Day 1 :
Keynote Forum
John G Thomas
West Virginia University, USA
Keynote: One health: Animals, humans and our globe, our mega-organism
Time : 10:00-10:30
Biography:
Professor John G. Thomas is recognized as an “International Educator and Global Microbiologist “ (www.globalbugs.com) having lectured in more than 43 countries while a clinical microbiologist in pathology, dentistry and medicine for 51 years, his research emphasizing biofilms and medical devices including endo-trachs and the connection between oral diseases, VAP and wound infections (“Intellectual Designâ€) with the recent integration of micro 3-D- bio Printing using bio-plastics and unique Prebiotics (Therapeutic Bacteria ) for intervention. He has over 50 publications, multiple book chapters, significant grant support, pending patents and over 100 posters/abstracts at national and international meetings. His sabbatical at Cardiff University, Wales, UK (2007) was a driving influence. He has been a member of the ADA Scientific Advisory Committee for the last 8years.As Faculty at 6 Universities during his career , he has received Alumni and University awards for research and International Student Mentoring ; retiring from WVU in 2013 after 23 years as Professor Emeritus , he presently is expanding his research/teaching utilizing the advanced resources of the Allegheny Health Network in Pittsburgh, PA, Carnegie–Mellon University and Mass. Gen. Hospital, Boston, MA.
Abstract:
We evolved to conquer and emerged with a concept that we were the focus of our environment, our world. The concept was doomed from the start and our disruption of the world we live in has forced us to re-evaluate or position, recognizing that there are 3 parts (Humans, Earth/Environment and Animals) and we collectively address Newton’s third law of Motion: For every action there is an equal and opposite reaction. Thus, balance or one health and its intrinsic buffering capacity is the key and humans have been collectively “Multi- Disruptiveâ€, particularly the untoward consequences of antibiotics. Human population will approach 9 billion shortly and with it an increase in low socio-economic habitat (Global Slums) with increased potential for disease transmission, so evident in the most recent Ebola episode in Africa. Old and new routes of emerging pathogens will be unmasked, perhaps 2-4 new ones per year. Animal diseases, zoonosis, particularly viral, will predominate as we realize that most global recent diseases have had an animal vector; over the last 3 decades approximately 75% of human diseases and have come from or through wildlife. Perhaps most significant will be the unpredictable impact of global warming , climate change and the subsequent increased range of vector-borne diseases and the pollution of water, a potential for wars. Microbes are central to all three and the unbalance now provides the opportunity for the “Perfect Microbial Stormâ€. There is ‘No Health Sanctuaries’ as humans have “collapsed their space†with animals and the environment/earth. Fungi offer a unique view as they encompass more animals/plants in a deadly fashion. Proactive surveillance is key-based on global collaboration using tools like Google Earth and GIDEON to maximize rapid computer inter-face and global diversity. New laboratory tools, like BARDOT, laser light scatter identification need incorporation to link international microbial libraries and unmask the other threatening potential, Bioterrorism.
Keynote Forum
Giovanni Matera
University of Catanzaro, Italy
Keynote: Successful Leishmania steering immunity cells and mediators to achieve a trade-off balance useful to host and parasite.
Time : 10:30-11:00
Biography:
Giovanni Matera obtained his M.D at University of Messina, Italy, in 1982 and PhD in Microbiology in 1987. Specialist in Infectious Diseases in 1994. From 1985 to 1986 Postdoctoral Fellow at the Medical University of South Carolina, Charleston, USA. From 1986 to 1988 Research Associate at the Dept. of Microbiology, University of Saskatchewan, Saskatoon, Canada. Instructor from 1990 to 2000 at the Chair of Microbiology, University of Catanzaro. From 2000 to 2006 Assistant Professor and since 2006 Associate Professor of Microbiology and Clinical Microbiology at the same University. Since 2008 Head of Simple Unit \"Parasitology\" at O.U. Clinical Microbiology, University of Catanzaro. Main lines of research: 1. Biological effects of bacterial endotoxins; 2. Physiopathology of sepsis and markers of systemic infections and endocarditis. 3. The mechanisms of antibacterial drugs; 4. Laboratory diagnosis of infectious diseases; 5.Clinical Parasitology. Author of over 80 articles published in reputed international journals of Microbiology and Infectious Diseases.
Abstract:
Parasitic diseases are very often chronic and during such long-lasting infections, host inflammatory and immune cells are quite unresponsive to stimulation by products from the infecting organism and other pathogens. Leishmania are hemoflagellate protozoa, which cause three types of illness (cutaneous, muco-cutaneous and often fatal visceral leishmaniasis) exhibiting a high prevalence and incidence worldwide. More informations are needed to explain the role of different branches of immune response in leishmaniasis pathogenesis, as well as to get a timely diagnosis, an accurate prognosis and a more effective therapy.rnOur contribution to understanding mechanism of Leishmania disease included in vitro model with live protozoa infecting human peripheral blood mononuclear cells, as well as the dosage of serum mediators in infected humans. In our approach with clinical specimens, both conventional and molecular tests were able to verify the presence of Leishmania spp. in cutaneous and mucous biopsies of suspected tegumentary leishmaniasis patients. In these subjects, levels of Th2, Treg cytokines, MCP-1 chemokine and CD25+ cells were statistically more elevated than in Leishmania-negative patients and healthy controls; on the other hand a decrease of TNFα, VEGF and EGF were found among samples obtained from Leishmania-positive group. We noticed the absence of allergic pathologies among parasitized patients.rnMoreover we sought to reproduce in vitro the early phase of the natural infection. Therefore human PBMC were challenged by Leishmania infantum or Leishmania major infective metacyclic promastigotes. Following 4 hours from L. major infection, differently from L. infantum, TNFα , IL-1β, IL-6 levels were significantly higher than controls. However, after 24 hours, promastigotes of both species stimulated significantly higher TNFα , IL-1β, IL-6 levels. A negative correlation was observed between the parasite concentration and the cytokine levels. Regarding MCP-1 release, at 24 hours, but not at 4 hours, an interesting dose-dependent effect was observed for both species. Nitric oxide levels, at 4 and 24 hours, were increased when lower parasite burden of both species was used; interestingly the higher promastigote concentration of L. major or L. infantum fails to stimulate nitric oxide levels. rnThe protozoal infection studied seems to reduce major Th1 response cytokines, while increasing both immune cells and mediators with regulatory/inhibitory effect, in both in vitro and in the clinical approaches used. Host might take advantage in reducing re-infections, as well as immunopathology/allergies. Leishmania species may exploit immunoregulatory mechanisms in early steps of infection in order to evade the host immune system and in the chronic phase to protect the earned host niche against other invaders. The clinical impact of our findings consists in the relevant number of mediators which could be proposed as potential diagnostic and prognostic biomarkers helpful to integrate clinical management of leishmaniasis.rnrnrn
Keynote Forum
Joanna S Brooke
DePaul University, USA
Keynote: Insights into Drug-Resistant Stenotrophomonas maltophilia, a Global Opportunistic Bacterial Pathogen.
Time : 11:20-11:50
Biography:
Joanna S. Brooke is an Associate Professor in the Department of Biology at DePaul University. She holds doctorate and masters degrees in Microbiology and Immunology from the University of Western Ontario, with focus on bacterial lipopolysaccharide assembly and bacterial cell ultrastructure, respectively. Her postdoctoral research at the University of Texas Southwestern Medical Center investigated the interactions of diphtheria toxin with its receptor. Her current research examines S. maltophilia and its biofilms. She also studies other potential bacterial pathogens. She has published 18 papers in peer-reviewed journals. She is a Guest Associate Editor for a Frontiers Research Topic on S. maltophilia
Abstract:
Stenotrophomonas maltophilia is a drug-resistant Gram-negative bacterial opportunistic pathogen found in moist environments in and outside the clinical setting. The World Health Organization has named S. maltophilia as a leading global drug-resistant pathogen in hospitals. Infections of S. maltophilia are of significant concern and associated with high mortality rates in the immunocompromised patient population. Various infections in humans are associated with S. maltophilia, most commonly those of the respiratory tract, and this bacterium forms biofilms on medical devices and living tissues. S. maltophilia can be a co-colonizer in polymicrobial biofilms found in cystic fibrosis patients. Our studies and others have shown that the biofilm is influenced by cell ultrastructure and culture growth conditions. Genome sequencing and analysis have revealed considerable genetic diversity among S. maltophilia strains and multiple molecular mechanisms used to thwart antimicrobial treatment. The rise in antibiotic resistance of S. maltophilia worldwide emphasizes the need to develop new strategies and therapies to combat this emerging opportunistic pathogen. Studying the colonization, biofilm formation, and virulence of S. maltophilia may help to identify suitable targets for pharmaceutical therapy. We will discuss the clinical challenges presented by this pathogen and new approaches being used to identify and treat S. maltophilia infections. rnrn
- Insights and Trends in Microbiology
Chair
John G Thomas
West Virginia University
Session Introduction
John G Thomas
West Virginia University, USA
Title: Unlearning the learned about wound management and biofilms: Reestablishing the microbial library within
Biography:
John G Thomas is recognized as an “International Educator and Global Microbiologistâ€; being lectured in more than 43 countries whiles a Clinical Microbiologist in Pathology, Dentistry and Medicine for 51 years. His research emphasizes bio-films and medical devices including endo trachs and the connection between oral diseases, VAP and wound infections (“Intellectual Designâ€) with the recent integration of micro 3-D- bio printing using bio-plastics and unique prebiotics (Therapeutic Bacteria) for intervention. He has over 50 publications, multiple book chapters, significant grant support, pending patents and over 100 posters/abstracts at national and international meetings. His sabbatical at Cardiff University, Wales, UK (2007) was a driving influence. He has been a member of the ADA Scientific Advisory Committee for the last 8years. As Faculty at 6 Universities during his career, he has received Alumni and University awards for research and International Student Mentoring; retiring from WVU in 2013 after 23 years as Professor Emeritus, he presently is expanding his research/teaching utilizing the advanced resources of the Allegheny Health Network in Pittsburgh, PA, Carnegie–Mellon University and Mass. Gen. Hospital, Boston, MA.
Abstract:
Chronic wounds are one of the most costly health care issues, affecting 6.5 million patients at an annual cost of US $25 billion, focusing on a growing aged population. Multiple interventions emphasizing silver gauze have not altered QOL issues or reduced significant collateral damage of MDR sepsis and C. difficile. In 2012, we introduced and evaluated the SMarT probiotics matrix concept, where a bi-phasic strategy harmonizes an ecologic approach of Minimal Intervention (MI) disruption while restoring microbial architecture and tissue engineering. The emergence of Probiotic therapy had been catalyzed by recent advances in microbiota and metagenomics, particularly dentistry where we initially evaluated probiotics in endodontic. In the discovery (in vitro) phase 2013/4, 3 pools of probiotics were evaluated for anti bio-film, anti-planktonic activity, ultimately creating a “designer symbiotic†combination against 3 wound pathogens: Staph. aureus, C. albicans and P. aeruginosa. In application (in vivo) phase (2015/16), the “designer symbiotic†is being evaluated using a rabbit wound mode infected with the same strains, complimenting bio-burden wound reduction and site closure/tissue regeneration bellow “critical colonization†with a combined silver dressing/silver gels “suite platform†enforcing “site-saturationâ€. The wound/tissue environment will be strengthened utilizing delivery of the symbiotic via biodegradable, bio-plastic scaffolding with barrier activity enhanced by form fitting gauze individually constructed with 3-D printing using plasma as the ink.
Biography:
Joanna S Brooke is an Associate Professor in the Department of Biology at DePaul University. She holds Doctorate and Masters’ degrees in Microbiology and Immunology from the University of Western Ontario, with focus on bacterial lipopolysaccharide assembly and bacterial cell ultrastructure, respectively. Her Post-doctoral research at University of Texas, Southwestern Medical Center investigated the interactions of diphtheria toxin with its receptor. Her current research examines S. maltophilia and its bio-films. She also studies other potential bacterial pathogens. She has published 18 papers in peer-reviewed journals. She is a Guest Associate Editor for a Frontiers Research Topic on S. maltophilia.
Abstract:
Stenotrophomonas maltophilia is a global human opportunist which is associated with infections that include those of the respiratory tract, bloodstream, soft tissue and bone, eye, heart and brain. S. maltophilia infection is of significant concern in immunocompromised patients and a high mortality rate has been reported. This bacterium is found in water, washed foods, plant roots and soils and animals. Hospital-acquired and community-acquired infections of S. maltophilia have been reported. Antimicrobial resistance surveillance monitoring networks worldwide report a steady rise in the number of drug-resistant strains of S. maltophilia recovered from patients. S. maltophilia is resistant to a wide range of antimicrobials, including beta-lactams, fluoroquinolones, aminoglycosides, polymyxins, macrolides, carbapenems, tetracylines, chloramphenicol and trimethoprim-sulfamethoxazole. Intrinsically drug-resistant strains of S. maltophilia have been recovered from environments outside of the clinical setting. New strategies are needed to prevent/challenge S. maltophilia infections. S. maltophilia forms biofilms on medical devices and on living tissues. One of the goals of our laboratory is to study the molecular mechanisms used by this pathogen to form biofilms and subsequently identify suitable targets for treatment strategies to prevent/inhibit S. maltophilia growth, biofilms, and cell survival. We have observed that S. maltophilia is able to form biofilms on polyvinyl chloride, polystyrene and glass. We have screened various chemicals and observed that the growth and biofilm formation of S. maltophilia can be hindered. We will report on recent studies that examine the effects of select chemicals on the growth, biofilm development and survival of S. maltophilia
- Track-2 Clinical Bacteriology
Session Introduction
Radjin Steingrover
Analytical Diagnostic Centre, Netherlands
Title: 5 year analysis of bacterial resistance in the Dutch Carribean Island of curacao and implications for antimicrobial stewardship
Time : 14:25-14:50
Biography:
Radjin Steingrover is a MD specialized in Clinical Microbiology and has recently joined ADC in Curacao to bring top-notch clinical microbiology to Curacao and Sint-Maarten, to coordinate antimicrobial stewardship and hospital hygiene and to implement tele-microbiology in collaboration with the Dutch Academic Medical Center. He has a background in Clinical Microbiology and international HIV-research. He is the head of the department of clinical microbiology at ADC and leads the department of Hospital Hygiene on Curacao. He has published papers in peer-reviewd journals and serves as an Editorial Board Member of a Dutch Journal on Infectious Diseases
Abstract:
Introduction: The Carribeannation of Curacao has experienced the burden of antibiotic resistance with outbreaks of MRSA and KPC. Curacao lacks comprehensive data on antimicrobial resistance to guide antibiotic poly-makers. This study presents an overview of the last 5years of susceptibility trends for the primary pathogens. The method of analysis facilitates the choice of empirical therapies and allows comparisons with The Netherlands, a country with which Curacao and its health-care system holds strong ties. Methods: The ADC laboratory information system was queried for all positive cultures from 2010 up to 2015. Susceptibility testing was done with the VITEK 2 AST cards with only a minority of tests having been done by disk diffusion. The results were filtered for bacterial results and the Curacao area. Identical to the Dutch resistance register (Neth Map) only the first isolate per patient per year was entered into the analysis to exclude bias from repeated sampling. Results: The results show extensive resistance overall. MRSA increases in blood cultures from 0% to 9%. The introduction of KPCpositive K. pneumoniae was shown as well as extensive overall resistance to all classes of antibiotics. Discussion: The high-rates of resistance are markedly different from the data in Neth Map. The data support the choice of a cephalosporin combined with an amino-glycoside as empirical sepsis regimens. The rise in MRSA and the introduction of KPC warrant an effective anti-microbial stewardship program to counter the spread and reduced option for antibiotic therapy.
Anand Ramasubramanian
University of Texas, USA
Title: Chips for antimicrobial drug discovery and diagnosis
Time : 14:50-15:15
Biography:
Anand Ramasubramanian is an Associate Professor of Biomedical Engineering and a Member of the South Texas Center for Emerging Infectious Diseases at the University of Texas at San Antonio (UTSA). His current research interests are in microbial bio-engineering and vascular mechano-biology. His lab focuses on developing microscale tools for understanding and combating infection and inflammation and in improving platelet storage modalities for transfusion. Prior to joining UTSA, he received his PhD in Bio-engineering from Rice University and Post-doctoral training in Chemical Engineering at UC Berkeley.
Abstract:
We are interested in the development of microscale technologies for applications in drug development and diagnostics for infectious diseases. We have developed a high-density microarray platform (‘chips’) consisting of nano-liter volumes of microbial pathogens on chemically modified glass slides using a robotic microarrayer. We have successfully grown 1200 individual cultures of 30 n-L volume on a standard glass slide consisting of either single or polymicrobial cultures of Candida albicans, Pseudomonas aeruginosa or Staphylococcus aureus as biofilms. These nano bio-films display morphological complexity, three dimensional architecture and drug resistance similar to conventional cultures in well-plates or flasks. I will demonstrate the suitability of the chip for single and combinatorial screening of small molecule libraries. I will also demonstrate an adaptation of the chip as a diagnostic tool for pathogen identification and antimicrobial susceptibility testing in clinical samples of MRSA. In summary, our chip platform cuts reagent use and analysis times, minimizes or eliminates labor intensive steps and dramatically reduces assay costs and thus opens a new chapter in microbial culture.
- Track-3 Clinical Virology
Session Introduction
Mervat G Elanany
Cairo University, Egypt
Title: Zoonotic endocarditis.Ten years experience: 2005-2015
Biography:
Mervat G A El-Anany is a Professor of Clinical Microbiology at Cairo University Medical School. She is also a Microbiology & Infection control Consultant. She is the Director of Infective Endocarditis Laboratory committee and is the Head of infection control team of medical hospital. The research activities included different clinical microbiology subjects including molecular typing of resistant organisms, diagnosis of brucellosis and diagnosis of infective endocarditis. He has published more than 30 papers in national and international journals.
Abstract:
Infective endocarditis (IE) is a life-threatening disease. Zoonotic bacteria can cause blood culture–negative endocarditis (BCNE). Zoonotic IE is prevalent in North Africa. The study aimed to diagnose IE caused by the zoonotic pathogens Brucella spp., Bartonella spp. and Coxiella burnetii in BCNE by PCR and serology. The study prospectively followed up all patients with suspected IE referred to the Endocarditis Service, Cardiology Department, Cairo University from February 2005 to February 2015. Three sets of blood culture were withdrawn on admission. Resected surgical material was cultured whenever available. Serologic testing was performed for detection of Brucella antibodies using standard agglutination test, IgG titers for Bartonella and IgG, IgM and IgA antibody titers for Coxiella burnetii using IFA on the sera of all referred patients. A patient was considered to have endocarditis caused by Brucella when antibody titers exceeded 1/320, Bartonella when IgG titers >1:800, and Coxiella when IgG phase I titer >1:800. Broad range bacterial 16S rRNA from blood culture bottles and surgical materials followed by sequencing for identification of positive cases was done. IE was classified as definite or possible in 400 patients; 50% of them had BCNE. Zoonotic endocarditis was diagnosed in 28(7%) patients including 16 cases of Brucella spp. 8 cases of Bartonella spp. and 4 cases of Coxiella burnetii. Zoonotic agents were a cause of 11% of BCNE. Zoonotic agents are important cause of IE in Egypt.
- Track-4 Parasitology And Mycology
Session Introduction
Moyra Machado Portilho
Oswaldo Cruz Institute, Brazil
Title: Usefulness of saliva samples for hepatitis B virus DNA (HBV DNA) detection
Biography:
Moyra Machado Portilho is a Biomedic with Master degree in Tropical Medicine Program from Oswaldo Cruz Foundation (Fiocruz, Rio de Janeiro, Brazil) and currently is a PhD student of the same institutional program. She has expertise on molecular diagnosis of HBV infection, conducting studies to evaluate the effectiveness of alternative fluids, such as saliva samples, for HBV diagnosis.
Abstract:
Worldwide, hepatitis B virus is responsible for 240 million chronic cases of infection and diagnosis is made by detection of antigens, antibodies and genome in sera samples. But HBV DNA can be detected in other biological fluids of infected individuals in smaller concentrations, such as seminal fluid, urine, oral fluid and tears. Saliva samples have been studied as alternative sample for molecular diagnosis of HBV since collection is cheaper, less invasive and less painful when compared to blood collection. Some studies have shown that the HBV DNA can be detected or quantified in saliva samples using different protocols (“in house†or commercial quantitative ones) where varied values of sensibility and specificity are observed, however no protocol had yet been described or optimized for use in saliva samples. So, in order to evaluate the usefulness of saliva samples for HBV DNA detection, extraction and detection methods for HBV DNA detection were investigated. In this study from our group it was possible to detect HBV DNA 200 copies/mL in artificially contaminated saliva samples using “in house†molecular methods and according to the literature, we could realize the importance of saliva collector for the success of PCR, since samples obtained by spontaneous salivation and commercial collectors with mechanical friction principle generates more satisfactory results in molecular studies, so we are now evaluating the efficiency of four different collection devices of oral fluid samples, a non commercial one (spitting) and three commercial assays.
Asho Ali
King Abdulaziz University, Saudi Arabia
Title: Association of gyrA mutation in Mycobacterium tuberculosis isolates with phenotypic ofloxacin resistance detected by resazurin micro-titer assay
Biography:
Asho Ali has completed her PhD at the age of 44 years from the Aga Khan University. She is Assistant Professor, Microbiology at the King Abdulaziz University, Jeddah, Saudi Arabia. She has extensive academic and research experience. Her research interests are infectious diseases; particularly tuberculosis, drug resistance in bacteria and molecular mechanisms of drug resistance. In her post-doctoral research work she has worked on molecular detection of drug resistance in Extensively Drug Resistant (XDR)-TB isolates. She has published more than 20 papers in reputed journals and has been serving as reviewer in many reputed journals.
Abstract:
Background: Ofloxacin (OFX) is an important second-line anti-tuberculosis drug for the treatment of MDR-TB. This study aimed to compare mutations in quinolone-resistance determining regions (QRDR) of the gyrA and gyrB genes of Mycobacterium tuberculosis (MTB) isolates with MICs of OFX determined by resazurin micro-titer assay (REMA) on OFX resistance in isolates. Methods: Thirty-nine MTB isolates collected during 2009 at the MTB bank were selected. This included 25 XDR, 3 MDR+OFX resistant and 11 OFX susceptible isolates (non-MDR). MICs for OFX were determined in duplicates by REMA method for the 39 MTB isolates as well as for control H37Rv strain. The presence of mutations in QRDR of the gyrA and gyrB genes was determined by sequencing. Then type of mutation identified on each codon was compared with MIC determined for OFX. The findings were also compared with the drug susceptibility results obtained by the proportion method. Result: Mutations were observed in the gyrA gene of 18 out of 28 OFX resistant MTB isolates. Frequency of mutations were 14% (n=4), 4% (n=1) and 39% (n=11) on codons 90, 91 and 94 respectively. In addition, two isolates showed concurrent mutation i.e. on 90 plus 91 and 90 plus 96 codons. Codon 94 showed more high level (4-8 μg/mL) OFX resistance as compared to codons 90 and 91. None of the OFX resistant isolates exhibited mutation in gyrB gene. Mutations were not observed in gyrA and gyrB gene in all the OFX susceptible as well as in control MTB strain. Agreement between phenotypic and genotypic OFX susceptibility testing was 64%. Conclusion: Results of this study supports the use of rapid, simple and inexpensive REMA method for OFX susceptibility testing particularly for MDR-TB isolates in resource limited settings. No significant association could be linked between type of mutated codon in gyrA gene and level of OFX resistance.