Significance of soil microorganisms with special reference to climate change

There are a large number of agronomic-ecological interactions that occur in a world with increasing levels of CO2, higher temperatures and a more variable climate. Climate change and the associated severe problems will alter soil microbial populations and diversity. Soils supply many atmospheric green house gases by performing as sources or sinks. The most important of these gases include CH4, CO2 and N2O. Most of the green house gases production and consumption processes in soil are probably due to microorganisms. There is strong inquisitiveness to store carbon (C) in soils to balance global climate change. Microorganisms are vital to C sequestration by mediating putrefaction and controlling the paneling of plant residue-C between CO2 respiration losses or storage in semi-permanent soil-C pools. Microbial population groups and utility can be manipulated or distorted in the course of disturbance and C inputs to either support or edge the retention of C. Fungi play a significant role in decomposition and appear to produce organic matter that is more recalcitrant and favor long-term C storage and thus are key functional group to focus on in developing C sequestration systems. Plant residue chemistry can influence microbial communities and C loss or flow into soil C pools. Therefore, as research takings to maximize C sequestration for agricultural and forest ecosystems - moreover plant biomass production, similar studies should be conducted on microbial communities that considers the environmental situations

Studies on nitrifying microorganisms in Cochin Estuary and adjacent coastal waters

This thesis entitled “Studies on Nitrifying Microorganisms in Cochin Estuary and Adjacent Coastal Waters” reports for the first time the spatial andtemporal variations in the abundance and activity of nitrifiers (Ammonia oxidizingbacteria-AOB; Nitrite oxidizing bacteria- NOB and Ammonia oxidizing archaea-AOA) from the Cochin Estuary (CE), a monsoon driven, nutrient rich tropicalestuary along the southwest coast of India. To fulfil the above objectives, field observations were carried out for aperiod of one year (2011) in the CE. Surface (1 m below surface) and near-bottomwater samples were collected from four locations (stations 1 to 3 in estuary and 4 in coastal region), covering pre-monsoon, monsoon and post-monsoon seasons. Station 1 is a low saline station (salinity range 0-10) with high freshwater influx While stations 2 and 3 are intermediately saline stations (salinity ranges 10-25). Station 4 is located ~20 km away from station 3 with least influence of fresh water and is considered as high saline (salinity range 25- 35) station. Ambient physicochemical parameters like temperature, pH, salinity, dissolved oxygen (DO), Ammonium, nitrite, nitrate, phosphate and silicate of surface and bottom waters were measured using standard techniques. Abundance of Eubacteria, total Archaea and ammonia and nitrite oxidizing bacteria (AOB and NOB) were quantified using Fluorescent in situ Hybridization (FISH) with oligonucleotide probes labeled withCy3. Community structure of AOB and AOA was studied using PCR Denaturing Gradient Gel Electrophoresis (DGGE) technique. PCR products were cloned and sequenced to determine approximate phylogenetic affiliations. Nitrification rate in the water samples were analyzed using chemical NaClO3 (inhibitor of nitrite oxidation), and ATU (inhibitor of ammonium oxidation). Contribution of AOA and AOB in ammonia oxidation process was measured based on the recovered ammonia oxidation rate. The contribution of AOB and AOA were analyzed after inhibiting the activities of AOB and AOA separately using specific protein inhibitors. To understand the factors influencing or controlling nitrification, various statistical tools were used viz. Karl Pearson’s correlation (to find out the relationship between environmental parameters, bacterial abundance and activity), three-way ANOVA (to find out the significant variation between observations), Canonical Discriminant Analysis (CDA) (for the discrimination of stations based on observations), Multivariate statistics, Principal components analysis (PCA) and Step up multiple regression model (SMRM) (First order interaction effects were applied to determine the significantly contributing biological and environmental parameters to the numerical abundance of nitrifiers). In the CE, nitrification is modulated by the complex interplay between different nitrifiers and environmental variables which in turn is dictated by various hydrodynamic characteristics like fresh water discharge and seawater influx brought in by river water discharge and flushing. AOB in the CE are more adapted to varying environmental conditions compared to AOA though the diversity of AOA is higher than AOB. The abundance and seasonality of AOB and NOB is influenced by the concentration of ammonia in the water column. AOB are the major players in modulating ammonia oxidation process in the water column of CE. The distribution pattern and seasonality of AOB and NOB in the CE suggest that these organisms coexist, and are responsible for modulating the entire nitrification process in the estuary. This process is fuelled by the cross feeding among different nitrifiers, which in turn is dictated by nutrient levels especially ammonia. Though nitrification modulates the increasing anthropogenic ammonia concentration the anthropogenic inputs have to be controlled to prevent eutrophication and associated environmental changes.

Production and Characterization of Lignin Peroxidases from Mangrove Ascomycetes

In this thesis, the production and characterization of ligninolytic enzymes using the fungi isolated from mangrove area are studied. The objective of the present work are isolation and screening of dye decolorizing micro-organisms from mangrove area, screening of the selected microorganisms for the production of lignin degrading enzymes, identification of the potent micro-organisms, characterization of the crude enzyme, lignin peroxidase, of the selected fungi—Aspergillus sp. SIP 11 and Penicillium sp. SIP 10 etc. This included the determination of the optimum pH, temperature, veratryl alcohol and H2O2 concentration. Besides the stability of crude LiP at different pHs and temperatures were studied. The immense applications, particularly in bioremediation, to which the lignin degrading micro-organisms could be used make this study important, the ascomycetes and deuteromycetes fungi, especially form the marine environment were studied with respect to their ligninolytic enzyme system making this study an initial step in unraveling the vast hidden potential of these microbes in bioremediation, the marine microbes are halophilic in nature which make them better suited to cope with the high salinity of industrial effluents thereby giving them added advantage in the filed of bioremediation. The thesis deals with the isolation and screening of lignin degrading enzyme-producing microbes from mangrove area. The identification of the most potent fungal isolates and characterization of LiP from these are also done.

L-glutaminase production by marine fungi

This study presents the L-Glutaminase Production by Marine Fungi. Enzymes are involved in all aspects of biochemical conversion from the simple enzyme or fermentation conversion to the complex techniques in genetic engineering. Enzyme industry is one among the major industries of the world and there exists a great market for enzymes in general. Food industry is recognized as the largest consumer for commercial enzymes (Lon sane and Ramakrishna, 1989). In industry, enzymes are frequently used for process improvement, for instance to enable the utilization of new types of raw materials or for improving the physical properties of a material so that it can be more easily processed. They are the focal point of biotechnological processe. The marine biosphere is one of the richest of the earth's innumerable habitats, yet is one of the least well characterized. The marine biosphere covers more than two third of the world's surface, our knowledge of marine microorganisms, in particular fungi, is still very limited (Molitoris and Schumann, 1986). The results obtained in the present study the following conclusions are drawn. Beauveria bassiana isolated form marine sediment has immense potential as an Industrial organism for production of L-glutaminase as an extracellular enzyme employing either submerged fermentnation or solid state fermentation

Characterisation and Distribution of Amino Acids in the Mangrove Sediments of Kochi

Mangrove swamps are unique inter-tidal wetland ecosystems found in sheltered tropical and subtropical shores.Mangrove sediments can be considered as large reservoirs of amino acids,which exist in several different forms,like free amino acids in the sediment micropores,as amino acids,peptides or proteins bound to clay minerals or as amino acids,peptides or proteins bound to humic colloids.Inorder to assess survival conditions of organisms of mangroves,it is important to understand stability of amino acids in the sediments.The amounts of amino acids present in sediment represent a balance between its synthesis and destruction by microorganisms.Thus amino acid analysis offers more insight into the processes of diagenesis,which changes the nature and characteristics of organic matter deposition and decomposition.

Process Optimization for Mass Production of Marine Yeast Candida sp. S 27 and its Nutritional Characterization

The main source of protein for human and animal consumption is from the agricultural sector, where the production is vulnerable to diseases, fluctuations in climatic conditions and deteriorating hydrological conditions due to water pollution. Therefore Single Cell Protein (SCP) production has evolved as an excellent alternative. Among all sources of microbial protein, yeast has attained global acceptability and has been preferred for SCP production. The screening and evaluation of nutritional and other culture variables of microorganisms are very important in the development of a bioprocess for SCP production. The application of statistical experimental design in bioprocess development can result in improved product yields, reduced process variability, closer confirmation of the output response to target requirements and reduced development time and overall cost.The present work was undertaken to develop a bioprocess technology for the mass production of a marine yeast, Candida sp.S27. Yeasts isolated from the offshore waters of the South west coast of India and maintained in the Microbiology Laboratory were subjected to various tests for the selection of a potent strain for biomass production. The selected marine yeast was identified based on ITS sequencing. Biochemical/nutritional characterization of Candida sp.S27 was carried out. Using Response Surface Methodology (RSM) the process parameters (pH, temperature and salinity) were optimized. For mass production of yeast biomass, a chemically defined medium (Barnett and Ingram, 1955) and a crude medium (Molasses-Yeast extract) were optimized using RSM. Scale up of biomass production was done in a Bench top Fermenter using these two optimized media. Comparative efficacy of the defined and crude media were estimated besides nutritional evaluation of the biomass developed using these two optimized media.

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.

Pigment production by marine Serratia sp. BTWJ8

The marine microorganisms are yet to be exploited as a source of natural pigments for probable utilization in various industries. Hence, in this study focus was made only on pigment producing marine bacteria for pigment production and evaluation of the same for some application besides development of an ideal bioprocess for subsequent indigenous production of the pigment using the same organism towards ultimate industrial application.

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.

Characterization of polyhydroxyalkanoates accumulating vibrios from marine benthic environments and production studies of polyhydroxyalkanoates by Vibrio sp. BTKB33

The present work deals with the characterization of polyhydroxyalkanoates accumulating vibrios from marine benthic environments and production studies of polyhydroxyalkanoates by vibrio sp.BTKB33. Vibrios are a group of (iram negative, curved or straight motile rods that normally inhabit the aquatic environments.The present study therefore aimed at evaluating the occurrence of PHA accumulating vibrios inhabiting marine benthic environments; characterizing the potential PHA accumulators employing phenotypic and genotypic approaches and molecular characterization of the PHA synthase gene. The study also evaluated the PHA production in V:'hri0 sp. strain BTKB33, through submerged fennentation using statistical optimization and characterized the purified biopolymer. Screening for PHA producing vibrios from marine benthic environments. Characterization of PHA producers employing phenotypic and genotypic approaches.The incidence of PHA accumulation in Vibrio sp. isolated from marine sediments was observed to be high, indicating that the natural habitat of these bacteria are stressful. Considering their ubiquitous nature, the ecological role played by vibrios in maintaining the delicate balance of the benthic ecosystem besides returning potential strains, with the ability to elaborate a plethora of extracellular enzymes for industrial application, is significant. The elaboration of several hydrolytic enzymes by individuals also emphasize the crucial role of vibrios in the mineralization process in the marine environment. This study throws light on the extracellular hydrolytic enzyme profile exhibited by vibrios. It was concluded that apart from the PHA accumulation, presence of exoenzyme production and higher MAR index also aids in their survival in the highly challenging benthic enviromnents. The phylogenetic analysis of the strains and studies on intra species variation within PHA accumulating strains reveal their diversity. The isolate selected for production in this study was Vibrio sp. strain BTKB33, identified as V.azureus by 16S rDNA sequencing and phenotypic characterization. The bioprocess variables for PHA production utilising submerged fermentation was optimized employing one-factor-at-a-time-method, PB design and RSM studies. The statistical optimization of bioprocess variables revealed that NaCl concentration, temperature and incubation period are the major bioprocess variables influencing PHA production and PHA content. The presence of Class I PHA synthase genes in BTKB33 was also unveiled. The characterization of phaC genes by PCR and of the extracted polymer employing FTIR and NMR analysis revealed the presence of polyhydroxybutyrate, smallest known PI-IAs, having wider domestic, industrial and medical application. The strain BTKB33 bearing a significant exoenzyme profile, can thus be manipulatedin future for utilization of diverse substrates as C- source for PHA production. In addition to BTKB33, several fast growing Vibrio sp. having PHA accumulating ability were also isolated, revealing the prospects of this environment as a mine for novel PHA accumulating microbes. The findings of this study will provide a reference for further research in industrial production of PHAs from marine microorganisms .

Biochemical effects of ethanol on oreochromis mossambicus (Peters)

The present study dealt with the haematological, biochemical and istopathological impacts of different sub lethal concentrations of ethanol on a euryhaline teleost Oreochromis In05.s‘ambicu.5' (Peters).Studies carried out using GC indicated an increase in blood ethanol oncentration of the fish which mainly arose due to fishes entering into a state of hypoxia which explains ethanol production as an ubiquitous “anaerobic” end product, which gets accumulated whenever metabolic demand exceeds the mitochondrial oxidative potential. The very low amount of ethanol detected in the control group ofO mossambicns was mainly due to the activity of microorganisms in the gut ofO. Nzossambicus.Oedcma observed in the present study, is a defense mechanism that reduces the branchial superficial area of the fish which comes in contact with the external milieu. These mechanisms also increase the diffusion barrier to the pollutant. Dilation of the blood vessels is due to increased permeability helping in the free passage of ethanol into the blood stream. Telangiectasis observed explains the state of asphyxia of the fish when subjected to ethanol toxicity indicating acute respiratory distress. Gill aneurysm observed indicates impaired respiratory function. This is related to the rupture of the pillar cells which results in an increased blood flow inside the lamellae, causing dilation of the blood vessel or even aneurysm of gill.The present findings warrant future studies to explore A'T'Pases as possible biomarkers of pollutant exposure in ecotoxicology. This study indicated that O. mossambicus when exposed for 7 and 21 days to ethanol was under tremendous stress and parameters employed in this study can be adapted for future investigations as biomarkers of damage caused by ethanol to aquatic organisms. The present study revealed that O. mossambicus is sensitive to sub lethal concentrations of ethanol.

Biodegradation of phenolic compounds in different ecosystems in Cochin

The purpose of this study is to determine the ability of specifically adapted bacteria to degrade phenol and to quantify the rate of biodegradation at. Different concentrations by mixed as well as individual isolates. Regular quantitative analysis of phenolics and aerobic phenololytic heterotrophs from five different ecosystems were done during 1990-1991, and the ability of microorganisms isolated from those areas, to utilize phenol, o-cresol and orcinol was also studied. In addition, data on environmental parameters like temperature, dissolved oxygen, salinity, pH, organic carbon and nutrients were also collected during the period of study The present study is one of its first kind in natural aquatic environment and has aimed to bring out some idea about the potential phenol biodegrades in such environments where the phenol concentration is beyond permitted level.

Ionomer Compatibilized Low Density Polyethylene - Tapioca Starch Blends Biodegradability and Photodegradability

Biodegradation is the chemical degradation of materials brought about by the action of naturally occurring microorganisms. Biodegradation is a relatively rapid process under suitable conditions of moisture, temperature and oxygen availability. The logic behind blending biopolymers such as starch with inert polymers like polyethylene is that if the biopolymer component is present in sufficient amount, and if it is removed by microorganisms in the waste disposal environment, then the base inert plastic should slowly degrade and disappear. The present work focuses on the preparation of biodegradable and photodegradable blends based on low density polyethylene incorporating small quantities of ionomers as compatibilizers. The thesis consists of eight chapters. The first chapter presents an introduction to the present research work and literature survey. The details of the materials used and the experimental procedures undertaken for the study are described in the second chapter. Preparation and characterization of low density polyethylene (LDPE)-biopolymer (starch/dextrin) blends are described in the third chapter. The result of investigations on the effect of polyethylene-co-methacrylic acid ionomers on the compatibility of LDPE and starch are reported in chapter 4. Chapter 5 has been divided into two parts. The first part deals with the effect of metal oxides on the photodegradation of LDPE. The second part describes the function of metal stearates on the photodegradation of LDPE. The results of the investigations on the role of various metal oxides as pro-oxidants on the degradation of ionomer compatibilized LDPE-starch blends are reported in chapter 6. Chapter 7 deals with the results of investigations on the role of various metal stearates as pro-oxidants on the degradation of ionomer compatibilized LDPE-starch blends. The conclusion of the investigations is presented in the last chapter of the thesis.

Alkaline Proteases From Bacteria Isolated From Cochin Estuary

Microorganisms distributed in the marine and brackish environments play an important role in the decomposition of organic matter and mineralisation in the system (Seshadri and lgnacimuthu, 2002). Estuary is one of the most productive ecosystems, at the same time one among the least explored ecosystems on earth, which has immense potential as a source of potent microorganisms that produce valuable compounds particularly, enzymes such as proteases. In this scenario, it is very appropriate to embark on finding novel alkaline protease producers from the estuarine system. The area where the present investigation was carried out is a part of the extensive estuarine system of South India viz. Cochin Estuary. There is meagre knowledge regarding the microbial composition, particularly the protease producers of Cochin Estuary. Hence, the present study has been undertaken with the objective of finding novel alkaline protease producing bacteria from Cochin Estuary