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http://inet.vidyasagar.ac.in:8080/jspui/handle/123456789/1087
2024-03-29T13:00:45ZMolecular identification and pathogenecity study of the Klebsiella pneumoniae isolated from fishes of West Bengal
http://inet.vidyasagar.ac.in:8080/jspui/handle/123456789/6204
Title: Molecular identification and pathogenecity study of the Klebsiella pneumoniae isolated from fishes of West Bengal
Authors: Das, Abhishek
Abstract: Fish health mainly depends upon the environmental conditions as well as on the farming
techniques. However, in present aquaculture scenario farmers are over-stocking their aquaculture
farms as well as they are providing excessive feed. The unutilized feed is detrimental to water
quality leading to the outbreak of various kinds of diseases. Bacterial infections in farmed fishes
due to Klebisella pneumoniae was not reported earlier. The thesis focuses on the molecular
identification and pathogenicity study of Klebsiella pneumoniae in fresh water fishes. Infected
fish samples viz. Labeo rohita, Oreochromis niloticus, Labeo catla, Cirrhinus mrigala, Anabas
testudineus and Clarias batrachus were collected from three different districts of West Bengal
viz. North 24 Parganas, Nadia, both east and west Burdwan. Bacterial strains were isolated from
liver, kidney and blood tissues of the infected fishes. Primarily bacterial strains were identified
using biochemical test and further 16S rRNA gene amplification was carried out for the
molecular identification of the isolates. The amplified products were sequenced and the sequence
was submitted to NCBI. Phylogenetic analysis of the isolated strains revealed the species
diversity and showed dispersion of bacterial strains to different location. Antibiotic sensitivity
assay was carried out to determine the multiple antibiotic resistance index of the isolated strains.
The MAR index was found to be 0.476 which showed that the strains were highly exposed to
antibiotics. The LD
50
value was found to be 1.05 X 10
6
CFU /fish. Histopathology of the liver
tissue revealed necrosis, vacuolation and disruption of hepatocytes whereas ultra-structural
alterations in the renal tubules and glomeruli were evident in the kidney tissue. The bacteria were
found to be β-hemolytic positive in nature as they have caused complete lysis of sheep blood on blood agar plates. Among the array of virulent genes screened the isolates were found to be
positive for fimA, fimH, mrkA, urea, ugeA, wabG, ecpRAB and entB. The amplified genes were
sequenced and the sequences were submitted to NCBI.
Various specific and nonspecific immune parameters of the challenged fishes were
studied at various time intervals to find out the immune response against this particular pathogen.
An elevated myloperioxidase activity, superoxide production, and lysozyme activity were
observed. However, the antiprotease and α-2 macroglobulin activities were found to be
downregulated. Alteration of immune related genes viz. C3, IL-1β and IL- 6 were also studied in
the internal tissues like kidney, muscle and liver. Early expression of C3, IL- 6 and IL-1β gene
was observed in the liver tissue however; late expression of all the genes were recorded in the
kidney tissue.
The thesis provides a detailed report of Klebsiella pneumoniae infection in farmed fishes
and also it could be concluded that, Klebsiella pneumoniae is an emerging pathogen which could
cause huge loss in aquaculture industry. A stringent surveillance is required to identify the
abundance of Klebsiella pneumoniae in aquaculture farms.2021-07-22T00:00:00ZEpitope Mapping of the Allergenic Proteins of the Pollen of Datura sp. and its implications for immunotherapy
http://inet.vidyasagar.ac.in:8080/jspui/handle/123456789/5887
Title: Epitope Mapping of the Allergenic Proteins of the Pollen of Datura sp. and its implications for immunotherapy
Authors: Bera, Barnali
Abstract: Allergic rhinitis, also known as hay fever is the most common of all allergic
diseases affecting approximately 400 million people worldwide. It is triggered
by allergens present in our immediate environment such as pollen, mould,
fungal spores, dust, mites, pet hair, etc. The role of pollen in the etiology of
nasobronchial allergy is now very well established. There are several tests to
reveal the specific allergens to which an individual is sensitive. Among these
skin prick test, intradermal test, intradermal or intracutaneous injection
inhalation, nasal or bronchial mucosa-provocation tests or the histamine release
test are some commonly employed to detect allergens (Stanley and Linskens,
1974). There are also certain immunoassay methods which include the radioallergo-sorbent-test
(RAST) developed by Wide et al.(1967) and a cheaper and easier method - enzyme linked immuno sorbent assay (ELISA) developed by Engvall and Pearlman (1971).
One of the most successful treatment for allergic rhinitis is immunotherapy,
also called hypo-sensitization. Identification of the actual allergenic
components of pollen, their characterization and epitope mapping of these
allergenic fractions is very essential for preparation of vaccines for successful
immunotherapy.
The present study was undertaken to identify the allergenic protein fractions of
the pollen of three related species of Datura (Datura metel, Datura inoxia and
Datura stramonium) and Epitope mapping of the allergenic proteins of the
pollen of Datura sp. in order to identify and characterize the binding sites of
antibodies which can aid in the development of new therapeutic vaccines for
successful immunotherapy. Thus the overall study involved the following:
Ultrastructure study of the pollen by Light microscopy, SEM and TEM
for proper identification of the airborne pollen to which a patient is
exposed to in his or her immediate vicinity and to study any specific
features of pollen morphology as well as the ultrastructure that might
have the potential to influence its allergenicity.
Extract, quantify and study the soluble protein profile of the pollen of
the three related species of Datura using standard methods and gel
electrophoresis and partial characterization of the proteins (determining
the molecular weights).
Study the variation in protein profile with maturity of pollen (before
and after anthesis).
Isolation of the individual protein fractions by gel filtration.
Identification of the allergenic protein fractions by performing
Ouchterlony Immunodiffusion and ELISA with blood plasma of Datura
sensitive patients.
Development of antibodies in Male LOBUND –Wistar rats and study
cross reactivity among the 3 species of Datura by Ouchterlony
Immunodiffusion and ELISA to identify the common proteins fractions.
Crosslinking coupled Mass Spectrometry for epitope mapping.
Detection and identification of antigen-antibody binding location with
high mass MALDI detection (high resolution mass spectrometry or
MS/MS techniques) and amino acid sequencing of the epitopes.
The pollens from the three species of Datura i.e. Datura metel, Datura inoxia
and Datura stramonium were collected and then processed by applying
different methodologies to evaluate the comparative protein concentrations of
the pollen on seasonal basis, SDS-PAGE protein profiles of the pollen of the
three species of Datura were also performed. Isolation of individual protein
fractions by gel filtrations, analysis of protein fractions by PAGE were also
carried out to obtain the individual proteins from the pollen. Later on,
identification of allergenic protein fractions was completed by employing
Immunodiffusion, ELISA and many other techniques. Cross reactivity among
these three species of Datura was also performed. Finally epitope mapping was
conducted to identify antigenic determinants of the allergenic protein fractions of the pollen of three species of Datura i.e. Datura metel, Datura inoxia and
Datura stramonium by MALDI/TOF.
An additional study on ultrastructure of pollen byy SEM and TEM was also
carried out to analyze the relativity of pollen structure with its allergenicity.2021-02-09T00:00:00ZStudies on the conservation and enhancement of biological diversity of economically and ecologically important ichthyofaunal resources of Paschim Medinipur district, West Bengal, India using Geoinformatics
http://inet.vidyasagar.ac.in:8080/jspui/handle/123456789/5377
Title: Studies on the conservation and enhancement of biological diversity of economically and ecologically important ichthyofaunal resources of Paschim Medinipur district, West Bengal, India using Geoinformatics
Authors: Kisku, Sujit; Patra, Bidhan Chandra
Abstract: Doctoral research work embodied in the thesis was studies on the conservation and enhancement of biological diversity of economically and ecologically important ichthyofaunal resources of Paschim Medinipur district, West Bengal, India using Geoinformatics.An extensive study carried out in the twenty nine Community Development Blocks in Paschim Medinipur district to record the present status of freshwater finfish fauna of this district.Also the habitat of the fish was of major concern as because fish biology and diversity depends on the perennial water bodies with permissible range of physicochemical parameters. In this present research work a total of 76 numbers of finfish species recorded. They belong to 8 orders and 23 families.Most fishes come under order cypriniformes followed by perciformes and siluriformes. Among the families cyprinidae shares 29 fish species followed by bagridae, channidae and ambassidae.As per the IUCN conservation status (Ver 3.1) among the recorded 76 finfish species 1 species is vulnerable, 7 near threatened, 60 are least concern, 2 data deficient and 6 not evaluated.Water parameters recorded using water analyzer kit mostly on the spot and water samples also brought to laboratory for further examination. Geological positioning of the sampling stations recorded by handheld GPS device.The data of aquatic parameters and GPS was plotted on spreadsheet and map generated. The fishes were collected directly from the water bodies, from the local fish market and from the fish farmers. The fish specimens were preserved after taking their photograph and then subjected to taxonomic categorization.The raw data was then utilised to see the diversity, richness, correlation with the help of different diversity indices in PAST software. Maps generated on sampled aquatic habitats. Geocoding of the satellite image data was done and outputs were performed by ERDAS imagine software.Georeferencing and ground truthing was done using ERDAS and Arc GIS software. The Community Development Blocks having permissible and near permitted range of physicochemical parameter of water bodies exhibit high fish diversity than the unfavourable zones.Fish diversity is highest in Daspur-I & II, Pingla, Ghatal and Sabang in comparison to the remaining Community Development Blocks. Correlation matrix shows positive and significant correlation among temperature, TDS, turbidity, conductivity and salinity and negative relation with pH, DO, OD and fish species.The study will help fish farmers about the culture and capture of fin fishes scientifically and judiciously in sustainable manner. It will help Policy makers to formulate policies to conserve the threatened fish species and fish habitat.It will also help researcher to work on in this field of diversity and conservation. It is established that the aquatic reservoirs like beels, jheels, lake and pond have greater potentiality and opportunity in this district to enhance the wild fish species.As a whole this research study will provide better management strategies in conserving threatened fish species and to raise ichthyofaunal diversity in Paschim Medinipur district, West Bengal, India.2020-02-10T00:00:00ZMicroscopical studied on olfactory apparatus of Mastacembelus armatus (Lacepede, 1800)
http://inet.vidyasagar.ac.in:8080/jspui/handle/123456789/5374
Title: Microscopical studied on olfactory apparatus of Mastacembelus armatus (Lacepede, 1800)
Authors: Jana, Shreya; De, Subrata Kumar
Abstract: Mastacembelus armatus (Lacepède, 1800) is a common spiny eel of South East Asia. The olfactory apparatus of M. armatus is studied under microscope to explore different subcellular elements of sensory components in relation to neurogenesis and neural degeneration, supported by macroanatomy, microanatomy, 3D surface topography analysis, transmission electron microscopy (TEM), TEM based energy dispersive X- ray microanalysis (EDX) respectively.The comparative analysis of variable subcellular elements has been correlated with the functional bioaccumulation of heavy metals within subcellular compartments of sensory cells. Apart from that this research contribution also addresses olfactory dysfunction of fish with various aspects of neural degeneration, effects heavy metal pollution respectively in lieu of variable neural diseases of higher vertebrates viz., Parkinson’s, Alzheimer’s, Huntington’s, incidental Lewy Body disease, etc.2020-02-10T00:00:00Z