Annual Congress on
Microbiology and Microbiologists

My basic research interest is to work on advanced techniques and methodology to study etiology of microbes, microbial products and enzymatic catalyzed reactions. In reference to that my interest of an area is to study various natural and synthetic sources which can be applied for biofilm eradication of various pathogenic bacteria. My other research interests are on study of microbial enzymes. For the same I have isolated and screened organic solvent tolerant lipase producing microorganisms. Immobilization of lipase was carried on nano carrier like magnetic nano-particles which applied for the different esterification reactions. Recently, I am working on different esterification reaction catalyzed by lipase immobilized on functionalized magnetic nano particles.

Statement of Problem: Biofilms are formed by different microorganisms where they communicate and develop in rigid film like structure. Cells in biofilms are deemed 100 to 1,000 times additional resistant to disinfecting agents and antibiotics than planktonic cells. Biofilms might form on a wide type of surfaces, together with living tissues, industrial or potable water system piping, indwelling medical devices, or natural aquatic systems. Methodology and Theoretical Orientation: In the present study Aloe vera, cabbage, lemon grass, Ginger, apple, olive, lemon, papaya and banana have been used to degrade the bacterial biofilms under laboratory conditions. Pseudomonas aeruginosa, Salmonella typhi, Staphylococcus aureus, Bacillus cereus and Escherichia coli usually form strong biofilm and similar group of microbes show resistant to antibiotics. Therefore, these five microorganisms were screened for biofilm formation individually and in different combinations. Minimum biofilm inhibitory concentrations (MBIC) of the plant and fruit extracts were determined and synergy between antibiotic and plant/fruit extract were checked. Conclusion and Significance: Particular attention is oriented nowadays towards the need for biofilm inhibiting compounds that are able to reduce or minimize infections; especially those caused by antibiotic-resistant bacterial strains. Synergy between antibiotics and natural extracts has exhibited anti biofilm activity against strong mixed biofilm. Minimum Biofilm Inhibition Concentration (MBIC) of plant extract with combination of antibiotic [ciprofloxacin + cabbage extract (inhibition- 69.12%) and ciprofloxacin + lemon grass extract (inhibition- 89.77%)] can remove of strong mixed biofilm. As well as MBIC of fruit extract with combination of antibiotic [ciprofloxacin + banana peel extract (inhibition-96.95%), ciprofloxacin + apple extract (inhibition-93.16%) and ciprofloxacin + lemon peel extract (inhibition- 61%)] can obtain removal of strong mixed biofilm compare to individual itself. This kind of synergistic approach can be further exploited for future therapeutic purpose however; it needs multiple kind of further investigations.

George Osei-Adjei, currently is a PhD student at the School of Medicine, Department of Biochemistry and Molecular Biology, Jiangsu University, China. His research focus is on transcriptional regulation mechanism in pathogenic bacteria. Our focus is identifying the virulence factors produced by the bacteria and to elucidate the mechanisms employed by the to cause human and fish diseases.

Vibrio parahaemolyticus, a common inhabitant of marine and coastal environments is a Gram- negative halophilic bacteria that causes seafood-associated gastroenteritis, wound infection and septicemia in humans. V. parahaemolyticus produces extracellular proteases that have been identified as potential virulence factors and are involved in the degradation of protein substrates to provide nutrients that aids the survival of the bacteria and enhances the spread of toxins. V. parahaemolyticus uses quorum sensing (QS), a density dependent signal transduction system to detect signal molecules called autoinducers. AphA and OpaR are the master QS regulators expressed at low cell density (LCD) and high cell density (HCD) respectively. Molecular biology techniques such as qRT-PCR, lacZ fusion, DNase I footprinting and electrophoretic mobility shift assay were employed to elucidate the mechanism of regulation. V. parahaemolyticus OpaR positively regulate the expression of the extracellular protease genes (VPA0459, VP1340, VPA0714, VPA0227, VPA0449 and VPA1071). Further, OpaR contributes to the activation of the extracellular protease genes in the hydrolytic activity against casein in skim milk powder. Data presented here provides insight into the pathogenic mechanism employed by V. parahaemolyticus.

Afaf Bastawy a fresh graduate from the faculty of pharmacy with excellent grades and a GPA 3.67. Moreover, my project was chosen as the best one in the whole faculty by a professional panel from Greenwich, Zewail, and Cairo universities.

Klebsiella pneumoniae is one of the significant causative agents for nosocomial infections. Rate of infections by this bacterium have increased dramatically and thus searching for quick and sensitive molecular methods for its detection became a must. Our research aims at validating a sensitive and specific PCR based method for diagnosis of infections caused by Klebsiella pneumoniae. Our work started by using online bioinformatics tools to find a unique gene that can be used for detection of Klebsiella pneumoniae and designing specific primer for its amplification. In silico studies results in choosing the rcsA gene, regulating colanic acid capsular biosynthesis activation protein A, as a unique gene for Klebsiella pneumoniae. Two sets of primer pairs (named K1 and K2) were designed to target the specific gene, followed by determining primers sensitivity and specificity in Klebsiella pneumoniae detection. Both primer pairs K1 and K2 were able to amplify targeted gene sequence in standard and clinical strains. K1 and K2 primer pairs were able to amplify target gene in diluted DNA template concentration up to 10-1 ng/µl, and 1 x 10-23 picogram/µl, respectively, indicating the higher sensitivity of primer pair K2. K1 primer pair worked well on bacterial culture and amplified target genes in bacterial cultures diluted to 101 CFU/ml. K1 primer pair failed to amplify genomic DNA template of common Gram negative uropathogens including Pseudomonas aeruginosa, Escherichia coli, and Proteus mirabilis, thus indicating its specificity. In conclusion, rcsA gene is a unique gene that can be used for detection of Klebsiella pneumoniae in clinical samples.

Azna Zuberi have passed her bachelors from biochemistry and masters from biotechnology. In Ph.D. she got enrolled under supervision of Prof Asad U Khan (Head of the medical microbiology lab, IBU, AMU, Aligarh). Her work was focused on one of the current emerging gene silencing tools i.e. CRISPRi. She explored the use of CRISPRi as a novel therapeutic approach against bacterial biofilm-mediated infections. During her Ph.D. tenure, she came across many techniques like cloning, biofilm quantification, qRT-PCR, microscopic techniques etc. and publish papers in high reputed journals (frontiers in immunology and frontiers in clinical and cellular microbiology). Further she also received many awards and prize during her academic tenure like Pre Doctoral Fellowship (JRF / SRF) from Department of Biotechnology, Govt. of India, New Delhi, India – 2014, first prize in poster presentation held in National symposium cum bioinformatics workshop, first prize in oral presentation at Biospark life science fest, AMU, Aligarh, 2018 and qualified the all India Graduate Aptitude Test in Engineering (GATE) conducted by the Indian Institute of Technology (IIT), India -2013.

Biofilm is a sessile bacterial accretion in which bacteria adapts different physiological and morphological behavior from planktonic form. It is the root cause of about 80% microbial infections in human. Among them, E. coli biofilms are most prevalent in medical devices associated nosocomial infections. The objective of this study was to inhibit biofilm formation by targeting luxS gene (involved in quorum sensing) as well as fimH gene (codes for the minor subunit of Type 1 fimbriae) using CRISPRi. To implement CRISPRi system, we have synthesized complementary sgRNA to target gene sequence and co-expressed with dCas9. Suppression of luxS and fimH was confirmed through qRT-PCR. Further their effect on biofilm inhibition was studied through crystal violet assay, XTT reduction assay, scanning electron microscopy, transmission electron microscopy, and confocal microscopic assay. Lastly, we conclude that CRISPRi system could be a potential strategy to inhibit bacterial biofilm through mechanism base approach.

Tuberculosis is a major global health problem. Unidentified or misdiagnosed cases & delay in receiving appropriate therapy increase the risk of transmission to contacts so investigation is important for intensified case finding. This study is determining the tuberculosis disease among household contacts of MDR-TB patients (Multiple Drug Resistant Tuberculosis) along with the molecular evidence of polyclonal TB and transmission of strains from index patient. Culture & drug susceptibility test of isolates from sputum were determined using liquid culture. Thin Layer Agar plate was also used to obtain isolated colonies of multiple strains in samples. Spoligotyping and reverse line blot hybridisation assay(RLBA) were then performed using culture DNA to determine spoligotype families and molecular drug resistant pattern respectively and results were analysed from 134 index patients and their 19 contacts having active MDR-TB (Total Contacts screened:305). Among the index & contact cases, CAS1DELHI, was found in 42.5%, 15.8%; EAI3IND in 21.6%, 21%; Beijing spoligotype families in 20% , 21% respectively. Samples from 39 index cases (29.1%) showed multiple MTB spoligotypes. We found disease prevalence among contacts, higher [6.22%(19/305)] than previous studies. Spoligotyping confirmed transmission in 18 contacts (94.7%) from their index case, one contact showed (EAI3IND) transmission from person outside of household, while index case showed clonal infection with Beijing spoligotype along with another infected contact. During pre-treatment & initial treatment phase, patient is potentially infectious and when investigated, contacts showed high prevalence of infection. Thus contact investigation and molecular typing is imperative for early detection & treatment to cease the chain of transmission, overall success in control programs.

Tuberculosis is a major global health problem. Unidentified or misdiagnosed cases & delay in receiving appropriate therapy increase the risk of transmission to contacts so investigation is important for intensified case finding. This study is determining the tuberculosis disease among household contacts of MDR-TB patients (Multiple Drug Resistant Tuberculosis) along with the molecular evidence of polyclonal TB and transmission of strains from index patient. Culture & drug susceptibility test of isolates from sputum were determined using liquid culture. Thin Layer Agar plate was also used to obtain isolated colonies of multiple strains in samples. Spoligotyping and reverse line blot hybridisation assay(RLBA) were then performed using culture DNA to determine spoligotype families and molecular drug resistant pattern respectively and results were analysed from 134 index patients and their 19 contacts having active MDR-TB (Total Contacts screened:305). Among the index & contact cases, CAS1DELHI, was found in 42.5%, 15.8%; EAI3IND in 21.6%, 21%; Beijing spoligotype families in 20% , 21% respectively. Samples from 39 index cases (29.1%) showed multiple MTB spoligotypes. We found disease prevalence among contacts, higher [6.22%(19/305)] than previous studies. Spoligotyping confirmed transmission in 18 contacts (94.7%) from their index case, one contact showed (EAI3IND) transmission from person outside of household, while index case showed clonal infection with Beijing spoligotype along with another infected contact. During pre-treatment & initial treatment phase, patient is potentially infectious and when investigated, contacts showed high prevalence of infection. Thus contact investigation and molecular typing is imperative for early detection & treatment to cease the chain of transmission, overall success in control programs.

Djassinra Tormal received her Ph.D in sciences of enviromment from university Ibn Tofail in 2016. After completion of her degree, she was appointed as a faculty fellow in the Department of Health and enviromment at the University of Ibn Tofail. She served as the Head of Study of antibacterial and antifugal activities of two medecinal plants growing wild in the Gharb region (Chenopodium ambrosiodes l and Rosmarinus officinalis l from 2014-2016. Her interests are focused on the use of microbiology to study the antibacterial and antioxidant effect of medicinal plant on Antibiotic Resistant Strains in 2016. Chemistry agro resources, polymers and process engineering.

In this study, the behavior of Xanthomonas fragariae, angular leaf spot of strawberry agent, was followed in the AB medium, enriched with nitrogen, phosphorus or with potassium, and in the soil of the Mamora forest with 14% to 28% of humidity in function of these fertilizer elements. The obtained results have shown that Na2HPO4 and NH4Cl, used, 0.01 and 0.05 mol/l, respectively as a phosphorus and nitrogen source, have a significant effect on the survival of Xanthomonas fragariae. By contrast, KCl, used as a source of Potassium, has no significant effect on the number of culturable cells. The three sources used NPK, 14% and 28% showed a great influence on the number of culturable cells of Xanthomonas fragariae, either increasing or decreasing. Potassium, at 28 to 14% of humidity, inhibited the rate growth of Xanthomonas, while the phosphorus and nitrogen stimulated its growth, greater than 28% of humidity than 14%. Similarly the bacterial growth was not affected during the incorporation of NPK at different concentrations in the soil of Mamora.