|Session 1: Antimicrobial Resistance: Challenges and existing Solutions Chaired by Prof. Andres Floto.|
|Name: Dr. Yizhi Xu|
Designation: Assistant Lecturer, Coventry University.
Abstract Title – The Current status and future perspective of antimicrobial resistance in China
China is experiencing a serious problem of antimicrobial resistance. The prevalence of resistant strains continues to increase or remain at a high level. It has brought a huge social economic burden throughout China. In this review, the antimicrobial resistance in China, which includes antimicrobial resistance in human patients, poultry and environment, will be investigated; the antimicrobial consumption in health care setting and food animal products will be studied; current antimicrobial resistance studies carrying out in China will be discussed; the relevant strategic measures for the control of surging antimicrobial resistance carried out in China will be reviewed and finally further research in antimicrobial resistance area will be explored.
|Name: Dr. Tharangani Herath |
Designation: Senior Lecturer, Harper Adams University, UK
Abstract Title - Anti-microbial’s in aquatic: Towards a responsible strategy
Farm and pet fish are rapidly growing industries in Indo-Asian sub-continent. Immersing infection diseases are one of the main threat to the growing industry and antimicrobials are commonly used to control infection in the farming practices. Human intervention to control and treat diseases that cause changes to mucosal biology/immunity and microbiota of the fish and aquatic ecosystem continuing to increase risk developing AMR, especially in the countries which are largely depend on antimicrobials to control infections. Improving our understanding on complete picture of host-pathogen-treatment with in aquatic ecosystems will allow us to determine points for intervention to provide better management strategies to tackle emergence of AMR.
|Name : Dr. Anas Abdulaziz |
Designation: Scientist, CSIR-National Institute of Oceanography.
Abstract Title - Species diversity and ecological significance of antibiotic resistance vibrios from coastal waters of India
Presence of multiple antibiotic resistant microorganisms in marine systems is increasingly a focus of concern as they pose potential health risk to humans and animals alike. The present study reports the antibiotic resistance profile, enzyme activities and presence of virulence genes in Vibrio spp. Isolated from coastal waters of India. Increased incidence of cholera outbreak, antibiotic resistance of causative organisms and health of coastal population are global concern as nearly 50 % of word population lives in coastal villages and cities. Nearly 14 % of Indian population also lives in districts along 7517 km long coast line, which spread along ten states and two union territories. The Vibrio spp isolated from coastal waters of India were distributed into eleven species such as V. cholerae, V. alginolyticus, V. hollissae, V. campbelli, V. fischeri, V. nereis, V. proteolyticus, V. mimicus, V. metschnikovii, V. furnissi. V. harveyi and V. proteolyticus. Surprisingly, more than 50 % of the isolates were extremely antibiotic resistant and showed a multiple antibiotic resistance index (MAR index) of 0.75 to 1.0. More than 90 % of isolates from Kerala and Tamil Nadu coasts showed MAR index of > 0.5, while it was nearly 80 % in the case of other coasts. Alarmingly, more than 80 % of Vibrio cholerae isolates from both Kerala and Tamil Nadu coast were extremely antibiotic resistance. Interestingly, the majority of these organisms does not harbour virulence genes. The high levels of enzymatic activities indicated the ecological significance of vibrios in marine environment. However, the increasing antibiotic resistance of vibrios isolated from coastal waters of India are alarming and the infections caused by them may be difficult to be controlled with the available chemotherapeutic tools.
|Name: Dr. Mahmoud Eltholth |
Designation : PhD student at City University of London,
Department of Economics-Health Economics
Abstract title - Assessing the economic impact of antimicrobial resistance: Model calibration and evaluation of policy interventions
Antimicrobials have been considered as an essential tool not only for combatting infectious diseases and saving human and animal lives but also for improving animal health and productivity. They are powerful drugs that prevent many deaths each year without which many invasive medical procedures would be much more dangerous. Many life aspects such as global trade, travel and security would be threatened by the emerging of resistant strains of microorganisms. Unfortunately, antimicrobials are used in a haphazard way that results in the development of antimicrobials resistance (AMR). Therefore, there is a growing concern about the efficacy of antimicrobials use in humans and livestock due to the emerging resistant strains of microorganism and consequently increasing costs of treatments and mortality rates in humans. The current approaches and measurements of cost and economic impacts of AMR and the programs to reduce their impacts are inaccurate and inadequate. The estimates for economic burdens of AMR are underestimated because these estimates are based roughly only on the incremental cost associated to the additional treatment of resistant over susceptible primary infection and other costs are ignored. The objective of this study was to model the channels and main contributors to AMR using models of contagion, network and propagation. A constructed model was used to estimate the economic impact of AMR and eventually identify the most effective policy interventions to alleviate the negative consequences of AMR. To do so, we first reviewed current approaches/methods to model contagion, propagation and networks, and also methods used to assess economic impact in similar contexts. Secondly, we constructed a stylized model of AMR progression. We will then calibrate the model and examine the cost implications using the AMR model. Finally, we will simulate how different intervention would affect AMR and its economic impact.
|Session 2:Clinical Development of Therapeutic Agents and Diagnostic Methods Chaired by Dr. N.K. Ganguly
|Name:Dr. Balaraman Madhan |
Designation : Principal Scientist, CSIR-Central Leather Research Institute, Sardar Patel Road, Adyar, Chennai 600020
Country : India
Abstract Title- Collagen based composite biomaterials to manage wound pathogens, and treatment of chronic wound.
Collagen is one of the most widely used biomaterial for various biomedical applications. Such collagen based composite materials are alternatives to traditionally used collagen scaffold. Recently we have developed Collagen hydrolysate composite scaffold (CHCS) showed good antimicrobial activity against the wound pathogens Staphylococcus aureus, Bacillus subtilis, and Escherichia coli due to the inherited antimicrobial activity of collagen hydrolysate. In vivo wound healing experiments with full thickness excision wounds in rat model demonstrated that wounds treated with CHCS showed accelerated healing when compared to wounds treated with collagen scaffold. Similarly, we have also developed collagen scaffolds loaded with silica microspheres containing plant based antimicrobial agents such as curcumin and capsacin.
|Name: Dr. Tariq sadiq|
Designation: Reader and Consultant Physician, Institute for Infection and Immunity, St George's, University of London.
Country : UK
Abstract Title - Enabling rapid personalised medicine and real-time antibiotic resistance surveillance for global and advanced health systems
Effective STI control is challenged by inadequate access to prompt diagnosis and treatment for patients and partners, poor community STI surveillance for targeting public health interventions and changing antimicrobial resistance patterns. Novel field based detection and sequencing technologies for diagnosis and antimicrobial resistance may address these issues by enabling immediate optimised treatment and partner notification, improved capacity to reach those disproportionately affected away from traditional settings of care as well as creating opportunities for monitoring outbreaks. To enable measurable gain from these advances, more thorough understanding of the underlying genomic determinants of AMR is required, while adaptation of technologies to suit clinical and public-health purpose in socially contextualized strategies will be necessary for deployment. This may ultimately give scope for reducing STI transmission and the burden of disease.
|Name : Dr. Anu Singh |
Designation:UGC WOMED Post-Doctoral Fellow at Jawaharlal Nehru University, New Delhi
Country : India
Abstract Title - Electrochemical Bio-sensing Platform to detect Biomarkers for Drug Resistance: Case study for H. pylori generated cancers.
Helicobacter pylori infection is known as class I carcinogen (as per WHO) that affects human-organ systems, generate gastric carcinoma, gastritis and peptic ulcer etc. These cancer cases can be prevented in high-risk populations through timely screening and eradication of H. pylori. The key to control this infectious disease is to provide a short course therapy and the post-exposure treatment in combination with easy relapse monitoring. However, H. pylori antimicrobial resistance is one of the critical issue faces during the disease diagnosis and selection of the treatment methodology; therefore it needs to be address immediately. Till date detection of H. pylori infection is restricted due to unavailability of universal/ubiquitous serological biomarkers owing to certain limitations including genetic diversity and unsuccessful expression of recombinant proteins. Therefore, present study is based on the application of recently reported specific serological biomarkers (analytes) as a potential candidate to detect H. pylori via development of a nano-molecular biosensing platform. This could facilitate faster and accurate diagnosis for assist appropriate treatment modalities. An electrochemical biosensing system has been designed via fabrication of carbon nano material onto ITO slides and functionalized with ligand. Differential Pulse Voltammetric (DPV) studies performed for this nano-molecular platform exhibited good electrochemical behaviour, specificity and selectivity. Performance characteristics of this biosensing system suggested further implications to real world samples and opportunity to fine-tune biomarker targeted assays to reduce the high incidence of gastric cancer.
|Name: Dr. Bethany Mills|
Designation : Research Fellow in Optical Imaging. University of Edinburgh
Abstract Title - Real-time optical imaging of bacteria for rapid and accurate diagnosis of infection in the clinic
Accurate diagnosis of bacterial infection is key to regaining control of antimicrobial stewardship and reducing the rate at which antimicrobial resistance (AMR) emerges. Current techniques for diagnosis rely on non-specific clinical symptoms and slow, insensitive laboratory investigations; which often result in the misuse or overuse of antimicrobials. During recent years the field of direct imaging of disease has rapidly expanded, largely due to improved detector technology and probe design. Optical imaging of suspected bacterial infection in-situ using appropriate bacteria-specific probes offers the prospect of direct identification of infection (with possibility of identifying Gram and AMR status) in real-time which may inform in subsequent treatment strategies. To this end a number of optical probes are under development; however none have yet been reported within the clinic.
Proteus (proteus.ac.uk) based at the University of Edinburgh, aim to produce novel optical Smart Probes and an optical endomicroscopy (OEM) imaging platform for real-time detection of infection within the distal lung of critically ill patients. The purpose of which is to enable appropriate treatment plans to be implemented and reduce the unnecessary use of antimicrobials. Results & Conclusions: A range of bacteria-specific optical Smart Probes have been developed and validated which demonstrate immediate specific detection of bacteria (according to Gram status) within complex microenvironments in the laboratory and in Phase 0/1 clinical trials. This is to our knowledge, the first use of optical probes for bacterial detection in humans. This technique has the potential to revolutionise patient diagnosis and subsequent treatment plans. We are currently exploring the possibility of using such probes as indicators of bacterial susceptibility or resistance to a particular antimicrobial in real-time at the bedside, with the aim of further promoting appropriate antimicrobial use.
|Session 3:Leveraging Novel Chemistry in the Design of new Antibiotics Chaired by Dr. T.S. Balganesh|
|Name: Dr. Rakesh Yadav|
Designation: Assistant Professor, Banasthali University
Department of Pharmacy, Banasthali University, Banasthali-304022, Rajasthan
Abstract Title - Synthesis characterization and evaluation of some newer biphenyl imidazo- [2,1-b][1,3,4]-thidiazole derivatives for amtimicobial activity
Synthesis, characterization and evaluation of some newer biphenyl imidazo-[2,1-b][1,3,4]-thiadiazole derivatives for antimicrobial activity In the present study, we have reported the synthesis of some novel heterocyclic derivatives comprising imidazole and 1,3,4-thiadiazole containing moiety. Imidazothiadiazoles are of interest because of their diverse biological activities and clinical applications. Reactions of biphenyl carboxylic acid with thiosemicarbazide in the presence of phosphorous oxychloride resulted in biphenyl containing 2-amino-1,3,4-thiadiazole which is then further subjected to condensation with α-bromoarylketone under reflux in dry ethanol. The structures of the newly synthesised compounds were characterized by various spectral techniques and screened for antibacterial activity against strains of Escherichia coli, Pseudomonas aeruginosa and Bacillus subtilis, and antifungal activity against Candida albicans, Saccharomyces cerevisiae and Aspergillus niger. The compounds exhibited moderate to good activity when compared with
|Name: Dr. Daniele Castagnolo |
Designation: Lecturer of Pharmaceutical Chemistry, King's College London
Country : UK
Abstract Title:Design and synthesis of BM212-SQ109 hybid derivatives active against drug-resistant bacteria.
Tuberculosis (TB) represents a leading cause of morbidity and mortality worldwide and it is the first infectious disease declared by the World Health Organization (WHO) as a global health emergency. The number of tuberculosis cases in Europe was estimated at 56 per 100000 population in 2013, the highest for nearly 30 years. In addition, the emergence in the last years of multi-drug resistant (MDR) and extensively-drug resistant (XDR) strains of Mycobacterium tuberculosis, which are resistant to almost all the medicines available, make the discovery and development of new drugs a priority. We recently designed, synthesised and biologically evaluated a novel class of pyrrole derivatives. The novel compounds have originally been designed as hybrids of the anti-tubercular drugs BM212 and SQ109 that showed common features and similar spatial conformations. Computational studies revealed a perfect superposition of the structures of SQ109 and BM212, and suggested that the introduction of bulky substituents and secondary amino moieties on the side chain of a pyrrole scaffold could lead to derivatives with improved anti-tubercular activities. Five novel compounds showed high anti-tubercular activities on Mycobacterium tuberculosis, and two of them proved to be highly active also against MDR-TB clinical isolates. The most active anti-tubercular derivatives showed minimal eukaryotic cell toxicity, turning out to be an excellent lead candidate for preclinical trials.
|Name: Dr. Vikas Singh |
Designation : Post Doctorate Research Associate, University of Cambridge
Country : UK
Abstract Title- Anti-bacterial strategies to target bacterial resistance.
Aminoglycosides are broad spectrum antibiotics, mainly prescribed in hospitals to treat severe infections. Aminoglycosides have been an essential component of the armamentarium in the treatment of life-threatening infections. However, their use is compromised by the emergence of resistant bacteria producing enzymes able to inactivate them. In some cases the levels of resistance reached the point that rendered them virtually useless. Among many known mechanisms of resistance to aminoglycosides, enzymatic modification is the most prevalent in the clinical setting. Here, we propose to screen a library of compounds to identify specific allosteric inhibitors of aminoglycoside phosphotransferases, enzymes that catalyze the addition of phosphate groups on aminoglycosides which alters the interaction with their rRNA target. Also, these inhibitors bind to small cavities to the enzyme thereby blocking its activity by perturbing its dynamics. After in vitro assays and structure-activity relationships, the efficiency of the most active inhibitors was evaluated on bacterial isolates resistant to aminoglycosides to determine if their presence restores the sensitivity to the antibiotic. Those inhibitors are indeed aimed to be co-administrated with the aminoglycoside antibiotic in patients infected by resistant bacteria. Altogether, these type of inhibitors may constitute an innovative way to treat infections due to multi- or extensively-drug resistant bacteria.
|Name : Dr. Gaurav Chauhan |
Designation: Project Scientist at Indian Institute of Technology Kanpur.
Country : India
Abstract Title -Nanohybrid drug delivery platforms to tackle antimicrobial resistance.
Albumin stabilized silver nanoparticles–clotrimazoleb-cyclodextrin hybrid nanocomposite for enriched anti-fungal activity in normal and drug resistant Candida cells Chauhan Gaurav*, Gupta Nikhil, Sehrawat Deepti, Sourav Kalra, Rath Goutam and Goyal K. Amit Nanotechnology unlocked distinctive platforms to move inside a hybrid therapeutic zone. Currently, nanometal technology is the targeted field with exceptional advantages. Exceptionally small size and dominance of the surface properties such as high surface charge has raised a great deal of interest. This work is designed to exploit an interesting mechanistic feature i.e. multiple therapeutic targets possessed by metal nanoparticles. In this study we selected silver nanoparticles (AgNPs), which possess well documented antifungal activity and a standard antifungal molecule i.e. “clotrimazole”. A hybrid of AgNPs and clotrimazole was aimed to tackle clotrimazole resistance. Clotrimazole was firstly included into a b-cyclodextrin cavity to render it water soluble; subsequently the drug loaded dextrin moiety is functionalized on the surface of bovine serum albumin (BSA) stabilized silver nanoparticles using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysulfosuccinimide (EDC/NHS) chemistry. BSA stabilisation was essential to shield the physical interaction of AgNPs with the clotrimazoleb-cyclodextrin complex that is otherwise experienced. Spectral and morphological characterization of the complex assures the synthesis of a hybrid metal complex. A cellular toxicity assay was performed to determine the toxic nature of the hybrid. This hybrid was then evaluated for its fungicidal activity on normal and clotrimazole resistant Candida cells. The toxicity and efficacy outcomes revealed a potent profile with a handy therapeutic window. Mechanistic explanations for this hybrid nature were supported by aggravated apoptotic cell percentage and reactive oxygen species production in both resistant and non-resistant cells. Cell cycle arrest studies further revealed G2/M phase cell cycle arrest, directing towards compromised cell membrane and DNA synthesis process equivalently in clotrimazole resistant cells.
|Session 5:Structure/ Chemical Biology Interface: Understanding Mutations at level of Protein Structure and Function (Chaired by Sir Tom Blundell)
|Name: Dr. Arun Prasad Panduranga|
Designation: Research Associate, Department of Biochemistry, University of Cambridge
Abstract Title -Knowledge based method for predicting the impact of mutations on protein stability.
The research project focuses on developing computational methods to understand the effects of mutation on the protein function and stability. The method analyses the variation of amino acid replacements occurring at specific structural environment that are tolerated within the family of homologous proteins of known 3-D structures and convert them into substitution probability tables. These tables are used to calculate the scores for the wild type and mutant proteins and the difference in the scores are used as a quantitative measure for predicting the protein stability. Here we report the improved and updated computer program SDM used to predict the effects of mutation on protein stability. The mutant stability predictions are validated using the widely used thermodynamic dataset and the results will be presented.
|Name: Dr. Lipi Thukral |
Designation: Scientist Fellow, Institute of Genomics and Integrative Biology.
Abstract Title - Peering into black-box of membrane dynamics for overcoming drug resistance mutations
Recognition of biomolecules is a fundamental control mechanism in eukaryotic cells. The precise targeting and versatility of these interactions such as, protein-protein, protein-ligand, and protein-membrane is the hallmark of several cellular processes that culminates into key biological functions. Therefore, an understanding of these processes on atomistic level would have significant biological and medical impacts. We use computational methods, in particular biomolecular simulations, to understand the following processes (a) mechanism of peripheral membrane proteins and followed structural changes associated with their membrane localization (b) identifying structural scaffolds existing in substrate proteins, that are targeted to chaperones, and (c) structural and dynamical information of GPCR’s upon ligand binding. In particular, the emphasis will be on molecular understanding of “membrane-mechanics” i.e., force induced membrane perturbations. Such stress propagation in membranes have clear implications in detailed characterization antimicrobial membrane attack and subsequently for development of new drugs.
|Session 6 :The Power of Genomics to Understand and tackle AMR Chaired by Dr. Tariq Sadiq/Timothy walker
|Name: Dr. Neelja Singhal |
Designation: Principal Investigator in SERB-DST project, University of Delhi South Campus
Country : India
Abstract Title -Comparative genomics of β-lactamases genes and promoters: A goldmine for rapid AMR detection and newer β-lactamases inhibitor.
β-lactam antibiotics have been a prime choice for treating a number of infectious diseases. However, their widespread & indiscriminate use has resulted in microbial resistance towards this important class of antibiotics. Bacteria hydrolyze these antibiotics using their intrinsic/acquired antibiotic modifying enzymes, the β-lactamases. Studies from our laboratory using comparative genomics of β-lactamase genes and their promoters in a large number of Y.enterocolitica and E.coli strains revealed that, though the promoters were conserved, point mutations were present in different β-lactamase genes. Similar observations were also made while compiling a database of β-lactamase genes. Identification of consensus sequences among the β-lactamase genes of different bacteria could be useful for developing rapid and simple methods for detection of pathogens harbouring these genes. Our studies also revealed that mutations at sites other than active site of the enzyme may create diverse local changes in the 3D structure of the enzyme which might affect its binding affinity with β-lactam antibiotics as well as β-lactamase inhibitors. These findings might be useful for designing better β-lactamase inhibitors with improved efficiencies in future. Currently, we are working on developing a rapid and simple Loop Mediated Isothermal Amplification (LAMP) test using β-lactamase genes for detection of Y.enterocolitica. Consensus sequences have been identified for designing six primers for LAMP test. If successful, the work would be extended to detection of other food-borne pathogens. We are also working to identify novel sequences in β-lactamase genes which can be used as ideal targets for designing newer β-lactamase inhibitors. These studies would surely help us make assessments of the true potential of β-lactamase genes to serve as markers for rapid detection of AMR and salvaging several β-lactam antibiotics by designing novel β-lactamase inhibitors.
|Name: Dr. Divakar K|
Designation : DST-INSPIRE Faculty, National Institute of Technology, NIT Warangal.
Country : India
Abstract Title - Unravelling antibiotic resistant genes from metagenome library of anaerobic microbiome.
Occurrence of antibiotic resistance in bacteria is steeply increasing, but the knowledge of antibiotic resistant genes (ARG) were limited due to limitations of culture techniques available for microbes from extreme environment. There is huge possibility for occurrence of antibiotic resistant genes (ARGs) from anaerobic/uncultivable microbiome. ARGs have potential to transfer horizontally among the bacterial species it can be between culturable and unculturable species. We have applied functional metagenomics, a culture-dependent isolation methods used to access ARGs from anaerobic microbiome. Effluent from anaerobic sludge treatment plant rich in anaerobic bacteria (culturable/unculturable) was selected as source for extracting metagenomic DNA (approximately ~40kb). The extracted ~ 40kb DNA fragments were not species specific (independent of type of microorganism present in the sludge), selection were made only based on the size of DNA. Sequence unbiased fosmid library was constructed using pCC2FOS fosmid vector and expressed using Maxplax phage expression on E. coli EPI 3000 cells against β-lactum antibiotics. Total of 43 colonies which showed positive for resistance against tested β-lactum antibiotics were selected further investigation. The fosmid DNA was purified from all the 43 clones, the sequence of fosmid DNA was determined using next generation sequencing on Illumina Nextseq500 platform. Sequence analysis using MG-RAST revealed the presence of 29 gene sequences associated to ARGs. Among 29 genes 6 were found to have closely associated to already reported well known ARGs. Remaining 23 genes were found to have very less sequence similarity with the reported ARGs and showed high sequence similarity with oxidoreductases. These 23 genes found to contain consensus sequence/bases that are present only in conserved regions found in many known ARGs/mobile elements. Our study provides broad view of occurrence of novel ARGs which are closely related to oxidoreductases in anaerobic microbiome.
|Name : Dr. Rahul Shrivastava|
Designation : Assistant Professor (Senior Grade), Jaypee Univ. of Information Technology Waknaghat, Distt. Solan (H.P.)
Country : India
Abstract Title - Identification of Novel Gene(s) Responsible for Mycobacterium fortuitum.
Drug Resistance Mycobacterium fortuitum is rapidly growing, nontuberculous mycobacteria (NTM) which is ubiquitously present in nature. It is one of the most important pathogenic species, representing more than 80% of clinical isolates of rapidly growing mycobacteria. To add up insult to injury, an obstruction is imposed on the way to its treatment as these organisms are evolving into a drug resistant species. Our study aims to identify the resistant gene(s) and the mechanism associated, and the subsequent use of these findings in establishing effective drug targets. As the genomic sequence of M. fortuitum is not known, random mutagenesis technology (using TnphoA) was employed. The plasmid (pRT291) containing transposon Tn5 was electroporated in M. fortuitum wild type strains which resulted in formation of 50 blue mutant colonies showing mutation. These mutants were further screened by β-galactosidase assay. 10 mutants (i.e. KM1 to KM10) having high β-galactosidase activity were shortlisted for drug profiling where Ciprofloxacin, Amikacin, Rifampicin, Isoniazid and Ethambutol were utilized as these are most commonly used for treatment of M. fortuitum related infections. The minimum inhibitory concentration (MIC) of mutant strain KM4 for Rifampicin was observed to increase by 4-fold whereas the strain KM5 was sensitive to it. All the mutants had same MIC as that of wild type when Isoniazid was used, but they were sensitive to Amikacin (4-fold decrease in MIC). Genomic analysis of these mutants was done by restriction digestion of their genomic DNA, cloning of the insert containing transposon into pUC19, followed by sequencing. Comparative genomic analysis of mutants revealed two hypothetical genes of M. fortuitum which may be used as potential drug targets.
|Name : Dr. R. Saranathan |
Designation: Scientist B, national Institute for Research in Tuberculosis (NIRT), Chennai.
Country : India
Abstract Title - Evolution confers elevated virulence to MDR Acinetobacter baumannii through inactivation of AdeN, a TetR-type regulator.
Emergence of MDR Acinetobacter baumannii strains in the Indian subcontinent harboring carbapenem-hydrolyzing OXA-type carbapenemases and metallo-β-lactamases is a major concern. Insertion sequence (IS) elements are one of the most pronounced phenomena that assist the pathogen in exhibiting resistance or enhanced virulence either by increasing the expression level through formation of hybrid promoters or by disruption of repressor protein through insertional inactivation that regulates expression of plethora of virulence factors. Methods: The present study attempted genome sequencing of a clinical strain (PKAB07) originating from India for the first time. Whole genome sequencing of PKAB07 was performed and analysed with special reference to the distribution of resistant determinants, IS elements and traits of invasive properties. Gene knockouts were also constructed following the methods basing the double cross over recombination. Results: In vitro studies revealed that PKAB07 is a hyper-invasive strain exhibiting maximum induction of early apoptosis in human epithelial cells. Whole genome analysis showed ISAba1 mediated insertions in nine genes at novel regions not reported hitherto. In addition, an array of 108 resistance-related genes found distributed throughout the genome and it also contained 21 two-component systems. The present study identified a naturally occurring ISAba1 insertion in adeN (negative regulator of drug efflux-pump AdeIJK) gene and intended to prove this experimentally and embarked on generating adeN knock outs in A. baumannii ATCC 17978 strain. We also speculate that AdeN may have more important physiological roles other than controlling efflux pumps. adeN knockout was checked for biofilm production and induction of apoptosis. Our results showed that adeN knockout strain abrogated the ability to form biofilms and furthermore, the complemented strain restored biofilms. Additionally when the wild type and adeN mutant strains were co-cultured with the A549 cells for 24 h, there was a drastic increase in the virulence potential of the adeN knockout strain that reduced the viability of A549 cells. Conclusion: This study results provides evidence that adeN could be a positive regulator in controlling the process of biofilm formation and a negative regulator of virulence, since adeN disruption in A. baumannii causes this bacteria to behave abnormally with elevated virulence properties.
|Name : Dr. P Arul Jose |
CSIR - Nehru Postdoctoral Research Fellow, Central Salt and Marine Chemicals Research Institute
Abstract Title -Revival of the natural products search from actinomycetes using genomic and metabolomics tools
Antibiotic drug discovery is an indispensable process to combat destructive ability of pathogenic microorganisms and emerging infectious diseases worldwide. According to the updated and expanded data on natural products, the utility of natural products as source of novel structures is still alive and the anti-infective area depends on natural products. The natural products that showed pharmaceutical importance are largely produced from primary and secondary metabolism of plants and microorganisms. Especially, actinomycetes are well-known for their ability to produce a wealth of natural compounds with enormous structural complexity and biological activities. About 90% of practically used antibiotics are originated from the actinomycetes and they are still considered as the chief natural antibiotic producers. Moreover, recent innovations in next-generation sequencing, genome editing and analytical methods ensures new avenues for the discovery of natural as well as nature inspired antibiotics of great biopharmaceutical significance. With this background, we have initiated a translational research program that was designed to discover new natural products from novel actinomycetes derived from previously unexplored environments such as salterns, deserts and marine systems. A total of 250 actinomycetes were obtained using a range of selective isolation methods, and they were assigned under different genera, Actinoalloteichu, Actinomadura, Micromonospora, Nocardia, Nocardiopsis, Nonomuraea, Streptomyces, Saccharopolyspora, and Pseudonocardia. The isolates were screened using modern genomics and metabolomics tools. In the genomic approach, the isolates were screened for presence of novel polyketide (PKS) and Non-ribosomal peptide synthases (NRPS). Simultaneously, the isolates were grown in a range of production media and the spent media were analyzed in HR-LC-MS, for secondary metabolic profiles. Based on the genomic and metabolic profile, prioritized isolates were being subjected for comprehensive whole genome studies and downstream natural products research.
|Name : Dr. Stephan Petrus Wilhelm de vries|
Designation : Research Associate at the Department of Veterinary Medicine - University of Cambridge.
Country : UK
Abstract Title - Tracking the dynamics of AMR genes within bacterial communities using chromosome conformation capture sequencing
Antibiotic resistance in bacteria can be the result of spontaneous mutations in targets of antibiotics, by altering the expression of genes that affect the function of the antibiotic or through the aquisition of transmittable genetic elements such as plasmids that contain resistance determinants. Metagenomics studies have revealed that the gut harbours a whole arsenal of antibiotic resistance genes, some of which are transferrable between related or even unrelated bacterial species. A current limitation of metagenomic sequencing is that it cannot accurately determine which species harbours antibiotic resistance genes. Studies based on traditional culturing techniques showed that plasmid-mediated antibiotic resistance determinants are transferred from enteric pathogenic to commensal E. coli species. Currently our understanding of the extend of transfer and destination of antibiotic resistance genes is limited. Here we hypothesise that the commensal microbiota serves as an importance reservoir of antibiotic resistance genes and which is potentially affected by antibiotic selection pressure. To invesigate this we recently established chromosome conformation capture (3C/Hi-C) sequencing technologies in our laboratory to identify co-localisation of chromosomal and plasmid DNA in bacteria. These technologies capture the localisation of plasmids by forming crosslinks between the chromosome and the plasmid(s) in intact bacterial cells using formaldehyde. After bacterial lysis, DNA is fragmented by restriction digestion and ligated under highly diluted conditions, forming proximity ligations between chromosomal and plasmid DNA, which are subsequently identified by Illumina sequencing. To demonstrate the potential of these techniques to track the movement of of antibiotic resistance genes, a panel of bacterial species was cultured individually, some of which containing plasmids, and mixed into mock microbial communities before 3C/Hi-C read-out. Bioinformatic analysis revealed accurate assignment of plasmids to their correct host species. Currently, we testing the potential of these techniques using mouse and pig faecal samples spiked with plasmid-containing pathogens.
|Name: Dr. Allesandro Lazdins|
Designation : Doctoral Researcher at the Institute of Microbiology and Infection, University of Birmingham.
Country : UK
Abstract Title -Displacement of stable bacterial plasmids by a self- transmissible pCURE plasmid as a means of reducing antibiotic resistance gene load.
Antibiotic resistance is becoming one of the greatest scientific and medical challenges of the 21st century. However, our arsenal to combat this surge is limited to a small number of different classes of antimicrobial drugs. The importance of plasmids carrying antibiotic-resistance genes as targets for potential therapies has not been fully exploited and technologies that specifically address the plasmid burden could provide alternative approaches. We have shown that by harnessing the innate biology of plasmid replication and maintenance functions we can stably repress replication and eliminate plasmids from a population. This concept has been taken further and using the broad-host range IncP1 plasmid RK2 backbone as the starting point, a conjugative curing plasmid was constructed. Experiments on solid and liquid media showed successful elimination of the test plasmid from a chosen population in the absence of any antibiotic selection. On the basis of these experimental results, we are now developing a safe to use and easily administered conjugative, broad-host range plasmids that can efficiently target antibiotic resistance carrying plasmids within the gastrointestinal tract of humans and animals, and improve the treatment options for infections caused by multi-drug resistant Enterobacteriaceae.
|Name : Dr. Timothy Walker|
Designation : Academic Clinical Lecturer, University of Oxford.
Country : UK
In 2014 the World Health Organization announced its END TB strategy. A target was set to reduce global TB incidence by 90% on 2015 rates by 2035, corresponding to fewer than 10 new cases per 100,000 population per year. To meet this target, it called for ‘new tools’, including a point of care test, to be ready by 2025. Whole genome sequencing (WGS) is one of the most exciting and disruptive technologies to enter stage for decades. The gap it promises to fill is perhaps greater for Mycobacterium tuberculosis than for any other pathogen because of the diagnostic delays its slow growth rate lead to. Indeed, in many low-income settings, drug susceptibility testing is not even performed due to the lack of infrastructure. The consequent use an ineffective combination of tuberculosis drugs can be disastrous, both at an individual level and at a public health level. I will discuss measures to define the Mycobacterium tuberculosis 'resistome' - a process of characterising all mutations in the genome as either compatible with drug susceptibility, or causative of drug resistance. Such a knowledge base is the necessary underpinning of any WGS-based clinical assay that could identify effective, bespoke treatment regimens, within a clinically useful timeframe. As portable genome sequencers are further developed, and as methods for processing clinical samples without the need for sophisticated microbiology laboratories are refined, these data have the potential to improve patient management from low to high-income settings. I will discuss where global efforts have got to, and what the remaining challenges are likely to be.
|Session 7 :HPI for insight into Disease Etiology and Vaccine Development Chaired by Dr. G.P.S. Raghava|
|Name : Dr. Sanjib Bhakta |
Designation : Programme Director & Admissions Tutor for MRes Microbiology and Strategic Dean (International and Partnership) at the Faculty of Science, University of London.
County : UK
Abstract Title - interdisciplinary approaches to tackle AMR in Tuberculosis.
Drug repurposing, wherein a drug is found to be effective for the treatment of diseases or ailments other than what it was primarily used for, has rapidly gained priority to tackle the high attrition rates that novel drug design faces.1-3 In such an attempt, the common non-steroidal anti-inflammatory drug (NSAID), carprofen was first discovered to be anti-tubercular specific in vitro.4 Whole-cell phenotypic analyses to determine the mode(s) of action of these drugs revealed a marked effect on mycobacterial cells, indicating multiple intra-cellular machineries to be involved in their endogenous mechanisms of action. Furthermore, carprofen inhibited efflux pump activity in mycobacteria with results at par with commercially available efflux pump inhibitors. It was found to a bactericidal drug that completely inhibited biofilm formation, bringing to light the requirement of active efflux systems for the development of biofilms. Additionally, transcriptomic analyses revealed regulation of several genes in M. tuberculosis implicated in key metabolic pathways, dormancy and resistance mechanisms that explain, in part, the information we have gathered through experiments thus far. Mutants of Mycobacterium aurum, resistant to carprofen are generated and gene sequence analysis is done on the target genes. As mortality and transmission rates of drug-resistant cases of tuberculosis (TB) increase, inclusion of these widely used over-the-counter drugs can be a major move forward in increasing the effectiveness of the current therapeutic regimens in TB.
|Session 8 :Systems Biology to Understand drug esistance Chaired by Dr. Anshu Bhardwaj
|Name : Dr. Sudipto Saha |
Designation: Ramalingaswami Fellow, Bose Institute, Kolkata.
Country : India
Abstract title - Towards cataloguing drug resistant- associated genes & its PPI networks.
Lab research focus is on functional interactomics, studying the protein-protein interaction(PPIs) interfaces and the networks. One of my running project is on a systematic discovery of linear motifs mediating PPIs. Developed a Linear Motif mediated Protein Protein Interaction Database (LMPID) for linear motifs that helps in PPIs and a Linear Motif Domain Interaction Prediction (LMDIPred) server for predicting peptides binding to SH3, WW and PDZ domains. Another ongoing project is to identify protein biomarkers for Asthma, an upper tract respiratory disease using label-free global proteomics from blood samples. Besides these, we study protein-protein interactions using affinity purification followed by Mass Spectrometry in Mtb-host protein-protein interaction. We focus on whether Mtb drug resistant gene mutations rewire the host-Mtb protein protein interaction network. For this, we have selected a few Mtb drug resistance genes and studying human macrophage-Mtb PPI networks in mutant and wild-type drug resistant genes. In summary, we use bioinformatics, proteomics, and protein-protein interaction networks to understand complex biological systems.
|Session 9:Drug Repurposing and Adjunct Therapies Chaired by Sir Tom Blundell
|Name : Wahajuddin |
Designation : Scientist, CSIR- Central Drug Research Institute Lucknow.
Country : India
Abstract Title-Investigation of the functional role of P-glycoprotein in limiting the oral bioavailability of lumefantrine with implication in drug resistance cases.
In the quest to explore the reason for the low and variable bioavailability of lumefantrine, we investigated the possible role of P-glycoprotein (P-gp) in lumefantrine intestinal absorption. An in situ single-pass intestinal perfusion study in rats with the P-gp inhibitor verapamil or quinidine and an ATPase assay with human P-gp membranes indicated that lumefantrine is a substrate of P-gp which limits its intestinal absorption. To confirm these findings, an in vivo pharmacokinetic study was performed in rats. The oral administration of verapamil (10 mg/kg of body weight) along with lumefantrine caused a significant increase in its bioavailability with a concomitant decrease in clearance. The increase in bioavailability of lumefantrine could be due to inhibition of P-gp and/or cytochrome P450 3A in the intestine/liver by verapamil. However, in a rat intestinal microsomal stability study, lumefantrine was found to be resistant to oxidative metabolism. Further, an in situ permeation study clearly showed a significant role of P-gp in limiting the oral absorption of lumefantrine. Thus, the increase in lumefantrine bioavailability with verapamil is attributed in part to the P-gp-inhibitory ability of verapamil. In conclusion, lumefantrine is a substrate of P-gp, and active efflux by P-gp across the intestine partly contributed to the low/variable bioavailability of lumefantrine.
|Name : Dr. Surekha k. Satpute |
Designation : Assistant Professor, Department of Microbiology, Savitribai Phule Pune University, Pune 411007, Maharashtra.
Country : India
Abstract Title - Anti-Biosurfactants Resistance in Biofilm producers: A Challenge of 21st century.
Medical devices industry is fairly large, intensely competitive and highly innovative. Devices and equipment’s used in most healthcare professionals needs to be safe. The increased microbial resistance to variety of antibiotics is major concern for medical practitioners. One of the modes by which bacteria exert this resistance is due their ability to develop biofilms. The inherent resistance of biofilms and their pervasive involvement in implant-related infections has raised demand for anti-adhesive biological agents. Lactobacilli spp. contributing an important genera of microbiota providing favourable opportunity to combat against pathogens. Biosurfactants (BSs) produced by Lactobacilli spp. do exhibit antimicrobial and anti-adhesive activities and therefore are highly effective against pathogens. We have successfully produced two different types of BSs from L. Acidophilus NCIM 2903 by growing in a simple fermentation medium. Both types of BSs displayed surfactant properties through various assays. Analytical characterization of BSs revealed glycolipid and glycolipoprotein nature. We aim to design biological coatings to cure biofilms on bioimplant materials. The formulation is under trials through microfluidics/lab-on-a-chip. This is the first report of microfluidic based approach to assess the effectiveness of BS formulation on biofilms through mimicking in-vivo. Work on different coating techniques like spin, plasma, dip coating is in progress to ensure the effective spreading on the surface of medical devices. Glycolipid type BS inhibits biofilm formation on contact lenses and urinary catheters effectively. We are further interested to predict the possible mechanisms involved in disruption of microbial biofilm mediated through BS produced by Lactobacillus strain. Our studies are significant and has tremendous medicinal and therapeutic perspectives. Anti-adhesive developed by us has commercial applications for several medical devices. This would prolong the life of the bioimplants as well as reduce the chances of opportunistic infections. From the commercial point of view, our research is extremely significant.