The facility houses the following essential instruments for studying host-pathogen interactions:

• Bio-safety Cabinets (Type II-A2, Type II-B2)
• -80^oC freezer
• Refrigerated Centrifuges and Shaker
• CO2 incubators
• Spectrophotometer
• Phase contrast microscope

• Used for handling live Risk Group - 2 pathogens. View more details
• Have tissue culture facility and provides options for studying host-pathogen interaction outcomes.
• Fully automated facility that maintains negative pressure, regulated temperature and humidity.
• Waste (both solid and liquid) and spill containment facilities are available.

A BSL-2 facility was set up in March, 2015 in Department of Bioscience and Bioengineering by Klenzaids Contamination Control Pvt. Ltd. In 2016, it was converted into a central facility. It is suitable for work involving Risk Group -2 pathogens that pose moderate hazard to personnel and the environment. Examples of agents typically worked with in a BSL-2 include equine encephalitis viruses and HIV, as well as Staphylococcus aureus, Salmonella etc.. It works efficiently to control generation of any aerosols outside the facility by filtering the incoming as well as outgoing air and maintaining a negative pressure. Apart from Bio-safety cabinets, the facility also houses all other necessary equipments to perform experiments which involves pathogen biology, host-pathogen interaction etc. Access to BSL-2 facility is strictly restricted and should be approved by BSL-2 user committee.

• For what kind of samples this facility is used?
  This is not an instrument but a facility which houses several instruments. Therefore the usage is not sample based but project based. This facility can be used for experiments that require handling of Risk Group -2 pathogens. A complete list of Risk Group -2 pathogens are given in following document: Risk Group 2 Pathogens.doc
• Who can use this facility?
  Ph.D students and post docs.
• Do you need any approval for using this facility?
  Yes, your project needs Institutional Biosafety Committee’s (IBSC) approval, followed by approval of the experimental protocols by the user committee of the facility.
• What sort of experiment one can do inside this facility?
  The facility has all the necessary equipments for conducting major experiments in infectious disease biology to study host-pathogen interactions.
• What sort of precautions one needs to take to work in this facility?
  All users must undergo a detailed training schedule before working independently inside the facility. Following completion of training the user have to sign an undertaking citing that he/she has been trained and now responsible for any safety related hazards the he/she faces.
• What happens if one violates the rules of the facility?
  The person is barred from entering the facility for indefinite period of time.

Surve MV, Anil A, Kamath KG, Bhutda S, Sthanam LK, Pradhan A, Srivastava R, Basu B, Dutta S, Sen S, Modi D, Banerjee A. (2016) Membrane Vesicles of Group B Streptococcus Disrupt Feto-Maternal Barrier Leading to Preterm Birth. PLOS Pathogens 12(9): e1005816. Patel Y, Mehra S (2017) Synergistic Response of Rifampicin with Hydroperoxides on Mycobacterium: A Mechanistic Study. Frontiers in Microbiology (under revision)

(a) Publications

• Kotta KB, Ghosh S, Mehra S. Potentiating the Anti-Tuberculosis Efficacy of Peptide Nucleic Acids through Combinations with Permeabilizing Drugs. Microbiol Spectr, 2022 10(1):e0126221.

• Ghosh, S., K.B. Cotta, A.A. Hande, M. Fernandes, and S. Mehra. PNA-mediated efflux inhibition as a therapeutic strategy towards overcoming drug resistance in Mycobacterium smegmatis. Microb Pathog, 2021. 151: p. 104737.

• Rai, D., and Mehra, S. The Mycobacterial Efflux Pump EfpA Can Induce High Drug Tolerance to Many Antituberculosis Drugs, Including Moxifloxacin, in Mycobacterium smegmatis: Antimicrob Agents Chemother. 2021; 65(11):e0026221.

• Suvarna, K., Salkar, A., Palanivel, V., Bankar, R., Banerjee, N., Gayathri J Pai, M., Srivastava, A., Singh, A., Khatri, H., Agrawal, S., Shrivastav, O., Shastri, J., & Srivastava, S. (2021). A Multi-omics Longitudinal Study Reveals Alteration of the Leukocyte Activation Pathway in COVID-19 Patients. Journal of proteome research, 20(10), 4667–4680.

• Suvarna, K., Biswas, D., Pai, M., Acharjee, A., Bankar, R., Palanivel, V., Salkar, A., Verma, A., Mukherjee, A., Choudhury, M., Ghantasala, S., Ghosh, S., Singh, A., Banerjee, A., Badaya, A., Bihani, S., Loya, G., Mantri, K., Burli, A., Roy, J., ... Srivastava, S. (2021). Proteomics and Machine Learning Approaches Reveal a Set of Prognostic Markers for
COVID-19 Severity with Drug Repurposing Potential. Frontiers in physiology, 12, 652799.

• Bankar,R., Suvarna, K., Ghantasala, S., Banerjee, A., Biswas, D., Choudhury, M., Srivastava, S. Proteomic Investigation Reveals Dominant Alterations of Neutrophil Degranulation and mRNA Translation Pathways in COVID-19 Patients (2021) iScience, Cell Press, 24(3), 101235.

• Banerjee, A., Gokhale, A., Bankar, R., Palanivel, V., Salkar, A., Robinson, H., Shastri, J. S., Agrawal, S., Hartel, G., Hill, M. M., & Srivastava, S. (2021). Rapid Classification of COVID-19 Severity by ATR-FTIR Spectroscopy of Plasma Samples. Analytical chemistry, 93(30), 10391–10396.

• Anil A, Apte S, Joseph J, Parthasarathy A, Madhavan S, Banerjee A. Pyruvate Oxidase as a Key Determinant of Pneumococcal Viability during Transcytosis across Brain Endothelium. J Bacteriol. 2021 Nov 19;203(24):e0043921. doi: 10.1128/JB.00439-21.

• Bhutda S, Ghosh S, Sinha AR, Santra S, Hiray A, Banerjee A. Differential Ubiquitination as an Effective Strategy Employed by the Blood-Brain Barrier for Prevention of Bacterial Transcytosis. J Bacteriol. 2022 Jan 18;204(1):e0045621. doi: 10.1128/JB.00456-21.

• Mistry, N., Bandyopadhyaya, R. & Mehra, S. Enhancement of Antimycobacterial Activity of Rifampicin Using Mannose-Anchored Lipid Nanoparticles against Intramacrophage Mycobacteria. ACS Appl. Bio Mater. 5, 5779–5789 (2022)
https://pubs.acs.org/doi/full/10.1021/acsabm.2c00796

• Pushpakaran A, Battaje RR, Panda D. Vitamin K3 inhibits FtsZ assembly, disrupts the Z-ring in Streptococcus pneumoniae and displays anti-pneumococcal activity. Biochem J. 2022 Jul 29;479(14):1543-1558. doi: 10.1042/BCJ20220077. PMID: 35789252

• Lin HJ, Battaje RR, Tan J, Doddareddy M, Dhaked HPS, Srivastava S, Hawkins BA, Al-Shdifat LMH, Hibbs DE, Panda D, Groundwater PW. Discovery of 2',6-Bis(4-hydroxybenzyl)-2-acetylcyclohexanone, a Novel FtsZ Inhibitor. Molecules. 2022 Oct 18;27(20):6993. doi: 10.3390/molecules27206993. PMID: 36296585; PMCID: PMC9610434

•A pte et al., An innate pathogen sensing strategy involving ubiquitination of bacterial surface proteins. Sci.Adv. 9, eade1851(2023). DOI:10.1126/sciadv.ade1851

(b) Conference presentations

• International Conference on Infectious Diseases and Immunopathology (IDIP-2021), Department of Biotechnology, Savitribai Phule Pune University, Pune (April 2021; Anirban Banerjee)

• BIO-CREST 2021, Amrita School of Biotechnology, Amritapuri, India (Feb., 2021; Anirban Banerjee)

● Athira Pushpakaran presented Vitamin K3, an inhibitor of Streptococcus pneumoniae with selective anti-FtsZ action at NIPER pharmacon held in November 10-12, 2022.

● Shweta Santra presented poster “Exploitation of Non-Canonical Autophagy by Streptococcus pneumoniae for prolonging intracellular persistence” at Integrating the molecular mechanistic and physiological diversity of autophagy, EMBO, Hungary, held on 27th June 2022.

● Battaje, R. R., Dhaked, H. P. S, & Panda, D. (2022). Interdomain cleft of FtsZ as a target for BTBenzo- 29, an inhibitor of Streptococcus pneumoniae cell division. Poster Presentation at the International Symposium on Recent Trends and Future Opportunities in Pharmaceuticals at National Institute of Pharmaceutical Education and Research, S.A.S. Nagar, Punjab, India held on November: 10-12, 2022.

● Kuldeep Sharma presented poster “Temperature-induced alteration in teichoic acid biosynthesis in Streptococcus pneumoniae” at Biological Transactions: From molecules to organisms (BTMO 2023), IISc Bangalore, held on January 18-23, 2023.

● Shruti Apte presented “Ubiquitination of bacterial substrates act as novel innate sensing strategy” at Pathogen Immunity and Signaling, EMBO conference, Saint Malo, France held on 4-8 April 2022.

● Shruti Apte presented “Ubiquitination of bacterial surface proteins aid in pathogen clearance” at All India Cell Biology Conference (AICBC), Banaras Hindu University, Varanasi, held on 20-22 January 2023.

● Sourav Ghosh presented “Ubiquitin-mediated tweezer like nanomachine protects the host from invasive bacterial infections” at Bacterial morphogenesis, survival and virulence: Dynamic genomes & envelopes, EMBO conference, Goa, held on February 6-10, 2023.

● Anirban Banerjee presented a poster at Bacterial morphogenesis, survival and virulence: Dynamic genomes & envelopes, EMBO conference, Goa, held in February 6-10, 2023.

The facility can only be used for projects that are approved by IBSC. Following IBSC's approval the protocol of different experiments needs to be approved by the user committee. Once the protocols are approved, the user (a PhD student or Post doc) will be trained and made aware of different hazards associated with working inside the facility. Following completion of training, the user will have to sign an undertaking that he/she has been trained and now responsible for any safety related hazards the he/she faces. The user must inform the facility supervisor in case of any accident or spillage at the earliest. If he/she feels sick, they need to inform the doctor about their work in BSL-2 facility and different kind of pathogens that are being handled inside the facility.

Online booking of slots are available through Qreserve software. At a time only 4 people can work inside the facility. A user can book the facility for a maximum of 3 h at a stretch. He/she can book it for another 3 h (depending on the length of the experiment), but with a gap of 1 h in between two consecutive 3 h slots to allow other users to perform minor work in between.

Project approval by IBSC and protocol approval by User committee is a must.