Development of Cell Culture Systems from Selected Species of Fish and Prawns

The present work deals with the development of primary cell culture and diploid cell lines from two fishes, such as Poecilia reticulata and Clarias gariepinus. The greatest difficulty experienced was the avoidance of bacterial and fungi contamination. Three types of cell cultures are commonly developed, primary cell culture, diploid cell lines and heteroploid cell lines. Primary cell culture obtained from the animal tissues that have been cultivated in vitro for the first time. They are characterized by the same chromosome number as parent tissue, cultivated in vitro for the first time, have wide range of virus susceptibility, usually not malignant, six chromatin retarded and do not grow as suspension cultures. Diploid cell lines arise from a primary cell culture at the time of subculturing. Diploid cell lines commercially used in virology are W1-38 (human embryonic lung), W1-26 (human embryonic lung) and HEX (Human embryonic kidney). Heteroploid cell lines have been subcultivated with less than 75% of the cells in the population having a diploid chromosome constitution. Tissue cultures have been extensively used in biomedical research. The main applications are in three areas, Karyological studies, Identification and study of hereditary metabolic disorders and Somatic cell genetics. Other applications are in virology and host-parasite relationships. In this study an attempt was made to preserve the ovarian tissue at low temperature in the presence of cryoprotectants so that the tissue can be retrieved at any time and a cell culture could be developed.

Kinetics of Bacterial Colony Growth by Laser Induced Fluorescence

The growth kinetics of an aerial bacterial colony on solid agar media was studied using laser induced fluorescence technique. Fluorescence quenching of Rhodamin B by the bacterial colony was utilized for the study. The lag phase, log phase, and stationary phase of growth curve of bacterial colony was identified by measuring peak fluorescence intensity of dye doped bacterial colony.

Semiconductor Oxides Mediated Photocatalytic Removal of Chemical and Bacterial Pollutants from Wastewater

School of Environmental Studies, Cochin University of Science and Technology

Studies on Production of Bacterial Xylanases

Xylanases with hydrolytic activity on xylan, one of the hemicellulosic materials present in plant cell walls, have been identified long back and the applicability of this enzyme is constantly growing. All these applications especially the pulp and paper industries require novel enzymes. There has been lot of documentation on microbial xylanases, however, none meeting all the required characteristics. The characters being sought are: higher production, higher pH and temperature optima, good stabilities under these conditions and finally the low associated cellulase and protease production. The present study analyses various facets of xylanase biotechnology giving emphasis on bacterial xylanases. Fungal xylanases are having problems like low pH values for both enzyme activity and growth. Moreover, the associated production of cellulases at significant levels make fungal xylanases less suitable for application in paper and pulp industries.Bacillus SSP-34 selected from 200 isolates was clearly having xylan catabolizing nature distinct from earlier reports. The stabilities at higher temperatures and pH values along with the optimum conditions for pH and temperature is rendering Bacillus SSP-34 xylanase more suitable than many of the previous reports for application in pulp and paper industries.Bacillus SSP-34 is an alkalophilic thertmotolerant bacteria which under optimal cultural conditions as mentioned earlier, can produce 2.5 times more xylanase than the basal medium.The 0.5% xylan concentration in the medium was found to the best carbon source resulting in 366 IU/ml of xylanase activity. This induction was subjected to catabolite repression by glucose. Xylose was a good inducer for xylanase production. The combination of yeast extract and peptone selected from several nitrogen sources resulted in the highest enzyme production (379+-0.2 IU/ml) at the optimum final concentration of 0.5%. All the cultural and nutritional parameters were compiled and comparative study showed that the modified medium resulted in xylanase activity of 506 IU/ml, 5 folds higher than the basal medium.The novel combination of purification techniques like ultrafiltraton, ammonium sulphate fractionation, DEAE Sepharose anion exchange chromatography, CM Sephadex cation exchange chromatography and Gel permeation chromatography resulted in the purified xylanase having a specific activity of 1723 U/mg protein with 33.3% yield. The enzyme was having a molecular weight of 20-22 kDa. The Km of the purified xylanase was 6.5 mg of oat spelts xylan per ml and Vmax 1233 µ mol/min/mg protein.Bacillus SSP-34 xylanase resulted in the ISO brightness increase from 41.1% to 48.5%. The hydrolytic nature of the xylanase was in the endo-form.Thus the organism Bacillus SSP-34 was having interesting biotechnological and physiological aspects. The SSP-34 xylanase having desired characters seems to be suited for application in paper and pulp industries.

Studies on biochemical genetics of the grey mullet Mugil cephalus linnaeus

The thesis deals with the results of an investigation on the "BIOCHEMICAL GENETICS OF MUGIL CEPHALUS" from Cochin, Madras and Orissa. It is presented under the following major headings: Introduction, Review of Literature, Materials and Methods, Results, Discussions, Conclusions, Recommendations, Summary and References.The introduction gives a brief account of historical and modern back ground on the stock concept in fisheries research and management, followed by the importance and potential role of biochemical genetics in the identification of natural units of fisheries management. In the review of literature published reports relevant to biochemical genetics with special reference to that of general proteins and enzyme systems of fish populations were considered. A detailed account of the source of experimental specimens, mode of collection, transportation, sample extraction, gel preparation/gel electrophoresis, buffer systems, staining procedures of proteins/enzymes, standardization of experiments, interpretation of electrophoretic data using basic formulae etc. are given in the materials and methods section. Four important conclusions were drawn on the basis of the results of the present investigation. Three recommendations were also made on the basis of evaluation of the results.

Studies on the effect of the organophosphorus pesticide Eka(Rlu) x EC 25 on the bacterial flora of Villorita cyprinoides var. cochinensis (Han1ey)

The thesis is Studies on the Effect or the Obganophosphorus Pesticide Ekalux(R) EC 25 on the Bacterial Flora or Villorita Cyprinoides Var.Cochinensis (Hanley). For the present investigation, the black clam Villorita gyprinoides var. cochinensis (Hanley), a most common clam genus present in this estuarine system has been selected as test organaism and Ekalux (R) EC 25 as toxicant. The aspects dealt with are 1. Total heterotrophic bacterial population, 2. Generic composition, 3. Hydrolytic enzyme producing bacteria, 4. Antibiotic resistance, 5. Heavy metal resistance, 6. The effect of pesticide concentration on the growth of the bacteria and 7. Effect of temperature, pH and sodium chloride on the growth and phosphate release of selected isolates.The samples for the experiment were collected from the Vembanad Lake, near Kumbalam Island during the period of September 1985 to May '86. The THB of the estuarine water and clams contained 6.5 x I04/ml and 2.975 x l06/g respectively, immediately after collection. Untreated water and clam samples showed enormous increase in THB from 0 hr population. The treated samples (water and clams) contained higher THB than 0 hr. In general, THB was observed to increase tremendously in the samples treated with pesticide when compared to their native flora. With reference to various concentrations of pesticides, THB recorded an increase with increase of concentration in water and clam samples.

Biochemical Genetics Of Selected Commercially Important Penaeid Prawns

‘Biochemical genetics of selected commercially important penaeid prawns‘ dloted was carried out by collecting samples from different important fishing ceatres of India and the practical work was carried out in the Research Centre of CMFRI laboratories attached with those places. On the whole, in crustacea little importance has been given so far in finding out tin genetic characteristics of different species, genetic variation within and between species and ontogenetic variations in lobsters, prawns and other crustaceans. Prawn is caunercially important group where very little attention had been given so far to find out the racial divergence which may exist in cufferent species. with the increased foreign exchange earning and consequent indiscriminate over exploitation of existing resources of prawns resulting in depletion of the marine rescurces, alternative ways and augmenting production has become essential. In this connection genetic manipulation of the broodstock will surely bring about the heterogenous characters to multiply production. In order to understand racial fragmentation of sane of the coumercially important prawns such as Pengeus ggdicus and Parggenagsis sgliferg the isozyme studies were carried out. Qatogenetic variation of g. indicus showed stage specific electrophoretic variation. Inter species variation studies was carried out for the closely aligned Penaeus species

Heterotrophic bacterial activity in selected aquaculture systems near cochin

The principal interest of the present investigation was to determine seasonal and vertical variation of chemoorganotrophic utilisation of glucose and sodium—acetate by the natural bacterial population in the aquaculture pond of Narakkal, Cochin using techniques which allow maintenance of the in situ gaseous concentrations during incubation. In addition salinity, dissolved oxygen, temperature, hydrogen—ion—.concentration, primary production, plant pigments and total bacterial concentration were determined seasonally and vertically because of their possible relationship to chemoorganotrophy.

Biochemical and molecular studies of cadmium resistance and metal biosorption in bacterial species isolated from cochin environment

Cochin, commercial capital of Kerala, located on the west-coast of South India has a large number of chemical and sea food industries. Earlier studies in the past indicated that these industries contribute to heavy metal pollution, particularly mercury, copper, and cadmium, in Cochin backwater. Hence, in the present study, it was desired to isolate cadmium resistant bacteria from effluent discharged by chemical industry with a view to develop an ideal bioremediation process for safe discharge of industrial effluent in to the nearby aquatic environment. Effluent from three industries, located in the industrial belt of Cochin, were collected from the discharge point and cadmium resistant bacteria were screened using standard microbiological techniques

‘Studies on Bacterial Indicators and Pathogen Vibrio parahaemolyticus in Cochin Backwater

Microbiological studies on the incidence, behaviour, activity and ecological implications of marine micro~organisms, particularly microbial pathogens in coastal waters and estuaries exhibit the increasing concern and awareness of environmental impacts on health and wealth. Marine microbiologists have been active in investigating on the distribution, kinds of organisms and their activity in the environment. However, informations on the effect of environment on the ecology or on the distribution (spatial/temporal) of microbial comunity and competition among groups inhabiting the ecosystem are sparE§L Estuarine environment are complex with respect to diversity of habitats, variation in physicochemical parameters and contamination by terrestrial bacterial species. Being the organisms of‘public health significance, ecological studies on total coliforms, faecal coliforms, faecal streptococci, §. ggli and X. parahaemolyticus have great relevance as studies of these types would provide a wealth of information to environmentalists and to fishery industry. In order to evalé%e the status, role and significance of potentially hazardous bacterial species in natural environment it is necessary to monitor the ecology of such organisms systematically in relation to physico-chemical parameters

Impact of climate change on hetrotrophic bacterial communities in the water and sediment of kongsfjord in norwegian artic

Considering the extent of warming in the artic region and the resultant changes in the dynamic marine enviornments there is a need to monitor the bacterial diversity in the fjord enviornments especially in terms of cultivable bacteria. The present study reports the diversity of cultivable hetrotrophic bacteria from the water and sediment samples of kongsfjord their growth responses to important enviornmental variables and ability to produce industrially important hydrolytic enzymes.

Growth enhancement of micro algae, Chaetoceros calcitrans and Nannochloropsis oculata, using selected bacterial strains

In natural systems phytoplankton interact with planktonic (free living) and attached epiphytic bacteria both synergistically and antagonistically. The specificity of the association with micro algae and bacteria differs in terms of adhesion mechanisms and metabolic cooperation. Present research was carried out to study the effect of bacterial isolates namely Bacillus sp. and Pseudomonas sp. from algal culture systems on the growth of micro algae such as Chaetoceros calcitrans and Nannochloropsis oculata. C. calcitrans (F= 15.34; P<0.05) and N. oculata (F=12.52; P<0.05) showed significantly higher growth, in treatments with Bacillus sp. and Pseudomonas sp when compared to control.