Production of Alkaline Protease by Free and Immobilised Cells of VIBRIO sp. Under Different Fermentation Systems and Its Application on Deproteinisation of Prawn Shell Waste for Chitin Recovery

This thesis presents a detailed account of a cost - effective approach towards enhanced production of alkaline protease at profitable levels using different fermentation designs employing cheap agro-industrial residues. It involves the optimisation of process parameters for the production of a thermostable alkaline protease by Vibrio sp. V26 under solid state, submerged and biphasic fermentations, production of the enzyme using cell immobilisation technology and the application of the crude enzyme on the deproteinisation of crustacean waste.The present investigation suggests an economic move towards Improved production of alkaline protease at gainful altitudes employing different fermentation designs utilising inexpensive agro-industrial residues. Moreover, the use of agro-industrial and other solid waste substrates for fermentation helps to provide a substitute in conserving the already dwindling global energy resources. Another alternative for accomplishing economically feasible production is by the use of immobilisation technique. This method avoids the wasteful expense of continually growing microorganisms. The high protease producing potential of the organism under study ascertains their exploitation in the utilisation and management of wastes. However, strain improvement studies for the production of high yielding variants using mutagens or by gene transfer are required before recommending them to Industries.Industries, all over the world, have made several attempts to exploit the microbial diversity of this planet. For sustainable development, it is essential to discover, develop and defend this natural prosperity. The Industrial development of any country is critically dependent on the intellectual and financial investment in this area. The need of the hour is to harness the beneficial uses of microbes for maximum utilisation of natural resources and technological yields. Owing to the multitude of applications in a variety of industrial sectors, there has always been an increasing demand for novel producers and resources of alkaline proteases as well as for innovative methods of production at a commercial altitude. This investigation forms a humble endeavour towards this perspective and bequeaths hope and inspiration for inventions to follow.

Effect of ambient gas on the expansion dynamics of plasma plume formed by laser blow off of thin film

A study has been carried out to understand the influence of ambient gases on the dynamics of laser-blow-off plumes of multi-layered LiF–C thin film. Plume images at various time intervals ranging from 100 to 3000 ns have been recorded using an intensified CCD camera. Enhancement in the plume intensity and change in size and shape occurs on introducing ambient gases and these changes are highly dependent on the nature and composition of the ambient gas used. Velocity of the plume was found to be higher in helium ambient whereas intensity enhancement is greater in argon environment. The plume shapes have maximum size at 10−2 and 10−1 Torr of Ar and He pressures, respectively. As the background pressure increases further (>10−2 Torr: depending on the nature of gas), the plume gets compressed/focused in the lateral direction. Internal structure formation and turbulences are observed at higher pressures (>10−1 Torr) in both ambient gases.

Application of Fermentation Techniques in the Utilization of Prawn Shell Waste

The present study aimed at the utlisation of microbial organisms for the production of good quality chitin and chitosan. The three strains used for the study were Lactobacillus plantarum, Lactobacililus brevis and Bacillus subtilis. These strains were selected on the basis of their acid producing ability to reduce the pH of the fermenting substrates to prevent spoilage and thus caused demineralisation of the shell. Besides, the proteolytic enzymes in these strains acted on proteinaceous covering of shrimp and thus caused deprotenisation of shrimp shell waste. Thus the two processes involved in chitin production can be affected to certain extent using bacterial fermentation of shrimp shell.Optimization parameters like fermentation period, quantity of inoculum, type of sugar, concentration of sugar etc. for fermentation with three different strains were studied. For these, parameters like pH, Total titrable acidity (TTA), changes in sugar concentration, changes in microbial count, sensory changes etc. were studied.Fermentation study with Lactobacillus plantarum was continued with 20% w/v jaggery broth for 15 days. The inoculum prepared yislded a cell concentration of approximately 108 CFU/ml. In the present study, lactic acid and dilute hydrochloric acid were used for initial pH adjustment because; without adjusting the initial pH, it took more than 5 hours for the lactic acid bacteria to convert glucose to lactic acid and during this delay spoilage occurred due to putrefying enzymes active at neutral or higher pH. During the fermentation study, pH first decreased in correspondence with increase in TTA values. This showed a clear indication of acid production by the strain. This trend continued till their proteolytic activity showed an increasing trend. When the available sugar source started depleting, proteolytic activity also decreased and pH increased. This was clearly reflected in the sensory evaluation results. Lactic acid treated samples showed greater extent of demineralization and deprotenisation at the end of fermentation study than hydrochloric acid treated samples. It can be due to the effect of strong hydrochloric acid on the initial microbial count, which directly affects the fermentation process. At the end of fermentation, about 76.5% of ash was removed in lactic acid treated samples and 71.8% in hydrochloric acid treated samples; 72.8% of proteins in lactic acid treated samples and 70.6% in hydrochloric acid treated samples.The residual protein and ash in the fermented residue were reduced to permissible limit by treatment with 0.8N HCI and 1M NaOH. Characteristics of chitin like chitin content, ash content, protein content, % of N- acetylation etc. were studied. Quality characteristics like viscosity, degree of deacetylation and molecular weight of chitosan prepared were also compared. The chitosan samples prepared from lactic acid treated showed high viscosity than HCI treated samples. But degree of deacetylation is more in HCI treated samples than lactic acid treated ones. Characteristics of protein liquor obtained like its biogenic composition, amino acid composition, total volatile base nitrogen, alpha amino nitrogen etc. also were studied to find out its suitability as animal feed supplement.Optimization of fermentation parameters for Lactobacillus brevis fermentation study was also conducted and parameters were standardized. Then detailed fermentation study was done in 20%wlv jaggery broth for 17 days. Also the effect of two different acid treatments (mild HCI and lactic acid) used for initial pH adjustment on chitin production were also studied. In this study also trend of changes in pH. changes in sugar concentration ,microbial count changes were similar to Lactobacillus plantarum studies. At the end of fermentation, residual protein in the samples were only 32.48% in HCI treated samples and 31.85% in lactic acid treated samples. The residual ash content was about 33.68% in HCI treated ones and 32.52% in lactic acid treated ones. The fermented residue was converted to chitin with good characteristics by treatment with 1.2MNaOH and 1NHCI.Characteristics of chitin samples prepared were studied and extent of Nacetylation was about 84% in HCI treated chitin and 85%in lactic acid treated ones assessed from FTIR spectrum. Chitosan was prepared from these samples by usual chemical method and its extent of solubility, degree of deacetylation, viscosity and molecular weight etc were studied. The values of viscosity and molecular weight of the samples prepared were comparatively less than the chitosan prepared by Lactobacillus plantarum fermentation. Characteristics of protein liquor obtained were analyzed to determine its quality and is suitability as animal feed supplement.Another strain used for the study was Bacillus subtilis and fermentation was carried out in 20%w/v jaggery broth for 15 days. It was found that Bacillus subtilis was more efficient than other Lactobacillus species for deprotenisation and demineralization. This was mainly due to the difference in the proteolytic nature of the strains. About 84% of protein and 72% of ash were removed at the end of fermentation. Considering the statistical significance (P

Studies on the exploratory trawl fishing and significance of selectivity in trawl fishing gear for conservation of marine fishery resources with special reference to the inshore waters off Mangalore

School of Industrial Fisheries, Cochin University of Science and Technology

Studies on the Effects of Bottom Trawling on the Benthic Fauna off Veraval Coast, Gujarat

Bottom trawling is one among the most destructive human induced physical disturbances inflicted to seabed and its living communities. The bottom trawls are designed to tow along the sea floor, which on its operation indiscriminately smashes everything on their way crushing, killing, burying and exposing to predators the benthic fauna. Bottom trawling causes physical and biological damages that are irreversible, extensive and long lasting. The commercial trawling fleet of India consists of 29,241 small and medium-fishing boats. The northwest coast of India has the largest fishing fleet consisting of 23,618 mechanized vessels, especially the bottom trawlers. However, attempts were not made to study the impact of bottom trawling along Northwest coast of India. The estimated optimum fleet size of Gujarat is 1,473 mechanised trawlers while 7402 commercial trawlers are operated from the coast of Gujarat. Veraval port was designed initially for 1,200 fishing trawlers but 2793 trawlers are being operated from this port making it the largest trawler port of Gujarat. The aim of this study was to investigate the effects of bottom trawling on the substratum and the associated benthic communities of commercial trawling grounds of Veraval coast. The study compared the differences between the samples collected before and after experimental trawling to detect the impacts of bottom trawling. Attempts were made to assess the possible impact of bottom trawling on:(i) the sediment characteristics (ii)the sediment heavy metals (iii) epifauna (iv) macrobenthos and (v) meiobenthos. This study is expected to generate information on trawling impacts of the studied area that will help in better management of the biological diversity and integrity of the benthic fauna off Veraval coast. An exhaustive review on the studies conducted around the world and in India on impact of bottom trawling on the benthic fauna is also detailed.In the present study, the bottom trawling induced variations on sediment organic matter, epifauna, macrobenthos and meiobenthos were evident. It was also observed that the seasonal/ natural variations were more prominent masking the trawling effect on sediment texture and heavy metals. Enforcement of control of excess bottom trawlers and popularization of semi pelagic trawls designed to operate a little distance above the sea bottom for off bottom resources will minimize disturbance on the sea bottom. Training and creating awareness in responsible fishing should be made mandatory requirements, to the coastal communities. They should be made wardens to protect the valuable resources for the benefit of sustainability. To protect the biodiversity and ecosystem health, the imminent need is to survey and make catalogue, identification of sensitive areas or hot spots and to adopt management strategies for the conservation and biodiversity protection of benthic fauna. The present study is a pioneering work carried out along Veraval coast. This thesis will provide a major fillip to the studies on impact of bottom trawling on the benthic fauna along the coast of India.

Cadmium Selenide Based Core-Shell Quantum Dots for Biosensing and Imaging Applications

The overall focus of the thesis involves the synthesis and characterization of CdSe QDs overcoated with shell materials for various biological and chemical sensing applications. Second chapter deals with the synthesis and characterization of CdSe and CdSe/ZnS core shell QDs. The primary attention of this work is to develop a simple method based on photoinduced charge transfer to optimize the shell thickness. Synthesis of water soluble CdSe QDs, their cytotoxicity analysis and investigation of nonlinear optical properties form the subject of third chapter. Final chapter deals with development of QD based sensor systems for the selective detection of biologically and environmentally important analytes from aqueous media.

Novel applications of cashew nut shell liquid in the polymer field

This research project aims at developing new applications for CNSL in the polymer field. Cashew nut shell liquid (CNSL) is a cheap agro-byproduct and renewable resource which consists mainly of substituted phenols. By using CNSL in place of phenol, phenol derived from petrochemicals can be conserved and a cheap agro-byproduct utilized.In this study CNSL based resin is prepared by condensing a mixture of phenol and CNSL with hexamethylenetetramine and the effect of P: F ratio and CNSL: P ratio on the properties of synthesized resin is studied. The adhesive properties of CNSL based resin in combination with neoprene rubber are investigated. The effect of varying the stoichiometric ratios between total phenol and formaldehyde and CNSL and phenol of the resin, resin content, choice and extent of fillers and adhesion promoters in the adhesive formulation are studied. The effect of resin on the ageing properties of various elastomers is also studied by following changes in tensile strength, elongation at break, modulus, tear strength, swelling index and acetone soluble matter. Crude CNSL and resins with different P: F ratios and CNSL: P ratios are incorporated into elastomers. Lastly, utility of CNSL based resin as binder for making particleboard is investigated.The results show that CNSL based resin is an effective ingredient in adhesives for bonding aluminium to aluminium. The resin used for adhesive fonnulation gives the best performance at 45 to 55 phr resin and a total phenol: formaldehyde of l:2.9. The resin when added at a rate of l5 phr improves ageing characteristics of elastomers with respect to mechanical properties. The reaction mixture of CNSL and hexa and the resin resulting from the condensation of CN SL, phenol and hexa can be used as effective binders for moulding particleboard.

Studies on the Back Water Oyster, grassostrea madra— sensis (Preston) of Cochin Harbour

A detailed study of the hydrography of the Cochin Backwaters, the habitat off crassostrea madrasensis has been carried out. Data pertaining to air temperature, water temperature, salinity, dissolved oxygen, turbidity and rainfall have been collected and presented. The temperature fluctuation was in the range of 5°C only and that of salinity between 1.1%o and 32.9%o. Fairly steady salinity has been recorded during the pre-monsoon period (February to May) and drastic declension during the monsoon period (June-September).Dissolved oxygen varied between 2.5 ml/l and 6.5 ml/l. Turbidity was highest in June (27.9 p.p.m.) and minimum (10.2 p.p.m.) in February. A detailed study on marine biofouling in the Cochin Backwaters has been made with special reference to primary film, settlement and growth of the fouling organisms such as hydroids, bryozoans, tube-dwelling polychaetes, barnacles and modiolus

Provenance, Sedimentation and Geochemistry' of the Modern Sediments of time Mud Banks Off the Centrai Keraia Coast, India

This thesis is an attempt to Provenence, Sedimentetion and Geochemistry of the Modern Sediments of the Mud Banks off the Central Kerela Coast, India. In the present doctoral work, an attempt has been made to study in detail the mud banks of central Kerala, i.e. of Narakkal, Saudi and Purakkad areas which are reported as permanent mud banks, since olden days. The studies have been conducted during the years 1985 and 1986. The important findings of the study is stated as clay mineralogical studies of the rivers, lake and mud bank sediments reveal that the dominant clay mineral is kaolinite followed by montmorillonite, illite and gibbsite. Geochemical analysis of the Vembanad lake and mud bank sediments show that the iron and manganese are widely distributed both in the lake and mud bank sediments

Bio-chemical changes associated with processing of shell fishes and flavour constituents of body meat and claw meat'of crab

In this thesis all these aspects are taken into consideration. Extensive studies were conducted on all aspects of processing of crabs, mussels and clams. The species taken for studies are commercially used ones namely Scylla sereta, perna viridis, and villorita cyprinoids. In Chapter 4.1 with regard to crab) the following aspects on their handling and processing are reported seasonal variation of chemical constituents, changes taking place during ice storage, freezing, canning etc. In Chapter 4._2 with regard to mussel, the relation between age (size) and chemical constituents, changes taking place during ice storage, freezing, canning etc. are reported and in Chapter 4.3 the changes taking place in clam muscle during icing and freezing are reported and the ame rebility of ice stored clams for canning purpose is reported.The interference of high concentration of glycogen in mussel and clam muscles during the colour development of ribose (Me-jbaum's method) is observed and remedial step are taken to minimise the interference.

Studies on the distribution and abundance of fish eggs and larvae off the south—west coast of India with special reference to scombroids

The broad objective of the present study is to present a synoptic picture of the distribution and abundance of fish eggs and the lmportant groups of fish larvae obtained off the SW coast of India. so as to delineate the spawning areas and seasones of the fish population. with special reference to the scombroid fishes. An attempt was also made to correlate the occurrence of certain categories of larvae and hydrographical factors like temperature and salinity. The present effort was a pioneering one in Indian waters. in as much as it involved systematic and seasonally repetitive collection of ichthyoplankton from a large stretch of our seas and mapping of their distribution and abudance.

Physical characteristics of the coastal waters off the south-west coast of india with an attempt to study the possible relationship with sardine mackerel and anchovy fisheries

The present study is aimed at observing the variations, in space and time, of see of the important hydrographic parameters such as sea water temperature, salinity and Resolved oxygen within the coastal waters along the south-west coast of Indiametween Ratnagiri (17°OO*N,73°20'E) and cape comorin ( 8°10'N,77°30*E). Specific data relating to the process of upwelling and sinking was collected mainly to evaluate the extent and intensity of the vertical mixing processes active in the area under study. The study also attempted possible correlations between the observed parameters and the occurrence and migrations of some of the major pelagic fishery resources such as sardine,mackerel and anchovy in the area under study

A Comparative Study of The Muscle Proteins of Fishes and Shell Fishes of Tropical Waters

This thesis is an attempt to make a comparative study of the composition of the muscle proteins of some commercially important species of fishes and shell fishes of our coast and their changes during preservation and processing. As a part of this the distribution of the major protein nitrogen fractions in several species of fishes and shell fishes was studied in detail.

Application of ocean colour data to study the oceanographic and atmospheric features off the southwest coast of India with special reference to upwelling

Satellite remote sensing is being effectively used in monitoring the ocean surface and its overlying atmosphere. Technical growth in the field of satellite sensors has made satellite measurement an inevitable part of oceanographic and atmospheric research. Among the ocean observing sensors, ocean colour sensors make use of visible band of electromagnetic spectrum (shorter wavelength). The use of shorter wavelength ensures fine spatial resolution of these parameters to depict oceanographic and atmospheric characteristics of any region having significant spaio-temporal variability. Off the southwest coast of India is such an area showing very significant spatio-temporal oceanographic and atmospheric variability due to the seasonally reversing surface winds and currents. Consequently, the region is enriched with features like upwelling, sinking, eddies, fronts, etc. Among them, upwelling brings nutrient-rich waters from subsurface layers to surface layers. During this process primary production enhances, which is measured in ocean colour sensors as high values of Chl a. Vertical attenuation depth of incident solar radiation (Kd) and Aerosol Optical Depth (AOD) are another two parameters provided by ocean colour sensors. Kd is also susceptible to undergo significant seasonal variability due to the changes in the content of Chl a in the water column. Moreover, Kd is affected by sediment transport in the upper layers as the region experiences land drainage resulting from copious rainfall. The wide range of variability of wind speed and direction may also influence the aerosol source / transport and consequently AOD. The present doctoral thesis concentrates on the utility of Chl a, Kd and AODprovided by satellite ocean colour sensors to understand oceanographic and atmospheric variability off the southwest coast of India. The thesis is divided into six Chapters with further subdivisions

studies on the clam meretrix casta(chemnitz)off cochin barmouth.

It is the object of the present study to contribute so much information as possible on the biology and economy of M.casta on the south west coast of India. It includes investigations on the growth of the species in three dimensions.