Pyocyanin (5-methyl-1-hydroxyphenazine) produced by Pseudomonas aeruginosa as antagonist to vibrios in aquaculture: overexpression, downstream process and toxicity

Pyocyanin is a versatile and multifunctional phenazine, widely used as a bio-control agent. Besides its toxicity in higher concentration, it has been applied as bio-control agents against many pathogens including the Vibrio spp. in aquaculture systems. The exact mechanism of the production of pyocyanin in Pseudomonas aeruginosa is well known, but the genetic modification of pyocyanin biosynthetic pathways in P. aeruginosa is not yet experimented to improve the yield of pyocyanin production. In this context, one of the aims of this work was to improve the yield of pyocyanin production in P. aeruginosa by way of increasing the copy number of pyocyanin pathway genes and their over expression. The specific aims of this work encompasses firstly, the identification of probiotic effect of P. aeruginosa isolated from various ecological niches, the overexpression of pyocyanin biosynthetic genes, development of an appropriate downstream process for large scale production of pyocyanin and its application in aquaculture industries. In addition, this work intends to examine the toxicity of pyocyanin on various developmental stages of tiger shrimp (Penaeus monodon), Artemia nauplii, microbial consortia of nitrifying bioreactors (Packed Bed Bioreactor, PBBR and Stringed Bed Suspended Bioreactor, SBSBR) and in vitro cell culture systems from invertebrates and vertebrates. The present study was undertaken with a vision to manage the pathogenic vibrios in aquaculture through eco-friendly and sustainable management strategies with the following objectives: Identification of Pseudomonas isolated from various ecological niches and its antagonism to pathogenic vibrios in aquaculture.,Saline dependent production of pyocyanin in Pseudomonas aeruginosa originated from different ecological niches and their selective application in aquaculture,Cloning and overexpression of Phz genes encoding phenazine biosynthetic pathway for the enhanced production of pyocyanin in Pseudomonas aeruginosa MCCB117,Development of an appropriate downstream process for large scale production of pyocyanin from PA-pUCP-Phz++; Structural elucidation and functional analysis of the purified compoundToxicity of pyocyanin on various biological systems.

Physiological and Genetic Diversity Studies on Regeneration of Santalum Album L.

S. album L. is the source of highly priced and fragrant heartwood which on steam distillation yields on an average 57 per cent oil of high perfumery value. Global demand for sandalwood is about 5000-6000 tons/year and that of oil is 100 tons/year. Heartwood of sandal is estimated to fetch up to Rs. 3.7 million/ton and wood oil Rs.70,000-100,000/ kg in the international market. Sandal heartwood prices have increased from Rs. 365/ton in 1900 to Rs. 6.5 lakhs/ton in 1999-2000 and to Rs. 37 lakhs/ton in 2007. Substantial decline in sandalwood production has occurred from 3176 tons/year during 1960-‘ 65 to 1500 tons/year in 1997-98, and to 500 tons/year in 2007.Depletion of sandal resources is attributed to several factors, both natural and anthropogenic. Low seed setting, poor seed germination, seedling mortality, lack of haustorial connection with host plant roots, recurrent annual fires in natural sandal forests, lopping of trees for fodder, excessive grazing, hacking, encroachments, seedling diseases and spread of sandal spike disease are the major problems facing sandal. While these factors hinder sandal regeneration in forest areas, the situation is accelerated by human activities of chronic overexploitation and illicit felling.Deterioration of natural sandal populations due to illicit felling, encroachments and diseases has an adverse effect on genetic diversity of the species. The loss of genetic diversity has aggravated during recent years due to extensive logging, changing landuse patterns and poor natural regeneration. The consequent genetic erosion is of serious concern affecting tree improvement programme in sandal. Conservation as well as mass propagation are the two strategies to be given due importance. To initiate any conservation programme, precise knowledge of the factors influencing regeneration and survival of the species is essential. Hence, the present study was undertaken with the objective of investigating the autotrophic and parasitic phase of sandal seedlings growth, the effects of shade on morphology, chlorophyll concentration and chlorophyll fluorescence of sandal seedlings, genetic diversity in sandal seed stands using ISSR markers, and the diversity of fungal isolates causing sandal seedling wilt using RAPD markers. All these factors directly influence regeneration and survival of sandal seedlings in natural forests and plantations.

Modification of Polypropylene and Polystyrene Using Nanosilica

The present work focuses on the modification of the commonly used thermoplastics, polypropylene and polystyrene using nanosilica preparcd from a cheap source of sodium silicate. Melt compounding technique has been used for nanocomposite preparation as it is simple and suited to injection moulding. Nanosilica in a polymer matrix provide significant enhancement in strength, stiffness and impact strength. Incorporation of silica particles in a polymer also improves its thennal stability. To achieve better dispersion of fillers in polymer matrices the mixing was done at different shear rates. The enhancement in material properties indicates that at higher shear rates there is greater interaction between particles and the matrix and it depends on filler concentration and type of polymer used. N anosilica is a useful filler in thennoplastic polymers and has been applied in automotive applications, electronic appliances and consumer goods.This thesis is divided into six chapters. General introduction to the topic is described in chapter 1. Salient features of polymer nanocomposites, their synthesis, properties and applications are presented. A review of relevant literature and the scope and objectives are also mentioned in this chapter.The materials used and the vanous experimental method and techniques employed in the study are described in chapter 2. Preparation of nanocomposites by melt blending using Thenno Haake Rheocord, preparation of samples, evaluation of mechanical and thennal properties using UTM, Impact testing and characterization using DMA, TGA and DSC and morphology by SEM are described.The preparation of nanosilica from a laboratory scale to a pilot plant scale is described in chapter 3. Generation of surface modified silica, evaluation of kinetic parameters of the synthesis reaction, scale up of the reactor and modeling of the reactor are also dealt with in this chapter.The modification of the commodity thennoplastic, Polypropylene using nanosilica is described in chapter 4. Preparation of PP/silica nanocomposites, evaluation of mechanical properties, thermal and crystallization characteristics, water absorption and ageing resistance studies are also presented.The modification of Polystyrene using synthesized nanosilica IS described in chapter 5. The method of preparation of PS/silica nanocomposites, evaluation of mechanical properties (static and dynamic), thermal properties melt flow characteristics using Haake Rheocord, water absorption and ageing resistance of these nanocomposites are studied.

Development of plant tissue culture of Plumbago rosea Linn. for enhanced production of secondary metabolites

P rosea syn. Indica belong to the family of plumbaginaceae, is an important medicinal plant, cultivated widely in India. The roots of these plant are generally used for medicinal purposes mainly as diuretic, germicidal, vessicant, and abortifacient. It is also used for anaemia, diarrhea, leprosy and common wart. The bark of the root contains orange yellow pigment named plumbagin, a crystalline substance, belongs to the class of naphthoquinone. Its chemical structure is 5-hydroxy 2-methyl 1,4naphthoquinone. Apart from P rosea, P zeylanica, P europea, Drosera and Drosophyllum also contains plumbagin. The most exploited source of plumbagin is, of course, P. rosea roots. The roots contain O.9mg/ g D.Wt. of plumbagin in the roots. These plants grow very slowly and the roots suitable for plumbagin extraction can be obtained only after several years of growth. The productivity of the plant is also rather poor. The focus of the present study was to develop alternative strategies to obtain plumbagin. The tissue culture of P rosea for micropropagation has been studied