6th Clinical Microbiology Conference October 20-22, 2016 Rome, Italy

Biography:

Maurizio Provenzano graduated as MD from La Sapienza University of Rome and obtained his specialization in Medical Oncology at the University of Milan and his PhD in Virology at the University of Padua. He is the Head of Research at the Department of Urology, University Hospital of Zurich and Lecturer at the Faculty of Medicine, University of Zurich. Since 2007 he has been member of the Cancer Network of Zurich. He has published more than 50 papers and has been serving as an editorial board member of reputed journals. He has received several acknowledgments for his efforts in cancer virology

Abstract:

Several studies associating BK polyomavirus (BKPyV) and prostate cancer (PCa) suggested that this virus may exert its oncogenic activity at early stages of cancer development. The BKPyV oncogene, the large T antigen (LTag), has frequently been detected in areas of proliferative inflammatory atrophy, which is considered a precursor lesion leading to prostatic intraepithelial neoplasia and overt PCa. In a recently updated systematic review, the presence of BKPyV was significantly higher in PCa tissues than in healthy control tissues, providing evidence for a link between BKPyV infection and cancer risk. In addition, recent original investigations highlighted an association between expression of the virus and the clinical course of PCa. For example, by studying immune responses elicited against BKPyV LTag, a significant association between LTag positive cancer lesions and a peculiar regulatory profiling has been observed in PCa patients with evidence of disease recurrence after surgical radical prostatectomy. Lastly, a study carried out in a larger cohort of patients undergoing radical prostatectomy revealed the IgG response against LTag as an independent predictor of disease recurrence. Although a full picture of the mechanisms potentially responsible for the involvement of BKPyV in PCa is not available yet, continuing work on this topic should help to refine the potential role of BKPyV in PCa patients, perhaps revealing unsuspected associations with the clinical course of this disease.

Biography:

Malgorzata G. Norton (nee. Mikolajczyk) received her M.S. degree in Veterinary Medical Sciences/Immunology from Louisiana State University, School of Veterinary Medicine studying Cat Scratch Disease. She is currently a Biologist at the U.S. Food and Drug Administration in the Laboratory of Plasma Derivatives, a position she’s held for over 10 years. She is involved in both regulatory review and research involving plasma-derived polyclonal immune globulin. Her current research involves protein-protein and protein-carbohydrate interactions using Surface Plasmon Resonance (SPR). Her collaborations span the fields of virology, bacteriology, immunology, and hematology.

Abstract:

Avian influenza continues to be a public health problem.  Vaccination may prevent or ameliorate the disease; however, it may not always be feasible to match the timing of the vaccine administration to the time of exposure, or additional therapeutics may be needed in the case of severe disease. Passive antibody therapy with polyclonal anti-influenza immune (hyperimmune) globulins may be an effective complementary therapy. Hyperimmune globulin therapy is already in place for several infectious agents and an anti-H5N1 Equine F(ab’)2 product has recently obtained EMA Orphan Drug status. In order to screen individual plasma donations for high neutralizing titers to pool into a hyperimmune product, a fast and reliable neutralizing antibody assay is needed. Currently, the hemagluttination inhibition (HAI) and microneutralization (MN) assays are used to detect neutralizing antibodies against Influenza viruses; however, each assay requires the use of live virus. We developed a Surface Plasmon Resonance (SPR) assay to measure anti-H5 HA antibodies for neutralizing activity which has no requirement for live virus. Briefly, biotinylated multimeric glycans containing sialic acid moieties were captured on a streptavidin-coated surface, acting as a model for influenza cell-surface receptors. Multimeric H5 HA recombinant protein micelles preincubated with a dilution sequence of antibodies or serum were then injected over the glycans.  The results of the antibody dilutions preincubated with H5 HA were compared to H5 HA only, and an IC50 was calculated. Using the IC50 measurement we could rank the mAb and polyclonal sera in order of neutralizing activity. The SPR assay may also be adapted to measure the neutralizing antibody against other infectious agents where the host receptor is known.

Biography:

A molecular virologist for over 30 years, and Professor in the Department of Cell and Tissue Biology at the University of California San Francisco Lenore Pereira has focused the last 16 years on the biology of human cytomegalovirus replication and pathogenesis at the uterine-placental interface. Dr. Pereira has published over 120 papers and invited reviews. Recently, her group in collaborative studies with Dr. Eva Harris at the University of California Berkeley identified ZIKV target cells and immune mechanisms that protect the placenta.

Abstract:

The Zika epidemic that began in Braziland has spread throughout the Americas has beenreported as definitively linked to severe birth defects – microcephaly, miscarriage and stillbirth.Detection of ZIKV RNA in the placenta and fetus and frequent intrauterine growth restriction suggestsinfection of the placenta leading to pathology.Our studies in primary cells from 14 placentas reveal that prototype and recently iso¬lated Nicaraguan ZIKV 2016 strains infect cells that express Axl, Tyro3 and TIM1 tyrosine kinase receptors, which modulate innate immune responses and mediate infection by ZIKV and the closely related dengue virus (DENV) infection in skin. Infected placental cells, including fetal-derivedamniotic epithelial cells (AmEpC), trophoblast progenitor cells (TBPC),placental fibroblasts (HPF),umbilical veinendothelial cells (HUVEC)and differentiating cytotrophoblasts(CTBs) showed cytopathic effects, expressed ZIKV envelope and non¬structural NS3 proteins, and titers of infectious progeny depended on cell type, receptors expressed,individual donors and gestational age.Indicative of infection route, the decidua (decidual cells, invasive CTBs), chorionic villi (HPF, Hofbauer cells, blood vessels) and amniochorionicmembranes (AmEpC,TBPC) expressed the receptors. Nonetheless, differential expression was foundin 26 biopsy specimens, in particular TIM1 was detected throughout pregnancy, but Axl was absent in late gestation. In summary, the results suggest that ZIKV could infect the pregnantuterus and spread to the placenta, fetus and amniochorionic membranes. The models of placental infection we have developed can be used to understand molecular mechanisms of ZIKV infection and assess the therapeutic potential of antibodies and small molecule inhibitors to block infection in the placental-fetal unit.

Biography:

Annika C Karlsson completed her PhD at the Karolinska Institutet in December 2000 and continued her postdoctoral studies at the Gladstone Institute of Virology and Immunology, University of California San Francisco. In 2005 she returned to Karolinska Institutet working as an independent research scientist, senior researcher and group leader. Dr Karlsson has published 38 original articles (12 as last author) in peer reviewed journals.

Abstract:

During chronic HIV-1 infection, CD8+ T cells display a loss of effector functions. These dysfunctiona CD8+ T cells also up-regulate inhibitory molecules, such as PD-1, 2B4, CD160, a process referred to as T cell exhaustion. Recently a new inhibitory molecule, T cell immunoglobulin and ITIM domain (TIGIT), expressed on T cells was shown to inhibit their function. In addition its co-stimulatory receptor CD226 and ligan PVR is severely altered in in chronic viral infections and human cancers. However, it remains to be identified if the TIGIT/ CD226/PVR axis is dysregulated during HIV infection and linked to CD8+ T cell exhaustion. We found an increased expression of TIGIT on bulk and HIV-specific CD8+ T cells that was linked to expression of PD-1, CD160 and 2B4. On HIV-specific cells the increase in TIGIT expression was coupled to a decreased expression of CD226. Furthermore, TIGIThi cells was less common in elite controller subjects, compared to treatment naïve and long-term treated subjects and the TIGIThi cells were linked to a decreased functional capacity (IFN-γ, TNF, granzyme B & CD107a). As TIGIT needs to bind its ligand, PVR, in order to inhibit T cell function, we next measured PVR expression on CD4+ T cells in blood and lymph node. PVR expression was elevated on CD4+ T cells, especially T follicular helper (Tfh) cells which is a major reservoir for HIV-1. In summary, the TIGIT/CD226/PVR axis is altered in HIV-infected subjects, a process which was linked to a decreased functional capacity as well as an upregulation of PVR on Tfh cells. These findings highlight the importance of the TIGIT/CD226/PVR axis in the context of HIV-infection and demonstrate an immune checkpoint barrier that potentially could hinder future cure approaches mediating cellular killing of HIV-1 infected Tfh cells.

Biography:

Fiona McPhee completed her D.Phil. in Organic Chemistry from Oxford University and postdoctoral studies on coxsackieviruses at the Max-Planck Institute (Martinsreid, Germany) and on HIV at the University of San Francisco (United States). She is the Discovery Head for Clinical Virology at Bristol-Myers Squibb where her current research focus includes HBV and HCV drug resistance. She has published more than 80 papers in peer-reviewed journals, over 80 communications at international scientific meetings, and has 7 patents.

Abstract:

The most prevalent Hepatitis C virus (HCV) genotype (GT) globally is GT-1 (46%) with GT-1b being the most common subtype (22%). In Europe, GT-1b accounts for approximately 50% of all HCV cases. Daclatasvir (DCV)-based regimens are approved in many countries globally, including Europe, United States and Asia, for the treatment of HCV. In patients infected with HCV, response rates to DCV-based regimens can depend on the regimen, GT, and pre-treatment nonstructural 5A (NS5A) resistance-associated polymorphisms (RAPs) to DCV. The effects of NS5A RAPs at L31 or Y93H on sustained virologic response (SVR) to treatment with DCV+asunaprevir (ASV; NS3 protease inhibitor) for 24 weeks or treatment with DCV+sofosbuvir (SOF; NS5B inhibitor) for 12 weeks were explored using pooled data from clinical studies in HCV GT-1b-infected patients. SVR with versus without baseline NS5A RAPs was compared by age (<65 vs.>65 years), cirrhosis status, and baseline viral load. Baseline NS5A sequences were available for 1224 GT-1b patients treated with DCV+ASV and 28 GT-1b patients treated with DCV+SOF. NS5A RAPs at L31 or Y93H were observed pretreatment in 4% or 11% of patients, respectively. The overall SVR rate in DCV+ASV-treated patients without NS5A RAPs at L31 or Y93H was 95% compared to 40% with these RAPs. SVR rates in DCV+ASV treated patients without RAPs was high irrespective of cirrhosis, age, or baseline viral load. The overall SVR rate in DCV+SOF-treated patients was 100%. These results will be discussed.

Biography:

Sheldon Stone is a Stroke Physician and Acute Care of the Edlerly Specialist with a research interest in infection control. He has been funded to carry out several national studies on evaluating the national hand hygiene campaign(doi: 10.1136/bmj.e3005), universal MRSA screening (DOI: http://dx.doi.org/10.1016/S1473-3099(15)00417-X), and the national RCT of the behaviourally designed Feedback Intervention Trial (FIT) for hand hygiene (PLoS ONE 7(10): e41617. doi:10.1371/journal.pone.0041617). Lead author of the ORION statement (www.idrn.org/php) and was secretary to the working party for national guidelines for prevention and control of Clostridium difficile infection . One wife, 3 children, 2 cats , 1 football team (Tottenham).

Abstract:

Background: Sepsis has a mortality rate of 40 %. This is halved if the evidence-based “Sepsis-Six” care bundle is implemented within 1h. UK audit shows low implementation rates. Interventions to improve this, often designed using plan-do-study-act (P-D-S-A) cycles, have had suboptimal effects. The aim of this presentation is to show how behavioural science tools were used to modify such an intervention that had achieved 60% implementation of Sepsis-Six with a view to increasing implementation to the hospital’s target of 95%. Methods: Factors influencing implementation were investigated using the TDF (Theoretical Domains Framework) to analyse interviews with 34 health professionals. The data was used to select modifications using the Behaviour Change Technique (BCT) Taxonomy and the APEASE criteria. Results: Five themes were identified as influencing implementation and guided intervention modification (1) “knowing what to do and why” (2) “risks and benefits” e.g. fear of harming patients through fluid overload versus belief in “Sepsis-Six”’seffectiveness (3) “working together” e.g. team collaboration versus doctor/nurse conflict (4) “empowerment and support” e.g lack of confidence of staff to challenge colleagues’ decisions not to implement (5) “staffing levels” e.g. shortages of doctors at night preventing implementation. The modified intervention consisted of two additional components (Sepsis-Six training for the Hospital at Night Co-ordinator; partnership agreement endorsing staff-engagement and collegial challenge) and modifications to two existing educational components Conclusions:This work demonstrates the feasibility of using the TDF and BCT Taxonomy to modify an existing Sepsis-Six implementation intervention and their compatibility with P-D-S-A cycle approaches to quality improvement.

Biography:

Jae Woong has completed his Mater at the age of 30 years from Jun-Nam Universityin Korea. He is working at Catholic-University in Korea at PhD course. His major is Molecular-biology and Viology.

Abstract:

Noroviruses (NoVs) is the dominant etiological agent of acute gastroenteritis in humans and recognized as a major ethiologic agent of nonbacterial acute gastroenteritis in all age groups worldwide. Furthermore, variants and recombinant strains of this virus are continuously emerging worldwide. These variants could be related to antigenic variations that alter viral transmission and immune systems in human bodies, thus influencing the patterns of viral activities. Therefore, studies of the genetic diversity and evolution of human NoV could provide important information that may prove useful for controlling human NoV infection. And the full genome sequence analysis of NoVs is important to be able to pursue of sporadic gastroenteritis in the world by NoV.

Here, we determined the full-length sequences of a recombinant NoV strain and uniqe NoV strain isolated from clinical samples in South Korea. Because these strains may result in hazardous NoV outbreaks in Korea, this information should prove to be valuable. The results from this study highlight the many challenges in the identification of new recombination strains and suggest that guidelines be applied for identifying newly emerging recombinant strains of NoV.

Biography:

Giada Frascaroli is part of Ulm University Medical Center, Germany

Abstract:

Human cytomegalovirus (HCMV) is an enveloped double stranded DNA virus member of the Herpesvirus family. HCMV has a very high seroprevalence among the human population and infects 50% to 100% of individuals depending on the socio-economic conditions. Although the course of infection is mainly asymptomatic in normally healthy individuals, HCMV infection in immunocompromised persons can lead to life-threatening complications, like gastrointestinal disease, hepatitis, or pneumonia. Importantly, HCMV is also the most frequent viral cause of malformations in newborns, leading to deafness or mental retardation. HCMV can infect a broad spectrum of cells in the human body including central cells of the immune system such as monocytes, dendritic cells and macrophages (Mφ). Despite their broad equipment of pathogen sensing and scavenging molecules, Mφ are susceptible to HCMV infection and support viral persistence and dissemination in the human body. Since Mφ are also a first line of defense against pathogens and key modulators of the innate and adaptive immune responses, these cells represent an essential switch between protection or viral persistence and pathogenesis. In this talk I will provide an overview about the manipulative strategies used by HCMV to escape the immune response explaining the viral proteins and mechanisms that impact 1) antigen presentation, 2) cell migration and 3) cytokine secretion. I will present several examples of the immunoevasive strategies used by HCMV to manipulate the immunological properties and functions of Mφ. Moreover, by introducing newly developed experimental systems and approaches, I will provide the last data showing the cross-talk existing between Mφ and essential cells of the innate immune system such as natural killer (NK) cells as well as cells of the adaptive immune system such as T lymphocytes.

Biography:

Qamar Gulzar is a part of Avalon University School of Medicine, Curaçao

Abstract:

Herpes simplex viral (HSV) keratitis is a common and serious external ocular infection leading to unilateral blindness primarily because of its recurrent nature. Despite considerable progress in understanding of the virus at cellular and molecular levels, the proper management of the disease in its different stages is still a dilemma particularly whether to use antiviral or steroids or both. The risk of using steroids with its attendant complications has to be weighed against the risk of progression of the disease if avoiding the use of steroids. This dilemma can be reduced to a considerable extent if basic principles of virology and pathogenesis are kept in mind. This article reviews current concepts of virological and clinical aspects of HSV keratitis to enable a broad understanding of the disease process. It is recognized several influential host factors including the fact that HSK is more common in men than women.It is observed that the ability of HSV to establish latent infection in sensory neurons and possibly cornea, but have as yet been unable to use this knowledge to prevent the disease limitations. Acknowledging limitations may further stimulate application of laboratory knowledge in coping with HSK which constitutes to present major challenge in terms of management.

Biography:

LuminiÅ£a Smaranda Iancu has completed his PhD at the age of 35 years from UMP “Gr. T. Popa” Iasi, Romania and postdoctoral studies from: Royal Free Hospital, London, University Hospital Groningen, The Netherland, and Biological Molecular Laboratory Innere Medizin II, Universt-sklinikum Freiburg, Deutschland. She is the head of Regional Center of Public Health Iasi, and of Microbiology Discipline from Faculty of Medicine and Vice rector of UMF “Gr. T. Popa” Iasi. She has published more than 50 papers, 11 of them in reputed journals.

Abstract:

Since 2009 in the Virology Laboratory of Microbiology Discipline we run Human Papilloma Viruses (HPV) genotyping tests by Linear Array HPV Genotyping test. In the first step we obtained WHO proficiency for our laboratory, after best results that we obtained for a panel with HPV samples. In a country in which the prevalence of endocol cancer is in the first place in Europe (23.9/100.000) we tested more than 500 woman for HPV genotype and our results show that 192/514 (37.4%) patients were positive for HPV (infected with single and with multiple HPV types). Most frequent types were: 16 (10.5%), 53 (5.44%), 51 (5.05%), 52 (4.08%) 18 (2.91%) and 31 (2.73%) (Virol J, 2011). In an other study we test HPV and HIV coinfection: DNA/HPV was detected in 18/40 (45%) of the HIV+ patients, and in 350/992 (35.2%) of the HIV- patients. We detect HIV being a risk factor for acquiring multiple HPV type infections. The eight most common HPV types for women below age 30, HIV+/- were: HPV 16, 18, 31, 51, 58, 68, 6 and 82 respectively (PLOS ONE, 2015). In the last years we were focused to test HPV infection related with different neck and head cancers; till know, even we tested more than 200 samples, we do not find a significant association between HPV and these types of cancers. An interesting cases was related with squamous cell carcinoma of conjunctiva, in which we find that 52 HPV type was involved (Indian J Ophthalmology, 2015).

Biography:

Sominina Anna has completed her PhD at the age of 29 years from the Institute of Experimental Medicine and postdoctoral studies from the Research Institute of Influenza. She was awarded the academic title of professor in1992. She is the head of WHO National Influenza Center and of the Laboratory of Biotechnology of the Research Institute of Influenza. She has published more than 120 papers in reputed journals.

Abstract:

The unusalrapid growth of influenza activity caused by A(H1N1)pdm09 viruses was registered simultaneously in number of geographically remote areas of Russia during the season 2015-2016.Intensity of epidemic was much higher in comparison with previous four years. Significant increase of incidence rate,hospitalization and number of lethal cases observed since week 3,2016.peak of epidemic registered already on the week 5.Such a rapid speard of the epidemic indicated high transmission level of the virus. Many factors can influence this process including changes in the antigenic properties of virus HA, certain changes in internal genes, as well as decrease of population immunity . Antigenic analysis of viruses circulated showed that all A(H1N1)pdm09 viruses were antigenically related to A/California/07/09. The contribution of the rest influenza and B viruses last season was insignificant: proportion of isolated influenza A(H1N1)pdm09, A(H3N2) and B viruses estimated as 93.2%, 2.2% and 4.5%. According to sequencing and phylogenetic analysis HA gene of all A(H1N1)pdm09 viruses belonged to 6B clade (A/South Africa/3626/2013-like), sub-clade 6B1 with substitutions S84N, S162N, I216T located outside of known antigenic sites. Substitution D222G revealed in HA of some H1N1pdm09 sequences from autopsy samples. Whole-genome sequencing of influenza A(H1N1)pdm09 viruses revealed specific mutations in internal genes NEP,NS1,NP,M1 and PA-X common to most of the strains (A. Komissarov et al.,2016). Decrease of population immunity level in pre-epidemic period (October 2015) registered in number of cities of Russia by determination of the GMT of antibiotics and percent of people seropositive to A(H1N1)pdm09 virus.

Biography:

Gavin Screaton is the Dean of Faculty of Medicine at Imperial College London. After completing his first degree from Cambridge he went on to Oxford to complete his medical studies and DPhil in General Medicine. He is a member of the MRC Strategy Board, Academy of Medical Sciences, Association of Physicians, a Fellow of the Royal College of Physicians and sits on funding committees at the Wellcome Trust and MRC.

Abstract:

Dengue is a rapidly emerging, mosquito-borne viral infection, with an estimated 400 million infections occurring annually. Of these approximately one quarter are clinically apparent or symptomatic. The majority of these result in a self-limited, but none the less unpleasant febrile illness, dengue fever. 1-5% of infections lead to a more severe disease, dengue haemorrhagic fever, which is characterized by a severe vascular leak, hypovolaemia and in extreme cases shock and haemorrhage. Dengue exists as four highly divergent serotypes differing in sequence by some 30-35%; infection with one serotype does not provide protection against the other three. In endemic areas serotypes frequently co-circulate and repeat infections are common. Interestingly, severe disease is much more common in secondary as opposed to primary infections, implying a role of the acquired immune system in disease pathogenesis. Understanding this immune enhancement of disease is crucial for the design of safe and effective vaccines. Through clinical collaborations in Thailand and Vietnam, we have been studying the immune response to dengue in cohorts of infected children. We have characterized 145 human monoclonal antibodies (mAbs) and identified a previously unknown epitope, the envelope dimer epitope (EDE) that bridges two envelope protein subunits that make up the 90 repeating dimers on the mature virion. We will describe the antibody response to the two virion surface glycoproteins prM and E and discuss the E dimer epitope (EDE), a novel site bridging the 90 basic head to tail envelope dimers making up the virion surface. The antibodies directed to the EDE are both potent and broadly neutralizing across the dengue serocomplex suggesting the EDE could be a target for a new generation of subunit vaccines against dengue.

Biography:

Maria Rita Gismondo is part of CLIMVIB – L.Sacco Fatebenefratelli University Hospital, Italy

Abstract:

On March 29, 2016, WHO Director-General declared the end of the Public Health Emergency of International Concern regarding the Ebola virus disease outbreak in West Africa. A total of 28 610 confirmed, probable, and suspected cases have been reported in Guinea, Liberia, and Sierra Leone, with 11 308 deaths since the onset of the Ebola outbreak (WHO) . The majority of these cases and deaths were reported between August and December 2014, after which case incidence began to decline as a result of the rapid scale-up of treatment, isolation, and safe burial capacity in the three countries. Seven countries (Italy, Mali, Nigeria, Senegal, Spain, the United Kingdom, and the United States of America) have previously reported a case or cases imported from a country with widespread and intense transmission. In order to effectively interrupt remaining transmission chains and manage the residual risks posed by viral persistence, WHO, as lead agency within the Interagency Collaboration on Ebola and in coordination with national and international partners, designed the phase 3 Ebola response framework. The first appeared in 1976 in 2 simultaneous outbreaks, one in what is now, Nzara, South Sudan, and the other in the Ebola virus causes an acute, serious illness which is often fatal if untreated. Ebola virus disease (EVD) Yambuku, Democratic Republic of Congo. The latter occurred in a village near the Ebola River, from which the disease takes its name. The diagnosis of VHF is based on 3 components: 1. History of exposure 2. Detailed clinical assessment 3. Laboratory investigations. Evidence from outbreaks strongly indicates that the main routes of transmission of VHF infection are direct contact (through broken skin or mucous membrane) with blood or body fluids, and indirect contact with environments contaminated with splashes or droplets of blood or body fluids. Avoiding transmission dictates strict adherence to standard precautions as well as droplet and contact precautions for health care, environmental, and laboratory workers. Moreover, while epidemiology during VHF outbreaks does not suggest airborne transmission, precautions should be taken to avoid procedures or protect health workers, family members and other patients during procedures that might aerosolize virus. The Ebola epidemic showed the need to be better prepared in order to face efficiently the next major health emergency. Elements such as coordination, risk assessment processes and intersectoral cooperation are paramount for a good preparedness planning. The Ebola outbreak also highlighted the need for action in areas which usually do not always get sufficient attention, such as border issues (exit and entry screening), medical evacuation, the mobilisation of specific expertise for EU and the affected countries, transport facilities for big amount of waste related to the laboratory and clinical activities in the EU, the sample sharing and contact tracing. A. shows international and Italian experience.

Biography:

Silvia Angeletti obtained the Degree in Biological Science at the University “La Sapienza” of Rome. She was specialist in Microbiology and Virology on November 1998. On October 2008 she obtained the Degree in Medicine and Surgery at the University Campus Bio-Medico of Rome, Italy. She is the Director of the Clinical Pathology and Microbiology Division at the University Hopsital Campus Bio-Medico and the Director of the Reaserch Unit of Clinical Pathology and Microbiology at the University Campus Bio-Medico of Rome, Italy. He has published more than 60 scientific papers in international journals and has been serving as a reviewer and an editorial board in reputed journal as Disease markers or PlosOne. Area of research interest are the following: epidemiology and diagnosis of infectious disease; phylogenetic analysis applied to viral and bacterial infections; Multidrug resistant (MDR) pathogens analysis in nosocomial settings; diagnostic and prognostic markers associated to infectious and chronic diseases; key indicators of laboratory quality.

Abstract:

Migrant populations are represented by group of people forced to migrate by the difficult conditions of life experimented in their Country of origin as a consequence of civil wars and political instability. The onset of civil war can led to the complete deterioration of the health infrastructure through the destruction of facilities, the shortage in health care personnel and medicines other than a lack of secure routes and transportation. These conditions joined with the deterioration of immunization programs induced the spread of communicable diseases like measles, poliomyelitis, meningitis through the migrant populations creating a fertile condition for the epidemic spread of unsual infections also within the refugee camps of the hosting Country. The refugees camps are a sort of community where almost 900 migrants and refugees stay for a medium of 15 months and for this reason the monitoring by a sentinel surveillance the people arriving so as their health status before and after the entrance in the camps it is noteworthy. At this aim we present data from the microbiological surveillance of migrants coming from Syria and Eritrea at their arrival in an asylum seekers Centre in Italy to define their health status and to evidence the need for strategic policy of assistance to people that have to travel within Europe facing many kilometers again. In conclusion the microbiological surveillance represents a useful action to understand refugees health status and to trace unusual microorganisms movement even carriers of antimicrobial resistance during migrants travelling.

Biography:

Niels Høiby MD DMSc a Professor of Medical Microbiology at the University of Copenhagen and at department of Clinical Microbiology at Rigshospitalet in Denmark. Prof. Høiby was the first president of the European Cystic Fibrosis Society and was the founder and Chairman of multiple societies, including the Scandinavian Society for Antimicrobial Chemotherapy, the ESCMID study group of Biofilms and the Biofilm Study Group of the Danish Society for Clinical Microbiology. He has published extensively in the area of chronic lung infection in CF and other biofilm infections. Prof. Høiby has won multiple awards and honors throughout his career.

Abstract:

Cystic fibrosis (CF) patients suffer from recurrent and chronic sinus- and lung infections due to the basic defect of the CFTR protein, which is a chloride channel. This leads to decreased volume of the paraciliary fluid in the airways and impaired mucus detachment which interfere with the mucociliary transport and the consequence is therefore defective host defense against bacterial infections. P. aeruginosa is causing the most important chronic lung infection in CF and it was the first biofilm infection which was described in human beings and the most well studied biofilm infection. The inflammatory response - dominated by polymorphonuclear leukocytes - to the chronic infection is the main cause of the lung tissue damage. The antibody response against the sinus biofilm infections is mainly s-IgA which reduce the inflammation, whereas IgG dominates against the lung biofilm infection and aggrevates the inflammation. The antibody response is used diagnostically. The current treatment is early aggressive eradication therapy of intermittent P. aeruginosa colonization to prevent chronic biofilm infection which is treated by chronic suppressive ’maintenance’ therapy to maintain the lung function for decades. Both systemic and inhaled antibiotics are used.

Biography:

Iolanda Francolini has obtained a degree in Industrial Chemistry in 2000 from the Sapienza University of Rome. In 2003, she was a visiting scientist at the Center for Biofilm Engineering, Montana, USA. In 2005, she obtained a Ph.D. degree in Chemical and Industrial Processes at the Sapienza University of Rome. She currently serves as a Lecturer in the Science and Technologies of Polymers at the Sapienza University of Rome and performs research on antimicrobial polymers. She has published more than 45 papers in reputed journals and has been serving as an editorial board member for International Journal of Molecular Science.

Abstract:

The impact for public health of medical device-related infections has received considerable attention over the last decade. Clinical signs of most of these infections clearly suggest that they are caused by microbes colonizing the implant surface and nearby tissues. The subsequent microbial biofilm formation aids the development of antibiotic resistant microrganisms and compromises the device functionality. A revision surgery is therefore frequently required adding healthcare costs to those already required for taking care of the infection. The enormous advances achieved in the last decades in polymer sciences together with therapeutic innovations hold the promise to meet the need of improving patient experience associated with device implants. Particularly, novel, high-performance polymer systems with antimicrobial or antifouling properties have been lately developed. The application of these materials as coatings for medical devices has demonstrated in some cases to offer a protection towards microbial colonization and biofilm formation. In this talk, an overview of innovative materials and technologies facing with biofilm-based medical device-related infections will be provided and how our group is addressing this issue will be presented.

Biography:

Trine Rolighed Thomsen has completed her PhD at the age of 32 years from Aalborg University and was appointed Associate Professor in Medical Microbiology in January 2012. She is affiliated to Aalborg University and the Danish Technological Institute and heads a research group. Main interests and research grants are on diagnosis, prevention and treatment of human infections, with a special focus on biofilm. She has published more than 40 peer-reviewed original papers and is active member of several danish and international boards and sicentific societies.

Abstract:

Recent evidence suggests that the microbial community, its spatial distribution and activity play an important role in the prolongation of treatment and healing of chronic infections. Standard bacterial cultures often underestimate the microbial diversity present in chronic infections. This lack of growth is often due to a combination of inadequate growth conditions, prior usage of antibiotics and presence of slow-growing, fastidious, anaerobic or unculturable bacteria living in biofilms. Thus, diagnosis of chronic infections is challenged by lack of appropriate sampling strategies and by limitations in microbiological testing methods. The purpose of this study was to improve sampling and diagnosis of chronic infections, especially considering the biofilm issue. Systematic and optimized sampling of various specimen types was performed. Extended culture, optimized DNA extraction, quantitative PCR, cloning, next generation sequencing and PNA FISH were applied on different types of specimens for optimized diagnosis. For further investigation of the microbial pathogenesis, in situ transcriptomics and metabolomics were applied. Molecular techniques detected a larger diversity of microorganisms than culture methods in several patients and a heterogeneous distribution of bacteria in various specimens from the same patient was evident. Data from a 2-year prospective study on prosthetic joint infections including 164 patient cases will be presented. Transcriptomic and metabolomic analyses indicated the important virulence genes and nutrient acquisition mechanisms of Staphylococcus aureus in situ. Our studies show that diagnosis of chronic biofilm related infections required multiple specimen types, standardized sampling, extended culture and molecular analysis.

Biography:

Claudia Vuotto has obtained her PhD in Biomedical sciences, Medicine and Public Health at Marche Polytechnic University Medical School. She won a fellowship at the Italian National Institute of Health and she is currently researcher at the Microbial Biofilm Laboratory of the Fondazione Santa Lucia in Rome. She has published 18 articles on international peer reviewed journals and 2 book chapters for Springer. She was in the Local Organizing Committee in the International Congress "Eurobiofilms 2009" and in the Scientific Committee of "Eurobiofilms 2015" as well as Scientific Secretary of the ESGB Meeting "Biofilm-based Healthcare-associated Infections: from Microbiology to Clinics"(2015).

Abstract:

The increasing number of anaerobic bacteria isolated from different clinical samples has led to a growing awareness of their contribution in acute and chronic infections. These bacteria, in fact, are able to become opportunistic pathogens after that a permissive environment within the host have been created or in presence of other predisposing factors. In these conditions, infections in which are involved anaerobic bacteria, often together with aerobes, arise. However, despite the increasingly frequent isolation of anaerobic species during infectious processes and the demonstrated ability of some of them to form biofilms, their involvement in the establishment of healthcare-associated infections (HAI) is still undervalued and thus little studied. The current knowledge about the implication of biofilm-forming anaerobic bacteria in some human infections will be examined, and the data acquired in our laboratory by means of Quantitative Biofilm Production Assay, Field Emission Scanning Electron Microscopy and Confocal Laser Scanning Microscopy will be presented. In particular, data obtained on the synergistic and/or competitive interactions within multi-species biofilms among different anaerobic species, including Clostridium difficile, Veillonella spp, Peptostreptococcus magnus, Porphyromonas gengivalis, will be summarized and discussed. Information on the involvement of some anaerobic species in causing difficult-to-treat infections as part of polimicrobial biofilms and on the ability of commensal microorganisms, including Lactobacilli, to antagonistically operate against anaerobic pathogens will be provided.

Biography:

Claus Moser is part of University of Copenhagen, Denmark

Abstract:

Biofilm infections are significant clinical challenges. They are frequent,can be related to foreign bodies or be tissue related, and are recalcitrant to host responses and antibiotic treatments. Furthermore, they can be primary focus for systemic spread, they are persistent and characterized for recurrent infections, especially since timing of termination of antibiotic treatment can be exceedingly difficult. The biofilm mode of growth can also render low virulent microorganisms into virulent strains due to the induced inflammation, which generates tissue damage.Microbiologically, biofilms are defined by a selfproduced matrix containing extrapolysaccharides, eDNA and proteins. Therefore, biofilms can be considered as independent compartments with distinct antibiotic PK/PD parameters. Adding to this, physiological gradients are observed in biofilms with significantly different nutrition and oxygen conditions in different zones of the biofilms.Diagnostically, biofilm growing microorganisms can be challenging to demonstrate by traditional culturing due to adherent and/or extremely slowly growing microorganisms (persisters/dormant types/viable but non-culturable microorganisms).Finally, susceptibility testing is only indirectly representative for the responsiveness of the biofilm to antibiotic treatments. Therefore, the first clinical guidelines for diagnosing, prevention and treatment of biofilm infections have been published in 2015. The present abstract aims at presenting novel therapeutic regimens published since the guidelines became available last year. The focus of the presentation will be on consequences of the biofilm as a distinct compartment, hyperbaric oxygen treatment, IgY gargling and human autologous leucopatches. The presented therapeutic possibilities are involved in ongoing clinical trials or used as adjuntive treatment of other medical syndromes.

Biography:

PalinaVyhouskaya has a master degree in LaboratoryMedicine, and currently she is a PhDstudent at the Jagiellonian University, Poland. She is anactive member of The Scientific Students Association of Laboratory Diagnosticians and Clinical Microbiology, where she experiences and practices modern research techniques in Medicine. She has been involved in various research programs, and her work was published in international journals.

Abstract:

Background
In therapeutic context, glycolytic pathway is an interesting field of research also for studying dental diseases. Metabolism of biofilm-forming Streptococcus mutans––main factor of caries progression––is based on glycolysis even in the presence of oxygen. In a such condition, an increase in the expression of genes coding key glycolytic enzymes, pyruvate kinase (PK) and phosphofructokinase (PFK), is observed. Modification of enzyme activity of PK and PFK gives an ability to inhibit bacterial growth and biofilm formation––a potential approach for caries prevention and therapy.

Material and methods

Activity of PK and PFK from double- and mixed-species biofilm was estimated spectrophotometrically. Biofilm assay was carried out according to Current Protocols in Microbiology using ATCC and clinical strains (Streptococcus mutans,Streptococcus sobrinus, Lactobacillus acidophilus,and Actinomycesviscosus). In this study, we measured the activity of PK and PFK in various pH values and formation time of double and mixed-species biofilms.

Results

The activity of PK and PFK increased after 6 and 12 hours of a double-species biofilm formation (1.43 mU/mg of protein vs. 1.52 mU/mg of protein) compared to 18 and 24 hourswhen we observed a slight decrease in the activity of the glycolytic enzymes (1.37 mU/mg of protein vs. 1.48 mU/mg of protein). In case offorming the mixed-cariogenic biofilm, the activity of glycolytic enzymes also grew after 18 and 24 hours of mixed-species biofilm formation and the differences were statistically significant (p<0.001).

Conclusions

The increase in the activity of glycolytic enzymes (PK and PFK) during the biofilm formation (due to the effect of low pH), can be explained by the need to overcome the inhibitory effect of acidification on the metabolic activity of the microorganisms in the biofilm. Bacterial cells adapt to new conditions better in mixed-species biofilm than in the mono- or double-species biofilms––the increase of theglycolysis rate associated with increased activity of glycolytic enzymes reflects this phenomenon very well.

Therefore, inhibition of glycolytic enzymes might be an essential step in the reduction of mixed-species cariogenic biofilm, what could be a useful tool in prevention of caries.

Biography:

Professor Bill Keevil completed his PhD at the University of Birmingham and postdoctoral studies at University of Southampton School of Biological Sciences. He is Director of the Environmental Healthcare Unit and Head of the Microbiology Group. Has published more than 200 papers in reputed journals and is a Chartered Biologist with 40 years of experience of Microbiology and Biofilms. Former Specialist Advisor to the House of Commons Science & Technology Committee; Fellow of the Royal Society of Biology, Fellow of the Royal Society of Medicine and Fellow of the American Academy of Microbiology. Winner of the Colgate Prize.

Abstract:

Horizontal gene transfer (HGT) conferring resistance to many classes of antimicrobials has resulted in a worldwide epidemic of nosocomial and community infections caused by multidrug-resistant microorganisms, leading to suggestions we are returning to the pre-antibiotic era. Whilst studies have focussed on HGT in vivo, this work investigates whether the ability of pathogens to persist in the environment, particularly on touch surfaces, may also play an important role. Here we show prolonged survival of multidrug resistant Escherichia coli and Klebsiella pneumoniae on stainless steel surfaces for several weeks. Plasmid mediated HGT of β-lactamase genes conferring resistance to third generation extended spectrum beta lactams and fourth generation carbapenems occurred to an azide-resistant recipient E. coli if donor and recipient cells were mixed together on contemporary stainless steel surfaces and in suspension but not on copper alloy surfaces. This transfer occured almost instantaneously, even when the surface was dry and the pathogens survived many days of dry contact, providing an environmental hygiene risk and a reservoir for the acquisition and dissemination of new antibiotic resistant strains. In addition, rapid death of both antibiotic-resistant strains and destruction of plasmid and genomic DNA was observed on copper and copper alloy surfaces which could be useful in the prevention of infection spread and gene transfer in the healthcare and public transportation environments, particularly where cleaning and disinfection practice is not 24/7.

Biography:

Margitta Dathe studied physics at the Humboldt University of Berlin and completed her PhD in 1978 from the Academy of Sciences of the GDR. Since 1999 she has been working as head of the peptide-lipid interaction research group of the Leibniz Institute of Molecular Pharmacology. Her research interest is focused on targeting, cellular uptake promoting and antimicrobial peptides. She has published more than 100 papers in reputed journals.

Abstract:

The synthetic cyclic hexapeptide cWFW (cyclo(RRRWFW)) is very stable towards proteolytic degradation, low pH and has a high activity against different Gram positive and Gram negative laboratory strains and clinical pathogens, including S. aureus and L. monocytogenes. With its amphipathic structure and high content of arginine residues, the peptide combines the prerequisites for membrane permeabilisation and membrane translocation as modes of action [1-3]. Using a number of techniques to study peptide interaction with bacterial and eukaryotic model membranes as well as with bacterial and eukaryotic cells, we could show that the activity of cWFW is based on a novel antimicrobial mechanism. Strong interactions with the bacterial membrane lead to reduction in membrane fluidity and disturbance of the native lipid matrix. The formation of distinct lipid domains is related to a severe disturbance in the positioning of functional proteins which finally leads to cell death. While the peptide does not enter the cytoplasm of bacteria, cWFW is rapidly internalized into human cells without decreasing cell viability. The combination of cell penetrating properties with high antimicrobial activity and the novel mechanism of action renders the cyclic hexapeptide an eligible compound with regard to the treatment of intracellular bacterial infections, as e.g. in the case of tuberculosis and pneumonia

Biography:

Alex Pym is an Investigator at AHRI (African Health Research Institute) in Durban, South Africa. Trained in Clinical Medicine, Infectious Diseases, Microbiology, and Mycobacterial Genetics he completed his PhD research at the Institute Pasteur, Paris, followed by post-doctoral studies at Stanford University. He has a broad research experience, and has published on Esx secretion systems, recombinant TB vaccines, and population genetics of M.tuberculosis. He subsequently worked in South Africa on clinical drug development of bedaquiline in MDR-TB patients. Now at AHRI, his laboratory is focussed on understanding the biological basis of resistance and persistence in M. tuberculosis and defining biomarkers of treatment response.

Abstract:

The continued advance of antibiotic resistance threatens the treatment and control of many

infectious diseases. This is exemplified by the largest global outbreak of extensively drug resistant

(XDR) tuberculosis (TB) identified at Tugela Ferry, KwaZulu-Natal, South Africa, in

2005. It is unclear whether the emergence of XDR-TB was due to recent inadequacies in TB control or other factors. Using whole genome sequencing on clinical and historical isolates we demonstrate the evolution of drug resistance over a 50 year period and identify a common mutational pathway responsible  for multiple episodes of de novo evolution. By combining association and correlated evolution tests with strategies for amplifying signal from rare variants, we were also able to identify new mutations associated with resistance. One of these was a loss-of-function mutations in ald (alanine dehydrogenase) which conferred resistance to D-cycloserine and emerged independent of antibiotic selection in the evolution of animal adapted strains of the M. tuberculosis complex. 

Biography:

Alexandre Thibodeau has completed his PhD at the University of Montreal and is currently doing postdoctoral studies at the NSERC industrial research chair in meat safety in Dr Ann Letellier laboratory at the Faculty of veterinary medicine of the University of Montreal. He is a specialist of Campylobacter jejuni and his postdoctoral research focuses mainly of C. jejuni and the chicken microbiome.

Abstract:

Antibiotic resistance development is a challengefor bothhumanand veterinary medicine. Enterotoxigenic Escherichia coli (ETEC: F4) associated withpost-weaning diarrhea (PWD) in pigs has developed resistanceagainstseveralantimicrobial families, leading to an increase use of colistin sulfate (CS) for the treatment of this disease.The objective of this study was to determine the efficiency of oral CS treatment in experimental PWD due to ETEC: F4 challenge and determine the effect of this therapeutic regimeninE. coli resistance apparition. In this study, 48 pigs were divided intofour groupsof12pigs each: challenged treated, challenged untreated, unchallengedtreatedand unchallenged untreated group. Fecal ETEC: F4, total E. coli population, CS-resistant E. coli shedding were evaluated. The MIC was carried out by microdilution method using a sterile 96-well polystyrene microplate. CS treatment resulted in areductionof fecal ETEC: F4 and E. coli population shedding butonly during the treatment period. However, CS treatment resulted in an increase in fecal shedding of CS resistant E. coli. Results indicated thatsomeE. coliisolateswereconfirmedresistant to CS. Our study is among the first to demonstrate that under controlled farming conditions, CSwas effective to reducefecal shedding of ETEC: F4 and total E.coli population in experimental PWD. However, CS treatment was associated with a selection pressure on E. coli. Further studies areneededinfield conditions, to better characterizeCS E. coli resistance dissemination in meat and in the environment.

Biography:

MLG Daroy is Scientist at the Research and Biotechnology of St. Luke’s Medical Center and Assistant Professor in the MS Molecular Medicine Program of the St. Luke’s College of Medicine-WHQ Memorial. She has published more than 20 papers on dengue, Japanese encephalitis, chikungunya, eye infections, dementia, diabetes, and coronary artery disease. She was Chair of the Board of Examiners of the Philippine Academy for Microbiology from 2013-2015, and authored a book chapter on Philippine microbiology research. Researches include dengue, chikungunya, diarrhea, CNS infections, pathogen genomics, antimicrobial resistance, plant antivirals, molecular diagnostics, and genetics of CVD, thyroid cancer, and dementia. Word Count: 100

Abstract:

The Philippines is a hotspot for the emergence of antibiotic-resistant “superbugs,” such as fluoroquinolone-resistant E. coli and carbapenemase-carrying Klebsiellapneumoniae. This paper reports on the use of next-generation sequencing and bioinformatics techniques for the the genome-wide analysis of genetic variation influoroquinolone-resistant E. coli, carbapenem-resistant K. pneumoniae, ESBL-positive K. pneumoniae, and a carbapenem-resistant Stenotrophomonasmaltophila. These bacteria were isolated in 2013-2014 from clinical specimens of patients confinedat St. Luke’s Medical Center in Manila, Philippines. De novo assembly, primary structural and functional annotations, gene search and SNV analyses performed on pooled reads identified several resistance determinants. Plasmid-borne New Delhi β-lactamase genes, blaNDM-1 and blaNDM-7, were found in K. pneumoniae, as well asstop codon mutations in the ompK35 and ompK36 porin genes.SNVs in the genomes of susceptible and fluoroquinolone-resistant E. coli were identified in scaffolds derived from pooled reads, which were then mapped against reference sequences of two susceptible and one fluoroquinolone-resistant E. coli. In silico filtering of SNVs using a subtraction strategy selected for variants that may be associated with fluoroquinolone resistance in 23 genes involved in transport, respiratory chain, oxidative stress, iron metabolism and bacterial cell death. The functional association of these variants and putative pathways offluoroquinolone resistance involving them were derived by mining gene annotations. This strategy of combining whole genome analyses with in silico identification of significant variants and searching gene ontologies for functional associations is useful to guide functional genomics studies on the development of antibiotic resistance in common clinical pathogens like E. coli and K. pneumoniae.

Biography:

Viktoryia has graduated  fromVitebsk State Medical University at the age of 23. She is the doctor in Vitebsk Regional Hospital. She has published more than 68 papers in reputed journals and other scientific papers.

Abstract:

Objective: to investigate the etiological structure and antimicrobial resistance of  pathogensisolated from patients with ventilator-associated pneumonia.
Materials and methods.We examined sputum of 49 patients with ventilator-associated pneumonia. The criterion for selection of patients was the duration of mechanical ventilation for at least 48 hours.
Results.Staphylococcus aureus was found in 33.3% specimens, Acinetobacter spp.  –in33.3%, Pseudomonas aeruginosa–in 15.4 %, Klebsiella pneumoniae- in 7.7%, Candida– in 12,8%, Streptococcus spp.–in 2,6% specimens.Staphylococcus aureus was sensitive to vancomycin in 100% of casesand  to amikacin in 38.5 % of cases. All strains  ofStaphylococcus aureus were methicillin-resistant. 100% of strains were resistant to ofloxacin, ciprofloxacin, levofloxacin, 92,3% - toclindamicin.

Isolates of Pseudomonas aeruginosawere sensitive to colistinin 100% of cases, to  amikacin in 16.6% of cases.All isolates of Pseudomonas aeruginosa were resistant to meropenem, imipenem, cefepime, ceftazidime, cefoperazone, ciprofloxacin.
46.2% of isolates of Acinetobacter spp. were sensitive to ampicillin in combination with sulbactam.92.3% of isolates of Acinetobacter spp. were sensitiveto cefoperazone-sulbactam. Acinetobacter spp. was resistant to amikacin, meropenem, imipenem, levofloxacin, cefoperazone, cefepime, ciprofloxacin.
All isolates of Klebsiella pneumoniae were sensitive to imipenem, 66.7% of Klebsiella pneumoniae- to meropenem and amikacin. Klebsiella pneumoniae was resistant to amoxicillin, cefotaxime, ceftazidime, ciprofloxacin, cefoperazone, ofloxacin in 100% of cases.
Conclusions.
1. Modern epidemiological feature of ventilator-associated pneumonia is the prevalence in the etiological structure of gram-negative microflora, represented mainly by Acinetobacter spp.
2. It is recommended to use vancomycin or linezolidagainst Staphylococcus aureus, colistin  againstPseudomonas aeruginosa,cefoperazone-sulbactam against Acinetobacter spp. and imipenem against Klebsiella pneumoniae.

 

Biography:

Demet Çelebi has complated university Ataturk University Biyology Department, MSC; Atatürk University Medical Faculty Microbiolgy, PHD; Atatürk University Medical Faculty Microbiolgy .Working now Ataturk University Veterinary Faculty. She has got studies about; Effect of Kefir upon the Performance, Intestinal Micro flora and Histopathology of Certain Organs in Laying Hens, Comparison of Surface Swab and Quantitative Biopsy Cultures Dependent on Isolated Microorganisms from BurnWounds, Prevalence of metallo-β-lactamase among Pseudomonas aeruginosa and Acinetobacter baumannii isolated from burn wounds and in vitroactivities of antibiotic combinations against these isolates and posters presentation four in Turkey, one in Cyprusandone in Germany about different microbiology topics.

Abstract:

Aim: The increase in antibiotic-resistant bacteria has dramatically revived the interest in plant products as alternative anti microbial agents to prevent the efficiency of pathogenic microorganisms. Our aim in this study is to show the anti microbial activities of cinnamon (Cinnamomum cassia), tarragon (Artemisiadra cunculus) and ginger (Zingiber officinale) oils against the clinically important bacteria and yeasts. Materials and method: The anti microbial activity of the cinnamon, tarragon and ginger oils were tested against Citrobacter spp., Klebsiella spp., Proteusmirabilis, Pseudomonas, MRKNS, Enterococcusspp., Staffilococcusaureus, Enterobacteraerugenes, Candida spp and E.coli. Essential oils of cinnamon,tarragon and ginger were extracted by steam distillation, final yield of extraction 0.5, the volume of EO extract was adjusted to 100 ml with sterile distilled water, thus obtaining the crude essential oil extracts used for anti microbial tests. Disc diffusion method (Kirby-Bauer) was used to show the anti microbial activity by measuring the zone diameters. Turbidity was visually adjusted to that of a 0.5 McFarland turbidity standard (1.5x 10 ~ CFU/ml) using sterile Mueiler-Hinton broth and after 24 hours. 0.1 ml for each oils, Sterile filter paper disks were prepared to a diameter of 6.35 mm and sterilized in a Pasteur-oven, (at 170~ for 2 hour). Volatile oils and extracts were sterilized by passing through 0.22 mm pore-size membrane filters and then 0.02 ml of the solution of volatile oils was pipetted into the center of each disk to achieve the desired potency. Disks were air-dried in a contamination free environment. Results:The cinnamon oil show an anti microbial activity against the tested micro organisms with different zone. Citrobacter 25mm, Klebsiella 18mm, P.mirabilis 23 mm, Pseudomonas 15mm, MRKNS 22mm, Enterococcus 70 mm, S.aureus 32 mm, Enterobacter 19 mm, Candidaspp 18 mm and E.coli 10 mm. Ginger and tarrogon oil wasn’t show an anti mirobial activity against bacteria and candida. Conclusion:.The cinnemon oil has a stronger anti microbial activity than the tarragon and ginger oils. Herbal essential oils are candidates to be alternatives in medical applications due to their anti-microbial effects.

Biography:

ÇiÄŸdem Eda Balkan has complated university acettepe University Biyology Department in 2009, MSC; Atatürk University Medical Faculty Microbiolgy 2011, PHD; Atatürk University Medical Faculty Microbiolgy 2016. She has got studies about;Investigation of Children Aged 0-5 in Erzurum Rotavirus and Adenovirus Frequency (2012), Turkish Society of Microbiology journal, Effects of Boiling Dairy Products on Human Brucellosis(2013) (Eurasian Journal of Medicine), Antibacterial Activity of Rose Thyme, Centaury and Ozone Oıls Against Some Bacterıas (2015) (Kafkas Journal of Medical Sciences) and six posters presentation four in Turkey, one in Cyprusandone in Germany about different microbiology topics.

Abstract:

In our study, our target is to review the bacterial drug resistance from blood samples of chickens in different antibiotic diets. We used 4 groups of 20 chickens. Each group was put on diet for 30 days with Tetracycline (30mg/kg), Penicillin (20mg/kg), Sulfonamid (30mg/kg) or Floroquinolone (40mg/kg) group of antibiotics. The sensitivity of E.coli, Stapycoccus spp. , Citrobacter spp. , Klebsiella spp. was observed in the mediums prepared from blood samples of the groups and compared with the results of previous studies. Compared with the previous sensitivity; it is observed that Trimethoprime resistance was present in E.Coli which passaged in the antibiotic feed chickens blood media. According to our results in this study, it was understood that bacterial resistance may be seen at the chickens when the chickens are feed with antibiotics due to that these antibiotics are available at their bloods.

Biography:

Neveen Saleh has completed her PhD at age of 34 years from Ain Shams University and has got postdoctoral studies from Massachusetts, Amherst (UMASS) University in Nanotechnology, Chemistry department. She is now amicrobiological researcher in National organization for drug control and research. She has published more than 10 papers in reputed journals.

Abstract:

Antibiotics have been widely used for the treatment of infectious disease owing to their ability to specifically target bacterial cells. However, aworld-wide increase in the disease/infections caused by Multi-Drug Resistant (MDR) bacteria has become a great threat to public health. Synthetic materials such as polymers and nanoparticles exhibit broad spectrum activity against bacterial species, but NPs lack specificity and can cause toxicity to mammalian cells leading to restrictions on their use. The combination of antibiotics and NPs is a promising therapeutic approach to combat MDR bacteria as it can reduce the requirement of high dosages as well as enhance their antimicrobial activities. Conventional methods for the treatment of MDR infections involve acombination of different antibiotics. However, they are susceptible to lose their efficacy due to increasing antibiotic resistance. Here, we hypothesize that combining the specificity of antibiotics and broad spectrum activity of NPs could yield synergistic combinations for antimicrobial therapy against MDR infections. However, engineering of NPs can increase the selectivity of combination therapy. In order to test this hypothesis, we used hydrophobic functionalized C12-AuNPs that can strongly interact with bacteria. C12-AuNPs in combination with fluoroquinolone antibiotics displayed 4-fold decrease in the MIC values against MDR E.Coli. Synergy arising from combination therapy was attributed to the ability of engineered NPs to act as efflux pump inhibitors. This was validated by theaccumulation of EtBr inside the bacterial cells upon incubation with NPs. Moreover, using proteomics analysis these cells exhibited altered expression of outer membrane proteins which are responsible for several drug resistance mechanisms such as efflux pump channels, porins, and lipoproteins.

Biography:

Marina Tediashvili graduated from Tbilisi State University.PhD workconductedat Eliava Institute of Bacteriophages inTbilisi andIvanovsky Institute of Virology in Moscow. PhD degree awarded by TSU in 1980. Postdoctoral studies:Institute of Medical Enzymology in Moscow and Cancer Reserch Center in Tbilisi. The major part of Dr.Tediashvili’s career has been linked to the Eliava Isntitute of Bacteriophages, where shecurrentlyruns the Laboratory for Microbial Ecology.Her scientific interests are Medical and Environmental Microbiology with main focus on phage research and phage therapy, development of biocontrol strategies of bacterial infections in humans, animals and plants.Dr.Tediashvili is professor of Microbiology at the University of Georgia and Tvilidiani Medical University in Tbilisi.

Abstract:

One Health concept, asa worldwide interdisciplinary strategy, targets the health issues linked to the increasing contacts between humans and animals, intensification and integration of agriculture and food production, expansion of international travel, bioterrorism threats, climate change, ecological stressors etc. Many infectious pathologies have been shared by humans and animals, some phytopathogenic bacteria can evolve as human opportunistic pathogens, the changing conditions in aquatic environments can trigger the development of more virulent strains of waterborne bacteria potentially pathogenic to humans. Bacterialvirusesarewidely spread in the biosphere, controlling the diversity and density of bacterial populations in various ecosystems. During up to 100 years history of phage research naturally occurring lytic phages have been found for majority of important pathogens of humans, animals and plants. Thisprovides a viable resource for development of effective phage-based control strategy consistent with “ One Health” approach. Emergence and re- emergence of life- threatening drug- resistant bacterial infections in the last two decades gave a new insight at the potential of bacteriophages as alternative to antibiotics, capable to eliminate or significantly reduce the use of chemical antimicrobials in various spheres of human activity. The presentation will cover the valuable experience in human phage therapy accumulated at the Eliava Institute of Bacteriophages as well as recent research developments aimed at phage application for control of bacterial diseases in livestock and aquaculture, for food safety and crop protection, also environmental decontamination.

Biography:

Rao Ph.D, worked as Professor of Micro biology ,Parasitology , immunology and Epidemiology in many Universities in India, China, Nepal, Libya, and Philippines . Currently I am working at Avalon University School of Medicine, Curacao, Netherland Antilles. I have more than 40 years of teaching and research experience. Supervised 3 students forPh.D, 8 students forM.Phil, 3 students submitted their Ph.D thesis to the Acharya Nagarjuna University, Guntur ,AP, India and waiting for degree. Authored 18 text books. Three Universities appointed me as their advisor and three universities with fellowships.

Abstract:

Warm countries are the worm countries". Parasitic infection is a global health problem especially in developing countries. Emerging parasitic diseases are those diseases that have been newly appeared or that have existed in the past but are now rapidly increasing in frequency, geographical range or both. Emerging zoonotic diseases hamper both human and animal health and cause economic loss. The ecosystem is increasingly and continually being put in to turmoil by human acts such as deforestation and global warming with the resultant alteration in the distribution and behavior of parasites and their vectors. Ecologic changes, demographic changes, behavioral changes, travel and immigration are also contributed in emerging and reemerging parasitic infection. These infections will continue to emerge and reemerge leading to unpredictable epidemics and challenges for the clinicians /scientists. Hence there is urgent need of surveillance and control. Dracunculus is almost eradicated but it is reemerging. African trypanosomiasis, Cutaneous Leishmaniasis, emerging as new out breaks. High level of innovation triggered on genomics, RNA biology,proteomics,genomics, nanotechnology,nanoscience, and animal models. The strategic focus is on technology development in the areas of bio therapeutics, cell therapy, vaccines, diagnostics, bio markers,advanced pharmaceutical sciences and clinical research for improvement of health and free from parasitic diseases. Challenges and risks in governance management certainly prevent the reemerging parasitic infection.

Biography:

Patricia Poeta, is Full Professor at UTAD, Portugal. PhD in Veterinary Medicine and specialist in Medical Microbiology. Coordinator of the Research Group of Antibiotic resistance and head of Medical Microbiology Laboratory. Director of several doctoral and masters’ theses. Also responsible for research projects funded by private companies and inter-university projects, teaches International courses and Master’s degree. Has published more than 100 articles in journals from the SCI, book chapters, communications at international scientific meetings. Received 16 awards.

Abstract:

The aim of this study is to synthesize and investigate the antibacterial properties of functionalized AgNPs-TET and AuNPs-TET complexes, evaluating their antimicrobial effect on antibiotic resistant bacteria with public health concerning. Strains tested were E. coli ST648 and S. aureus ST398, both resistant to tetracycline, and the control strains E. coli K12 and S. aureus ATCC 25923, susceptible to tetracycline. MIC and MBC tests were performed on LB medium. Control cultures without nanoparticles and/or tetracycline were included in all experiments. Our results are supported by innumerous previous studies reporting that AgNPs are exquisite bactericides and, as AuNPs, they both are suitable antibiotic carriers (Brown et al., 2012; Li et al., 2014; Wang et al., 2014). Nevertheless, direct comparisons between our results from other studies are difficult because the physicochemical properties of nanoparticles used differ, since the functionalization of nanoparticles in this study was made in aqueous phase instead of silica matrix, and the protocols to assess antimicrobial activity diverge among studies. We compared the potential of AuNPs and AgNPs used as tetracycline delivery systems and showed evidences that both agents succeed on inhibiting bacterial growth. Also, AgNPs functionalized with tetracycline had an inhibitory and bactericidal ability even against Gram-negative and Gram-positive tetracycline-resistant bacteria. The functionalization of nanoparticles with antibiotics could be a potent prospective therapeutic method to overcome infections caused by antibiotic-resistant bacteria. Future studies should focus on functionalization of nanoparticles with other antibiotics, testing their antimicrobial efficiency against a broader spectrum of pathogens, prevention on biofilm formation on in vivo and in inert substrates.

Biography:

Ségolène Caboche has completed his PhD in Computer Sciences in 2009 from Lille University and postdoctoral studies from European Bioinformatics Institute (UK) in 2009-2010. Her research interest is focused on high-throughput sequencing data. She is the author of 11 international publications and a regular reviewer for several high-impact internantional journals. She joined the Transcriptomics and Applied Genomics team in 2011 and co-advised PhD students in bioinformatics.

Abstract:

Recent advances in high-throughput sequencing (HTS) technologies open new perspectives in diagnostic microbiology (management of infected patients) and in epidemiology of infectious diseases. The Whole-Genome Sequencing application has significantly increased our knowledge of microorganism genomes and has facilitated investigations in clinical microbiology, but from concept to routine use, some bottlenecks still need to be unlocked. One of these is the step of data analysis which requires bioinformatic expertise, needs to be reproducible and is often time consuming. Here we present MICRA, a freely available on-line pipeline to quickly and automatically characterize microbial genomes from HTS data. MICRA requires only the raw sequencing data (fastq) as input and then automatically finds the most closely related genomes based on sequence similarity (called reference genomes) and establish lists of discrepancies in term of genes and plasmids content and in term of variations compared to reference genomes. Optional modules allows the user to obtain Venn diagrams to quickly compare the gene content and/or the variations list between several strains. Another module identifies drug susceptibility and resistance genes and generates an e-antibiogram of the analyzed strain. All the steps included in MICRA were optimized in time. For example, for a PGM Ion Torrent run containing more than 2 million of 200-bases reads, the pipeline requires only 15 minutes to generate a full analysis. Diagnostic microbiology and epidemiology of infectious diseases, will benefit from such pipeline as it defines a new standard of genome-based diagnosis combining results from a former time-consuming and heterogeneous multiple step method.

Biography:

Rossella Colomba Lelli has completed his PhD at the age of 24 years from Bologna University, Italy and postdoctoral studies from Pisa University, School of Veterinary Medicine. She is the Laboratory Head of Epidemiology Dpt, IZSAM, a Ministry of Health Gov. organization. She has been working long time on Italian foreign animal disease, i.e. FMD, West Nile Disease, Contagious Bovine Pleuropneumonia, Bluetongue and Brucellosis epidemics. She has experience on research coordination on animal infectious diseases and food safety, good laboratory practices and Quality Assurance. She has published more over 150 papers in international scientific journals.

Abstract:

In Italy, the Veterinary Public Health concept, linked to “Human-animal-ecosystems interface”, International Cooperation and Contribution to the economy of Countries, is connected with the history of the Veterinary Medicine Faculties’ curricula, the Veterinary Services and their organization, under the umbrella of Ministry of Health. This is the main reason to have the consumers as the main focus of the Veterinary activities. At international level, the concept of Veterinary Public Health has evolved over time, and today is considered as "the sum of the contributions to the physical, mental and social development of people through the knowledge and application of veterinary science" (WHO, 1999). On the subject of Cooperation, Sustainability and Public Health, it is necessary to take into account the EXPO 2015 event, and the activities of international organizations such as the WHO, the FAO and the World Organization for Animal Health (OIE). These organizations are focusing their mandate on the defense of Health worldwide, on the interrelation Man-Animal-Environment, and on the aim to protect the public health and the economy in the poorest Countries to also protect the health and the economy of the developed Countries. It is made a brief analysis of the activities that the Istituto Zooprofilattico Sperimentale "G. Caporale” (IZSAM), Italy, has put in place for International Cooperation during his mandate, as a Government Institute of the Ministry of Health. The main focus are infectious diseases, food safety, microbiological studies and research.

Biography:

Waleed Al momani has completed his PhD in 2005 from Kings College London University. Currently he is an associate professor of diagnostic microbiology at department of allied medical sciences, Balqa; Applied University, Jordan. He has published more than 27 papers in reputed journals and has been serving as a reviewer of international peer reviewed journals.

Abstract:

The field of coordination chemistry has registered a phenomenal growth during last few decades. It is well known that precious metals have been used for medicinal purposes for at least 3500 years. At that time, precious metals were believed to benefit health because of their rarity, but research has now well established the link between medicinal properties of inorganic drugs and specific biological properties. The current study was designed to explain the synthesis and characterization of the lanthanide (III) nitrate complexes with N-(2-hydroxynaphthalen-1-yl) methylene) nicotinohydrazideschiff base and to evaluatethe antibacterial and the antioxidant activities of the schiff base and it's lanthanide ion complexes.Antimicrobial activity of the Lanthanide (III) nitrate complexes with N-(2-hydroxynaphthalen-1-yl) methylene) nicotinohydrazideschiff base was estimated by minimum inhibitory concentration (MIC, µg/mL) using a micro-broth dilution method for different clinical isolates such as Eschereshia coli and Enterococcus faecalis. Our present study has shown that moderate antimicrobial activity exists against both ligand and its complexes. There was no significant difference between Gram-positive and Gram-negative bacteria towards the tested ligand and its complexes. The results obtained herein indicate that the ligand and its complexes havea considerable antibacterial activity

Biography:

Lucio Souza Gonçalves is Associate Professor of Periodontology at the Estacio de Sá University (Post-Graduate Program – Master and PhD) and Research Affiliate in Oral Microbiology at the Federal University of Rio de Janeiro (UFRJ - Brazil). He received his Dental Degree (1988), Master’s degree in Periodontology (2000), and PhD in Oral Microbiology (2006) from the UFRJ - Brazil. He also completed a Postdoctoral Research Fellowship at the University of Otago (2014) in a collaborative research with Dr. Gregory J. Seymour and Dr. Nicholas Heng on a study involving oral microbiome in HIV-infected Brazilian children using an Ion Torrent sequencing platform.

Abstract:

The periodontal microbiota is composed of a highly complex bacterial multispecies community organized in biofilms. The composition of the periodontal microbiota in HIV- infection has demonstrated a higher prevalence of periodontal pathogens in non-HIV-infected than HIV-infected individuals. However, microorganisms not commonly associated with periodontitis have frequently been detected in subgingival sites of HIV-infected patients, including Staphylococcus epidermidis, Candida albicans, Enterococcus faecalis, Clostridium difficile, Mycoplasma salivarium, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumanni, Dialister pneumosintes and Entamoeba gingivalis. In particular, E. faecalis has been more prevalent in the subgingival microbiota of HIV-infected patients with reduced levels of TCD4+ lymphocytes (<200 cells/mm3), suggesting that HIV-related immunodeficiency can provide appropriate conditions for the colonization and growth of opportunistic pathogens that are unusual in the oral microbiota. In addition, it has been demonstrated that detectable plasmatic HIV viral load (PHVL) in HIV-infected patients is associated with elevated levels of known periodontal pathogens such as Prevotella nigrescens, Tannerella forsythia, and Eikenella corrodens, as well as Campylobacter concisus, Capnocytophaga gingivalis, and Dialister pneumosintes in the subgingival biofilm, and that these associations are not related with CD4+ T lymphocytes levels. Another interesting microbiological finding is that HIV can be found in the subgingival biofilme. In a recent study, the detection and quantification of HIV in subgingival biofilme of 20 individuals with detectable PHVL, 40% showed detectable subgingival HIV viral load (SHVL). On the other hand, all patients with undetectable PHVL demonstrated also undetectable SHVL. This can be influenced by the CD4+ T lymphocytes levels.

Biography:

Zhiheng Pei obtained PhD in Microbiology and Immunology and did residency in Anatomic Pathology and fellowship in Molecular Pathology at Vanderbilt University School of Medicine. He is currently a tenured Associate Professor of Pathology at New York University School of Medicine and Staff Physician at the Department of Veterans Affairs New York Harbor Healthcare System. In the past several years, he has been evaluating and pioneering a new concept - microecological disease in several NIH-sponsored projects involving cancers in the mouth, esophagus and stomach as well as disease in tonsils.

Abstract:

For unclear reasons, the incidence of esophageal adenocarcinoma (EA) arising out of Barrett’s esophagus (BE) and reflux esophagitis (RE) has risen more than 600% in the United States since the 1970s. Although specific host factors might predispose one to disease risk, such a rapid increase in incidence must be predominantly environmental. The widespread use of antibiotics since 1950s could have contributed to this drastic change. Antibiotic exposure prior to 1980s could have unintentionally eradicated Helicobacter pylori which plays a protective role against EA, BE, and RE. Both human and animal studies suggest that exposure to antibiotics changes the colonic microbiome in favour of obesity. Case control studies showed microbiome is altered in the distal esophagus in patients with reflux disorders including EA. The microbiome in esophageal diseases is more diversified than in controls. This effect is not only seen in the esophagus but also observed in the mouth and stomach. Overall, esophageal diseases tend to be associated with depletion of Gram-positive bacteria and enrichment of Gram-negative bacteria. The relative abundance of Streptococcus, the most abundant Gram-positive bacteria in the foregut, tends to decrease along the disease progression from normal to reflux, Barrett’s esophagus, and adenocarcinoma, in both the mouth and esophagus. Microbiome alteration often extended to the mouth and sometimes to the stomach and rectum. The altered microbiome could play a more direct role in the initiation and progression of reflux disorders than H. pylori or obesity by in situ induction of chronic inflammation and/or activation of carcinogens.

Biography:

Liying Yang obtained Doctor of Medicine from Jiamusi University School of Medicine in China and Master of Science in Clinical Research from New York University. Currently, she is Assistant Professor of Medicine at New York University School of Medicine. She serves as Principal Investigator and Co-investigator on a number of NIH-sponsroed projects studying the role of microbiome in HIV infection and cancers and has published more than 30 papers in reputed journals.

Abstract:

To identify microbial targets for treatment of HIV enteropathy, we contrasted microbiota composition between HIV-1-infected patients and HIV-negative controls in the esophagus, stomach, and duodenum as well as the mouth using a universal 16S rRNA gene survey and correlated the findings with HIV serostatus and peripheral blood T-cell counts. HIV infection was associated with an enrichment of Proteobacteria and depletion of Firmicutes in the proximal gut. In particular, environmental species Burkholderia fungorum and Bradyrhizobium pachyrhizi colonized the duodenum of HIV patients who had abnormally low blood CD4+ T-cell counts but were absent in HIV-negative controls or HIV patients whose CD4 counts were normal. The two species coexisted and exhibited a decreasing trend proximally towards the stomach and esophagus and were virtually absent in the mouth. Their abundance was inversely correlated with CD4 counts but not viral load. The colonization of the duodenum by environmental bacteria reflects loss of colonization resistance in HIV infection. Their correlation with CD4 counts suggests that compromised immunity could be responsible for the observed invasion by exogenous microbes. These findings provide unprecedented insight into a mechanisms guiding future efforts to develop therapeutic interventions in HIV infection, such as antibiotic treatment aimed at eradicating the species that are associated with an increased gastrointestinal opportunistic infections, or alternatively, combined with probiotics to introduce the species that are associated with a decreased the diseases risk.

Biography:

After spending some years as formal employee of Federal Government Laboratories in Canada Kamaleldin Said has moved to McGill University in Montreal where he completed his PhD in molecular microbiology and genomics. He then continued at McGill to work on projects involving molecular typing and comparative genomic hybridizations of pathogens shortly before accepting a contract position at Qassim University, College of Pharmacy as assistant professor. He has published significangly on the development and validation of molecular markers for host- and organ-specific strain typing as well as genome-wide expression profilings for vaccine and therapetuic gene candidates. He is now a visiting scholar at Systems Biology, Carleton University and has many projects underway and many manuscripts ready for publicaton.

Abstract:

Despite enormous efforts, the molecular mechanisms of specialization and emergence of invasive methicillin-resistant Staphylococcus aureus (MRSA) in a clonal background has been quite elusive. In an effort to contain the devastating outbreaks, significant reseach has been conducted that included many contraverses and inconsistent reports. We have previously shown that a reason for this to occur when genotypes used for vertical analysis were determined based only on the DNA pattern of the marker irrespective of its function, the host, and the strain factors. Furthermore, major genome based subtyping processes often fail to recognize minor differences in clonal genomes. We have established in recent years through multicenter collaborations that the use of adaptation-sensitive repeats in subtyping followed by subsequent genome wide expressions yielded relavant data on host and clonal differentiations. In this study, applicatoin of clfA- R-domain genotyping of clinical Staphylococcus aureus isolates revealed 14 different repeat types (RTs) designated, A to L, and Q and X while eight sequenced strains belonged to 3 types A, C, and Q. The most dominant types were D (24 isolates, 57 copies), X (19 isolates, 52 copies), B (13 isolates, 60 copies), E (11 isolates, 55 copies), and F and Q (8 isolates 49, 47 copies each). Interestingly, the genotypes showed long-variable and short-clonal copy numbers reflective of the length and functional properties of clfA in human-specific and animal-associated lineages, respectively. Furthermore, presence of resistant phenotypes of the global genotype X among human isolates suggested potential transmission of bovine lineage to human hospital. Thus, we have shown the dominant types as well as the potential transmission lines of MRSA in a tertiary care hospitals. Future vertical analysis and genome-wide sequence expression profiling of host-specific lineages would reveal potential candidates for vaccine and therapeutic developments.

Biography:

Maria Hedberg completed her PhD in 1995 at the Karolinska Institute, Stockholm, Sweden where also the postdoctoral studies took place. From 2005 she has been situated as a researcher at Umeå University, Sweden. Main research interests are anaerobic bacteria in health and disease and antimicrobial resistance. In 2010 she became Associate Professor of Experimental Bacteriology at Umeå University. She has published more than 50 papers in peer reviewed journals and are at present serving as an editorial board member of the journal Anaerobe.

Abstract:

Prevotella sp. are strictly anaerobic Gram-negative rods constituting a substantial part of the normal human micro-flora. About 50 species have been described at present, of which a great part are of oral and upper airways origin, while others have been isolated from the gastrointestinal and urogenital tract, and some species origin from animal sources. Prevotella sp. often occur in opportunistic infections and dysbiosis-associated diseases, but may also be involved in severe infections and various virulence factors have been demonstrated. The new species Prevotella jejuni was discovered from a biopsy taken from the small intestine of a child with coeliac disease. Three different strains of the species were isolated and phylogenetic analysis based on 16S rRNA gene sequence analysis revealed a close relationship between them. When grown on blood agar plates the three P. jejuni strains were pigmented and weakly or strongly hemolytic. They were able to form saccharolytic and proteolytic enzymes and by flow cytometry they were shown to bind to human intestinal carcinoma cell-lines in suspension. Scanning electron microscopy (SEM) and transmission electron microcopy (TEM) displayed formation of aggregates in which tube-like structures were connecting between individual cells and outer membrane vesicles (OMVs) could also be observed. Prevotella sp. are in general considered to be susceptible to bile but in the case of P. jejuni the growth was stimulated by sub-inhibitory concentrations of bile, suggesting an adaption to the hostile milieu of the small intestine. The strains have been subjected to whole genome sequencing (WGS) using 454-pyro-sequencing technology.

Biography:

Su has completed residency training in Anatomic Pathology at the age of 31 years and in Clinical Pathology two years later; and obtained the Master degree from the Institute of Biomedical Engineering, National Cheng Kung University, Taiwan when he was 32 years old. At present, he is the attending physician of Departments of Anatomic Pathology and Clinical Pathology, Buddhist Dalin Tzu Chi Hospital, and the Associate Professor of Departments of Laboratory Medicine and Pathology, Tzu Chi University, Taiwan. He has published more than 40 papers in reputed journals and has been serving as an editorial board member of repute

Abstract:

Many patients with tuberculosis (TB) are seropositive for human herpesvirus type 8 (HHV-8), and many patients with primary effusion lymphoma have high levels of HHV-8 DNA in their effusions. However, the status of HHV-8 in the effusions of patients with TB remains unclear. Blood samples were collected from 129 patients with pulmonary TB and 129 age- and sex-matched healthy controls. Forty of the TB patients had pleural or peritoneal effusions, and 38 of these effusions were available. Both blood and effusion samples were analyzed for lymphocyte and monocyte counts and/or HHV-8 antibodies and DNA. TB patients with or without effusions had significantly greater HHV-8 seropositivity (P = 0.009) and titers of HHV-8 antibodies (P = 0.005) than healthy controls. The seropositivity and blood titers of HHV-8 antibodies were similar in TB patients with and without effusions. Among TB patients with effusions, similar percentages had seropositive plasma and seropositive effusions. Plasma samples of 6 TB patients, but none of the healthy controls, were positive for HHV-8 DNA (P = 0.03). TB patients with or without effusions had lower blood lymphocyte counts and higher blood monocyte counts than healthy controls (P < 0.0001 for both). TB patients with effusions had significantly lower blood lymphocyte counts than those without effusions (P = 0.035). TB patients with and without effusions had similar and greater HHV-8 seroprevalence compared with healthy controls. However, TB patients with effusions had lower blood lymphocyte counts than those without effusions.

Biography:

A molecular virologist for over 30 years, and Professor in the Department of Cell and Tissue Biology at the University of California San Francisco Lenore Pereira has focused the last 16 years on the biology of human cytomegalovirus replication and pathogenesis at the uterine-placental interface. Dr. Pereira has published over 120 papers and invited reviews. Recently, her group in collaborative studies with Dr. Eva Harris at the University of California Berkeley identified ZIKV target cells and immune mechanisms that protect the placenta.

Abstract:

Human cytomegalovirus (HCMV) is the leading viral cause of birth defects, including neurological deficits, impaired hearing and vision loss. We previously reported that epithelial cells in amniotic membranes of placentas from newborns with intrauterine growth restriction and underlying congenital infection contain HCMV proteins in cytoplasmic vesicles. Here we immunostained amniotic membranes from diagnosed symptomatic congenital infection and preterm deliveries and detected viral proteins and aberrant epithelial cell morphology in cases of virus transmission. Primary amniotic epithelial cells (AmEpC) infected with pathogenic viral strainsdysregulated stem cell proteins, and viral replication was gestational age dependent. In contrast to highly differentiated retinal pigment epithelial cells, infected AmEpC failed to make intact viral assembly compartments and instead formed diffusely localized multivesicular bodies, proliferated and survived for months releasing progeny without plaque formation. Explants of amniochorionic membranes infected ex vivo upregulated IFN- in surrounding epithelial cells. Infected AmEpCs produced IL-6 and upregulated the anti-apoptotic proteins survivin and Bcl-xL by mechanisms dependent and independent of the activated signal transducer and activator of transcription 3. Both survivin and Bcl-xL were expressed by control and infected amniotic membranes in utero, suggesting an opportune environment to sustain persistent infection. Interventions that target signaling pathways contributing to HCMV persistence in AmEpC could reduce the viral load and inflammation in the fetal compartment and improve outcome.

Biography:

Alexandre Thibodeau has completed his PhD at the University of Montreal and is currently doing postdoctoral studies at the NSERC industrial research chair in meat safety in Dr Ann Letellier laboratory at the Faculty of veterinary medicine of the University of Montreal. He is a specialist of Campylobacter jejuni and his postdoctoral research focuses mainly of C. jejuni and the chicken microbiome.

Abstract:

Campylobacter jejuni is a foodborne pathogen mainly associated to chickens, reaching caecal concentration up to 109 CFU/g. Scarce reports mention clinical signs associated to C.jejuni chicken colonization, making it a bacteria acting like a commensal. One aspect of our work focuses on the characterization of the ability of C.jejuni to colonize the chicken caecum. C. jejuni chicken strains were characterized for their ability to autoagglutinate, to be attracted to mucins and to adhere or invade chicken primary caecal cells; these phenotypes having individually been linked to the ability of strains to colonize chickens. In parallel, a microarray was developed to screens the strains for genes that could be linked to colonization. Strains were also typed by comparative genomic fingerprinting. We confirmed in vivo that strains possessing different phenotypes also possessed different abilities to compete for the colonization of the chicken gut. This ability could be associated with genes such as genes coding for arsenical resistance. Representative strains possessing the best competition abilities were also used in a serie of in vivo chickens studies to assess the efficiency of control methods to lower C. jejuni chicken caecal loads. Limited results were obtained, showing the complexity of the intestinal microbial ecosystem. The effect of the colonisation of these strains on the chicken microbiota was also assessed and we observed small disturbance of some bacterial populations. Our strain collection is currently being sequenced and further characterized in vitro to deepen our understanding of C.jejuni chicken colonization.

Biography:

Wasa Alibe Ahmed currently a PhD student at the University of Canterbury, Christchurch, New Zealand has a B.sc in Applied Microbiology and M.sc Medical Microbiology from Abubakar Tafawa Balewa University, Bauchi, Bauchi State, Nigeria and Modibbo Adama University of Technology, Yola, Adamawa State, Nigeria respectively, he has few papers published in journals and is currently working towards developing immunodiagnostic biosensors.

Abstract:

This experiment was carried out with the aim of formulating growth media using organic waste materials (banana peel and maize cob) from the environment. Banana peel and maize cob were collected from different locations within Gombe main market from various banana and maize sellers. The collected materials were air dried, grinded into fine particles using mortar and pestle and then sieved with 0.8mm sieve size. Bacteria isolates (Escherichia coli, Staphylococcus aureus, Shigella sp, Salmonella spp, Klebsiella sp and Pseudomonas aureginosa) were collected from Federal Teaching Hospital Gombe and confirmed using relevant biochemical tests and the fungi isolates were isolated from spoilt bread and orange using potato dextrose agar (PDA) and the formulated media. Proximate analysis carried out, show that the banana peel contained 18.56% moisture, 3.05% Ash, 7.20% Fat, 16.54% Protein, 15.42% crude fibre and 45.23% Carbohydrate, while the corresponding values for maize cobis 10.14% moisture, 2.86% Ash, 2.20% Fat, 14.18% Protein, 13.26% Crude fibre and 59.36% Carbohydrate. The bacterial isolates were sub-cultured onto commercially prepared nutrient agar medium and the formulated banana peel and maize cob media. The colonial characteristics of both the commercially prepared agar and the formulated media were compared for each isolate. Total bacterial count was carried out and the result showed 2.50x104, 2.12x104 and 1.95x104 on nutrient agar, 1.50x104, 1.10x104 and 1.20x104 on banana peel agar and1.85x104, 1.40x104 and 1.70x104 on maize cob agar for E. coli, Staphylococcus aureus, and Shigella sp respectively. Visible growth 0.67x104 of Salmonellawas observed on maize cob only, whileno visible growth for Klepsiella spp. and Pseudomonas aeureginosa was observed on both the banana peel and maize cob agar. Fungi were also isolated from spoilt bread and orange using commercially prepared potato dextrose and the formulated banana peel and maize cob media. Three fungi were isolated and identified; the cultural characteristics of each fungus on each medium were compared. The fungi isolated and identified with their total plate count were as follows: Aspergillus niger (3.0x104, 5.0x104 on banana peel and potato dextrose agar respectively with no visible growthobserved on maize cob agar), Rhizopus stolonofer (4.0x104, 2.2x104, 4.7x104 on banana peel, maize cob and potato dextrose agar respectively) and Saccharomycse cerivisiae (2.9x104, 1.5x104 and 3.6x104 on banana peel, maize cob and potato dextrose agar respectively). From these results, it can be proposed that banana peel agar could be used as an enrichment medium for the growth of fungi, while maize cob agar could be more useful inbacterial growth medium.

Biography:

Adnan Saeed is a PhD student at the age of 34 years Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw Poland

Abstract:

Human actinomycosis, is rare a chronic, suppurative granulomatous infectious disease, has been recognized for over a century, caused by microorganisms of genus Actinomyces species. Presently, Actinomyces species comprises 42 species, 20 of them are relevant to human medicine. Actinomycosis is defined as a hard mass-type lesion with a specific histopathological structure. There are a large number of case reports of actinomycosis in the literature, but in most cases, diagnosis has been based solely on clinical and histopathological findings. The goal of the study was to determined the chemical composition of polysaccharide antigens extracted from A. israelii and generate monoclonal antibody reactive with the polysaccharide to understand their role in pathogenicity. Polysaccharide were extracted from dry bacterial cell mass by using trichloroacetic acid (TCA) and enzymes (DNas, RNse and protease). Further were purified by ion-exchange chromatography (DEAE Sephadex A25) and gel filteration (TOYOPEARL HW 55 s). Composition and structure of polysaccharide was determined by gas liquid chromatography-mass spectrometry GLC-MS. Monoclonal antibodies were generated by the hybridoma technique. The ELISA method was carried out for evaluation the specificity of monoclonal antibody to the polysaccharide antigen. Then quantitative immunoprecipitation test has been performed. The experimental infection of mice with A. israelii was performed and investigated in histological study. Detection of the bacterial strain within the mouse tissues was done by immunohistochemical test. Scanning electron microscopy showed a variation of the branched shape of A. israelii with filamentous character of slime. Polysaccharide of A. israelii consists of glucose, galactose and mannosamine in the molar ratio 1,5,10 respectively. Two hybridomas 101 and 121 producing mAbs against polysaccharide antigen were IgM class. Quantitative microimmunoprecipitation test showed that monoclonal antibody precipitated polysaccharide antigen of A. israelii. Immunohistochemical test with monoclonal antibodies identified A. israelii infection in the liver mouse tissue. Monoclonal antibodies with polysaccharide used in immunohistochemical assays could serve as tools for diagnostic purposes in vitro approaches.

Biography:

Alessandra Lo Presti has completed her PhD in Medical Microbiology, Immunology and Infectious Diseases at the University “Tor Vergata” of Rome. She is expert in epidemiology and phylogeny of infectious diseases and practices the research activity at the Department of Infectious, Parasitic, and Immune-Mediated Diseases of the National Institute of Health, Rome. She has published about 77 papers in reputed journals and is in the editorial board member for some journals. Moreover she is the Principal Investigator of a Project that aims to evaluate the interactions between gut microflora and immune system at the cross-road of the pathogenesis of Inflammatory Bowel Diseases and Irritable Bowel Syndrome.

Abstract:

Forty-eight Syrian migrants, upon their arrival in Italy, were accommodated at the asylum seekers centre of Castelnuovo di Porto. They received a physical examination and were subjected to microbiological surveillance by blood, rectal, pharyngeal and nasal swabs collection and delivering to the Clinical Pathology and Microbiology Laboratory of the University Campus Bio-Medico of Rome. All refugees resulted negative for HBV, HCV and HIV infections. In swabs a large number of unusual gram-negative bacteria species were isolated, such as Pseudomonas putida, Pseudomonas monteilii, Pseudomonas fulva, Pseudomonas moselii, Aeromonas veronii, Aeromonas caviae, Aeromonas hydrophila, Acinteobacter guilloviae, Acinteobacter lowffii; Acinetobacter johnsonii; Acinteobacter tjernbergae; Pantoea agglomerans; Pantoea calida. Among isolates, strains resistant to carbapenems, ESBL producers and methicillin resistant were found. The microbiological surveillance performed represents a useful action to understand refugees health status and to trace unusual microorganisms movement even carriers of antimicrobial resistance during migrants traveling.

Biography:

R.A.N.Ranathunga, Senior Medical Officer at Medicine Unit, Base Hospital Kuliyapitiya, has completed her MBBS in 2010 from the Faculty of Medicine, University of Kelaniya. She is currently reading for MPhil at University Kelaniya. She has six international publications on health related topics and more than ten communications both locally and internationally

Abstract:

Urinary Ttract Infection (UTI) is one of the common bacterial infections worldwide. Management is mainly with antibiotics and the initial choice of antibiotics is mainly decided according to recommended guidelines or by personal preference and experience. The main objective was to determine the prevalence of UTI in suspected patients, common pathogen and antibiotic sensitivity pattern, at the medical wards in Base hospital Kuliyapitiya, Sri Lanka. This study was conducted for 6 months. Each and every patient with clinically suspected UTI was recruited and samples collected for full report and culture just after the recruitment. The initial step of the study was conducted in Base Hospital Kuliyapitiya. Total of 94 subjects were recruited and the prevalence of UTI among suspected patents was 68.5% with 37.7% positive culture reports. More than the half (54.1%) got at least one co- morbidity and diabetes mellitus is leading the list. The commonest pathogen was Escherichia coli (80.6%). The commonest empirical antibiotic given was Co-amoxyclave and its sensitivity is 61.9%, followed by ciprofloxacin which is sensitive only in 25% of the cultured samples. The most sensitive antibiotic was Nitrofurantoin which was found to be sensitive in 83.9% of the cases which are culture positive. In conclusion, clinical suspicion was accurate in 68.5% of cases with UTI. The commonest pathogen causing community acquired UTI in Base Hospital Kuliyapitiya is Escherichia coli and the most sensitive drug is Nitrofurantoin. Ciprofloxacin which is commonly prescribed as empirical therapy is sensitive on in 25% of cases.

Biography:

Prabha U Krishnan, MBBS, MD, Dip NB(Micro), FRCPath, DTM&H(Lond) is a Senior Consultant in Laboratory Medicine and the Deputy Head of the National Public Health Laboratory, Singapore. She is als an Adj. Associate Professor  and the Lead for Microbiology at the Lee Kong Chian School of Medicine, Singapore. Her areas of special interest are multi-drug resistant pathogens and nosocomial infections.

Abstract:

The local epidemiology of CPE has been evolving over the last decade; since 2010, this 1500-bed-hospital has adopted an expanding screening program for CPE detection.  The high-risk screening strategy has been replaced by a  combined proactive (selective on-entry screening) and reactive (contact-tracing) strategy for early identification of asymptomatic carriers. Between 2010 and 2015, the number of CPE detected on screening has increased from one to 31, genotypes from one to eight and enterobacterial species  from three to 14.  Laboratory protocols too have evolved over this period. A rectal swab (or stool sample) was initially processed using the CDC-TSB-Enrichment-Method with 10-µg ertapenem disc. Since 2013, to improve turn-around-time and sensitivity, samples were plated onto chromogenic agar (chromID^TM CARBA, bioMérieux) and colonies with characteristic colours were investigated further. From April 2016 the selective plate has been changed to chromID^TM CARBA-SMART Agar to improve detection of OXA-48-type CPE. Species identification is by MALDI-ToF-MS (Bruker Daltonics) and antimicrobial susceptibility testing by VITEK-2 (bioMérieux Vitek, Inc.), interpreted according to revised (June 2010) breakpoints set by CLSI. A few isolates harbouring OXA-48-type and IMI-1 carbapenemases were not detected with this protocol. The genotype of meropenem non-susceptible isolates was determined using a multiplex RT-PCR assay targeting Class A (KPC, IMI, GES, SME), Class B (NDM, IMP, VIM) and Class D (OXA_-48-type) carbapenemases.  No one protocol is ideal for the early detection of all CPEs; an algorithm to investigate all Enterobacteriaceae with meropenem MIC  > 0.25 mgs/L as proposed by Hrabák J. et al could lead to a more sensitive CPE detection workflow

Biography:

Abstract:

In Slovakia vaccination against measles was introduced in 1969. Firstly it began with monovalent (1969 - M), then we used bivalent (1987 – MM) and trivalent (1992 –MMR) vaccine. Our aims were to analyse vaccination coverage by MCV1 and MCV2 and their impact on measles incidence at national and regional levels. We made retrospective review of measles vaccination reported from 2000 to 2015. Cohort studies  evaluating measles vaccination coverage in 24-months old children born between 1996 and 2013 (MCV1) and 10-years-old children born between 1987 to 2003 (MCV 2) were conducted. The data were obtained from the databases of the Epidemiological Information System of the Slovak Republic and from the regular annually controls in Slovakia, 2000-2015. Since 1999, measles has been eliminated, only imported cases were reported in our country. The vaccination coverage remained at the highest levels (98.0-99.9%). In the last children cohorts (born in 2011, 2012, 2013) MCV1 decreased from 96.8% (2011) to 93.9% (2013) and MCV2 to 97.6% (2003) at national level. MCV1 at regional level was from 90.1% (2013) to 98.2% (2011) and MCV2 from 95.8% to 99.1% (2003). Children born in 2013 - only 40 (50.7%) districts reached level 95.0% and more, 31 (39.2%) reached from 90% to 94.9% and 8 (10.1%) districts had less that 90.0% vaccination coverage. Decrease of MCV2 in adolescents (2003) below 90.0% only 2 (2.5%) districts. Our analysis showed positive impact of vaccination against measles. Increasing of anti-vaccination activities and risk populations (Romany population, migrants) are the main factors affecting the vaccination coverage.

This  work was supported by the Slovak Research and Development Support Agency under the Contract No. APVV-0096-12 (EPIBIOMAT).

Biography:

Abstract:

Over the past several decades epidemiological situation in Slovakia was affectedd by vaccination using combine M-M-R vaccine (against measles, mumps and rubella). Obligatory measles vaccination started in Slovakia in 1969, against rubella in 1984 and mumps vaccination in 1987 with bivalent vaccine (against measles and mumps). In Slovakia trivalent vaccine (M-M-R) has been used since 1992. Objectives of our work were to describe the impact of vaccination strategy on mumps incidence at national level and to assess the risk factors. The data were collected from the Epidemiological Information System of the Slovak Republic (EPIS). Until 1988 a few thousands cases of parotiditis per year occurred in Slovakia. After introduction of the vaccination in 1987, a declining trend and gradual transfer of the disease to the older age groups were observed. Several hundreds of cases were  reported from 1988 to 1998, only a few dozens of cases occurred until 2006. Between 2007 and 2012 only 2 – 5 cases were reported in the EPIS. Change occurred in 2013 with incidence from 218 (4.03 / 100,000) to 1707 (31.49 / 100,000) in 2015. In the long term vaccination coverage against measles, mumps and rubella according to the years of birth is 99%.  Last controlled cohorts (2011, 2012, 2013) showed a decline in vaccination coverage to 96% at national, under 95% at regional level. M-M-R vaccine applied to the children (MCV1) and adolescents (MCV2) plays a significant role in these diseases prevention and elimination. Risk factors infulencing mumps outbreaks are a decrease in vaccination coverage and risk population groups (little children and minority population).

This  work was supported by the Slovak Research and Development Support Agency under the Contract No. APVV-0096-12 (EPIBIOMAT).

Biography:

Luis Guzman J., has worked in the area of natural products, especially in the Research and Development of Bioactive Products. He is a profesor at the Faculty of Health at the University of Talca, Chile and external consultant of Fraunhofer Chile Research in the area of Nanomedicine.

Abstract:

Bacteria demonstrate impressive effectiveness in adapting to changing environmental challenges. In contrast to most drugs in other medical fields, antibacterial drugs aim at shifting targets and, hence, lose their effectiveness over time. A novel approach for antibacterial compounds is the utilization of ionic liquids, formerly known as molten salts, constitute one of the hottest areas in chemistry these days.

Different N-styryl imidazolium salts with different alkyl size chain (1, 6 and 8 carbon atoms) were synthesized using microwave radiation, under solvent-free condition. The resulting ionic liquids were cooled at room temperature and washed with EtOAc to remove the starting reagents and concentrated under high vacuum to afford N-styryl methyilimidazolium ([SMIM]Cl), N-styryl hexilimidazolium ([SHIM]Cl) and N-styryl octylimidazolium ([SOIM]Cl) chlorides.

The antibacterial activity was tested against S. aureus, E. coli, S. epidermidis and S. pyogenes bacterial strains, using a modified Kirby-Bauer method and the minimal inhibitory concentration (MIC) was determined using concentration of the ionic liquids ranging from 7.8 μM to 1 mM.

Motivated by secure and eco-friendly protocols we reported the synthesis of different N-styryl imidazolium salts employing microwave radiation under solvent-free condition as a Green Chemistry approach. The resulting ionic liquids were soluble in MeOH and H₂O, showing a good antibacterial activity with MIC ranging from 62.5 μM to 15.6 μM depending on the bacteria studied, also a positive correlation was found between the large of the alkyl size chain (1 and 6 carbon atoms) and the antibacterial activity.

Bacteria as S. aureus, S. epidermidis, E. coli and S. pyogenes plays a widespread role in a variety of infections combined with accentuated antibiotic resistance, making these organism a significant threat to the industry and medical community, that is why that the antibacterial properties of the N-Styryl alkylimidazolium ionic liquids with specific properties as different alkyl size chain and the incorporation of a cation in the structure along with the utilization of eco-friendly synthesis, does to these compounds a promising tool for a new class of antibacterial agents.

Biography:

Nataliya Piletska is a final year medical student of King’s College London due to begin her foundation year placement in Oxford University Hospitals Trust this autumn. She has a special interest in microbiology and paediatrics, intending to continue participating in research alongside her work. Claire Stewart graduated with distinction from the King’s College London School of Medicine and has a BSc degree in International Health from University College London. She is a Paediatric Trainee at the London School of Paediatrics, currently working at the Evelina Children’s Hospital in Paediatric Surgery. She has published 10 papers, presented in 33 conferences worldwide and recently won the Georgia Fieldsend International Medical Research Award.

Abstract:

The purpose of this audit was to investigate the use of antibiotics in the management of the most common surgical emergency, acute appendicitis. Data was compiled from all appendicectomies performed over a 12-month period at the Evelina London Children’s Hospital, from Jan 2014-15. In the sample of 101 patients, 66 had been diagnosed with acute appendicitis. Of these patients, both electronic and paper-based information was collected regarding their antibiotic treatment. The most common bacteria cultured was Escherichia Coli, followed by Pseudomonas aeruginosa, then mixed anaerobes. In a third of cases, no swabs were taken. 34% of swabs taken grew no bacteria. On average patients received a total of 5 days of intravenous followed by 4 days of oral antibiotic therapy. Co-amoxiclav and Gentamicin were the most frequently used antibiotics, commonly in combination, often alongside Metronidazole as ‘triple therapy’ (Table 1). This audit studies the bacteriological epidemiology of acute appendicitis, which antibiotics are most frequently prescribed during the admission and on discharge, their route, duration, frequency and any change to the regimen either indicated by bacteriological swab results or clinical picture. The aim is to guide and improve guidelines and indications for antibiotic management.

Biography:

Mariela Srednik is doing her PhD at the University of Buenos Aires (UBA). She is veterinarian and has completed her Specialization in Quality and Food Control and her Master’s in Biotechnology at UBA. She is Assistant Teacher in Microbiology at the Faculty of Veterinary Sciences at UBA. She did several internships at Université de Montreal, Canada. She has published 3 papers, one in a national journal and two in reputed international journals; these past months, she had submitted 2 more manuscripts in collaboration with the Université de Montréal.

Abstract:

Bovine mastitis causes important economic losses in the dairy industry. Staphylococcus aureus and coagulase-negative staphylococci (CNS) are commonly isolated from bovine mastitis. β-lactams and macrolides-lincosamides (ML) antibiotics are frequently used in intrammamary therapy. CNS can be considered reservoir of antimicrobial resistance genes. The gene blaZ confer resistance to some β-lactam antimicrobials whereas the mecA and mecC genes confer resistance to all β-lactams. The genes ermA, ermB, ermC, mefA, msrA, mphC and lnuA confer resistance to ML antimicrobials. S. aureus resistance to β-lactams in Argentina is 23.1%. Among 80 S. aureus isolates, 32 (40%) were positive to blaZ gene and 15 (18.75%) were carriers of ML resistant genes: ermB and mefA (n=4), ermB (n=2) mefA (n=3), ermB, ermC and mefA (n=1), ermA, ermB, ermC and mefA (n=1), ermB, mefA, lnuA and msrA (n=1), ermC, ermB and lnuA (n=1), ermA (n=1) and lnuA (n=1). Among 90 CNS isolates, 12 (13.3%), 4 (4.4%), and 1 (1.1%) were positive for blaZ, mecA and mecC genes, respectively. Both blaZ and mecA genes were only found in one isolate whereas 6 (6.7%) isolates were resistant to ML antimicrobials via the following genes: ermC (n=1), ermB and ermC (n=2), ermB, ermC and mphC (n=1), mphC (n=1), and mphC and mrsA (n=1). The recently described mecC gene has been detected by PCR in a few CNS of animal origin only around Europe. We describe here for the first time a mecC positive isolate of CNS from bovine mastitis in Argentina. Identification of mastitis pathogens is important for selecting appropriate antimicrobial therapy. Reservoirs of antimicrobial resistance genes are potential threat to public health

Biography:

Miss Pimwan Thongdee has completed my PhD at the age of 28 years from Thammasat University, Thailand. I am a product specialist, i+MED Laboratories Company Limited. I have published 3 papers in reputed journals before I have completed my PhD.

Abstract:

Prostaglandin D₂ (PGD₂) is the most crucial prostanoid produced in the brain and involved in pain responses. Moreover, PGD₂ is a key factor derived from malaria within erythrocytes and might influence to parasite growth. The aim of the study was to preliminarily investigate the role of PGD₂ in malaria pathogenesis. Blood samples were collected from patients with Plasmodium vivax and Plasmodium falciparum (moderate and high parasitemia:1,000-50,000, and >50,000 /ml, respectively) infections, patients with fever associated with other infections, and healthy subjects of both genders and all age groups. PGD₂ concentrations were determined using Prostaglandin D₂-MOX express EIA kit (Cayman Chemical, USA).

Median (range) of plasma PGD₂ concentrations in patients with fever associated with other infections, patients with P. vivax and patients with P. falciparum infections, and healthy subjects (control) were 60 (11 - 525), 34 (22 - 130), 28 (16 - 38), 22 (13 - 75), and 16 (6 - 30) pg/ml, respectively. The median (range) plasma PGD₂ concentrations in patients with fever associated with other infections was significantly higher than those with P. vivax (p < 0.05), P. falciparum infections with moderate parasitemia (p < 0.0001), and healthy subjects (p < 0.0001). The concentrations in patients with P.vivax infection was significantly higher than those with P. falciparum with moderate parasitemia (p < 0.0001), and healthy subjects (p < 0.0001). Results of this preliminary study may suggest at least in part, an involvement of PGD₂ in fever-associated infections including malaria. Confirmation this finding is required with a larger sample size.

Biography:

Jakub Lenart is 29 years old PhD student from faculty of science, Charles University in Prague. He works in the Laboratory for Biology of Secondary metabolism, Institute of Microbiology of the Czech Academy of Sciences and Biotechnology and Biomedicine Center of the Academy of Sciences and Charles University in Vestec. Since 2012 he is member of Czechoslovak Society for Microbiology.

Abstract:

Staphylococcal ABC-F proteins Vga(A)LC and Msr(A) confer resistance either to lincosamides (L), streptogramins A (SgA) and pleuromutilines or to 14- and 15-membered macrolides and streptogramines B, respectively. All these antibiotics bind near peptidyl transferase centre in 50S ribosomal subunit and inhibit translation. Previous experiment has shown that 15 amino acid stretch of the Vga(A) interdomain linker is important for the antibiotic specificity of Vga(A) proteins. Substitutions L212S, G219V, A220T and G226S, clustered in the stretch, were responsible for the changes in resistance to SgA and L. It has been demonstrated in vitro, that ABC-F proteins mediate antibiotic resistance through interaction with ribosomes (Sharkey,2016). However, in our in vitro experiments, we confirmed that both proteins – Vga(A)LC , Msr(A) co-localize with membrane fractions of the cells. Therefore, it may be assumed that the proteins cooperate specifically with the ribosomes on the membrane, or, alternatively, the resistance mechanism is more complex, including also a cooperation of ABC-F proteins with a transmembrane partner. Vga(A)LC and Msr(A) might influence binding of lincomycin and erythromycin to the ribosomes in a similar way. Therefore, the functional interference of these two proteins is expected. To test this hypothesis we determined the resistance to lincomycin and erythromycin in the strain expressing both proteins. We found that activity of Msr(A) was not inhibited by Vga(A)LC as we expected but by the presence of subinhibitory concentration of lincomycin. Thus, usage of lincomycin may help in suppression of erythromycin resistance, conferred by Msr(A).

Biography:

Lae-Hyung Kang has in a doctoral course from Catholic University School of Medicine. He has completed his a Master´s degree from Catholic University School of Medicine. Mainly his study is waterborne viruses.

Abstract:

Norovirus is a major cause of viral gastroenteritis and a common cause of foodborne and waterborne outbreaks. Norovirus outbreaks are responsible for economic losses, most notably to the public health and food industry field. Norovirus has characteristics such as low infectious dose, prolonged shedding period, strong stability, great diversity, and frequent genome mutations. Besides these characteristics, they are known for rapid and extensive spread in closed settings such as hospitals, hotels, and schools. Norovirus is well known as a major agent of food-poisoning in diverse settings in South Korea. For these reasons, nationwide surveillance for norovirus is active in both clinical and environmental settings in South Korea. Recent studies have reported the emergence of variants and novel recombinants of norovirus. In this review, we summarized studies on the molecular epidemiology and nationwide surveillance of norovirus in South Korea. This review will provide information for vaccine development and prediction of new emerging variants of norovirus in South Korea.

Biography:

Kichan Lee has completed her PhD from School of Veterinary Medicine, Seoul National University. She is a researcher of Bacterial diseases division, Animal and Plant Quarantine Agency. She has studied on bacterial diseases of animals, and antimicrobial resistance of bacteria isolated from diseased animals and livestock products.

Abstract:

Tetanus is an acute, often fatal, infectious diseases caused by tetanospasmin, a neurotoxin of the anaerobic spore-forming bacteria Clostridium tetani. Tetanus spores are commonly introduced into tissues through wounds and they convert under anaerobic conditions to a vegetative toxin-producing form. We report here a case of C. tetani infection in a horse. A seven-year-old thoroughbred mare had laceration on the fetlock of right hind leg two weeks prior to her death. She had a raised tail, extended head and neck, and was salivating profusely.The third eyelids were prolapsed and covered the eyes. Almost all voluntary movement was impossible bacause of hyper-responsiveness to tactile nad auditory stimuli. The necropsy was performed, and there were no specific lesions. C. tetani was cultured from wound swab of the fetlock of right hind leg. Based on the characteristic clinical sings of generalized tetanus and detection of C. tetani from wound, a diagnosis of tetanus associated with C. tetani was made.

Biography:

Leona Zieglerováis 25 years old PhD student from Faculty of Science, Charles University in Prague. She works in the Laboratory for Biology of Secondary metabolism, Institute of Microbiology of the Czech Academy of Sciences and Biotechnology and Biomedicine Center of the Academy of Sciences and Charles University in Vestec. Since 2015 she is member of Czechoslovak Society for Microbiology.

Abstract:

Glycopeptide antibiotics are used as last choice antibiotics for treatment of infections, caused by gram-positive bacteria. But, increasing occurrence of glycopeptide resistant strains in recent years, rapidly decreases the range of treatment options of the bacterial infections. The glycopeptide resistance may be connected to expression of van resistance gene cluster, which alters the binding site of the glycopeptides. VanZ, one gene, from this cluster, expressed in Enterococcus faecalis confers specific resistance to teicoplanin without expressed other genes from this cluster (Arthur et al, 1995). VanZorhologs, forming big VanZ like protein family, are present in many bacterial genomes and their function is unknown. In this study, we compared the ability of two orthologous genes,vanZfrom the vanRSHAXYZ cluster (vanZTei) and vanZ from the genome of Enterococcus faecium (vanZg,), to confer resistance to glycopeptides, when expressed in Staphylococcus aureus RN4220. We have shown that VanZg, but not VanZTei, decreases 4 times sensitivity of S.aureustoteicoplanin. Rate of resistance depends on the level of induction of gene expression.Both genes were not able to confer resistance to vancomycin or other non-glycopeptide antibiotics; however, they effect decrease bacteria sensitivity to dalbavancin: new clinical using lipoglycopeptide. We tested antibacterial activity of four novel semisynthetic lipoglycopeptides against S.aureus, expressing vanZ. Expression of vanZ decreased sensitivity of S. aureus RN4220 to three lipoglycopeptides. All glycopeptides that decreased their activity against S.aureus had lipophilic tails. This suggests, that decreased sensitivity to glycopeptides, caused by vanZ expression, may be associated with the presence of lipid tail in the structure of antibiotic.

Biography:

I have completed my PhD at the age of 29 years in Molecular Parasitology Unit, Graduate Program in Biomedical Sciences, Faculty of Allied Health Sciences, Thammasat University, Thailand. I have published 3 papers in reputed journals.

Abstract:

Cystatins are the inhibitors of papain-like cysteine proteases and serve various important physiological functions including modulation of normal proteolytic processes, balance of the host–parasite interaction, and defense against pathogens. This research was conducted to characterize the molecular and biochemical properties of a type I cystatin (Stefin-2) in the liver fluke F. gigantica. The cDNA encoding FgStefin-2 had a size of 887 bp and contained a 351 bp open reading frame. The bacterial expression and purification resulted in highly pure rFgStefin-2 with an expected molecular mass of 13-14 kDa. The rFgStefin-2 was used for cysteine protease inhibition assays and polyclonal antibody production. The polyclonal antibody was used to study the distribution of native FgStefin-2 in 2- and 4-week-old juveniles and used in immunoblots. Immunohistochemical analysis showed that FgStefin-2 is located in several tissues of the parasite including the prostate gland, gut epithelium and intrauterine eggs. The polyclonal antibody reacted with rFgStefin-2, CW extract and ES product of the adult parasite, but did not cross-react with CW extracts of other trematodes. Mouse antisera raised against rFgStefin-2, rFgMDCd10, rFgStefin-1 showed no cross-reactivity in immunoblots to the different recombinant cystatins. The purified rFgStefin-2 exhibited inhibitory activity against cysteine proteases (cathepsins B and L) and the proteolytic activity of ES product, CW extracts from metacercariae and adult parasites. We have demonstrated that FgStefin-2 is able to inhibit cysteine proteases (cathepsins B and L). In the further analysis we would like to investigate the potential host-affecting functions such as immunomodulation.

Biography:

Ying Shun Zhou has completed his PhD at the age of 29 years from Sichuan University. And now he works in the Southwest medical university and is the director of pathogenic microbilogy labrotary. He has published more than 10 papers in reputed journals and has been serving as a member of Chinese Medical Association.

Abstract:

Acinetobacter baumannii is an opportunistic pathogen of hospital-acquired infection. The aim of this study was to analysis the 𝛽-lactamases genes prevalence in Acinetobacter baumannii isolated from a hospital in the southwest of China. In this study, 87 A.baumannii strains isolated from a hospital in southern of Sichuan province that were tested for antibiotic susceptibility performed by determination the MIC, molecular genotyping by multilocus sequence typing (MLST), and more importantly, the prevalence of 𝛽-lactamasesgenes (including blaOXA-23, blaOXA-24, blaOXA-58, blaGIM, blaSPM, blaSIM, blaNDM, blaCTX-M, blaSHV and blaTEM) were studied by PCR method. Antibiotic susceptibility test against 13 commonly used antibiotics indicated that most of the stains showed multidrug-resistance. And the result of MLST analysis showed that there were 5 STs in the 87 A. baumannii isolates, and a majority of them was ST195 (69/87). The PCR amplication and sequencing results illustrated that most of these isolates were positive to blaOXA-23 and blaTEM genes, and the prevalence of them were 74.71% and 93.10% respectively. In addition, 1 positive strain for the blaGIM, 1 positive strain for the blaSIM, 1 positive strain for the blaSHV and 2 positive strains for the blaSPM were found. However, all of the isolates were negative to the rest 𝛽-lactamases genes detected including blaOXA-24, blaNDM and blaCTX-M . In conclusion, the 87 A. baumannii isolates in this study showed severe drug resistance, and the main 𝛽 -lactamases genes carried were blaOXA-23gene and blaTEM gene.

Biography:

Abstract:

Food contaminated with multiple antibiotic-resistant S. aureus can be a major threat to the public health.  The purpose of this study was to isolate S. aureus from different food sources, determine their antimicrobial susceptibility as well as detection of mecA gene among some resistant isolates. Out of 125 samples from food of animal origin samples, 19 S.aureus isolates were recovered, and the antimicrobial susceptibility testing showed high resistance against kanamycin, penicillin G, oxacillin, erythromycin and tetracycline. All the tested isolates were multiple drug resistant (MDR). Eight out of 19 (15.2%) isolates were phenotypically resistant to oxacillin as well as they were carriers for mecA gene.

Biography:

Abstract:

Background: Peritonitis is currently one of the serious complications of continuous ambulatory peritoneal dialysis (CAPD), leading to considerable morbidity and mortality.We reviewed the incidence and the etiology of CAPD-associated peritonitis from January 2009 to December 2014.

Material/methods:Peritoneal fluid samples were cultured on appropriate solid and liquid media after centrifugation.Enumeration of WBC was done using standard counting chamber. The identification of the microorganisms was performed using standard methods,the API systems and the automated VITEK2 system (BioMérieux,MarcyL’Etoile,France).

Results:A total of105 dialysates were positive int the 6-year study period. Six of the positive samples were polymicrobial. Gram-positive organisms accounted for 49.5% of the infections of which staphylococci were the commonest (44.2%). Gram-negative bacteria were found in 39%of the positive samples, anaerobic bacteria in 6.7% and fungi in 4.8%.

Conclusions: Staphylococci   are the most common agents of continous ambulatory peritoneal dialysis(CAPD) peritonitis. Prompt identification of the causative agents is essential for the appropriate management of microbial peritonitis in patients on CAPD.

Biography:

Angeliki Mavroidi has completed her PhD at the age of 30 years from National and Kapodistrian University of Athens, Greece and postdoctoral studies from Imperial College London, UK. She has published 23 papers in reputed journals. She is a member of the European Society of Clinical Microbiology and Infectious Diseases (ESCMID).

Abstract:

Colistin is often used as salvage therapy for the treatment of infections caused by multidrug-resistant K.pneumoniae. The aim of the present study was the determination of colistin resistance rates among carbapenemase-producing (CP Kp) and the characterisation of colistin-resistant (COL-R) CP Kp recovered from bloodstream infections during May 2011 - December 2015 in a Greek hospital. Identification of the isolates to the species level and antibiotic susceptibility testing were performed by the MicroScan ® (Siemens Healthcare, PA, USA). The MICs of imipenem, meropenem and colistin were additionally determined by the Etest method, according to the interpretive criteria of the Clinical and Laboratory Standards Institute (CLSI, 2014). Phenotypic screening for carbapenemase production was performed by the modified Hodge test and the boronic acid/EDTA combined-disk test. DNA extraction was performed using the QIAcube (Qiagen, Düsseldof, Germany). Carbapenemase-encoding genes were detected by PCR. During the study period, COL-R represented 30% of 95 CP Kp isolates, while a rapid increase was observed in the incidence of COL-R CP Kp from 2014 to 2015 (25% to 55%, respectively). The majority of COL-R CP Kp (69%) was recovered from patients in the ICU. The COL-R CP Kp carried the blaKPC (n=16), blaOXA-48 (n=9), blaOXA-48+blaNDM (n=3) and blaNDM (n=1) genes. The first COL-R CP Kp carrying the blaOXA-48 and blaNDM were isolated on February 2015. The alarming increase in the colistin resistance rates and the spread of different carbamenemase genes among CP Kp recovered from bloodstream infections further complicates the therapy and infection control measures for combating this organism.

Biography:

Chemist and Master in Science of the University of Talca, currently is a PhD student from the University of Talca. His work is focus in Natural Products Chemistry and wine analysis. He has published several papers and has participed in different investigation project.

Abstract:

The grape production worldwide has been estimated in around of 60 millions tonnes per year. About 80% of the grape produced is utilized for winemaking, also in this process are produced grape waste that consists in around 20% of the weight. Currently, Chile is one of the main wine producers in the world, nevertheless, the wine industry produced a lot of grape waste that is a potential source of natural products, that can be used for different purposes. Nowadays, near 90% of the bacterial infections of the skin and soft tissue are produced by Gram-positive bacteria, taking advantage of the large amount of grape waste and the potential of this residue, a chemical characterization of a grape waste extract and the antibacterial activity against Gram-positive bacteria was made. A large amount of phenolic compounds were found in extract methanolic and extract methanolic/water as kaempherol, derivates of cinnamic acids, galic acid and cafeic acid. Determinated for HPLC-DAD. Among the studied bacteria, the extract of grape waste showed around of 50% of bacterial inhibition against Staphylococcus epidermidis compared with the reference compound (clindamycin). This study shows that the pomace extracts count on antibacterial activity on S. epidermidis. Thus the extract of grape waste has potential and can supply an opportunity replace some drug.

Biography:

Summaiya Mulla at present holding the post of professor and  head in department of Microbiology since 2004. She has established state reference laboratory for leptospirosis diagnosis and been awarded for contribution in leptospirosis control and diagnosis by health minister in 2006. She has established swine flu molecular laboratory at GMC Surat have been awarded for control of swine flu by health secretory in 2011. She has developed state  reference laboratory for tuberculosis She has done more than 30 paper publication in national and international journals. She has presented oral as well as poster presentations at different international conferences held across the globeShe has participated as delegate in many national and internal workshops and conferences.

Abstract:

Re-establishing the cut-off titre for IgM ELISA using ROC (Receiver Operating Characteristic) curve in South Gujarat region because cut off titre for these serological tests needs to be re-confirmed at timely intervals keeping in mind the endemicity of Leptospirosis in the region. For  determining  the  ROC  curve   and  the   cut  off  titre,  30 healthy population from  each geographic region like districts Tapi, Navsari and valsad, 30 patients with diseases other than Leptospirosis as well as 30 known Leptospirosis cases was taken from South Gujarat region and tested for Panbio ELISA for leptospirosis. All the data obtained by testing was entered into SPSS (Statistical Package for Social Sciences) software for ROC analysis. Analysis showed best cutoff value for valsad  and  navsari 14.5 panbiounits and for tapi 16.5 panbiounits as being higher than kit base cutoff value of 11 panbiounits. Using the cutoff value recommended by the manufacturer (11 Panbio units), the sensitivity on paired sera is high (90.8%) and may be used to rule in patients with suspected leptospirosis. However, the poor specificity of the test (55.1%) suggests that using this test in the clinical setting could lead to over diagnosis of leptospirosis and a high frequency of false positivity. Using the high cutoff value generated by ROC curve analysis (15 Panbio units), the overall accuracy of the test was improved. We  recommend  as WHO to use such an assay for the diagnosis of leptospirosis if financial resources permit requires area -specific evaluation to determine its clinical use before implementation.

Biography:

Hayam M Hamouda is part of NODCAR, Egypt

Abstract:

One of the major concern is the increasing incidence of multidrug-resistant organisms, particularly extended spectrum beta lactamases producers (ESBLs) among diabetic foot infection known to increase the duration of hospital stay, cost of management as well as morbidity and mortality. In This study organisms found in diabetic foot infection DFI samples were,2 Aerococcus viridians, 3 Enterococcus raffinosus, 4 Enterococcus avium, 6 Rhizobium radiobacter,7 Staphylococcus sciuri . antibiotic susceptibility test aganist producing extended-spectrum β-lactamases bacteria was done. Also, in The present study detecting the gene responsible for (ESBLs) resistance of CTX-M-type of ESBLs in DFI bacteria and sequencing it.

Biography:

Nwachukwu, Michael I. Is a Senior Lecturer and the Head, Department of Microbiology and Industrial Microbiology, Imo State University, Owerri, Nigeria. He holds a Ph.D Degree from Rivers State University of Science and Technology, Port Harcourt, Nigeria. He is a member and the Institutional Coordinator of Nigeria Society for Microbiology (NSM). He has published more than 40 papers in reputed journals and His research interests are in the areas of Environmental Microbiology, Ecology and Public Health.

Abstract:

Study to determine the antibacterial effect of ethanolic extract from leaf, root and seed coat of Moringa oleifera on bacterial agents of infantile diarrhea was carried out. This was determined using disc diffusion and well-in-Agar antimicrobial screening methods. A total of sixty nine suspected infants (0-5years) were screened using standard microbiological methods. Bacterial agents isolated and identified were E. coli and Salmonella sp. While E. coli was isolated from 17(60.71%) subjects, Salmonella sp. was isolated from 11(39.29%) subjects. Results show that more infants of three years old had diarrhea while those at the age of five years were the least. Male infants were infected more than female infants. Well-in-Agar antimicrobial screening method showed appreciable inhibitory effect on both E. coli and Salmonella sp. than disc diffusion. Furthermore, ethanolic extract from leaf exhibited more antimicrobial action on the two isolates followed by extract from seed, then extract from the root while extract from seed coat that showed antimicrobial effect at a very high concentration was the least. Phytochemical analysis showed that alkaloid and anthraquinone were present in the four parts of the plant analysed. Glycoside, steroid and terpenoid were present in the seed coat, flavonoid was absent in the leaf and root, saponin was absent in the seed, tannin was only present in the leaf. Above results indicate that extract from most parts of Moringa oleifera have curative effect hence can be used for curing diseases such as diarrhea.

Biography:

Wolfgang R.Heizmann has completed his MD in 1982 from the University of Tübingen and became assistant Professor for Medical Microbiology in 1988 and full Professor in 1994 from the University of Tübingen, Germany. He was head of microbiology in several laboratories and is now director of Orgamed Consulting, a company advising hospitals in the field of microbiology and hospital hygiene as well as specialised software solutions. He has published more than 100 papers and is editor of several textbooks.

Abstract:

The avoidance of nosocomial infections is the main goal of hospital hygiene. There are two prominent sources of pathogens causing infections: exogenous and endogenous. An example of an exogenous infection is the acquisition of a pathogen from other patients or from the environment. To combat this source, guidelines and “bundles” of measurements are able to reduce the number of cases. However, endogenous nosocomial infections e.g. resulting from the microbiome of the gut are much more challenging. In the area of rising numbers of multiresistant bacteria (MRB), especially Gram-negative species producing ESBL’s or carbapenemases, the chance to acquire an infection caused by one of these organisms is also increasing. Many of the MRB’s are Enterobacteriaceae and reside in the gut of apparently healthy people. Admission to the hospital e.g. caused by a diabetic foot, may result in an in endogenous nosocomial infection by these MRB’s. It is now accepted that early adequate antimicrobial therapy is able to decrease mortality. However, even in the area of rapid methods for identification of bacteria and detection of the most important resistance mechanisms, in many cases results of susceptibility testing needs up to 72h. Therefore it is of utmost importance to know the risk in a given patient for the possibility of a MRB infection. Consequently it seems rational to screen for colonisation with MRB’s not only patients in so called risk groups, but all patients at least at admission. The question arises which method is best and at what costs.

Biography:

Amaresh Das had completed his PhD from Calcutta University, India and did postdoctoral researchat the Department of Biochemistry and Molecular Biology, University of Georgia. Currently, he works as a microbiologist in the Reagents and Vaccines Services Section, Foreign Animal Disease Diagnostic Laboratory, NVSL, STAS, VS, APHIS, USDA, Plum Island Animal Disease Center, Orient Point, New York. He has published more than 20scientific papers in reputed peer reviewed journals and wrote several chapters in books and proceedings. He also serves as reviewerfor many reputed peer reviewed journals

Abstract:

The real time polymerase chain reaction (qPCR) has now been widely used as one of the mainstream diagnostic assays for rapid detection of infectious diseases in animals due to its sensitivity and specificity. However, the interpretation of qPCR results can be challenging and must be interpreted in conjunction with the history, clinical signs and the evidence of disease. Performing qPCRassays involves multiple steps including sample preparation, nucleic acid extraction and amplification. Thisstudy aims at critical evaluation of each step based on our experience working with multiple animal disease viruses including avian influenza virus (AIV), classical swine virus (CSFV) and Capripoxvirus (CaPV). Diagnostic specimens were collected from experimentally and naturally infected animals including swabs (multiple types), blood and tissues (multiple types). Critical factors that influence the performance of qPCR include: inefficient release of the virus from specimens (sample preparation), co-purification of naturally occurring PCR inhibitors (extraction) and PCR failure (amplification). The foremost challenge has been the false negative results due to PCR inhibition. Indigenous (beta-actin) or exogenously addedcontrols (reverse transcribed RNA) were used to detect PCR inhibitors. The highest level of PCR inhibitorswere detected in fecal samples (AIV-infected specimens) followed by blood, tissues and swabs (AIV- and CSFV-infected specimens). Commercial extraction kits failed to completely remove PCR inhibitors from the clinical specimens. Commercial protocols were modified by adding an extra high-salt (NaCl-EDTA) washing step and the modified protocols were found to efficiently remove PCR inhibitors from the clinical specimens and subsequently improved the diagnostic sensitivity (DSe) of the qPCR assays. The choice of PCR kit(DNA polymerase) also had a large impact on the DSe (CaPVqPCR) due to the differences in their tolerance against the PCR inhibitors.

Biography:

Astra Vitkauskiene has completed her PhD in Medical Sciences at Kaunas University of Medicine, Institute for Biomedical Research in 2008. She is a Head of Department of Laboratory Medicine in Lithuanian University of Health Science and Physician-microbiologist, Head of Laboratory of Microbiology in Hospital of Lithuanian University of Health Science Kaunas Clinics. She has published more than 50 papersin reputed journals.

Abstract:

Despite the advances in hospital care and the introduction of a wide variety of antimicrobial agents, Pseudomonas aeruginosa (P. aeruginosa) continues to be a common cause of nosocomial infections and are important pathogens which causes problems clinically as a result of its high resistance to antimicrobial agents. Treatment of P. aeruginosa infections is usually difficult and mortality is high. In our study we found that 53.7% of clinical P. aeruginosa strains were resistant to carbapenems. Carbapenem-resistant strains more frequently were resistant to majority of tested antibiotics (to ceftazidim, piperacillin, ciprofloxacin, gentamicin, amikacin), comparing to carbapenem-sensitive strains (P<0,001). 41 (56.0%) of carbapenem resistant strains had MIC value higher than 16 µg/ml; 6 (14.6%) stains out of these have shown to be high-level ceftazidime resistance with MIC >64 µg/ml. Cephalosporin with β-lactamase inhibitor combination inhibited 53 out of 73 carbapenem-resistant P. aeruginosa at concentration less than 2 µg/ml. We found 9 (13.4%) fully resistant strains with MIC >32 µg/ml and 58 (86.6%) intermediately resistant strains with MICs range between 2 and 16 µg/ml to aztreonam. Our study has shown that more than 50% (n=28) of ciprofloxacin resistant isolates exhibited MIC values above 16 µg/ml. Tobramycin had activity against 56% (n=41) of tested isolates which twenty four were inhibited at MIC below 1µg/ml. However more than 65% (n=21) of tobramycin resistant strains had MICs above 16 µg/ml. β-lactam resistance was caused by chromosomal mechanisms (AmpC +/- OprD) in 54 isolates out of 73 (74%). In 16 (22%) of P. aeruginosa strains the presence of a carbapenemase was found and three isolates were ESBL produces.

Biography:

Lisa Lindesmith is a Research Specialist collaborating with Dr. Ralph Baric since 1999 to study the molecular mechanisms regulating norovirus evolution, susceptibility, and protective immunity. Major contributions include: i) Identifcation of FUT2 (histoblood group antigen expression) as a major norovirus susceptibility allele; ii) demonstration that GII.4 noroviruses are evolving by epochal evolution in response to human herd immunity; iii) first to map neutralizing blockade epitopeselucidating the molecular mechanisms which allow GII.4 norovirus escape from herd immunity, iv) first to identify strategies to increase the breadth of blockade immune responses by multivalent vaccination in mice and humans.

Abstract:

Noroviruses (NoVs) are the leading cause of human acute gastroenteritis. Strains within the GII.4 genotype drive pandemic levels of infection every 2-3 years. Each pandemic strain correlates with evolution in the major capsid protein and emergence of a GII.4 strain with distinct antigenic and ligand binding properties. GII.4 strain emergence and extensive antigenic heterogeneity among the >40 additional NoV genotypes are primary obstacles to development of an efficacious vaccine. Multivalent virus-like particle (VLP) vaccination shows promise for overcoming these challenges. Volunteers vaccinated simultaneously with GI.1 and GII.4C VLPs generated broad cross-genotype blockade antibody responses, a surrogate measurement for protective immunity. Importantly, breadth of blockade antibody response extended to novel GII.4 VLPs that had not circulated prior to sample collection, indicating that vaccination may provide protection from emergent strains.The breadth and uniformity of the blockade antibody response across antigenically diverse GII.4 strains suggests that immunization primarily activated a memory antibody response to multiple GII.4 strains. Antigenic cartography and epitope-specific blockade-of-binding assays support this finding. These results are in contrast to the observed strain-specific secondary antibody response to the GI.1 vaccine component and identify pre-exposure history, mediated by host genetics, as a key determinant to NoV vaccine response.

Biography:

Antoni Bennasar-Figueras completed his PhD at the Universitat de les Illes Balears (UIB, Spain) and postdoctoral research at the Gesellschaft für Biotechnologische Forschung (GBF, Germany). He is Profesor of Microbiology and member of the Microbiology Research Group at the UIB. Nowadays, he has specialized in the application of Next-Generation Sequencing (NGS) technologies to the genome and comparative analysis of several microorganisms (Pseudomonas, Mycobacterium and Streptococcus). He has published more than 30 articles in well-known journals and has been serving as an editorial board member of recognized journals like Environmental Microbiology, Systematic and Applied Microbiology, The ISME Journal or PLoS One.

Abstract:

The species classified as non-tuberculous mycobacteria (NTM), or rapid growing mycobacteria (RGM), are widely distributed in the environment and some of them are considered as emerging opportunistic pathogens. Nosocomial infections caused by NTM are usually difficult to treat due to their resistance to antibiotics or other external factors. The next-generation sequencing (NGS) technologies are opening new frontiers to different fields, and clinical microbiology is not an exception. The main objectives of this work imply the genome sequencing of NTM isolates (especially type strains), the identification of gene families, functional characterization, comparative analysis applying several clustering algorithms and the description of the core-pangenome of NTM. Briefly, the results obtained showed different rates of genome evolution and exclusive genes for each species (pangenes). Interestingly, the pangenome analysis of NTM has revealed also the presence of toxin-antitoxins (TA) systems in several of the strains compared. Curiously, most of the TA systems discovered in NTM are also present in M. tuberculosis. Furthermore, a brand new TA system has been discoverd. The toxic potential of the proposed toxins and its neutralization with the hypothetical antitoxins have been tested in vitro. All together, contributes to improve the NTM evolution knowledge, as well as to gain a better understanding of the mechanisms underlying their ability to adapt to different ecological niches; i.e., the resistome, the toxin-antitoxin systems or their ability to form biofilms. All these aspects affect the human’s lifestyle. Definitively, the new NGS based information may lead to important breakthroughs, both in biotechnology and clinical microbiology.

Biography:

Latre Buntaran has completed his Clinical Microbiologist Specialist in 1996 at University of Indonesia and post doctoral studies from Hasanudin University at 2012. He is the head  of microbiology department as well as head of infection control committee at Mayapada and Bethsaida Hospitals. He has published 4 papers in reputed journals and as a speaker in more than 200 national symposium and several international congress.

Abstract:

Community Acquired Methicillin Resistant Staphylococcus aureus (CA-MRSA) is a strain of MRSA that can cause infections in patients in the community, in which these patients had no previous risk factors for MRSA infection and the patient received 72 hours prior to infection when admitted to hospital.This study aims to determine and compare the characteristics of epidemiological, clinical, and molecular biology of CA-MRSA with HA-MRSA.

           Of the 311 S.aureus isolates collected from 2 hospitals (RSAB Harapan Kita and RS Siloam Kebun Jeruk) during the period 2009 to 2011, the prevalence of MRSA is 6% and consists of CA-MRSA (2%) and HA-MRSA (4%) , the pattern of infection as follows : SSTI (skin and soft tissue infections) : 56%, UTI (urinary tract infection) : 17%, RSA (acute rhino sinusitis) : 11%, Pneumonia : 6%, febrile observation : 5% and ILO (wound infection) : 5%. The third-generation cephalosporins and quinolones are the antibiotics most used in this study. These third-generation cephalosporins are resistant to all isolates of MRSA (CA-MRSA and HA-MRSA), whereas quinolone resistant to HA-MRSA, but still sensitive to CA-MRSA. The use of antibiotics against infections by S.aureus of 311 isolates showed that the use of antibiotics : inappropriate : 57%,  appropriate : 43%, adequate : 43%, inadequate : 57%, oral : 79%, parenteral : 21%, original : 36% and copy product : 64%.                

          Futhermore, 11 strains of Staphylococcus aureus were performed by PCR,  in which, there is one strain of Community-Acquired MRSA (CA-MRSA) with SCCmec type II, 3 strains of Hospital-Acquired MRSA (HA-MRSA) with SCCmec type IV, and two strains of Hospital-Acquired MSSA (HA-MSSA) and five strains of Community Acquired MSSA (CA-MSSA) that do not contain mecA genes and SCCmec. From the three strains of one strain of CA-MRSA and two strains HA-MRSA containing plasmid pUB110. vraA present in 91% of the 11 strains,  vraF : 36% , vraG : 45%, and vraR : 36%. Noteworthy, strains without pUB110 contained in a relatively high frequency of 75% in vraR as well as vraF, and 70% in vraA compared to strains with pUB110 : 60% in vraG.

Biography:

Niels Nørskov-Lauritsen, M.D., Ph.D., and D.M.Sc., is a senior consultant and associate professor in clinical microbiology at Aarhus University Hospital, Denmark. He completed his medical education at Aarhus University and received his Ph.D. in microbiology at the University of Copenhagen, Denmark, and his D.M.Sc at the University of Aarhus. Scientific contributions encompass classification and identification of Haemophilus and Aggregatibacter species. He is a member of the Subcommittee on the Taxonomy of Pasteurellaceae under the International Committee on Systematics of Prokaryotes. He is a principal investigator at Aarhus University, focusing on cystic fibrosis microbiology, antimicrobial resistance, and taxonomy of Pasteurellaceae.

Abstract:

Non-hemolytic variants of Haemophilus haemolyticus are difficult to differentiate from Haemophilus influenzae, despite a wide difference in pathogenic potential. We characterized a challenging set of 60 clinical strains by multi-locus sequence analysis (MLSA) and near-full length 16S rRNA gene sequencing. MLSA unambiguously allocated all study strains to either of the two species, while identification by 16S rRNA sequence was inconclusive for three strains. Notably, the two methods yielded conflicting identifications for two strains. Most of the "fuzzy species" strains were H. influenzae that had undergone complete deletion of the fucose operon. Such strains, which are untypeable by the H. influenzae MLST scheme, have sporadically been reported and predominantly belong to a single branch of H. influenzae MLSA phylogenetic group II. We also found evidence of interspecies recombination between H. influenzae and H. haemolyticus within the 16S rRNA genes. Full-genome sequences are presently being analysed.

Biography:

Vaclava Adamkova is the Head of the Department of Clinical Microbiology and Antibiotic Centre of the Institute of Medical Biochemistry and Laboratory Diagnostics of the General University Hospital and of The First Faculty of Medicine of Charles University in Prague and senior consultant. She is the specialist in the field of clinical microbiology and bacteriology. She has long been interested in the identification of infecting agents in critically ill patients, especially in respect of infections of the skin and soft tissue and intra-abdominal infections. She´s co-author of recommended procedures of above mentioned area of clinical microbiology. She´s board member of Society for medical microbiology of the Czech Medical Association of J.E. PurkynÄ›, board member of Society of Infectious Diseases of the Czech Medical Association of J.E. PurkynÄ› and board member of the Czech Surgical Infection Society.

Abstract:

Intraabdominal candidiasis (IAC) is the predominant type of invasive candidiasis after candidemia. IAC is associated with mortality rates around 25–60 % . The majority of epidemiological studies on Candida are focused only on bloodstream infections. Nevertheless, the role of blood cultures has a limited application in patients with abdominal candidiasis. IAC, which includes peritonitis and intraabdominal abscesses, may occur in around 40 % of patients following repeat gastrointestinal (GI) surgery, GI perforation, or necrotizing pancreatitis. Candida is reported to be isolated in 41 % of upper gastrointestinal (GI) sites, 35 % of small bowel, 12 % of colorectal, and less than 5 % of appendicular sites in. Candida spp. has been reported as the second most frequent pathogen cultured in peritonitis patients Major risk factors for Candida peritonitis include hollow organ perforation, abdominal and thoracic surgery, surgical drains in situ, intravenous and urinary catheters, severe sepsis, and extensive Candida colonization. For many years, there has been debate over the importance of Candida isolated from the sites of intraabdominal infection. The organism is a part of normal flora of the gastrointestinal tract and its isolation is often difficult to interpret. Unfortunately, the pathophysiologic importance of Candida isolation from the abdominal space is by far not clear in many clinical situations. Generally, infection is suspected when the organism is cultured from samples obtained intraoperatively or directly from an intraabdominal collection. When Candida is cultured from subsequently obtained drainage fluid samples, colonization is a possibility

Biography:

Roby P. Bhattacharyya completed his MD and PhD from the University of California, San Francisco in 2007trained in Internal Medicine and Infectious Diseases at Massachusetts General Hospital. He is currently an Assitant in Medicine in the MGH Division of Infectious Diseases, an Instructor at Harvard Medical School, and a researcher at the Broad Institute of MIT and Harvard. His research interests focus on both basic and applied aspects of antibiotic resistance in bacteria.

Abstract:

Rapid antibiotic administration remains our most effective weapon against bacterial pathogens, but the emergence of antibiotic resistance poses serious challenges. Current culture-based susceptibility determination is too slow for early evidence-based antibiotic selection, while newer genotypic methods have limited ability to predict phenotypic antibiotic resistance. RNA detection offers a “semi-phenotypic” assay with the potential to identify antibiotic susceptibility more rapidly, agnostic to the genetic basis for resistance. Susceptible bacteria enact different transcriptional programs than resistant ones within minutes of antibiotic exposure, independent of resistancemechanism. We report RNA-Seq analysis of the key drug-resistant pathogens Acinetobacter baumanii and Klebsiella pneumoniae treated with three key antibiotics from distinct classes: ciprofloxacin, gentamicin, and meropenem. These studies reveal transcripts whose expression levels clearly distinguish susceptible from resistant organisms within minutes of drug exposure. We validate these transcriptional signatures for multiple antibiotic classes against clinical isolates, including a “test set” of multidrug-resistant strains from the US Centers for Disease Control, using a simple, rapid commercial RNA hybridization assay (NanoString) to robustly distinguish susceptible and resistant clinical isolates within hours. A susceptibility metric derived from these transcriptional assays correctly grouped isolates in 148 of 151 antibiotic susceptibility tests (98% accuracy); 2 of the 3 incorrectly grouped isolates were within one dilution of the breakpoint MIC. We have previously shown proof-of-concept that this 8-hour assay may be applied to a positive blood culture with minimal sample processing. In principle, this rapid phenotypic assay for antibiotic resistance can be extended to any pathogen-antibiotic pair, without foreknowledge of resistance mechanism.

Biography:

Haiying Liu has completed her PhD at the age of 35 years from Second Military Medical University. She is the director of Clinical Laboratory, Guangzhou Women and Children's Medical Center, Guangzhou Medical University, Guangzhou, Guangdong, China.

Abstract:

      Group B streptococcus (GBS) is a leading infectious cause of neonatal morbidity and mortality in many countries, while the disease burden in China is almost blank. We investigated the epidemiology of invasive GBS infection to appraise the necessity of implementation of appropriate interventions.  Data for infants less than 3 months old with culture-positive GBS were retrospectively collected from three large urban tertiary hospitals in Guangdong, southern mainland China from Jan 2011 to Dec 2014. Of the 127206 live births in the three hospitals, 70 cases of culture-confirmed invasive GBS infection were identified, with the overall incidence of 0.55 per 1000 live births (95% CI 0.44-0.69). The incidence among infants less than 6 days was 0.39 per 1000 live births (95% CI 0.29-0.51) and increased significantly from 0.12 per 1000 live births in 2011 to 0.58 per 1000 live births in 2014 (P < 0.05). The incidence remained stable among infants aged 7 days through 89 days during the study period (mean, 0.16 per 1000 live births). Eighty-four additional GBS cases were confirmed in the study hospitals but were born elsewhere. Among the total 154 cases, 9 deaths (5.8%) occurred before discharge, and 18 cases (12.4%) experienced neurological sequelae. All 154 isolates were susceptible to penicillin, ampicillin, and vancomycin, while frequently resistant to erythromycin (75·6%) and clindamycin (38·5%). In the 68 isolates tested, serotype III (76.5%) was the most commonly identified, followed by Ib (14.7%), V (4.4%), and Ia (4.4%).  This study concluded that incidence of neonatal invasive GBS infection in southern mainland China was higher than previously anticipated and increased significantly during the study period, especially early-onset disease. Standardized preventive measures against GBS should be adopted.

Biography:

Abstract:

The first step of the pathogenesis of cholera which is due to Vibrio cholerae (V. cholerae) have to attach onto enterocyte. Enterobacteriaceae usually use fimbriae or outer membrane protein (Omp) as an adhesive protein. Up to nowno researchers   have reported that the adhesive proteins V. cholerae which found in the pili or Omp of the bacteria. If the adhesive molecules have been found and subsequently   the research can be continued with the aim is to determine whether V. cholerae adhesion molecule if used as an antigen has the ability   in producing antibody and it can prevent the leaking of fluid from intestinal cells in mice.

Fimbriae protein isolation methods using bacterial pili cutter. Omp taken from V. cholerae cells by which the fimbriae have been shearedfrom the cell and solubilize   using the solvent of N-Octyl-Glycopyranoside (NOG) 0.05%. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used to monitor the protein profile. Screening of adhesive molecules by using the hem agglutination method. Confirmation of adhesion molecule is done by calculating index of adhesion which use enterocyte. For detecting the activity of adhesive molecule V. cholerae for   producing antibody and which can to prevent the leaking of fluid from intestinal cells in mice using a Mice Legated Ilea Loop (MLIL) test and cholera toxin (ct) sub unit B as adjuvant.

Research results have clarified that the adhesion molecules of V. cholerae have been found in fimbriae as   well   as   in OMP and turned out to have identicalMW  37.8kDa, but under natural conditions OMP adhesive has   MW 75.6 kDa. The test results protectives adhesive protein of 37.8 kDa MW fimbriae with adjuvant ct subunit B produce antibodies that can prevent the leaking of fluid into the lumen of the small intestine of mice.

Protein adhesives with MW 37.8 kDaV. cholerae in the future may be can to be developed as acellular vaccine candidate cholera.

Biography:

André Ricardo Araujo da Silva has completed his PhD at the age of 35 years from FIOCRUZ-National Institute of Infectology-Brazil. He is the coordinator of Scientific Program of Medicine Course-Federal Fluminense University (UFF)-Brazil and leads the Laboratory of Teaching of Prevention and Control of Healthcare-associated infections. He has published more than 15 papers in reputed journals and is a member of International Federation of Infection Control.

Abstract:

Nosocomial infections (NI) represents a serious problem in all parts of the world. Data from Center for Disease Control-CDC-USA estimates 722,000 cases and 75,000 deaths in 2011, as direct consequences of NI. In the European Union there’s about 4.1 million cases/year with 37,000 deaths. NI are also frequently related to antimicrobial resistance (AMR). Without an global plan to reduce AMR, 10 million of deaths are expect in 2050 due to infections by MDR. The main NI are pneumonias (associated or not to mechanical ventilation), surgical site infections, urinary tract infections, gastrointestinal infections and bloodstream infections. Implementation of infection control programs (IPC) in hospitals is strongly recommended bt World Health Organization (WHO) and it’s compulsory by law in several countries. Between 20-30% of NI cases are preventable through institution and maintenance of IPC. For example, it’s possible to reduce BSI rates in until 70%, with adoption of a package of measures (bundles) to prevent them. Some key components are necessary to implement a successfull IPC as: organisation of infection control at the hospital level, bed occupancy staffing, workload and employment of pool or agency nurses, avaibility of and ease of access to materials and equipment and optimum ergonomics, appropriate use of guidelines, education and training, auditing, surveillance and feedback, multimodal and multidisciplinary prevention programmes that include behavioural change, engagement of champoins and positival organisational culture. In conclusion there’s an urgency of global efforts to implement and mantain IPC in all countries, together with governmental and civil society suport.

Biography:

B. Y. Chin received her degrees in Physiology and Toxicological Sciences from the Department of Environmental Health Sciences at Johns Hopkins University, Baltimore, Maryland. She continued her research at the Department of Surgery at Beth Israel Deaconess Medical Center in Boston, Massachusetts after completing her post-doctoral fellowship at Pacific Northwest National Laboratory, U.S. Department of Energy in Richland, Washington. Dr Chin also had a joint Faculty appointment at Harvard Medical School since 2006. She has published over 33 peer review journal articles and is an active member on 3 editorial boards. Currently, she is a Professor of Medical and Health Sciences at the International Medical University.

Abstract:

Inflammation and immunity result in a wide range of disease processes, including chronic

obstructive pulmonary disease, ischemia-reperfusion injury, atherosclerosis, vascular thrombosis and sepsis. Heme oxygenase-1 (HO-1) is a key enzyme that is indispensable for the temporal and spatial regulation of host response and, together with its essential metabolite carbon monoxide (CO), is crucial for maintaining homeostasis, inhibition of inflammation and the preservation of function and life. Of the numerous physiologic effects observed with CO, in the last 5 years, it has become apparent that CO has been ascribed an additional novel, yet innate role as a “bactericidal agent”. Its role in the maintenance of homeostasis remains intact, however, the designation necessitates the paradoxical induction of the inflammatory response and binding to hemoproteins in order to restore physiological balance and sustain life. In this presentation we will review and discuss recent reports that highlights the paradoxical use of CO as a host defense molecule agent against pathogens.

Biography:

Luigina Romani received her Ph.D. and M.D. degrees from the University of Perugia, where she also trained in basic immunology and transplant immunology. She is currently Professor of Pathology and a faculty member of the University Medical School at the University of Perugia. Luigina Romani is internationally recognized in the area of antifungal immunity — a field in which her major interest is on the comprehension of innate mechanisms of antifungal immunity that lead to the activation of protective and non-protective adaptive immunity. She has been recognized “Fellow" of the American Academy of Microbiology and elected to the American National Academy of Sciences. Luigina Romani pioneered studies on the role of T cells first (early 80’) and then of the different Th cell subsets (early 90’) in experimental fungal infections. She introduced several novel scientific and translatable concepts which have become popular in the field of medical mycology. Examples are the concept of protective tolerance, the basis of immunotherapy, the use of dendritic cells to develop fungal vaccine, the pathogenetic role of inflammation in infection, the discovery of tryptophan metabolites as potential antifungal strategies and, more recently, the use of functional genomic to predict individual risk for fungal infections in transplanted patients

Abstract:

An increased understanding of the importance of microbiota in shaping the host’s immune and metabolic activities has rendered fungal interactions with their hosts more complex than previously appreciated. It is now clear that a three-way interaction between host, fungi, and microbiota dictates the types of host-fungus relationship. Indeed, microbial dysbiosis predisposes to a variety of chronic fungal infections and diseases at local and distant sites. We have explored metagenomics for the purpose of deciphering the contribution of the microbiota to fungal commensalism and parasitism. By correlating changes in metabolite profiles with microbiota metagenomic composition, we have defined a functional node by which certain bacteria species contribute to host-fungal symbiosis and mucosal homeostasis in the gut. The trpyptophan metabolism pathway is exploited by commensal bacteria and the mammalian host to increase fitness in response to Candida albicans by inducing resistance and tolerance mechanisms of the antifungal immunity. Similarly to the gut, the lung microbiome also has a profound impact on lung immunity and resistance to pulmonary infection. We have explored in both human and mice airways the dynamics and diversity of prokaryotic population as a key alarm of upcoming Aspergillus fumigatus infection, a threatening infection in lung or hematopoietic transplanted patients. Using 16S rRNA sequencing, a substantial bacterial diversity was observed in intact or Aspergillus-infected mice. At the taxa level, the Firmicutes decreased significantly both in the lung and BAL upon infection, while the Proteobacteria significantly increased. A similar bacterial diversity was observed in BAL samples from a cohort of patients undergoing allogeneic hematopoietic transplantation with and without positive Aspergillus diagnosis. These data suggest that the study of the human microbiota in the trans-omics era, with a focus on metagenomics and metabonomics, is providing novel insights into the microbial regulation of host immune resistance to fungi. This work is supported by the Specific Targeted Research Project FUNMETA (ERC-2011-AdG-293714).

Biography:

Harrison Magdinier Gomes is a scientist from Oswaldo Cruz Foundation - RJ - Brazil. He Worked on Leprosy and Tuberculosis. Between 2014 and 2015 he was a visiting scientist at the University of Paris (Team of Dr. C. Sola),) He was in charge of the new research project on the improvement of Pyrazinamide and Second- Line drug resistance molecular testing of the Mycobacterium tuberculosis. As an expert in high throughput genotyping on Luminex and method design for public health and infectious diseases control and as biologist, one important project was deciphering the population of tubercle bacilli in Brazil.

Abstract:

Pyrazinamide is used as a first line drug in the treatment of tuberculosis. It should be converted by the Mycobacterium tuberculosis (Mtb) pyrazinamidase enzyme into pyrazinoic acid, as the active metabolite. Mutations in the pncA gene are related to Mtb pyrazinamide resistance, and seem to appear within the entire gene, hence, until now, sequencing was the single method to detect the pyrazinamide resistance at the genetic level. Recently Miotto et al., identified 280 pncA genetic variants and very high confidence mutations were found only in pyrazinamide resistant strains (85%). The goal of the current study is to develop a molecular method to detect mutations related to pyrazinamide resistance in the pncA gene, based on a microbead-based high-throughput suspension array format. To amplify the pncA gene, we use the DPO concept (Dual Priming Oligonucleotides) followed by a nested PCR with three conventional pair of primers that divides the gene into three fragments. We designed for the high confident mutations 30 wild-type/mutant probes couple. The assay was developed using pyrazinamide-resistant reference (sequenced) DNA isolates provided by the SRL in Stockholm. Out of 30 probes couples, 26 got significant signal/noise threshold to distinguish between wild-type and mutant alleles. The codon 175 presents two possible mutations that are related to resistance (ATG wt, GTG mut 1 and ATA mut 2) and based on currently probe only mut 2 was properly detected. Our model to detect pyrazinamide resistance still needs to be optimized to increase signal/noise level for some probe couples, and to detect new mutations. The detection of pyrazinamide resistance in this model will be associated to TB-SPRINT and TB- SPRINTplus our assays that allow detecting 1st and 2nd line drug-resistance associated genes.