Microbiology Training Module
The Microbiology Division of CoE-NPT is committed to investigate the folkloric medicinal plants of Gandhamardan, Odisha as potential therapeutic agents against severe chronic microbial infections. Particularly, our group focused upon the discovery and design of potential drug candidates (from medicinal plants) as therapeutic agents against multidrug resistant (MDR) ESKAPE (Enterococcus faecalis, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumanni, Pseudomonas aeruginosa, and Enterobacter sp.) pathogens with special reference towards biofilm dynamics. The Microbiology Division also strongholds on the synergistic potential of plant-derived phytocompounds with the conventional antibiotics for improved antibacterial and antibiofilm activities against ESKAPE pathogens. This introductory training module focuses upon the basic training in the field of basic microbiological techniques and standard operating procedures (SOPs) for the maintenance of bacterial pathogens.
Objectives:
- The introductory training module will be supported by theoretical knowledge and Hands-on Practical classes on principles of media preparation, sterilization techniques, maintenance of bacterial cultures under asceptic conditions, morphological and biochemical characterization of bacterial cultures, antibiotic susceptibility techniques, determination of antibacterial and antibiofilm activities. The participants will also be able to interact with Experts and discuss their doubts on the different techniques used in the proposed training module.
Basic Microbiological Techniques
• Introduction to good laboratory practices
• Preparation of media
• Inoculation of bacterial strains
• Maintenance of bacterial culture (Pour plate technique, Quadrant streaking, Bacterial slant, Cryopreservation)
Biochemical and Molecular characterization of Bacterial cells
• Morphological identification of bacteria (Gram staining)
• Biochemical characterization (Indole test, Methyl Red test, Catalase test, Urease test)
• Molecular characterization (16S rRNA sequencing)
Evaluation of Antibiotic susceptibility
• Disk diffusion assay
Screening and evaluation of antibacterial activities
• Agar well diffusion assay
• Minimum inhibitory concentration (MIC) assay
• TTC-based cell viability assay
Determination of antibiofilm potential of samples against bacterial pathogens
• Congo Red Agar (CRA) assay
• Qualitative Biofilm formation assay (Tube Method)
• Quantitative Biofilm formation assay (Crystal violet staining method)
• Exopolysachharides (EPS) production assay
Microscopic observation of Biofilms
• Light microscopic analysis of biofilm matrix
• Fluorescence microscopic analysis of biofilm matrix
Training Program Modules
Module-1
Introduction to basic microbiology, Scope and relevance of Microbiology, Good laboratory practices and maintenance of asceptic conditions, Familiarization with the Microbiology laboratory, standard operating procedures (SOPs) to be maintained in the laboratory, and troubleshooting
Module-2
Preparation of media, Sterilization techniques, inoculation of bacterial strains, maintenance of bacterial cultures (Pour plate method, quadrant streaking, bacterial slant, cryopreservation)
Module-3
Morphological (Gram staining method) and Biochemical identification methods (Indole test, Methyl Red test, Catalase test, Urease test), Molecular characterization technique (16S rRNA sequencing)
Module-4
Antibiotic susceptibility test (disc diffusion method), Screening for determination of antibacterial activities (Agar well diffusion assay), Determination of minimum inhibitory concentration (MIC), TTC-based cell viability assay, Calculation of MIC and sub-MIC values
Module-5
An introduction to Biofilms in bacterial pathogens, Role of biofilms in antibiotic resistance, Basic techniques to detect biofilm formation, Biofilm formation assay, Biofilm inhibition assay
Module-6
Effect of medicinal plants on biofilm formation in bacterial pathogens using Congo red agar (CRA) assay, qualitative and quantitative biofilm formation assay (Tube method and CV staining method)
Module-7
Exopolysachharides (EPS) production assay, Detection of biofilms in bacterial pathogens using microscopic methods (Light and fluorescence microscopic methods)
Evaluation of trainees and certification
Evaluation of the training will be done and certificate will be issued to the successful candidates for the course..
