Antibiotic Resistance, Biofilm and Bacterial Pathogenesis

Academic Qualifications

1. Ph.D. (1990):Biochemistry(Microbiology), Banaras Hindu University.
   Title: Studies on certain virulence properties of Aeromonas hydrophila.
2. M.Sc. (1985): Biochemistry, Banaras Hindu University.
3. B.Sc. (1982): Botany, Zoology Chemistry (Hon’s), Banaras Hindu University.

Academic Positions:

1. Scientist F, (June 2, 2012 – Contd.), Scientist E, (June 2, 2007- June 1, 2012); Scientist D/ Associate Professor/ Reader, (June 2, 2003-June 1, 2007.), Institute of Life Sciences, Bhubaneswar-751023, India;
2. Scientist C, (November 4, 1997-May 31, 2003); Scientist E-I, (August 28, 2000-November 30, 2002, on deputation); Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram–695014, India;
3. Assistant Professor, (February 17, 1997–October 31, 1997), SBSPG Institute of Biomedical Sciences and Research, Dehradun–248161, India;
4. Senior Research Associate, (August 1, 1996–February 16, 1997); Research Associate, (April 4, 1991–May 31, 1996); Senior Research Fellow and Junior Research Fellow, (March 1, 1986–February 28, 1991); Research Fellow, (September 17, 1985–February 28, 1986), Department of Microbiology, Institute of Medical Sciences. Banaras Hindu University, Varanasi – 221 005, India

Training:

1. September 29, 2008- November 21, 2008, Microarray based analysis of Vibrio cholerae (Supervisor: Prof. Rita R. Colwell), Maryland Pathology Research Institute, University of Maryland, College Park, MD
2. August 5, 2002-September 13, 2002: PCR, gene cloning, expression, purification of protein, fermentation, mice immunization and subsequent challenge and survival study (Supervisor: Prof. Dr. G.S. Chhatwal, Department of Microbial Pathogenesis and Vaccine Research, GBF, Mascheroder Weg 1, 38124 Braunschweig, Germany).
3. December 12, 2000-February 6, 2001: Monoclonal antibody production using Hybridoma technique and tissue culture assay (Supervisor: Dr. T. Ramamurthy, National Institute of Cholera & Enteric Diseases, Calcutta).
4. March 22, 1998-July 12, 1998: Isolation and purification of plasmid DNA, homologous recombination, Colony and Western Hybridization (Supervisor: Prof. James B. Kaper, Center for Vaccine Development, University of Maryland School of Medicine, Baltimore, Maryland).
5. February 3, 1998-March 21, 1998: Molecular Biology including isolation and purification of chromosomal DNA, PCR, AFLP, RFLP, PFGE, Ribotyping, Dot-Slot, and Southern Hybridization (Supervisor: Prof. Rita R. Colwell, Center of Marine Biotechnology, UMBI, Baltimore, Maryland).

Research Experience: Twenty five years excluding PhD (5 years).

Research Guidance- PhD students: 07; in progress 04; MSc Thesis: 08

Membership of Professional Associations / Societies:

1. Life Member: Association of Microbiologist of India
2. Life Member: Indian Immunology Society
3. Life Member: Indian Association of Medical Microbiologists
4. Founder Member : Biotech Research Society of India
5. Full Member: American Society for Microbiology

Reviewer’s of Scientific Journals: Ad-hoc reviewer of numbers of national/ international scientific journals.

Ad-hoc Referee of PhD Examination:

1. University of Kerala, Trivandrum
2. Banaras Hindu University, Varanasi
3. University of Delhi, Delhi
4. Jadavpur University, Kolkata

Ad-hoc Reviewer of Research Projects:

1. Department of Science and Technology, New Delhi
2. Department of Biotechnology, New Delhi

Antibiotic Resistance, Biofilm and Bacterial Pathogenesis

Antibiotic Resistance, Biofilm and Bacterial Pathogenesis

Original peer-reviewed articles:

1. Singh DV, Sanyal SC. Haemolysin and enterotoxin production by Aeromonas caviae isolated from diarrhoeal patients, fish and environment. Journal of Diarrhoeal Diseases Research 1992; 10:16–20.
2. Singh DV, Sanyal SC. Enterotoxicity of clinical and environmental isolates of Aeromonas spp. Journal of Medical Microbiology 1992; 36:269–272.
3. Singh DV, Sanyal SC. Production of haemolysin and its correlation with enterotoxicity in Aeromonas spp. Journal of Medical Microbiology 1992; 37:262-267
4. Singh DV, Sanyal SC. Production of chitinase by enterotoxigenic Aeromonas species isolated from clinical and environmental sources. Journal of Diarrhoeal Diseases Research 1992; 10:213–216
5. Singh DV, Sanyal SC. Biochemical characteristics and enterotoxicity of Aeromonas species isolated from man and environment. Journal of Diarrhoeal Diseases Research 1992; 10:231-234.
6. Singh DV, Sanyal SC. Haemagglutinating activity, serum sensitivity and enterotoxicity of Aeromonas spp. Journal of Medical Microbiology 1993; 38:49–53.
7. Singh DV, Dubey RS, Sanyal SC. Adherence of haemagglutinating and non-haemagglutinating clinical and environmental isolates of Aeromonas. Journal of Diarrhoeal Diseases Research 1993; 11:157-160.
8. Singh DV, Sanyal SC. Antibiotic resistance in clinical and environmental isolates of Aeromonas spp. Journal of Antimicrobial Chemotherapy 1994; 33:368–369.
9. Tikoo Anjali, Singh DV, Sanyal SC. Influence of animal passage on haemolysin and enterotoxin production in Vibrio cholerae O1 biotype El Tor stains. Journal of Medical Microbiology 1994; 40:246–251.
10. Singh DV, Sanyal SC. Relationship between enterotoxicity and multiple drug resistance in Aeromonas spp. Journal of Diarrhoeal Diseases Research 1995; 13:172–175.
11. Shukla BN, Singh DV, Sanyal SC. Attachment of non-culturable toxigenic Vibrio cholerae O1 and non-O1 and Aeromonas spp. to the aquatic arthropod Gerris spinolae and plants in the River Ganga, Varanasi. FEMS Immunology and Medical Microbiology 1995; 12:113–120.
12. Singh DV, Tikoo Anjali, Sanyal SC. Production of the new cholera toxin by environmental isolates of Vibrio cholerae non-O1. Journal of Medical Microbiology 1996; 45:31–34.
13. Singh DV, Tikoo Anjali, Sanyal SC. Haemolysin produced by Vibrio cholerae non-O1 is not enterotoxic. Journal of Medical Microbiology 1996; 45:35–39.
14. Tikoo Anjali, Singh DV, Shukla BN, Sanyal SC. Development of an improved synthetic medium for a better production of the new cholera toxin and its immunological relationship with the toxin produced by Vibrio cholerae O139 strains. FEMS Immunology and Medical Microbiology 1996; 14:67–72.
15. Singh DV, Sanyal SC. Production of haemolysin and enterotoxin by Aeromonas jandaei and Aeromonas trota after animal passage. Journal of Diarrhoeal Diseases Research 1996; 14:274–279.
16. Singh DV, Tikoo Anjali, Sanyal SC. Candidate live oral cholera vaccine strains produce a new cholera toxin. Current Science 1996; 70:237–238.
17. Singh DV, Tikoo Anjali, Sanyal SC. Cholera toxin zonula occludens toxin and accessory cholera enterotoxin gene negative Vibrio cholerae non-O1 strains produce the new cholera toxin. Current Science 1996; 70:1041–1043.
18. Singh DV, Sanyal SC. Enterotoxicity, haemolytic activity and antibiotic susceptibility of Aeromonas eucrenophila strains isolated from water and infected fish. Indian Journal of Experimental Biology 1997; 35:144–147.
19. Singh DV, Sanyal SC. Enteropathogenicity of Aeromonas jandaei and A. trota. FEMS Immunology and Medical Microbiology 1997; 17:243–250.
20. Singh DV, Sanyal SC. Virulence patterns of Aeromonas eucrenophila isolated from water and infected fish. Journal of Diarrhoeal Diseases Research 1999; 17:37-42.
21. Singh DV. Virulence factors in Aeromonas veronii bv veronii. Journal of Diarrhoeal Diseases Research 1999; 17:90-91.
22. Singh DV. A putative heat-labile enterotoxin expressed by strains of Aeromonas media. Journal of Medical Microbiology 2000; 49:685-689.
23. Singh DV. Production of heat labile enterotoxin by strains of Aeromonas veronii bv veronii. Journal of Health Population and Nutrition 2000; 18:44-48.
24. Singh DV, Matte MH, Matte GR, Ziang S, Sabeena F, Shukla BN, Sanyal SC, Huq A, Colwell RR. Molecular analysis of Vibrio cholerae O1, O139, non-O1, and non-O139 strains: clonal relationships between clinical and environmental isolates. Applied and Environmental Microbiology 2001; 67(2):910-921. (IF: 3.688)
25. Sabeena F, Thirivikramji G, Radhakutty G, Indu P, Singh DV. In vitro susceptibility of Vibrio cholerae O1 biotype El Tor strains associated with an outbreak of cholera in Kerala, Southern India. Journal of Antimicrobial Chemotherapy 2001; 47:361-362. (IF: 3.490)
26. Singh DV, Isac SR, Colwell RR. Development of a hexaplex PCR assay for rapid detection of virulence and regulatory genes in Vibrio cholerae and Vibrio mimicus. Journal of Clinical Microbiology 2002; 40(11):4321-4324. (IF: 3.565)
27. Bhanumathi R, Sabeena F, Isac SR, Radhakutty G, Singh DV. Characterization of a toxigenic Vibrio cholerae O139 strain belonging to a new ribotype and isolated from a diarrheal patient. Journal of Clinical Microbiology 2002; 40(12):4779-4781. (IF: 3.565)
28. Bhanumathi R, Isac SR, Shukla BN, Singh DV. Molecular characterization of Vibrio cholerae O139 Bengal isolated from water and the aquatic plant Eichhornia crassipes in the River Ganga, Varanasi, India. Applied and Environmental Microbiology 2003; 69(4):2389-2394. (IF: 3.820)
29. Singh DV, Bhanumathi R, Colwell RR. Characterization of a cholera toxin-gene negative clinical strain of Vibrio cholerae O139. Journal of Clinical Microbiology 2004; 42:1381. (3.439)
30. Mantri CK, Mohapatra SS, Ramamurthy T, Ghosh R, Colwell RR, Singh DV. Septaplex PCR assay for rapid identification of Vibrio cholerae including detection of virulence and intSXT genes. FEMS Microbiology Letters 2006; 265(2):208-214. (IF: 2.068)
31. Mohapatra SS, Ramachandran D, Mantri CK, Singh DV. Characterization of the genetic background of Vibrio cholerae O1 biotype El Tor serotype Inaba strains isolated in Trivandrum, Southern India. Journal of Medical Microbiology 2007; 56(2):260-265. (IF: 2.091)
32. Ramachandran D, Bhanumathi R, Singh DV. Multiplex PCR for detection of antibiotic resistance genes and the SXT element: application in the characterize Vibrio cholerae. Journal of Medical Microbiology 2007; 56(3):246-251. (IF: 2.091)
33. Patra Minakshi, Pandey SK, Ramamurthy T, Singh DV, Das SK. Characterization of cytotoxin producing Aeromonas caviae (strain HT10) isolated from a sulfur spring in Orissa, India. Letters in Applied Microbiology 2007; 44:338-341. (IF:  1.623)
34. Mohanty A, Kar P, Mishra K, Singh DV, Mohapatra N, Kar SK, Dash AP, Hazra RK. Multiplex PCR assay for the detection of Anopheles fluviatilis species complex, human host preference, and Plasmodium falciparum sporozoite presence, using a unique mosquito processing method. American Journal of Tropical Medicine & Hygiene 2007; 76(5):837-843. (IF: 2.183)
35. Thomas S, Ramachandran D, Remani B, Singh DV. Detection and molecular characterization of Vibrio cholerae O1 Inaba biotype El Tor strain in Kerala, S India. World Journal of Microbiology and Biotechnology 2008; 24:433-434. (IF: 0.945)
36. Mohapatra Harapriya, Mohapatra SS, Mantri CK, Colwell, RR, Singh DV. Vibrio cholerae non-O1, non-O139 strains isolated before 1992 from Varanasi, India are multiple drug resistant, contain intSXT, dfr18 and aadA5 genes. Environmental Microbiology 2008; 10(4):866-873. (IF: 4.707)
37. Mohapatra SS, Ramachandran D, Mantri CK, Colwell RR, Singh DV. Determination of relationships among non-toxigenic Vibrio cholerae O1 biotype El Tor strains from housekeeping gene sequences and ribotype patterns. Research in Microbiology 2009; 160: 57-62. (IF: 2.154)
38. Mohanty A, Swain S, Singh DV, Mahapatra N, Kar SK, Hazra RK. A unique methodology for detecting the spread of chloroquine-resistant strains of Plasmodium falciparum, in previously unreported area, by analyzing anophelines of malaria endemic zones of Orissa, India. Infection, Genetics and Evolution 2009; 9(4):462-467. (IF: 3.223)
39. Mohapatra SS, Mantri CK, Mohapatra Harapriya, Colwell RR, Singh DV Analysis of clonally related environmental Vibrio cholerae O1 El Tor isolated before 1992 from Varanasi, India reveals origin of SXT-ICEs belonging to O139 and O1 serogroups. Environmental Microbiology Reports 2010; 2(1):50-57. (IF: 3.076)
40. Mantri CK, Mohapatra SS, Colwell RR, Singh DV. Sequence analysis of Vibrio cholerae orfU and zot from pre-CTXΦ and CTXΦ reveals multiple origin of pre-CTXΦ and CTXΦ. Environmental Microbiology Reports 2010; 2(1):67-75. (IF: 3.076)
41. Turabe Fazil MHU, Kumar Sunil, Subbarao N, Pandey HP, Singh DV.  Homology modeling of a sensor histidine kinase from Aeromonas hydrophila, Journal of Molecular Modeling 2010; 16(5):1003-1009. (IF: 1.871)
42. Mantri CK, Mohapatra SS, Singh DV. Effect of storage and sodium chloride on excision of CTXФ or pre-CTXФ and CTXФ from Vibrio cholerae O139 strains. Infection, Genetics and Evolution 2010; 10: 925-930. (IF: 3.086)
43. Turabe Fazil MHU, Bhanumathi R^, Pandey HP, Singh DV. Characterization of Vibrio cholerae O139 belonging to multiple ribotypes and isolated from diarrhoeal patients in Kerala, southern India.  Infection, Genetics and Evolution 2011; 11: 454-459. (IF: 3.128)
44. Mohapatra SS, Mantri CK, Fazil MHUT, Singh DV.  Vibrio cholerae O1 biotype El Tor strains isolated in 1992 from Varanasi, India harbored El Tor CTXФ and classical ctxB on the chromosome-I and classical CTXФ and classical ctxB on the chromosome-II. Environmental Microbiology Reports 2011; 3(6): 783-790. (IF: 3.232)
45. Turabe Fazil MHU, Kumar Sunil, Former Rohit, Pandey HP, Singh DV.  Binding efficiencies of carbohydrate ligands with different genotypes of cholera toxin B: molecular modeling, dynamics and docking simulation studies. Journal of Molecular Modeling 2012; 18(1): 1-10. (IF: 1.797)
46. Isac Sree Renjini, Nair, GB, Singh DV. Purification and characterization of cytotoxin produced by a clinical isolate of Vibrio cholerae O54 TV113. Applied Biochemistry and Biotechnology, 2012; 167(4): 809-823. (IF:1.943)
47. Fazil MHUT, Kumar Sunil, Subba Rao N, Selvaraj C, Singh SK, Pandey HP, Singh DV. Comparative structure analysis of two proteins belonging to quorum sensing system in Vibrio cholerae. Journal of Bimolecular Structure and Design 2012; 30(5):574-584. (IF: 4.986)
48. Soni DK, Singh RK, Singh DV, Dubey SK. Characterization of Listeria monocytogenes isolated from water of the River Ganga, human and milk in Varanasi, India. Infection, Genetics and Evolution 2013; 14: 83-91. DOI: 10.1016/j.meegid.2012.09.019 (IF: 3.128)
49. Kar Sarita, Panda Sasmita, Sharma Savitri, Singh DV, Das Sujata, Sahu SK, Das Taraprasad. Comparison of type of species and antibacterial susceptibility profile of staphylococci isolated from normal healthy conjunctiva and ocular infections. Asia-Pacific Journal of Ophthalmology, 2013; 2(6): 365-371. (OnlineFirst 26.03.2013).
50. Panda Sasmita, Kar Sarita, Choudhury R, Sharma S, Singh DV. Development and evaluation of hexaplex PCR for rapid detection of methicillin, cadmium/ zinc and antiseptic resistant staphylococci, with simultaneous identification of PVL-positive and- negative Staphylococcus aureus and coagulase negative staphylococci. FEMS Microbiology Letters, 2014, 352:114-122. (IF: 2.046)
51. Soni Dharmedra, Singh Major, Singh DV, Dubey SK. Virulence and genotypic characterization of Listeria monocytogenes isolated from vegetable and soil samples. BMC Microbiology 2014, 14: 241. DOI: 10.1186/s12866-014-0241-3 (IF: 2.98)
52. Mohapatra SS, Mantri CK, Bhotra Tilothama, Singh DV. Characteristics of Vibrio cholerae O1 isolated from water of the River Ganga, Varanasi, India. Indian Journal of Medical Microbiology 2015, 33: 507-515. (IF: 1.037)
53. Soni Dharmedra, Singh DV, Dubey SK. Pregnancy-Associated listeriosis: virulence and genotypic analysis of Listeria monocytogenes from clinical samples. Journal of Microbiology 2015, 53: 653-60. (IF: 1.439)

Review articles – peer reviewed:

1. Singh DV. Hexaplex PCR for rapid detection of virulence factors. Expert Review in Molecular Diagnostics 2003; 361:781-784. (IF: 2.425)
2. Singh DV, Mohapatra Harapriya. Application of DNA based methods in typing Vibrio cholerae strains. Future Microbiology 2008; 3(1):87-96. (IF: 1.976)
3. Fazil MHUT, Singh DV. Vibrio cholerae infection, novel drug targets and phage therapy.  Future Microbiology 2011; 6(10):1199-1208. (IF: 3.819)
4. Choudhury Ranginee, Panda Sasmita, Singh DV. Emergence and dissemination of antibiotic resistance: a global problem. Indian Journal of Medical Microbiology, 2012; 30(4), 384-390. (IF: 0.988)

Chapters in Books/ Encyclopedia:

1. Singh DV. Vibrio cholerae. In: Encyclopedia of Medical Genomics and Proteomics. J. Fuch, M. Podda (Eds.), Informa Healthcare, 2005, pg. 1307-1311.
2. Mohapatra H, Mohapatra SS, Mantri CK, Singh DV. Horizontal gene transfer and ICEs: a perspective with reference to SXT ICEs in Vibrio cholerae. In: New Horizon in Biotechnology. Pandey A, Larroche C, Soccol CR, Dussap C-G (Eds.), Asiatech Publishers, Inc, New Delhi, 2008, pp388-403.
3. Fazil MHUT, Kumar Sunil, Singh DV. In silico analysis of sequence variations in cholera toxin B subunit and their binding with carbohydrate ligands. In: Current Topics in Biotechnology and Microbiology, Dhingra HK, Jha, PN, Bajpai, P  (Eds.), LAP Lambert Academic Publishing AG & Co. KG, Dudweller Landstr, Germany, 2011, pp 168-177.  (ISBN 978-3-8443-2975-9)
4. Choudhury Ranginee, Panda Sasmita, Sharma Savitri, Singh DV. Staphylococcal infection, antibiotic resistance and therapeutics. Chapter 10, In: Antibiotic Resistant Bacteria – A Continuous Challenge in the New Millennium, Maria Pana (Ed.), InTech Publication, Croatia, 2012, pp 247-272, ISBN: 978-953-51-0472-8, InTech, Available from: http://www.intechopen.com/books/antibiotic-resistant-bacteria-a-continuous-challenge-in-the-new-millennium/staphylococcal-infection-antibiotic-resistance-and-therapeutics
5. Singh DV, Bhotra T. 240-Cholera Disease. In: Reference Module in Biomedical Sciences, Michael Caplan (ed.), Elsevier Inc, San Diego, USA, 2014, pg 1-12.  http://dx.doi.org/10.1016/B978-0-12-801238-3.00142-2

Articles in Conference Proceeding:

1. Singh DV, Sanyal SC. Virulence factors of Aeromonas spp. In: Recent Advances in Toxinology, Gopalakrishnakone P, Tan CK editors, National University of Singapore 1992;1:291–307.
2. Thomas S, Praveen Kumar PP, Wilson PA, Bibin T, Singh DV. Molecular monitoring of Vibrio cholerae in the aquatic environment of Kerala. In: Environment and Health, eds. Natarajan P, Paulose CS, Bright Singh IS, Pillai Raji K. Rajiv Gandhi Chair in Contemporary Studies, Cochin University of Science and Technology, Cochin. 2007, pg. 67-78.

National and International Conferences/Symposium: Presented research papers, delivered invited talks and chaired session in fifty four National/ International conferences, seminars, workshops and symposium.

Antibiotic Resistance, Biofilm and Bacterial Pathogenesis

Tilothama Bhotra JRF (DBT) M.Sc. in Biotechnology from Ravenshaw University Email: tilothama [dot] bhotra [at] gmail [dot] com	Sasmita Panda JRF (DST Project) M.Sc. in Biotechnology from Utkal University Email: cometoasmi97 [at] rediffmail [dot] com
Smrutiti Jena JRF (DST-INSPIRE) M.Sc. in Biotechnology from Ravenshaw University Email: smrutittijena [at] ymail [dot] com	Moon Moon Das DBT, JRF B.Sc Zoology, Gauhati University M.Sc Biotechnology, Gauhati University Email:

Antibiotic Resistance, Biofilm and Bacterial Pathogenesis

FUNDING AGENCIES
DST: Department of Science and Technology, Government of India
ICMR: Indian Council of Medical Research, Government of India

In association with international collaborators, we have successfully shown the clonal relationships between clinical and environmental isolates of V. cholerae indicating aquatic environment serving as a reservoir and transmission of the disease cholera.

Our group is the first to report occurrence of SXT ICE, dfr18, sulII, strAB and aadA5 genes in environmental V. cholerae O1, and non-O1, and non-O139 strains isolated before 1992 from Varanasi, India, showing sequence homology with other bacterial species suggesting acquisition of SXT ICEs and antibiotic resistance genes from other bacteria.

Nucleotide sequence analyses of the core region genes, orfU and zot, V. cholerae showed pre-CTX^ETΦ and CTX^ETΦ lineage with V. cholerae El Tor and pre-CTX^ClassΦ, pre-CTX^CalcΦ, and CTX^CalcΦ with classical V. cholerae O1 and O139.  The observation showing loss of the complete and/or partial loss of phage array arranged in tandem lead to understand the emergence and evolution of variant of V. cholerae strains.

We have also developed number of multiplex PCRs for rapid detection of virulence genes, serogroups, SXT and antibiotic resistance genes. We also reported new ribotypes of V. cholerae O1 and O139 strains. V. cholerae O1 carrying Haiti ctxB allele and belonging to multiple pulsotype was found causing outbreak in Silvassa, India.

Pulsed-field gel electrophoresis of SXT-positive V. alginolyticus strains showed genetic heterogeneity, plasticity of their host’s genome and in their adaptation to the environment.

While working on staphylococcus, we have developed a multiplex PCR for rapid detection of antibiotic, cadmium/zinc and antiseptic resistance in PVL-positive and -negative S. aureus and CoNS.

Analyses of Staphyloccocus haemolyticus from ocular infection showed multiple drug resistance including resistance methicillin that possesses mecA belonging to SSCmec V type cassette but belonging to multiple STs and pulsotypes.

S haemolyticus strains were found to produce biofilm and matrix belongs mostly to eDNA and/or protein.