According to 2005 report of World Health Organization (WHO), approximately 0.807-1.221 billion humans suffers from ascariasis, 604-795 million from trichuriasis and 576-740 million from hookworm infections worldwide.

Participation means different things to different people. They way participant is defined largely depends upon the context and back ground in which participation is applied. While the Economic Commission for Latin America (1973) considers contribution by the people to public programmes to the complete exclusion of any involvement in the decision making process as participation. Cohem and Uphoff (1977) maintain that participation tools includes the peoples involvement in the entire decision making process. Like Cohem & Uphoff, FAO (1982) emphasizes participation in all the decision making processes but by the people’s organization and through self organized action. Paul (1987) introduces the concept of enhancement of well-being in terms of values cherished by the communities as being the ultimate objective of participation.

Agriculture is world’s single largest employer and probably the most weather dependent of all human activities.  Climate and its variability have been and continue to be, the principal source of fluctuations in global food production. Throughout history, extreames of heat and cold, droughts and floods, and various form of violent weather have wretched havoc on the global agricultural production. With the unprecedented increase in population over past few decades, clearly the mankind faces undertaking task to ensure food and nutritional security for all, considering reduced per capita land, reduced availability of water, depleting biodiversity and need to preserve ecology and environment. In India, of the total geographical area of 328 m ha, about 175 m ha (53 % of total) are categorized as degraded land, of which 150 m ha are subjected to serious water and wind erosion (Singh et al. 1981). Crops are sown in about 180 m ha annually. Since it seems very much impossible to bring more area under cultivation we need to achieve greater efficiency in the natural resources use while conserving the environment by placing greater emphasis on understanding and exploiting climatic resources for the benefits of agriculture.