21 resultados para Biochemical and molecularcharacterization

em Cochin University of Science


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A modified atmosphere may be defined as a packaging or storage of a perishable product in an atmosphere other than that of air. A modified atmosphere (MA) applies to food packaged products changes continuously throughout the storage period. The pearl spot (Etroplus suratensis) is an important brackish water fish belonging to the family Cichlidae. The present work was carried out to see the effect of modified atmosphere packaging on the shelf life fresh pearl spot stored in ice to extent the shelf life. The objectives of the present study are to study the suitability of Thermoformed Trays for modified atmosphere packaging, to standardize the most suitable gas mixture for modified atmosphere packaging pearl spot based on sensory evaluation, to find out the effect of modified atmosphere packaging in comparison to air packaging, to study the biochemical, microbiological, sensory and textural characteristics during storage, to study the safety concern regarding the Clostridium botulinum during modified atmosphere packaging, to find out the most suitable chemical quality indices for modified atmosphere stored pearl spot

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Division of Marine Biology, Microbiology and Biochemistry, School of Marine Sciences, Cochin University of Science and Technology

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Department of Marine Biology, Microbiology and Biochemistry, Cochin University of Science and Technology

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School of Industrial Fisheries, Cochin University of Science and Technology

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The present investigation was envisaged to determine the prevalence and identify the different Salmonella serovar in seafood from Cochin area. Though, the distribution of Salmonella serovars in different seafood samples of Cochin has been well documented, the present attempt was made to identify the different Salmonella serovars and determine its prevalence in various seafoods. First pan of this investigation involved the isolation and identification of Salmonella strains with the help of different conventional culture methods. The identified isolates were used for the further investigation i.e. serotyping, this provides the information about the prevalent serovars in seafood. The prevalent Salmonella strains have been further characterized based on the utilization of different sugars and amino acids, to identify the different biovar of a serovar.A major research gap was observed in molecular characterization of Salmonella in seafood. Though, previous investigations reported the large number of Salmonella serovars from food sources in India, yet, very few work has been reported regarding genetic characterization of Salmonella serovars associated with food. Second part of this thesis deals with different molecular fingerprint profiles of the Salmonella serovars from seafood. Various molecular typing methods such as plasmid profiling, characterization of virulence genes, PFGE, PCR- ribotyping, and ERIC—PCR have been used for the genetic characterization of Salmonella serovars.The conventional culture methods are mainly used for the identification of Salmonella in seafood and most of the investigations from India and abroad showed the usage of culture method for detection of Salmonella in seafood. Hence, development of indigenous, rapid molecular method is most desirable for screening of Salmonella in large number of seafood samples at a shorter time period. Final part of this study attempted to develop alternative, rapid molecular detection method for the detection of Salmonella in seafood. Rapid eight—hour PCR assay has been developed for detection of Salmonella in seafood. The performance of three different methods viz., culture, ELISA and PCR assays were evaluated for detection of Salmonella in seafood and the results were statistically analyzed. Presence of Salmonella cells in food and enviromnental has been reported low in number, hence, more sensitive method for enumeration of Salmonella in food sample need to be developed. A quantitative realtime PCR has been developed for detection of Salmonella in seafood. This method would be useful for quantitative detection of Salmonella in seafood.

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Vibrio are important during hatchery rearing. aquaculture phase and post-harvest quality of shrimps. Vibrio spp are of concern to shrimp farmers and hatchery operators because certain species can cause Vibriosis. Vibrio species are of concern to humans because certain species cause serious diseases.With the progress in aquaculture, intensive systems used for shrimp aquaculture create an artificial environment that increases bacterial growth. To maintain the productivity of such an intensive aquaculture, high inputs of fish protein have to be employed for feeding together with high levels of water exchange and the massive use of antibiotics/ probiotics / chemicals. It seems that the combination of these conditions favours the proliferation of vibrios and enhances their virulence and disease prevalence. The risk of a microbial infection is high, mainly at larval stages. The effect and severity are related to Vibrio species and dose, water, feed, shrimp quality and aquaculture management.Consumption of seafood can occasionally result in food-bome illnesses due to the proliferation of indigenous pathogens like Vibrio.Of the l2 pathogenic Vibrio species, 8 species are known to be directly food associated. Strict quality guidelines have been laid by the importing nations, for the food products that enter their markets. The microbiological quality requirement for export of frozen shrimp products is that V.cholerae, V.parahaemolyticus and V. vulnificus should be absent in 25g of the processed shrimp (Export Inspection Council of India, 1995). The mere presence of these pathogenic Vibrios is sufficient for the rejection of the exported product.The export rejections cause serious economic loss to the shrimp industry and might harm the brand image of the shrimp products from the countiy.There is a need for an independent study on the incidence of different pathogenic vibrios in shrimp aquaculture and investigate their biochemical characteristics to have a better understanding about the growth and survival of these organisms in the shrimp aquaculture niche. PCR based methods (conventional PCR, duplex PCR, multiplex-PCR and Real Time PCR) for the detection of the pathogenic Vibrios is important for rapid post-harvest quality assessment. Studies on the genetic heterogeneity among the specific pathogenic vibrio species isolated from shrimp aquaculture system provide; valuable information on the extent of genetic diversity of the pathogenic vibrios, the shrimp aquaculture system.So the present study was undertaken to study the incidence of pathogenic Vibrio spp. in Penaeus monodon shrimp hatcheries and aquaculture farms, to carry out biochemical investigations of the pathogenic Vibrio spp isolated from P. monodon hatchery and. aquaculture environments, to assess the effect of salt (NaCl) on the growth and enzymatic activities of pathogenic Vibrio spp., to study the effect of preservatives, and chemicals on the growth of pathogenic Vibrio spp. and to employ polymerase chain reaction (PCR) methods for the detection of pathogenic V ibrio spp.Samples of water (n=7) and post-larvae (n=7) were obtained from seven Penaeus monodon hatcheries and samples of water (n=5), sediment (n=5) and shrimp (n=5) were obtained from five P. monodon aquaculture farms located on the East Coast of lndia. The microbiological examination of water, sediment, post-larvae and shrimp samples was carried out employing standard methods and by using standard media.The higher bacterial loads were obtained in pond sediments which can be attributed to the accumulation of organic matter at the pond bottom which stimulated bacterial growth.Shrimp head. (4.78 x 105 +/- 3.0 x 104 cfu/g) had relatively higher bacterial load when compared to shrimp muscle 2.7 x 105 +/- 1.95 x 104 cfu/g). ln shrimp hatchery samples, the post-larvae (2.2 x 106 +/- 1.9 x 106 cfu/g) had higher bacterial load than water (5.6 x 103 +/- 3890 cfu/ml).The mean E.coli counts were higher in aquaculture pond sediment (204+/-13 cfu/g) and pond water (124+/-88 cfu/ml). Relatively lower Escherichia coli counts were obtained from shrimp samples (12+/-11 to 16+/-16.7 cfu/g). The presence of E.coli in aquaculture environment might have been from the source water. E.coli was not detected in hatchery waters and post-larvae.

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The present work is focussed mainly on the utilization of this weed-biomass on a biochemical and biotechnological basis. Before designing scientific and systematic utilization of any given biomass, the detailed analysis of its chemical componets is essential. Hence, as the preliminary part of the experimental works, samples of Salvinia were analysed for its chemical constituents.Before designing scientific and systematic utilization of any given biomass, the detailed analysis of its chemical componets is essential .The composition of the substrate contributes much to the nutritive value of mushrooms. Hence, alterations in the nutritive value of mushrooms (in terms of total carbohydrates, proteins, lipids and minerals) in response to Salvinia as substrate were analyzed.Substrate after mushroom harvest (spent substrate) can be utilized for various purposes such as cattle feed, as a source of degradative enzymes, as a substrate for other mushrooms and as garden manure. But studies are limited with regard to the utilization of Pleurotus spent substrate as garden manure. So the value of spent substrate as an organic supplement and its multidimensional impacts on soil chemical status, soil microbial population dynamics and plant growth (Amhurium andreanum) were carried out.Major findings of this work have got much relevance in designing measures to utilize different types of plant biomass, especially aquatic weeds, with the aid of a powerful biological tool, the lignocellulolytic fungus, Pleurorus

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The group cyanobacteria includes a large number of organisms characterised by a low state of cellular organization. Their cells lack a well defined nucleus. Cell division is by division of the protoplast by an ingrowth of the septum. These organisms are characterised generally by a blue green colouration of the cell, the chief pigments being chlorophyll-a, carotenes, xanthophylls, C phycocyanin and C phycoerythrin. The product of photosynthesis is glycogen. These organisms lack flagellate reproductive bodies and there is a total lack of sexual reproduction. They are also unique because of the presence of murein in the place of cellulose (cell wall) and the absence of chloroplast, mitochondria and endoplasmic reticulum. Just like bacteria some of them possess Plasmids and can fix atmospheric nitrogen. In the present study growth kinetics, heavy metal tolerance, tolerance mechanisms, heavy metal intake, and antibacterial activity of §ynechocystics salina Wislouch - a nanoplanktonic, euryhaline, Cyanobacterium present in Cochin back waters has been carried out for the potential biotechnological application of this organism. _§; salina occur as small spherical cells of 3n diameter (sometimes in pairs) with bluish green colour. The species is characterised by jerky movement of the cells and is structrually similar to other cyanobacteria

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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

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Actinomycetes are gram-positive, free-living, saprophytic bacteria widely distributed in soil, water and colonizing plants showing marked chemical and morphological diversity. They are potential source of many bioactive compounds, which have diverse clinical effects and important applications in human medicine. In the present work, we have studied some of the physiological and biochemical characteristics of 36 actinomycete strains isolated from the shola soils of tropical montane forest; a relatively unexplored biodiversity hotspot. Ability of actinomycetes isolates to ferment and produce acids from various carbohydrate sources such as innositol, mannose, sorbitol, galactose, mannitol, xylose, rhamnose, arabinose, lactose and fructose were studied. Almost all the carbon compounds were utilized by one or other actinomycete isolates. The most preferred carbon sources were found to be xylose (94.44%) followed by fructose and mannose (91.66%). Only 41.76% of the isolates were able to ferment lactose. The ability of actinomycetes isolates to decompose protein and amino acid differ considerably. 72.22% of the isolates were able to decompose milk protein casein and 61.11% of the isolates decompose tyrosine. Only 8.33% of the strains were able to decompose amino acid hypoxanthine and none of them were able to decompose amino acid xanthine. Potential of the actinomycetes isolates to reduce esculin, urea and hippurate and to resist lysozyme was also checked. 91.66% of the isolates showed ability to decompose esculin and 63.88% of the isolates had the capacity to produce urease and to decompose urea. Only 25% of the isolate were able to decompose hippurate and 94.44% showed lysozyme resistance

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The thesis is comprised of seven chapters. Chapter 1 gives a general introduction to marine actinomycetes; Chapter 2 gives an account on the morphological, biochemical and physiological characterization of marine actinomycetes. Comprehensive description of molecular identification and phylogenetic analysis of actinomycetes is dealt with in Chapter 3. The antimicrobial property with special reference to antivibrio activity is described in Chapter 4. Chapter 5 explores the melanin production ability of marine actinomycetes, characterization of melanin and evaluation of its bioactivity. Chapter 6 illustrates the study on chitinolytic Streptomyces as antifungal and insecticidal agents. Summary and Conclusion of the study is presented in Chapter 7, followed by References and Appendices.The present study provides an insight into the various actinomycetes occurring in the sediments of Arabian Sea and Bay of Bengal. Streptomyces was found to be the dominant group followed by Nocardiopsis. Eventhough generic level identification is possible by traditional phenotypic methods, species level identification necessitate a polyphasic approach including both phenotypic and genotypic characterization. Antibiotic production coupled with biogranulation property helped in the effective utilization of the actinomycetes for the control of vibrios. Melanin from Streptomyces bikiniensis was proved to be a promising antioxidant and photoprotectant. Marine actinomycetes were found to be a good source of hydrolytic enzymes and the chitinolytic isolates could be explored as biocontrol agents in terms of antifungal and insecticidal property. The present study explored the potential of marine actinomycetes especially Streptomycetes as a promising source of bioactive molecules for application in aquaculture and pharmaceutical industry.

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Hypoxia in neonates can lead to biochemical and molecular alterations mediated through changes in neurotransmitters resulting in permanent damage to brain. In this study, we evaluated the changes in the receptor status of GABAA in the cerebral cortex and brainstem of hypoxic neonatal rats and hypoxic rats supplemented with glucose and oxygen using binding assays and gene expression of GABAAa1 and GABAAc5. In the cerebral cortex and brainstem of hypoxic neonatal rats, a significant decrease in GABAA receptors was observed, which accounts for the respiratory inhibition. Hypoxic rats sup- plemented with glucose alone and with glucose and oxygen showed, respectively, a reversal of the GABAA receptors, andGABAAa1 and GABAAc5 gene expression to control. Glucose acts as an immediate energy source thereby reducing the ATP-depletion-induced increase in GABA and oxygenation, which helps in encountering anoxia. Resuscitation with oxygen alone was less effective in reversing the receptor alterations. Thus, the results of this study suggest that reduction in the GABAA receptors functional regulation during hypoxia plays an important role in mediating the brain damage. Glucose alone and glucose and oxygen supplementation to hypoxic neonatal rats helps in protecting the brain from severe hypoxic damage.

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The problem investigated is on the haematological aspects of two freshwater pulmonate snails, Indoplanorbis exustus (Deshayes),and Lymnaea acuminata f.rufescens (Gray). An important aspect of the present investigation is to emphasize the utilization of freshwater organisms as models for research directed at understanding the basic biomedical problems that remain unresolved. Another aspect is to demonstrate how haemolymph can be treated as a tissue because of late, it has been shown that several parameters of blood can be taken as reliable indicators for diagnostic purposes, and also to monitor environmental pollution. The various haematological parameters studied are total haemocyte number,packed cell volume, haemoglobin, and inorganic as well organic constituents in three size groups of both the snail species. The effect of copper toxicity was measured in terms of total haemocyte count, and the activity pattern of selected phosphatases and transaminases.The study concluded that enzyme activity levels can be taken as reliable indicators to monitor pollution. Age is a factor that determines several of the physiological, biochemical and metabolic activities. This study also indicates that haemolymph can be taken as an organ system to study the various changes taking place at organ systems levels.

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Development of organic molecules that exhibit selective interactions with different biomolecules has immense significance in biochemical and medicinal applications. In this context, our main objective has been to design a few novel functionaIized molecules that can selectively bind and recognize nucleotides and DNA in the aqueous medium through non-covalent interactions. Our strategy was to design novel cycIophane receptor systems based on the anthracene chromophore linked through different bridging moieties and spacer groups. It was proposed that such systems would have a rigid structure with well defined cavity, wherein the aromatic chromophore can undergo pi-stacking interactions with the guest molecules. The viologen and imidazolium moieties have been chosen as bridging units, since such groups, can in principle, could enhance the solubility of these derivatives in the aqueous medium as well as stabilize the inclusion complexes through electrostatic interactions.We synthesized a series of water soluble novel functionalized cyclophanes and have investigated their interactions with nucleotides, DNA and oligonucIeotides through photophysical. chiroptical, electrochemical and NMR techniques. Results indicate that these systems have favorable photophysical properties and exhibit selective interactions with ATP, GTP and DNA involving electrostatic. hydrophobic and pi-stacking interactions inside the cavity and hence can have potential use as probes in biology.

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In the present study, the effects of 5-HT, GABA and Bone Marrow Cells infused intranigrally to substantia nigra individually and in combinations on unilateral rotenone infused Parkinsonism induced rats. Scatchard analysis of DA, DA D1 and D2 receptors in the corpus striatum, cerebral cortex, cerebellum, brain stem and hippocampus showed a significant increase in the Brain regions of rotenone infused rat compared to control. Real Time PCR amplification of DA D1, D2, Bax and ubiquitin carboxy-terminal hydrolase were up regulated in the brain regions of rotenone infused rats compared to control. Gene expression studies of -Synuclien, cGMP and Cyclic AMP response element-binding protein showed a significant down regulation in Rotenone infused rats compared to control. Behavioural studies were carried out to confirm the biochemical and molecular studies.Our study demonstrated that BMC administration alone cannot reverse the above said molecular changes occurring in PD rat. 5-HT and GABA acting through their specific receptors in combination with bone marrow cells play a crucial role in the functional recovery of PD rats. 5-HT, GABA and Bone marrow cells treated PD rats showed significant reversal to control in DA receptor binding and gene expression. 5-HT and GABA have co-mitogenic property. Proliferation and differentiation of cells re-establishing the connections in Parkinson's disease facilitates the functional recovery. Thus, it is evident that 5-HT and GABA along with BMC to rotenone infused rats renders protection against oxidative, related motor and cognitive deficits which makes them clinically significant for cellbased therapy. The BMC transformed to neurons when co-transplanted with 5-HT and GABA which was confirmed with PKH2GL and nestin. These newly formed neurons have functional significance in the therapeutic recovery of Parkinson’s disease.