Team: Miss Manasi Naik (STRIP Student, Bacteriology Department), Mrs. Dipti Kolte (STA & Co-PI, Bacteriology Department) and Dr. Anuya Rege [Principle Investigator (PI), Bacteriology Department]. 
Funding Agency: Haffkine Institute (STRIP Fee)
Duration: December 2024 – May 2025.
Abstract:
This in vitro study demonstrated the significant Antimicrobial and Antibiofilm efficacy of Citric acid against a broad range of clinically relevant bacteria including Gram-positive as well as Gram-negative and fungal pathogens in both mono- and poly-microbial settings. The combinatorial treatments of citric acid with ciprofloxacin for bacterial strains and fluconazole for fungal strains showed enhanced Antimicrobial and Antibiofilm activity compared to individual agents, especially in dual- and mixed-species biofilms which suggests a synergistic interaction. In conclusion, Citric acid, alone and in combination with standard antibiotics, holds a promising agent as a cost-effective and broadly active Antimicrobial and Antibiofilm compound. Future studies, including mechanistic investigations, are suggested to translate these findings into clinical and industrial applications.

2. Study of Biodegradation of Azo Dye (Congo Red) by Bacterial and Fungal Isolates from Textile Effluent Contaminated Soil & Water Samples.

Team: Miss. Swarangi Kalvitkar [Short Term Research Internship Program (STRIP) Student, Bacteriology Department] and Mrs. Jayashree More [Principle Investigator (PI), Bacteriology Department]. 
Funding Agency: Haffkine Institute
Duration: December 2024 – May 2025
Abstract: The extensive use of azo dyes, such as Congo Red, in the textile industry has led to serious environmental concerns due to their toxic, carcinogenic, and non-biodegradable nature. This study aims to explore the potential of bacteria and fungi isolated from textile effluent-contaminated soil and water to biodegrade Congo Red dye. Soil and water samples were collected from industrial zones in Ambernath and Badlapur, Maharashtra. A total of 40 bacterial and 20 fungal isolates were obtained through enrichment and serial dilution techniques. These isolates were screened for dye-degrading ability using qualitative (decolourisation assay) and quantitative (UV-Visible spectrophotometer and multimode reader) methods. Seven bacterial and 9 fungal isolates showing significant decolourisation were selected for further analysis. Bacterial isolates were characterized morphologically, biochemically (using Bergey’s Manual), and genetically through 16S rRNA sequencing, identifying the genus Pseudomonas. Fungal isolates were identified morphologically and microscopically, with 18S rRNA sequencing confirming the genus Aspergillus. Optimization studies on selected bacterial strains revealed that neutral to alkaline pH (7–9), moderate temperatures (25–32°C), and lower dye concentrations (20–50 mg/L) enhanced biodegradation efficiency. Phytotoxicity tests using Vigna radiata demonstrated reduced toxicity in treated dye solutions, confirming the effectiveness of microbial degradation. The results highlight Bioremediation using these microorganisms may provide an environmentally safe, economical, and efficient alternative to conventional physical and chemical treatment methods for textile wastewater management.

3. Study of Biodegradation of Anthraquinone Dye (Reactive Blue 19) by Bacterial and Fungal Isolates from Textile Effluent Contaminated Soil & Water Samples.

Team: Mr. Sidhhesh Gavkar [Short Term Research Internship Program (STRIP) Student, Bacteriology Department] and Mrs. Jayashree More [Principle Investigator (PI), Bacteriology Department].
Funding Agency: Haffkine Institute (STRIP Fee)
Duration: December 2024 – May 2025
Abstract:
Textile industries are a major contributing factor to soil and water pollution due to the release of untreated effluents in the environment. The dyes lost during the dyeing process are the main pollutants in these effluents. Among the many classes of dyes, anthraquinones are non-degradable and thus, resistant in nature. This has negative impact on soil quality and aquatic life. Various different methods have been employed to degrade these dyes. Bioremediation, that is the use of microorganisms, is a very Aspergillus effective and eco-friendly method of dye degradation. This study focuses on the dye degradation potential of bacterial and fungal isolates obtained from textile effluent contaminated soil and water samples on the anthraquinone dye – Remazol Brilliant Blue R (Reactive Blue 19). Initially, 40 bacterial and 20 fungal isolates were obtained from the collected samples. These isolates were visually observed for decolourisation of dye by employing the decolourisation test. Based on the observations, the dye degrading isolates were selected for further quantitative analysis using UV-Visible spectrophotometer and multimode reader. Gram staining results showed majority of the isolates to be gram negative in nature and, through biochemical tests, many of the isolates were found to be capable of fermenting various sugars. Phytotoxicity study on Vigna radiata demonstrated that bacteria decolourized broth supported radicle growth as opposed to the untreated dye solution. Based on absorbance readings, isolates showing prominent biodegradation potentials were selected for gene extraction. From sequencing procedures, Pseudomonas spp. and Aspergillus spp. were identified to be the Reactive Blue 19 degrading strains. This research supports the potential of microbial degradation of dyes in textile effluents as a sustainable approach for negating harmful effects of such contaminants on plant and animal life. The biodegrading isolates obtained can be experimented further for large scale bioremediation of textile dye effluents.

4. Assessment of Antibacterial and Antibiofilm effect of Ascorbic Acid- an In vitro study.

Team: Miss Saloni Akharkar (STRIP Student, Bacteriology Department), Mrs. Dipti Kolte (STA & Co-PI, Bacteriology Department) and Dr. Anuya Rege [Principle Investigator (PI), Bacteriology Department]. 
Funding Agency: Haffkine Institute (STRIP Fee)
Duration: November 2023 – March 2024
Abstract:
In the present in vitro study, Ascorbic Acid was evaluated for its Antibacterial activity against 6 bacteria (standard strains) namely, E. coli (ATCC-25922), S. enterica (NCIM-5256), S. typhi (NCTC-786), E. faecalis (ATCC-29212), MRSA (ATCC-25923), S. aureus (NCTC-3750)and their 3 Mixed Cultures using MIC, MBC and Agar Well Diffusion methods. Further, their effect on bacterial biofilms was also detected using crystal violet staining method. The antibiofilm activity of Ascorbic Acid was noted in the order of Mixed Culture-III > MRSA > S. typhi > Mixed Culture-II > E. faecalis > E. coli > S. aureus > S. enterica. Furthermore, Ascorbic Acid + Ciprofloxacin combination acted synergistically against the biofilms of E. faecalis, MRSA and S. aureus. Besides, In the Time Course Study with Ascorbic Acid, S. enterica, S. typhi and MRSA were found to be inhibited around 3hr., whereas, E. faecalis, S. aureus, Mixed Culture-II and Mixed Culture-III were inhibited around 6hr. Furthermore, E. coli and Mixed Culture-I were inhibited around 24hr. Thus, Ascorbic Acid exhibited broad spectrum Antibacterial as well as Antibiofilm potential by inhibiting Gram Positive as well as Gram negative bacteria and their mixed cultures.

5. In vitro Evaluation of Antimicrobial and Antibiofilm Potential of Ascorbic Acid.

Team: Miss Disha Thakur (STRIP Student, Bacteriology Department), Mrs. Dipti Kolte (STA & Co-PI, Bacteriology Department) and Dr. Anuya Rege [Principle Investigator (PI), Bacteriology Department]. 
Funding Agency: Haffkine Institute (STRIP Fee)
Duration: November 2023 – March 2024
Abstract:
In the present in vitro study, Ascorbic Acid was evaluated for its Antimicrobial effect against 6 Clinical isolates namely, K. aerogenes, K. pneumoniae, P. aeruginosa, P. mirabilis, P. vulgaris, S. flexneri and mixture of all bacteria as well as 2 Fungi namely, A. niger (ATCC-16404) & C. albicans (ATCC-10231) and Mixture of C. albicans & Bacteria using MIC, MBC and Agar Well Diffusion methods. Besides, Ascorbic Acid was further evaluated for its effect on microbial biofilms using Crystal Violet staining method. The antibiofilm activity of Ascorbic Acid was noted in the order of P. aeruginosa > P. mirabilis > Mixed Culture-I > K. pneumoniae > K. aerogenes > S. flexneri. However, it did not show any Anti-biofilm activity against P. vulgaris at any concentrations tested. Ascorbic Acid also exhibited the Antibiofilm activity against C. albicans and Mixed culture-II (C. albicans + MixedBacteria) at RT as well as at 370 C and the activity was found to be concentration dependent. Furthermore, combination Ascorbic Acid + Ciprofloxacin acted synergistically against the biofilms of K. aerogenes, K. pneumoniae and S. flexneri, whereas, combination Ascorbic Acid + Fluconazole acted synergistically against the biofilms of C. albicans. Likewise, combination Ascorbic Acid + Ciprofloxacin+ Fluconazole acted synergistically against the biofilms of Mixed Culture-II. Besides, the Time Course Study with Ascorbic Acid revealed inhibition of all clinical isolates and their mixture around 3 hours, whereas, it displayed inhibition of C. albicans and Mixed Culture-II around 48 hours and 6 hours respectively. Thus, Ascorbic Acidcan be used for combating the microbial biofilms-related menaces.

6. Biodegradation of PAHs (Naphthalene and Anthracene) by bacteria Isolated from Oil-Contaminated Soil Sample.

Team: Miss Nishitta Nayak [Short Term Research Internship Program (STRIP) Student, Bacteriology Department] and Mrs. Jayashree More [Principle Investigator (PI), Bacteriology Department]. 
Funding Agency: Haffkine Institute
Duration: January 2024 – June 2024
Abstract:
Petroleum hydrocarbons' pervasive use in various sectors has led to significant soil pollution, necessitating effective remediation strategies. This study investigates the degradation of polycyclic aromatic hydrocarbons (PAHs), focusing on Naphthalene and Anthracene, by isolating and identifying hydrocarbon-degrading bacteria from petroleum-contaminated soils. A total of 45 bacterial isolates were identified using Bergey's Manual, with the majority being Gram-negative rods capable of fermenting various sugars and producing hydrogen sulfide. The biodegradation capabilities of these isolates were demonstrated through qualitative assays, such as the Ring Fission test, and quantitative assays using Multimode Reader and HPLC analysis, which showed significant degradation of Naphthalene and Anthracene over an incubation period. The identified bacterial strains, including Enterobacter cloacae, Chromobacterium violaceum, Enterobacter aerogenes, and Klebsiella pneumoniae, exhibited substantial potential for bioremediation. These strains were not only efficient in breaking down complex PAH structures but also showed adaptability to various environmental conditions typically found in contaminated sites. The metabolic pathways utilized by these bacteria for PAH degradation were explored, providing insights into the mechanisms of microbial bioremediation. This study underscores the potential of microbial bioremediation as a sustainable solution for managing hydrocarbon contamination in industrial areas. The findings suggest that these bacterial strains could be further developed and optimized for large-scale bioremediation applications, offering an eco-friendly and cost-effective alternative to conventional methods. However, further investigation is needed.

7. In vitro Evaluation of Antibacterial & Antibiofilm Potential of Alum (Sphatika) and it’s Bhasma (Sphatika Bhasma) against Clinical Isolates.

Team: Miss Mrudali Ambre (STRIP Student, Bacteriology Department), Mrs. Dipti Kolte (STA & Co-PI, Bacteriology Department) and Dr. Anuya Rege [Principle Investigator (PI), Bacteriology Department]. 
Funding Agency: Haffkine Institute (STRIP Fee)
Duration: January 2023 – June 2023
Abstract:
In the present in vitro study, antimicrobial activities of Alum (Sphatika) and Sphatika Bhasma were evaluated against total 6 clinical isolates, viz., K. aerogenes, K. pneumoniae, P. aeruginosa, P. mirabilis, P. vulgaris, S. flexneri and a mixed culture, using various assays such as MIC, MBC and Agar Well Diffusion Method. Further, their effect on bacterial biofilms was also detected using crystal violet staining method. The antibiofilm activity of Alum was noted in the order of P. mirabilis > P. aeruginosa > P. vulgaris > K. pneumoniae > S. flexneri > Mixed culture > K. aerogenes, whereas, antibiofilm activity of Sphatika Bhasma was noted in the order of P. aeruginosa > Mixed culture > P. vulgaris > S. flexneri > K. pneumoniae > P. mirabilis > K. aerogenes. Furthermore, AL + SB combination acted synergistically against the biofilms of clinical isolates. Moreover, Alum and Sphatika Bhasma have revealed Antimicrobial and Antibiofilm potential against mono as well as mixed cultures in present in vitro study.

8. Determination of Antimicrobial & Anti-Biofilm activities of Alum and it’s Bhasma- An In vitro Comparative Study.

Team: Miss Susan Martin [Short Term Research Internship Program (STRIP) Student, Bacteriology Department], Mrs. Dipti Kolte (STA & Co-PI, Bacteriology Department) and Dr. Anuya Rege [Principle Investigator (PI), Bacteriology Department]. 
Funding Agency: Haffkine Institute (STRIP Fee)
Duration: January 2022 – June 2022
Abstract:
With an approaching antibiotic crises due to the emergence and extensive spread of antimicrobial resistance among bacterial agents, as well as to increasing number of patients with chronic and bacterial biofilm-associated infections, the naturally occurring molecules may become new sources of antibacterial and antibiofilm drugs for clinical usage. Hence, in the present in vitro study, Antimicrobial potential of Alum as well as its Bhasma- Sphatika Bhasma was evaluated against total 10 organisms, viz., 4 Gram Negative, 4 Gram Positive and 2 Fungi using various assays such as MIC, MBC/MFC and Agar Well Diffusion method. Alum as well as Sphatika Bhasma revealed broad spectrum Antibacterial activities inhibiting gram positive as well as gram negative organisms and moderate antifungal activities. Hence, Antibiofilm potential of Alum and Sphatika Bhasma was further evaluated against total 8 bacterial biofilms using crystal violet staining method. Antibiofilm effect of Alum was found to be in the order of S. lutea > P. aeruginosa > E. coli > MRSA > S. typhi > B. subtilis > K. pneumoniae > E. faecalis, whereas, Antibiofilm activity of Sphatika Bhasma was noted in the order of P. aeruginosa > S. lutea > E. coli > K. pneumoniae > S. typhi > MRSA > B. subtilis > E. faecalis. Alum as well as Sphatika Bhasma also exhibited good Antioxidant potential which was estimated using DPPH method. Antimicrobial, Antibiofilm and Antioxidant effects of Alum and Sphatika Bhasma could be attributed to their Phenolic and Flavonoid contents. Overall, Sphatika Bhasma exhibited better Antimicrobial, Antibiofilm and Antioxidant activities than Alum.

9. Isolation and Identification of Hydrocarbon Biodegrading Bacteria from Soil Samples.

Team: Miss Sakshi Ajay Gupta [Short Term Research Internship Program (STRIP) Student, Bacteriology Department] and Mrs. Jayashree More [Principle Investigator (PI), Bacteriology Department]. 
Funding Agency: Haffkine Institute (STRIP Fee)
Duration: January 2022 – June 2022
Abstract:
Isolation of hydrocarbon degrading bacteria specifically BTEX degrading bacteria was carried out from oil Greece contaminated soil samples collected from different locations of Western Ghat region of Maharashtra. 87 bacterial isolates were isolated using Bushnell Haas media. Qualitative assay was performed for the 87 isolates using Methylene blue as a redox indicator and 52 isolates were selected that showed hydrocarbon degrading activity in the form of color change from blue to colorless. Gram staining and Biochemical tests was performed for all the selected isolates and also morphological characteristics were recorded. Quantitative assay was performed using multimode reader and UV-Vis Spectrophotometer by taking different concentrations of BTEX. Finally 11 isolates were selected that showed significant reduction in absorbance using UV-VIS spectrophotometer. Isolates showing best results were further used for DNA extraction and among these two samples were will undergo 16S rRNA Sequencing. BLAST was run for the sequences obtained after 16S rRNA sequencing, the organism present in both the samples were identified to be Enterobacter.

10. To determine Neutraceutical Potential and Antimicrobial Potential of Powder of Dried Hibiscus Flowers. 

Team: Mr. Atsih Dongarkar [Short Term Research Internship Program (STRIP) Student, Chemical Testing Department], Mrs. Suvarna Pachpore [Principle Investigator (PI), Chemical Testing Department], Mrs. Dipti Kolte (STA, Bacteriology Department) and Dr. Anuya Rege (Co-PI, Bacteriology Department). 
Funding Agency: Haffkine Institute (STRIP Fee)
Duration: November 2021 – March 2022
Abstract:
Nutraceuticals, in broad, are food or part of food playing a significant role in modifying and maintaining normal physiological function that maintains healthy human beings. These nutraceuticals help in combating some of the major health problems of the century such as obesity, cardiovascular diseases, cancer, osteoporosis, arthritis, diabetes, cholesterol etc. Hibiscus rosa-sinens is a medicinal plant member belonging to the family Malvaceae. Hibiscus extracts have been used for ages in Ayurveda to cure many ailments. A numerous types of tree flowers are present in nature and few of them are being utilized traditionally as food and medicine by the localities of India. However, many flowers are still unexplored as Nutraceutical. Hence, powder of dried flowers of H. rosa-sinensis was subjected to estimation of various Nutraceuticals and to evaluate its Antimicrobial potential in the present in vitro study.
The present work has shown presence of Vitamin B1, Vitamin B2, Vitamin B6, Anthocyanin and phytochemicals such as Resins, Saponin, Flavonoids, Anthraquinone, Steroids and Carbohydrate in the powder of dried Hibiscus Flowers. Besides, two extracts namely, Aqueous and Acidified prepared from the powder of dried Hibiscus Flowers were tested for their Antimicrobial potential against E. coli, K. pneumoniae, P. aeruginosa, S. typhi, E. faecalis, Methicillin Resistant Staphylococcus aureus (MRSA) and C. albicans using MIC, MBC/MFC and Agar Well Diffusion Methods. However, in the present in vitro study, none of the extracts were effective against any of the 7 organisms tested. Thus, present study reveals Neutraceutical potential of powder of dried Hibiscus Rosa-sinensis flowers.

11. Synthesis of Substituted Triazol and Its Biological Activity.

Team: Miss Anchita Goyal [Short Term Research Internship Program (STRIP) Student, Chemical Testing Department], Mr. Manoj Patil [Principle Investigator (PI), Chemical Testing Department], Mrs. Dipti Kolte (STA, Bacteriology Department) and Dr. Anuya Rege (Co-PI, Bacteriology Department)
Funding Agency: Haffkine Institute (STRIP Fee)
Duration: January 2017 - May 2017
Abstract:
The main aim of the project was to synthesize three triazole compounds and check for their biological activities. The compounds were synthesized by a long alkyne- azide 3+2 cycloaddition reaction. The three synthesized Triazol compounds were subjected to MIC determination against E. coli, P. aeruginosa, S. aureus and E. faecalis and the MIC was found to be more than 320 μg/ml. Furthermore, the antimicrobial activity of 3 compounds were evaluated against E. coli, P. aeruginosa, S. aureus, E. faecalis, C. albicans and A. niger by Agar well diffusion method, wherein, compound I at 1 mg/ml concentration showed moderate activity against E. faecalis and weak activity against E. coli, C. albicans and A. niger. However, none of the compounds revealed free radical scavenging activity. From the biological activity point of view E. faecalis sets an impact on anti-bacterial activity as the zone of inhibition as compared to that of standard is in a positive and good range.

12. Antibacterial, Antifungal and Anti-Cancer Activity of Indian Medicinal Plants.

Team: Miss Shraddha Rane [Short Term Research Internship Program (STRIP) Student, Bacteriology Department] and Mrs. Jayashree More [Principle Investigator (PI), Bacteriology Department]
Funding Agency: Haffkine Institute
Duration: January 2016 - June 2016
Abstract:
The plants Acorus calamus (AC) and Myristica fragrans (MF) were included for the present study. Extraction of plants was carried out using Soxhlet apparatus with organic solvents. Furthermore, oil extraction was accomplished using oil extractor. Oil samples were subjected to GC-MS analysis to identify the components. Plant extracts did not show any activity in the preliminary work. Hence, both the oil samples were included for the present in vitro study. The extracted essential oils were screened against 16 bacterial standard strains, both grams positive and negative. The antifungal activity was tested on 5 standard fungal strains and 6 environmental isolates. Furthermore, the cytotoxicity of the two samples was determined by the MTT assay against Vero and MCF 7 cell lines and result is expressed as IC50 value.
It was observed that the A. calamus essential oil showed inhibition of gram negative bacteria only, whereas, M. fragrans essential oil showed activity against gram negative as well as positive organisms. Both the oil samples exhibited antifungal activity against all standard as well as environmental fungal isolates, except M. fragrans oil which showed no effect on A. solanni. Thus, the samples showed moderate antibacterial activity, whereas, prominent antifungal activity was noted. The oil samples showed cytotoxic effect on both MCF 7 and Vero cell lines. The IC50 values of A. calamus and M. fragrans oils were lower on MCF 7 cell line than that on Vero cell line. It can be concluded that both these oils show potential to be used as anti-cancer as well as anti-fungal agents. However, further investigation is needed.