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Chaudhary Sarwan Kumar Himachal Pradesh Agriculture University, Palampur

Himachal Pradesh Krishi Vishvavidyalaya (renamed as Chaudhary Sarwan Kumar Himachal Pradesh Krishi Vishvavidyalaya in June, 2001) was established on 1st November, 1978.The College of Agriculture (established in May, 1966) formed the nucleus of the new farm University. It is ICAR accredited and ISO 9001:2015 certified institution. The Indian Council of Agricultural Research has ranked this University at eleventh place among all farm universities of the country. The University has been given the mandate for making provision for imparting education in agriculture and other allied branches of learning, furthering the advancement of learning and prosecution of research and undertaking extension of such sciences, especially to the rural people of Himachal Pradesh. Over the years, this University has contributed significantly in transforming the farm scenario of Himachal Pradesh. It has developed human resources, varieties and technologies and transferred these to farming community enabling the State to receive the “Krishikarman award” of Govt. of India four times in row for food grain production among small states of the country. Today, the State has earned its name for hill agricultural diversification and the farming community has imposed its faith in the University.

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Author: Dhial, Kritika × Start date: 2018 × Thesis Type: M.V.Sc. ×
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    ThesisItemOpen Access
    SELECTION AND EVALUATION OF PHAGE DISPLAY PEPTIDES AGAINST PASTEURELLA MULTOCIDA
    (CSKHPKV, Palampur, 2018-07-24) Dhial, Kritika; Sharma, Mandeep
    Haemorrhagic septicemia is an acute fatal septicaemic disease of cattle and buffaloes and is caused by P. multocida serotype B: 2 in India. Despite its economic importance, there is no specific field level diagnosis for this disease. The pathogenicity of the organism is associated with various virulence factors such as the capsule, lipopolysaccharides, adhesins, toxins, siderophores, sialides and outer membrane proteins. They facilitate the colonisation and invasion of the host tissue. These surface antigens could be a target for both therapeutics as well as diagnostics. Keeping this in mind, the current study was planned to select ligands in the form of peptides using phage display peptide library against major structural components of the Pasteurella multocida and characterize them by using phage ELISA. Ph.D.-12 phage display library was used to select phage peptides. The library titer was 1.8 × 1011 pfu/ml. This heterogenous mixture of phages carrying diverse peptides as ligands was amplified to a final concentration of 2.1x1013 pfu/ml. These amplified phages were then subjected to the alternate selection/subtraction methodology of panning using suspension method in which alternate rounds of positive selection against P. multocida and negative selection against Haemophilus influenzae and Actinobacillus lignieresii were performed. For round 1 of positive selection, 100μl of 2.1x1013 pfu/ml phages were employed and 5.8 × 106 pfu/ml of eluted phages were recovered. For negative selection of round 1 these recovered eluted phages were amplified and 1013 pfu/ml phages were employed. Around 2.8 × 1011 pfu/ml phages remained unbound which were re-amplified to carry out round 2 of positive selection. 1.3 × 1011 pfu/ml were eluted and amplified for round 2 of negative selection to the concentration 1013 pfu/ml and 1.4 × 1012 phages were left unbound. After completing round 3 of alternate selection/subtraction, 4.8 × 1012 pfu/ml were recovered. This study showed that with every round of selection, the non-binding and non-specific phages reduced and the pool of selectively binding phages to the P. multocida with each amplification increased. Further to analyse the affinity of selectively binding phages indirect phage ELISA was carried out. Out of a total of 48 phages, 16 clonal phages were selected for indirect phage ELISA. Out of these 16, five clonal phages viz. B6, B3, B7, B5 and A5 bound their target with high intensity giving higher OD values at 450 nm. These did not or bound poorly to closely related bacteria as the OD values were close to the negative control. The phages have been submitted for sequencing to further characterize them for their structural and functional attributes. This study concludes that it possible to select peptides against the intended target and that such peptides could be used for specific diagnosis of pathogens or as antimicrobial therapeutics after thorough characterization.