Keynote: The Clinical Impact of Rapid Nucleic Acid Amplification Tests for Detection of M. Tuberculosis
Phyllis Della-Latta completed her PhD in Basic Medical Science, Molecular Biology, from New York University, NYC. Dr. Della-Latta is a diplomate, American Board of Medical Microbiology and Public Health and holds the academic position of Professor of Clinical Pathology & Cell Biology in Medicine at Columbia University, College of Physicians and Surgeons. She is the Director of the Clinical Microbiology Service at Columbia University Medical Center, NY-Presbyterian Hospital, NYC. She has over 160 publications in peer reviewed journals and book chapters and has delivered over 120 invited presentations both nationally and internationally.
The global burden of tuberculosis (TB) is enormous, impacting industrialized as well as developing countries. In order to achieve the goal of the 2013 World TB Day Towards Zero TB theme, fast tracking detection of Mycobacterium tuberculosis (MTB)\ and drug resistant strains are critical in breaking the chain of TB transmission. Cases of TB are on the decline in the USA; however, New York City has three times the average USA cases. Although culture remains the gold standard for MTB detection as per the guidelines of the Centers for Disease Control and Prevention, nucleic acid amplification tests (NAAT), namely the Xpert MTB/RIF (Cepheid) or the MTD (Gen-Probe), are highly recommended as critical assays for TB diagnosis. The advantages to the Xpert MTB/RIF are simultaneous MTB detection and resistance to rifampin, which is used as a marker for multi-drug resistance. The Clinical Microbiology Service at Columbia University Medical Center, a major teaching hospital in NYC, has utilized NAATs for rapid detection of MTB in both pulmonary and non-pulmonary acid-fast smear variable specimens. In addition, the automated AutoGenomics BioFilmChip Microarray technology is used to detect MTB resistance to rifampin and isoniazid and to differentiate Mycobacterium bovis from MTB. Gene sequencing using 65 kDa heat shock protein (hsp65) primers is employed for identification to species level of all mycobacteria. New paradigms for the laboratory diagnosis of pulmonary and extrapulmonary TB, particularly acid-fast smear negative cases, are being introduced due to the implementation of these highly sensitive and specific molecular based assays.
Duncan Dartrey Adams
University of Otago
Keynote: Microbial triggers of Autoimmune Diseases
Duncan Adams entered Medicine with a view to doing research on the aetiology of asthma. However, Dean Sir Charles Hercus apprenticed him to work under Dr HD Purves on use of radioactive iodine in thyroid research. Attacking the cause of Grave’s disease, Adams and Purves discovered the thyroid-stimulating auto antibodies. Later, in Adams’ MRC Autoimmunity Research Unit, John Knight solved the genetics of the autoimmune diseases, which was confirmed at the molecular level by Alan Ebringer, who has discovered two microbial triggers, making it likely that all autoimmune diseases have microbial triggers and will be preventable by discovering and vaccinating against them. Duncan Adams has published more than 138 papers and chapters in medical science journals and books
Microbial triggers. It is now clear that autoimmune diseases are caused by faulty immune responses to specific microbial infections, Alan Ebringer leading the search for these triggers.
Forbidden Clones. Following Niels Jerne’s epochal realization that antibodies are not built on a template of antigen, but are pre-formed in myriad diversity, awaiting selection by an antigen that fits, Macfarlane Burnet realized that it is not antibodies that are selected but the cells that make them, and that these are the lymphocytes. Burnet proposed that each lymphocyte is covered in multiple copies of a single antigen receptor to form an immunological clone, with millions of cells in a clone and millions of different clones in a person. This is his Clonal Selection Theory of the immune response. Furthermore, he realised that multiplying lymphocyte will mutate, and if the mutation involved is in a V (variable region) gene, a new clone will be formed. Hence, his Forbidden Clone Theory, postulating that unlucky mutations in V genes cause the autoimmune diseases. This theory has been amply confirmed in detailed studies of Graves’ disease, which is caused by B lymphocyte forbidden clones, whereas T lymphocyte forbidden clones cause Type1 diabetes.
The H Gene Theory of Adams and Knight explains all the features of the genetics of the autoimmune diseases by postulating that Histocompatibility antigens, major, minor and H-Y, dictate the immune repertoire by deleting nascent complementary clones, and so altering the risk of occurrence of the various autoimmune diseases.
Specific immunotherapy. Because unlucky somatic mutations in multiplying lymphocytes cause the autoimmune diseases, they are unlikely to recur in regenerating immune repertoires following immune ablation and autologous bone marrow cell reconstitution, as shown by Englert et al with cases of lethally severe systemic scleroderma.
After isolation of its auto antigen, an autoimmune disease should be curable by selective destruction of its forbidden clone by making and using a cytotoxic auto antigen complex.
Prophylaxis. This will be possible by identifying and vaccinating against microbial triggers of the various autoimmune diseases. Johannes Salk’s anterior poliomyelitis vaccine has prevented the leg paralyses that must have been a rare autoimmune complication of the virtually universal infections with the virus occurring in poliomyelitis epidemics.
Ebringer has shown that rheumatoid arthritis, ankylosing spondylitis and multiple sclerosis have microbial triggers (Ankylosing Spondylitis and Klebsiella, Springer London 2012). A particular challenge is to find schizophrenia’s microbial trigger, probably a virus, and vaccinate against it to prevent this distressing disease.
Keynote: A New and effective Treatment for Dengue Fever
Professor Max Reynolds has over half a century of global experience in scientific research, business and product development.
His commitment and determination to exploring biotechnology and discovering new practical uses for it has led to many advances, including the development of 98alive™, which is produced by 98 Alive Pty Ltd of which Professor Reynolds is also the Managing Director.
Professor Reynolds has previously lectured at leading Australian university, Griffith University in Brisbane, Queensland and has held the position of Director of the Program in Australasia Botanical Medicine for Population Health.
For over the last 50 years plus the scientific world has been trying to find a way to stop Dengue Fever
without much success until now.
The problem has been to kill all 4 different strains of the virus which cause this disease without injuring the patient in the process.
There have been some major expensive failures in the arena of vaccine production.
Now we have a specialist Australian based company who has developed, from a plant extract, a compound which kills all 4 strains and is safe for human use.
Professor Max Reynolds and his team have carried out all 3 Phase Studies successfully over the last 8 years and have most recently achieved, in a Double Blind 540 patients trial at the University of Airlangga in Indonesia, have achieved a 96.6 % kill rate of all 4 virus within 3 days and at the same time increased the CD4 (helper cells) and CD8 (killercells) immune elements within the patients by 64.4 %.
These results have been achieved by the use of a plant extract from the Melaleucaalternifolia tree, a native of Australia, from which they have removed, by a patented system, 90% of the mono terpenes and made the active compounds safe for human oral consumption.
The compounds dissolve the outer envelope which covers the virus and then proceeds to kill the virus directly.
With this drug Dengue will be a disease of the past as soon as it reaches the worlds medical centers where Dengue is of major importance.
Hamid Yahay Hussain
Dubai health authority
Keynote: Ebola Virus Disease outbreak - Gaps in response, mandate actions and potential threatens
Member of editorial Board of Middle east journal of internal medicine, Middle east journal of age and aging, middle east journal of Alzheimer and psychiatry,
Member of different international, regional and national associations e.g. APHA, IEA, MENAPF, EMAME, American college of occupational and environmental medicine, American college of Epidemiology , American academy of family medicine, Canadian College of family physician, international family physician association(Wonca). Qualified with PhD family and community medicine, Higher diploma respiratory Diseases Edge hills University UK, fellowship of institute for international education/ New York, Master course of Epidemiology and biostatistics / American University of Beirut, training certificates on Public Health ( north Carolina, Johan Hopkins and Harvard) . Supervised more than 20 PhD theses, 10 MSc Thesis, achieved about 50 research work and contribute to CDC textbook on Ethics in Public Health
Background: Ebola virus disease (EVD), formerly known as Ebola hemorrhagic fever, is a severe, often fatal illness in humans, have a case fatality rate of up to 90%, occur primarily in remote villages in Central and West Africa, near tropical rain forests.
Objectives: To study the incidence and mortality rates of EVD outbreak .To identify main threatens and opportunities.
Methodology: MEDLINE and CINAHL electronic databases were searched using a pre-defined search strategy. Additional references from the bibliographies of retrieved articles were also reviewed and experts in the area were contacted
Results: The chronological reporting of EVD cases and mortalities, it revealed that the diseases reported first time on 1976 with high incidence rate mortalities in Sudan and Congo, the table shows as well that frequent EVD outbreaks took place at different African countries in the years 2000, 2003, 2007, 2008 and 2012. the current diseases outbreak(2014) showed that almost 1975 cases has been reported in Guinea, Liberia, Nigeria, and Sierra Leone out of which 1069 case were died according to the last WHO updating on 11th of august
Conclusion: EVD starting to be beyond control in the focus areas in Africa (Guinea, Liberia, Nigeria, and Sierra Leone), and new trend has just start to has potential international dissemination and threatens.
Key Words: EVD outbreak, Gaps analysis, Mandate action
Keynote: Ankylosing spondylitis (Klebsiella), rheumatoid arthritis (Proteus) and multiple sclerosis (Acinetobacter) are autoimmune diseases caused by bacteria.
Professor Alan Ebringer graduated in medicine in Melbourne in 1962. He has published over 300 papers.
Over the last 40 years, his group at King’s College in London, has been studying the bacterial causes of several autoimmune diseases, using as a model “molecular mimicry” which explains how streptococcal infections cause rheumatic fever.
His first book “Ankylosing spondylitis and Klebsiella” (Springer 2011) describes “molecular mimicry” between HLA-B27 and the bowel microbe Klebsiella.
His second book “Rheumatoid arthritis and Proteus” (Springer 2012) demonstrates “molecular mimicry” between HLA-DR1/4 and the urinary microbe Proteus thereby explaining why this disease occurs more frequently in women.
Over the last 20 years, his group has shown “molecular mimicry” between myelin, and the soil, skin and nasal microbe Acinetobacter. These results suggest that this microbe is involved in multiple sclerosis and bovine spongiform encephalopathy (“mad cow disease”).
Molecular mimicry between self-antigens and bacteria is the link that provides a scientific explanation for some autoimmune diseases.
Ankylosing spondylitis occurs predominantly in individuals who possess HLA-B27. The bowel microbe Klebsiellacontains sequences resembling HLA-B27.
Rheumatoid arthritis occurs predominantly in individuals who possess HLA-DR1/4. The urinary microbe Proteus contains sequences which resemble HLA-DR1/4.
Multiple sclerosis occurs predominantly in individuals who suffer from sinusitis. The maxillary sinus microbe Acinetobactercontains sequences which resemble myelin. Anti-myelin autoantibodies are thought to be involved in the pathology of multiple sclerosis.
These specific bacterial discoveries provide a new approach to therapy.
Track 1: Novel Insights in to Clinical Microbiology
Track 2: Microbiology-Current Research
Track 3: Antimicrobials and Disinfectants
Track 4: Infection Control & Immunity
Track 5: Clinical Infectious Diseases
Chris JLM Meijer
Vrije Universiteit Medical Center, (VUMC), Netherlands
Ross University School of Veterinary Medicine, West Indies
Amin A Fadl
University of Wisconsin
Title: Post-transcriptional modification of RNA: Effect on biology and virulence of Salmonella
Amin A Fadl obtained his PhD from University of Connecticut, USA and completed a postdoctoral training at the University of Texas Medical Branch. Currently, he is an Assistant Professor of Microbiology at the University of Wisconsin-Madison. His research focuses on the molecular pathogenesis, immune and inflammatory responses, and host-pathogen interaction of Salmonella. He has published more than 46 papers in reputed journals.
GidA together with another protein known as MnmE catalyze the tRNA modification required for correct gene translation. Our study have shown that GidA and MnmE complex together to modulate virulence genes in Salmonella as indicated by in vitro experiments, animal model and a global transcriptome and proteome analyses. Mice immunized with the gidA, mnmE and gidA/mnmE mutants were protected against a lethal dose of wild-type. The mechanistic basis of such protection was identified to be humoral and cellular immunity with the humoral immune response potentially being the main mechanism of protection. The gidAB operon includes the gidA and gidB genes. The gidB encodes a methyltransferase enzyme responsible for methylation of 16S ribosomal RNA in Escherichia coli. Deletion of Salmonella gidB gene indicated a compromised overall bacterial fitness, significantly reduced motility and showed a filamentous morphology under the stress of nalidixic acid. Most importantly, deletion of gidB conferred high-level resistance to the aminoglycoside antibiotics. Additionally, transcriptional repressor AsnC, located upstream of the gidAB operon, is thought to negatively regulate the gidAB operon post-transcriptionally. Therefore, we investigated the role of AsnC and the environmental factors affecting gene expression in the gidAB operon using transcriptional and proteomic analyses. Taken together, these data indicated that GidA, GidB and MnmE enzymes play a significant role in modulation of biological and virulence characteristics in Salmonella under stress conditions. Further, these studies indicated that the gidAB operon is regulated at post-transcriptional level by glucose and stressful conditions as well as the AsnC protein.
Canadian Food Inspection Agency
Title: Strategy for developing a molecular sub typing tool for a food borne bacterial pathogen using a whole genome analysis approach: the case of Salmonella Enteritidis
Dele Ogunremi is a research scientist at the Canadian Food Inspection Agency, Ottawa Laboratory Fallow field, and is a trained veterinarian with doctoral and postdoctoral training in Molecular Biology and Immunology. He graduated with DVM (1984) and MVSc (1986) from the University of Ibadan, Nigeria and PhD from the University of Saskatchewan, Canada (1993), where he also completed a postdoctoral training (1996). His research interests include the application of genomics sequencing technology to detect, identify and characterize food borne microbial pathogens. He has generated, assembled and characterize Salmonella and Listeria genomes. Recently he completed the development of genomics based single nucleotide polymorphism genotyping test for Salmonella Enteritidis. He played a lead role in the establishment of the Pulse Field Gel Electrophoresis at the Canadian Food Inspection Agency. His research work has been protected by patents granted in Canada, United States and New Zealand.
Differentiating between outbreaks of food borne infections and sporadic occurrences requires a highly discriminatory laboratory tool for pathogen subtyping which has remained a difficult goal for a highly clonal organism such as Salmonella Enteritidis (SE). Because of the high degree of similarities among isolates of SE, developing a reliable and informative sub typing tool requires the availability of comprehensive information on the organism such as that provided only by whole genome analysis. By examining core and accessory genome compartments of SE for genetic diversities, markers and traits that distinguish related strains from unrelated strains can be defined. The proportion of SE genome constituting the core compartment is large, i.e., >95% of the entire chromosome, and diversity among isolates are limited to an average of 600 single nucleotide polymorphisms (SNPs) between two apparently, unrelated strains. The accessory genome include mobile genetic elements especially prophages, which are made of remnants or full sequences of infecting bacteriophages that have since integrated into the SE genome. It was found six prophages to be present consistently and showing little or negligible diversity among 12 SE genomes including the reference Salmonella Enteritidis P125109 phage type. The presence and composition of another eight prophages or remnants provided considerable diversity for delineating isolates into groups of SE subtypes. The use of SNPs in SE genomes assembled with the aid of the reference P125109 genome and the composition of prophages could provide a complementary and synergistic framework for differentiating SE subtypes for the purpose of regulatory intervention during food borne outbreaks.
Ross University School of Veterinary Medicine
St. Kitts, West Indies
Title: Bovine trichomoniasis in beef cattle in Wyoming, USA
Dr. Chaoqun Yao completed a MD and PhD from Tongji Medical University and University of Georgia, respectively. He pursued post doctoral training at Washington State University and University of Iowa where he was promoted to Associate Research Scientist. He was a tenure-track Assistant Professor at University of Wyoming with a concomitant appointment of Parasitologist at the Wyoming State Veterinary Laboratory. He is currently an Associate Professor at Ross University School of Veterinary Medicine. He has published more than 45 papers inpeer-reviewed journals and has been serving as an editorial board member and ad hoc reviewer for many journals.
Tritrichomonasfoetus, a trichomonad protozoan residing in the reproductive tract of cattle and the large intestine of the domesticated cat, causes trichomoniasis with very different clinical manifestations among these animals. Bovine trichomoniasis is a venereal disease and has been endemic in the USA since the 1930s. Currently 26 states including Wyoming impose regulations on the disease. The major purpose of the present study was to gauge the disease trend in Wyoming since enforcement of the state rules. A retrospective study was performed to include all accessions between 2000 and 2010with producers’ residency in Wyoming collected by the Wyoming State Veterinary Laboratory and the Wyoming Livestock Board. A questionnaire surveyed all Wyoming beef cattle producers to identify herd-level risk factors. Individual prevalence of T. Foetus in bull populations declined from 1.7% to 0.2%, which was linearly correlated with year (R=0.717, P=0.009). At the same time an accumulative positive rate of 9.7% and 4.5% was found among 93 cows/heifers with a history of abortion and 22 aborted fetuses, respectively. Furthermore, between 2007 and 2010, average herd prevalence was 2.2% with 15 of the 23 counties having at least one positive herd.T. foetus infections were found to be significantly (P0.05) associated with neighbouring positive herd(s), grazing on public allotments and comingling with other herds. Based on these data an integrated approach was developed to control and eventually eliminate bovine trichomoniasis, which is suitable for all herds in which live cover breeding is used regardless of geographic regions.
Vrije Universiteit Medical Center, (VUMC), Amsterdam
Title: From cytology to full molecular cervical screening
Chris Meijer received his MD in 1972 at the Leiden University Medical Center (LUMC), The Netherlands, and was awarded with his PhD during his medicine study in 1971 at the Vrije Universiteit Medical Center, (VUMC) Amsterdam. He is board certified as surgical pathologist (1977 Leiden) medical immunologist since1981. He was staff pathologist at the Dept. of Pathology of LUMC till 1980, and staff member and head of the Department of Immunopathology at the SSDZ, Delft, from 1980-1982. From 1983-2009 he was chairman and director of the Dept of pathology of the VUMC, Amsterdam. He is and has been chairman or member of numerous national and international scientific and strategy committees advising in the field of oncology and pathology. He received several awards for his scientific work.
Presently he works at the Dept. of Pathology, VUMC, Amsterdam heading a research group aiming to elucidate molecular aspects of HPV infections leading into transforming cervical intra-epithelial neoplasms and cancer and focusses on the role of promotor methylation of tumor suppressor genes involved in carcinogenesis. As such he is conducting several large clinical trials evaluating the role of HPV-self sampling and HPV testing as primary screening tool in optimising cervical cancer screening programmes and the role of methylation markers in cervical screening and the management of HPV positive women
Evidence will be given why HPV testing should be used as primary screening test instead of cytology. It will be shown that a negative HPV test protects better than a negative smear against cervical cancer and the most advanced precursor lesion for cervical cancer i.e. CIN3. In addition guidelines for clinical validation of HPV tests will be discussed. Since a considerable number of HPV positive women have a transient HPV infection triage of HPV pos women is necessary to detect in the group of HPV positive women those who have disease i.e cervical (pre)cancer. This is presently done by cytology and/or HPV 16/18 genotyping. Principles for HPV triage testing will be discussed. New developments in triage testing are dual staining of cervical cells for Ki67/p16 and molecular testing by methylation marker. The use of HPV testing on self-collected cervico-vaginal specimens and direct molecular triage testing with methylation markers on the left overs of these self-collected samples opens the way to full molecular screening. Finally the way how The Netherlands is implementing HPV testing in population based screening including the possibility for women to use HPV-self-sampling will shortly be discussed
ACURE Biotechnology, Inc
Title: IL17 Pathway Involves Moderating Pulmonary Hypertension, a common complication of COPD, in Statins Therapy in Smoking Rats
Xin Wang gained her MD at the age of 21 years from Hebei Medicine University in China. She specialized as neurologist to study cerebrovascular diseases. The project, stroke rehabilitation, awarded the First Prize of excellent paper in the Third Annal National Neurologic Rehabilitation meeting. After ten years clinical medicine practicing, including clinical research, she pursued PhD on Medical Science from Institute on Aging and Adaptation, Graduate School of Medicine, Shinshu University in Japan. During PhD course, she discovered an acute leukemia associated brain-specific gene and was the first one who studied its function at molecular & cellular biology level. The first postdoctoral training in NIH/NCI-Frederick in US, gave her an opportunity to complete a publication studying mouse model related to cardiovascular dysfunction that had been pointed as 'must reads' by Faculty1000. In Dental School & Medical School of University of Maryland, Baltimore, Xin studied head & neck cancer and prostate cancer with human specimen, cancer cell lines and animal models. Her summary of a project about tumor microenvironment as primary author gained Minority Scholar Award in AACR meeting. She is the CEO and CSO in biotechnology company, ACURE TECHNOLOGY, Inc. that she is the founder, to develop and discover stem cell therapeutic products based on her extensive medical research experiences. She has published more than 20 peer-reviewed papers in scientific journals.
Smoking and atmospheric pollution are major causes of COPD affecting public health. Lack of effective treatments leads to COPD and later on causes Pulmonary Hypertension. Statins, lipid lower drugs, are reported to be beneficial to COPD patients. However, the mechanism remains unknown. Here, we establish a smoking model with rats. Increase of mean linear intercept and decrease of mean alveolar number are seen in smoking rats (P<0.01). Neutrophils, lymphocytes, especially macrophage pullulate in bronchoalveolar lavage fluid. Infiltration of immune cells is substantially observed in the lungs of smoking rats. IL-17, RORγt and IL-21 positive cells manifold in bronchial epithelium and alveolar septa and walls of smoking rats in immunohistochemistry staining. The mRNA expression of IL-17, RORγt and IL-21 are increased in smoking group (P<0.01). The impair of lung function of smoking rats is demonstrated by significant subtraction of forced expiratory volume in 0.3 second vs. forced vital capacity, decrease of lung dynamic compliance and increase of resistance of inspiratory and expiratory. Pulmonary artery hypertension in smoking rats is determined by dramatically elevating pulmonary artery pressure. Statins therapy reverses increase of neutrophils, especially macrophage, but not lymphocytes. Expression of IL-17, RORγt and IL-21 in lung of smoking rats is significantly reduced by Statins at both protein and mRNA level (P<0.05). Surprisingly, although pathological changes and damage of lung function caused by smoking are slightly improved, the mPAP is largely reduced by Statins therapy in smoking rats. This study suggests that IL-17, IL21 and RORγt play an important role in the progression of COPD in smoking rats. Statins can relieve the PAH in smoking rats, probably through reducing the expression of IL17 and its mediators. Statins have a great potential in application for COPD patients complicating with pulmonary hypertension.
Singapore Eye Research Institute
Title: Breaking down barriers to promote effective antibiotic action
Roger W Beuerman is currently Senior Scientific Director of the Singapore Eye Research Institute, Professor of Neuroscience and Behavioral Disorders at DUKE-NUS School of Medicine and Adjunct Professor of Ophthalmology, Yong Loo Lin, School of Medicine at the National University of Singapore, and Head of Translational Research, and National Distinguished Professor (FidiPro) of Ophthalmology at the University of Tampere Medical School in Finland. His focus has been on understanding eye disease for the development of new diagnostics and therapeutics for infectious disease.
Gram negative bacteria are a particular therapeutic issue as they mutate readily, and this tendency is especially common for Pseudomonas aeruginosa resulting in clinically resistant strains. To combat this problem some antibiotics are used at much higher concentrations to combat Pseudomonas. Part of the issue of admitting antibiotics into these bacteria and killing gram negative bacteria lies in the protective outer membrane, which is overlain with lipopolysaccharide, LPS. Small, branched peptides that interact with various sites in the LPS assembly with the results that the LPS change its structural mode is being developed. It is hypothesized that this may allow greater penetration of antibiotics into the bacteria. To test this idea, the method of fractional inhibitory concentration index was used to test gatifloxicin and a peptide B2099. It was found that the FICI was about 0.53 suggesting that there was synergistic action between the peptide and the gatifloxicin. Next, this combination was tested in a mouse model of corneal infection using P. aeruginosa ATCC9027 with B2099 (0.5 mg/ml), gatifloxicin (3 mg/ml-the standard ophthalmic dose), and a combination of B2099 (0.5 mg/ml) and gatifloxicin (1.5 mg/ml-half the usual dose). The result showed that the combination was significantly more active than either drug alone. Further, it was shown that B2099 interacts with lipid A to allow penetration of the gatifloxicin and the effect extends to E. coli and K. pneumonia. This unique interaction may provide a new method for killing this difficult group of pathogens.
Polish Mothers Memorial Hospital Research Institute
Title: Genetic variability of HCMV strains isolated from Polish pregnant women, their fetuses and newborns
Wioletta Wujcicka has completed her Ph.D. in medical sciences at the age of 28 years from Medical University of Lodz, in Poland. She was an assistant at the Polish Academy of Sciences in Lodz. She is an assistant professor at the Department of Fetal-Maternal Medicine and Gynecology at the Polish Mother’s Memorial Hospital Research Institute in Lodz. She is a member of several scientific societies including the European Society of Clinical Microbiology and Infectious Diseases as well as the Polish Society of Human Genetics. Her scientific papers are related to genetic background of congenital infections with Toxoplasma gondii and HCMV.
Human cytomegalovirus (HCMV) is the most common cause of intrauterine infections worldwide. HCMV genome is the largest among human herpesviruses, and nucleotide variability was reported for about 20 viral ORFs. Considering highly polymorphic region within glycoprotein B (gB), four main gB1, gB2, gB3 and gB4 genotypes, and three rare non-prototypic variants: gB5, gB6 and gB7, were identified.
In this study we searched for HCMV genotypes in pregnant Polish women, their fetuses and newborns. The impact of the genotypes on both the transplacental transmission of the virus and disease outcome in the offsprings was estimated as well.
The study included eighteen pregnant women, their nine fetuses and five neonates with confirmed HCMV infections. Serological status of the patients was assessed by CLIA and ELFA tests. HCMV DNAemia in various clinical samples and the viral genotypes were determined by real-time PCR assays for UL55 gene.
The infections during pregnancy were caused by gB1, gB2, gB3 and gB4 viral genotypes. Fourteen (77.8 %) of pregnant women were infected with single gB2, gB3 or gB4 genotypes of HCMV and four (22.2 %) with multiple gB1-gB2 or gB2-gB3. Maternal genotypes of HCMV influenced the viral genotypes causing congenital infections in their offspings (P≤0.050). A half of fetuses and newborns were singly infected with gB1, gB2 or gB3 viral genotypes and the other half – with multiple gB1-gB2 or gB2-gB3. The infections with single and multiple HCMV genotypes were found in asymptomatic as well as symptomatic disease. The gB3 genotype was related only to the symptomatic cytomegaly.
Monica Cartelle Gestal
Title: Tyrosine - silica biostructures for improved drug delivery against S. aureus
Monica Cartelle Gestal has completed her PhD at the age of 26. Since then she has been awarded with the Barrie the la Maza postdoctoral fellow and two different Marie curie awards. The first, a FP7 for her project to study quorum sensing system, in the second she was part of a study group to develop new antibiotic targets. Now she is a Professor and P.I and she wants to join together her two passions, antibiotic resistance and quorum sensing.
Despite the technological progress, the rate of efficient antimicrobial drug development remains low. Due to the increasing microbial resistance rates, novel therapeutic approaches for increasing the efficiency of current drugs are needed. Here, we report the synthesis, characterization (FT-IR, XRD, SEM) and bioevaluation of a novel tyrosine-silica hybrid nano-biostructure, for the improved delivery of erytrhomycin and clotrimazol antibiotics, used in the treatment of Staphylococcus aureus infections. Antimicrobial effect was quantified by a dilution method in order to establish the MIC (minimal inhibitory concentration) of tested antibiotic loaded nanostructures. In vitro biocompatibility was assessed through fluorescence microscopy. The prepared nano-biostructure showed IR bands which were assigned to Si–O–Si (stretch mode). The prepared bio-active nanostructures have significantly improved the anti-staphylococcal activity of erytrhomycin and clotrimazol, as demonstrated by the drastic decrease of the MIC of the respective antibiotics loaded in the nanostructure. These results, correlated with a good biocompatibility of this porous structure, tested on cultured human diploid cells highlights the possibility of using this nano-carrier for the delivery of the antimicrobial substances in lower active doses, thus reducing their cytotoxicity and related side-effects.
Title: Invitro antifungal susceptibility testing of 5 antifungal agents againstdermatophytic species by CLSI(M38-A) micro dilution method
Gadangi Indira has completed her PhD on comprehensive study of dermatophytosis of Warangal district, A.P from Kakatiya University and working on a major research project funded by UGC at present. She is the HOD of Department of Microbiology in Pingle Govt. U.G and P.G College at Warangal. She has published more than 15 papers in reputed journals, and given proceedings of both national and international seminars.
Cases of dermatophytoses have increased over the past few decades. In the last few years, a number of newer less toxic antifungal drugs have become available for clinical use. The increased use of antifungal, often for prolonged periods, has led to the recognition of the phenomenon of acquired antifungal resistance among previously susceptible strains or species and to the increased incidence of infections with less common species. Our study mainly focused on the in vitro susceptibility of clinical isolates of dermatophytes. The microbroth dilution method was performed according to CLSI standards. In the present study, antifungal susceptibility testing was done by micro dilution method of dermatophytes against 5 antifungal agents namely, ketoconazole (imidazoles) fluconazole, itraconazole (triazoles), griseofulvin and terbinafine and their activity against significant number of strains, representing a wide spectrum of dermatophyte species is assessed.
Dermatophytic strains: A total of 119 strains of dermatophytes belonging to 10 species were tested. They were T. rubrum (n=40), T.mentagrophytes (n=19), T. violaceum (n=15), M. gypseum (n=12), E. flocossum (n=9), M. audouinii (n=8), T. schoenleinii (n=5), M. canis (n=5), T. tonsurans (n=4) and T. verrucosum(n=2). The MIC ranges of all the 119 isolates of dermatophytes tested for antifungal susceptibility showed that terbinafine had the lowest MIC range of 0.001 to 0.64 μg/ml followed by ketoconazole at a MIC range of 0.01-3.84 μg/ml. The itraconazole showed a MIC range of 0.082-20.45 μg/ml whereas the griseofulvin and fluconazole showed a highest MIC range of 0.32-5.12 μg/ml. The MIC50 of terbinafine was low at 0.02 μg/ml followed by ketaconazole 0.24 μg/ml.The MIC50 of itraconazole and griseofulvin was 1.28 μg/ml. The highest MIC50 with 2.56 μg/ml was recorded for fluconazole. The MIC90 of terbinafine was low at 0.32 μg/ml followed by ketaconazole with 1.92 μg/ml. The MIC90 Itraconazole was 2.50 μg/ml and for griseofulvin it was 2.56 μg/ml. The highest MIC90 of flucanozole was high at 10.24 μg/ml. In our study, we observed that terbinafine had the lowest MIC values compared to ketoconazole, itraconazole, griseofulvin and fluconazole.
St. Lukes Medical Center
Title: Molecular Analysis of Viral Pathogens Causing Diarrhea in Filipino Children
Maria Luisa G 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 20 papers on dengue, Japanese encephalitis, eye infections, genetic markers of various diseases, and microbiology. She is Chair of the Board of Examiners of the Philippine Academy for Microbiology and authored a chapter of a book on Philippine microbiology research. Current researches include dengue, chikungunya, diarrhea, CNS infections, pathogen genomics, plant antivirals, molecular diagnostics, dengue POC, and genetics of CVD, diabetes, thyroid cancer, and dementia.
Rotaviruses and noroviruses are two important etiologic agents of acute diarrhea in the world, with an especially pronounced impact in the developing regions of Asia and Africa. Rotaviruses are non-enveloped RNA viruses, with an outer capsid composed of the VP7 glycoprotein and the VP4 attachment protein, coded by gene segments 9 and 7, respectively. These proteins are the basis of a binary system of classifying rotaviruses into G and P genotypes, respectively. Two vaccines against rotavirus target the most prevalent genotypes, G1P, G2P, G3P, and G4P but not emerging genotypes such as G9 and G12. Noroviruses are single-stranded RNA viruses that are classified into 5 genogroups (GI-GV), of which I , II and IV are found in humans. At present there is still no vaccine against noroviruses. This study was performed to determine the genotypes of both rotaviruses and noroviruses in 105 samples collected from children who presented with diarrhea at 2 hospitals in the Philippines in 2010-2013. Thirty-six samples (34.38%) were positive for rotavirus group A by enzyme immunoassay and RT-PCR, and 25 (23.81%) were positive for norovirus by RT-PCR. Rotavirus genomic RNA from 34 samples was electropherotyped using PAGE which identified 8 electropherotypes in 26 samples. Predominant genotypes in rotavirus-positive samples were G1P with 22 (61.1%) of the samples, followed by G2P with 6 (16.6%), G3P with 5 (13.8%), G9P with 2 (5.55%) and G4P with 1 (2.7%). The norovirus- positive samples were made up of 3 (12%) belonging to genogroup I, and 22 (88%) to genogroup II. The latter group includes the Sydney strain presently causing outbreaks in different countries worldwide. Amplified DNA from the gene segment 9 of rotavirus and VP1 gene of norovirus were sequenced by Sanger dideoxy method. Phylogenetic analysis showed clustering of our isolates with those isolated in 2005-2012 from the Asia-Pacific region.
Vasantha Kumari Neela
Universiti Putra Malaysia
Title: Could clinical Stenotrophomonasmaltophilia be a potential pathogen in clinical setting?
S. maltophilia which is an environment pathogen has recently emerged as nosocomial pathogen. In clinical setting, it usually infects immunocompromised patients by forming biofilm in the lines and catheters, thereby causing systemic infections. Although S. maltophilia are associated with a wide range of infections, bacteraemia/sepsis and respiratory illness are the most common cause of mortality. Its intrinsic resistance to most antibiotics and problems associated with in vitro susceptibility testing makes the therapeutic success always a challenge. In clinical setting, it’s always a debate whether S. maltophilia is a true pathogen or only a colonizer with multiple drug resistance property. We investigated the pathogenic potential of S. maltophilia isolated from various clinical presentations and compared with environmental strain. Virulence potential of S. maltophilia will be explained during the presentation.
Africa Health Research Organization
Title: Efficacy of combined Antiretroviral Therapy (cART) in Hepatitis B and C-associated Hepatocellular Carcinoma-A systematic review of randomized controlled studies
Introduction: Hepatocellular carcinoma (HCC) ranks the fifth most common cancer among men and the seventh among women globally. It is aggressive with hepatitis fibrosis or cirrhosis being the most common end-stage. The well know risk factors includes chronic infection with Hepatitis B virus (HBV) and hepatitis C virus (HCV) while habitual betal gum chewing and secretan have been implicated. The most effective measure to prevent or reduce the incidence of HCC is by utilizing antiviral agents. This study is to review combined antiretroviral therapy (cART) in HBV & HCV-associated confirmed HCC.
Methods: A systematic review of randomized controlled trials published in Pub Med and Clinical Trials Sites will be carried out. Citation of included studies will be checked to identify additional studies not identified in the electronic searches. It will include any randomized controlled studies that considered combined ART in HBV & HCV associated HCC. Data extraction and analysis will be performed. The extracted data will be discussed, decision documented and, where necessary the authors of the studies will be contacted for clarification.
Ethics & Dissemination: Ethical approval is not required given that this is a protocol for a systematic review. The finding of this study will be presented at the 12th Clinical Microbiology Conference in Spain and also published in peer reviewed journal. Updates of the review will be conducted every two years to inform and guide healthcare practice.
Institut Pasteur de Tunis
Title: A ten-year evolution of a multidrug-resistant tuberculosis (MDR-TB) outbreak in an HIV-negative context, Tunisia (2001-2011)
After obtaining his doctorate in Veterinary Medicine (1988) at the Tunisian National School of Veterinary Medicine, Helmi Mardassi moved to the University of Montreal (Canada) where he completed a master degree in Microbiology and Animal Pathology. In 1996, he obtained his PhD degree in Molecular Virology at the Institut Armand-Frappier (University of Quebec, Canada), and next joined the Biotechnology Research Institute of Montreal for a post-doctoral training. Actually he is leading a research unit focusing on molecular epidemiology, evolution and genetics of Mycobacterium tuberculosis. He has published more than 26 papers in reputed journals.
Basically, almost, if not all, multidrug-resistant tuberculosis (MDR-TB) outbreaks described thus far have expanded among HIV-positive patients. Here we describe the emergence and expansion, through a 10-year period, of a MDR-TB outbreak among non-institutionalized, HIV-negative, and apparently healthy young individuals.
Since its identification in September 2001 to June 2011, the outbreak involved 45 individuals (mean age 29.7 yrs; 88.9% male), 17 (37.78%) of which were cured, while 9 (20%) have died. Failure and relapse cases represent 13.33% and 8.89%, respectively.
The Haarlem3-ST53 outbreak strain evolved mainly as an 11-banded IS6110 RFLP profile (77.77%), while the remaining (22.23%) strains exhibited one additional band (12-banded). The majority (95.55%) of the outbreak-associated strains displayed the same MIRU-VNTR24 profile. The 12-banded outbreak strains, though less frequent, were significantly more associated with smear positivity [OR=4,333 (0,992-18,938); P = 0.043]. However, there were no significant differences between the 11- and 12-banded strains in terms of epidemiological and treatment outcome parameters
Drug susceptibility testing coupled to mutational analysis of drug resistance-conferring genes, revealed that initial transmission involved a strain that is simultaneously resistant to isoniazid, rifampicin, ethambutol, and streptomycin. Secondary resistance to pyrazinamide and second-line drugs was further acquired. Despite frequent cases of non adherence and treatment discontinuation, only one strain evolved to the XDR phenotype.
Taken together, the data indicate that MDR-TB outbreaks can successfully evolve and cause substantial death rates among HIV-negative young patients. The study also suggests that despite the concourse of favorable behavioral factors, transition to the XDR phenotype is unlikely to be systematic.
The Federal Polytechnic
Title: Antimicrobial, phytochemical and haemolytic evaluation of Kamue Local Concoction in vitro
Comfort Vandu is a chief medical laboratory scientist in charge of the laboratory unit, medical centre, Federal Polytechnic Mubi, in Adamawa state, Nigeria. She had her degree (AIMLT) in 1994 and post graduate studies in 2007 (FMLSCN). She obtained a higher diploma in business administration and management studies in 2004, was awarded the “Fellow Business Specialist and Entrepreneur” in 2011 because of her contributions to business and entrepreneurship in her community; in this regard she is the co-ordinator of the ENACTUS Federal Polytechnic Mubi Team. Her interest, commitment and determination to clinical research are deterred by financial/gender constraints and administrative bottle necks, she has one publication and two papers presented nationally.
Aqueous extracts of the barks of Azadirachta indica, Parkia biglobosa, Anogeissus leiocarpus, Vitellaria paradoxa, Kaya senegalensis, red potash and their combination, (a Kamue antityphoid traditional medicine), were investigated for antimicrobial activities on Salmonella typhi, Eschericia coli and Pseudomonas species using the disk diffusion method. E. coli, Pseudomonas spp. and S. typhi showed strong vulnerability to A. leiocarpus and A. indica extracts. The highest activity was with A. leiocarpus in E. coli, Pseudomonas spp. and S. typhi with comparable activity to the standard antibiotics. Phytochemical analysis showed the presence tannins, saponins and anthraquinones among others. Elemental analysis of the red potash showed the presence K and Na in concentrations of 0.424 and 0.163 ppm, respectively. The haemolytic analysis conducted with rat’s whole blood, showed agglutination of RBC with V. paradoxa, A. leiocarpus, K.senegalensisandthe concoction at concentrations of 63 and 125 mg. This study showed that, “Kamue antityphoid concoction” is an effective antibiotic.
Armenian National Agrarian University
Title: Comparative analysis of bacterial enumerations in fecal samples of patients with Familial Mediterranean fever disease by culture-based and phylochip techniques
Astghik Pepoyan has completed her PhD in 1990 and DSc in 2002 at the Institute of Biochemistry at NAS RA. She is the head of the Food Safety and Biotechnology department at Armenian National Agrarian University. She is also President of the International Association for Human and Animals Health Improvement, and the scientific consultant at Neo Lab Diagnostic Center, Yerevan, Armenia. She has about 200 publications in reputed journals and serving as an editorial board member of repute.
Background: Cultural analysis is widely applied method for the bacterial identification and differentiation in majority of Armenian clinical laboratories. It is commonly used for the quantitative assessment of gut E. coli, Lactobacillus spp. and Bifidobacterium spp. in gut microbiota of patients with familial Mediterranean fever (FMF) disease during the probiotic-therapies.
Main goal of current investigation was the comparison of results on numbers of isolates of Escherichia, Lactobacillus spp. and Bifidobacterium spp. in gut microbiota of Armenian FMF patients by the clinical culture-based and phylochip techniques.
Methods: For the assessment of bacterial numbers the clinical cultivation and phylochip™ analysis was used.
Results and Conclusion: The results of investigations revealed compliance between cultural and phylochip analysis for the identification of bacterial counts for E. coli and Lactobacillus spp. Despite of a variety of not cultivated lactobacilli, total of these isolates in gut microbiota of investigated patients was comparable with the totals of the isolates described in the course of the cultural analysis. No relationship found between the culture and phylochip results for bifidobacteria.