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Title of Thesis BIOCHEMICAL STUDIES OF ALGAE FROM INLAND WATERS OF BALOCHISTAN Author(s) MUDASSIR ISRAR Institute/ University/ Department Details UNIVERSITY OF BALOCHISTAN/BIOCHEMISTRY Status (Published/ Not Published/ In Press etc) Published Date of Publishing 1995 Subject Biochemistry Number of Pages 253 Keywords (Extracted from title, table of contents and abstract of thesis) Biochemical Studies, Algae, Fresh water algae, Algal flora, Chara vulgaris Linneaus, Cladophora fracta (Dillw) Kutz, Spirogyra webri Kutz, Oscillatoria subbrevis Suchmidle, Ash content, Lipids, Carbohydrates, Proteins, Inorganic elemental composition, Fatty acid, Hydrocarbons, Terpene, Sterols, Pennisetum, Abstract Increase in human population and limited sources of food, compelled the scientists to investigate new sources for food. The present study comprises of ecological, biochemical and chemical studies on fresh water algae of Balochistan. The composition and seasonal distribution of fresh water algae collected from twenty localities of Balochistan were investigated. 729 species with 120 genera were identified. These belonged to five divisions of algae species showed a distinct seasonal pattern. Water samples were analyzed for their chemical characteristics and for the levels of radioactivity. The correlation of these chemicals with the algal flora has been established. Among the dominant algae four species were selected for the biochemical and chemical studies namely Chara vulgaris Linneaus, Cladophorafracta (Dillw) Kutz. Spirogyra weberi Kutz..,belonged to green algae while Oscillatoria subbrevis Schmidle, belonged to blue green algae. The ash content, lipids, carbohydrates, proteins and inorganic elemental composition of these algae were determined. Significantly high ash, lipids, carbohydrates and protein were found in Spirogyra weberi which proved it, to be a good source of food. These content were high in green algae than those found in blue green algae. A variety of saturated and unsaturated fatty acids, hydrocarbons, terpene and sterols have been reported for the first time from these algae. These were identified quantitatively and qualitatively through different spectroscopic techniques including EI, HR-MS, GC-MS, IHNMR, 1HNMR and 13CNMR and 2D experiments. Quantitative and qualititative micro, macroelements and heavy metal concentration of these four algae were analyzed. Significantly high calcium content were found in these algae as compare to other elements. Antifungal activity of the ethanolic algal extracts was carried out against ten fungal strains. Chara vulgaris, Cladophora fracta and Oscillatoria subbrevis exhibit strong antifungal activity, while Spirogyra weberi showed no activity against any of the fungi tested. The antibacterial activity of the four ethanolic algal extracts has been carried out in vitro against gram positive and gram negative bacteria. The algal extracts showed no antibacterial activity against these bacteria. Brine shrimp toxicity test was carried out on all the four crude ethanolic algal extracts. No toxicity was found in any algae. Allelopathic effects of these algal extracts were tested towards the Pennisetun seeds. It was found that C. vulgaris, C. fracta and S. weberi inhibit the growth' of Pennisetun seeds at very high concentration. O. subbrevis was found to be growth promoter at al concentrations. The dried four algae were baked in bread and fed to wistar rats, their growth and weight were observed. The rats fed with S. weberi showed good growth and attained a significant increase in body weight with reference to control rats. The hypotensive and hypertensive pharmacological activity of all algal extracts were carried out on wistar rats C. vulgaris and C. fracta lowered the blood pressure, respiratory and heart rate by 30%, while other two algae were found to be ineffective. Table of Contents Chapter Page Certificate i Acknowledgement ii List of Contents iii List of Tables v List of Figures vii ABSTRACT ix I INTRODUCTION 1 1. 6 Description of Localities and their Climatic Conditions II REVIEW OF LITERATURE 21 III MATERIAL AND METHODS 32 A 1. 32 Ecological Studies Biochemical and Chemical Investigation 2. Ash Contents 34 3. Carbohydrates 35 4. Proteins 36 5. Lipids 37 6. Phytochemical Investigations 38 6.1 Instrumentation 39 6.2 Fatty Acid and Hydrocarbons 40 6.3 Saponification and Esterification 41 6.4 Acetylation 41 6.5 Sterols and Terpenes 42 6.6 Micro and Macro Elements & Heavy Metal Concentrations 43 PHARMACOLOGICAL SCREENING 8. Anti Fungal Activity 43 9. Anti Bacterial Activity 44 10. Toxicity Bioassay 45 11. Allelopathic Bioassay 46 12. Nutritional Studies 47 13. Anti Hypertensive Effect 47 Ecological Survey 111 Chapter IV A 1. B C V Biochemical and Chemical Investigation 2. Ash Cotents 164 3. Carbohydrates 166 4. Proteins 167 5. Lipids 168 6. Phytochemical Investigations 6.1 Fatty Acids Hydrocarbons & Sterols of Chara vulgaris 170 6.2 Fatty Acids Hydrocarbons Sterols & Terpenes of Cladophora fracta 181 6.3 Fatty Acids Hydrocarbons and Sterols of Spirogyra weberi 197 6.4 Fatty Acids Hydrocarbons and Sterols of Oscillatoria subbrevis 206 6.5 Micro and Macro Elements & Heavy Metal Concentrations 215 PHARMACOLOGICAL SCREENING 7. Anti Fungal Activity 220 8. Anti Bacteriall Activity 222 9. Toxicity Bioassay 224 10. Allelopathic Bioassay 229 11. Nutritional Studies 232 12. Anti Hypertensive Effect 236 References 238