Executive Summary : | Objective: The most effective and affordable formulation will be developed from C. variabilis (ATCC PTA 12198) as anti-virulent agent against V. cholerae. A mixture of fatty acids will be optimized for different experiments. Screening of other potential microalgae species will be done. Study on the shelf life of formulation; its generation study for the induction of resistance in V. cholerae with different parameters. Mode of action study- in the presence of microalgal formulation, transcription mechanisms will be studied in V. cholerae. Expression of virulence genes (upregulation/downregulation) will be studied in the presence of formulation in RT¬PCR. Encapsulation of the formulation with a suitable matrix. In vivo testing for the efficacy of formulation from C. variabilis and generation study.
In vivo will be done in CSIR-IMTECH where animal ethical committee clearance is available. In BALB mice gut assay will be done for fluid accumulation and colonization and cell anatomy study (at CSIR-IMTECH). The formulation will be checked against other enteropathogenic bacterial strains causing diarrhoea such as Enterotoxigenic E. coli, Vibrio paraheamolyticus, Shigella spp by ELISA (ETEC, Shigella sp.) and type three secretion system (V. parahaemolyticus) study. Presence of proteins, micro & macro-nutrients parameters like zinc, iron, iodine, antioxidants, vitamin B, vitamin C, vitamin E, etc. in the C. variabilis will be analyzed from third-party laboratory and formulation will be developed as nutraceutical. |