Day 3 :
The University of Notre Dame, Australia
Time : 10.00AM
Professor George L. Mendz has an MSc from the University of Barcelona (Spain), and a PhD from the University of NSW. He has been Lecturer at the College of Natural Sciences, the University of Puerto Rico (San Juan); Research Associate at the School of Chemistry and the Department of Biochemistry, the University of Sydney; ARC Senior Research Fellow and Associate Professor at the School of Biochemistry and Molecular Genetics, the University of NSW; Associate Professor at the Department of Bacteriology, the University of Bordeaux II; and Associate Professor at the School of Medical Sciences, the University of NSW.
Background: There is limited knowledge of the community structure of the vaginal microbiome of pregnant women. This work aimed to provide a more complete description the vaginal microbiome of pregnant women in late pregnancy, and to investigate differences between the microbiota of healthy gravidae women and those who experienced complications during pregnancy, in particular women giving birth at term or preterm. rnMethods: Upper vagina swabs were obtained from 225 women of Caucasian, Asian, Indian, Middle East, and Pacific Island racial backgrounds during the third trimester of pregnancy. Participating women were administered a questionnaire including demographic data and summarising the history of the pregnancy; 132 women of five ethnic backgrounds had no complications during the pregnancy. The identity of the taxa present was determined by ultrafast sequencing of the V1V3 regions of the 16S rDNA gene of DNA extracted from the swabs. The sequence data were analysed with MOTHUR and statistical analyses were performed employing various bioinformatics tools.rnResults and Discussion: The relative abundance of some phyla was different in pregnant and non-pregnant (from other studies) women. Multidimensional scaling analyses showed the bacterial populations of women without complications to cluster in four vaginotypes driven by four Lactobacillus species. This organisation was partially dependent on the racial background of the woman and was altered by complications during pregnancy. A CLUSTER analysis showed a fine structure in these vaginotypes created by taxa belonging to the genera Escherichia, Atopobium and Prevotella. The participants were stratified into two groups: with or without complications during pregnancy. Using univariate analysis, 9 genera were found significantly correlated with genital infections. PERMANOVA analyses yielded significant differences between the microbiomes of both groups
University of Tulsa, USA
Time : 10.30AM
Mohamed K. Fakhr, is currently working as an Associate Professor of Molecular Microbiology in the Department of Biological Science at the University of Tulsa, USA. After obtaining his Ph.D. from Oklahoma State University in 2002, Dr. Fakhr, moved to North Dakota State University where he worked as a Postdoctoral Research Associate then a Research Assistant Professor at the Department of Veterinary and Microbiological Sciences. In 2008, Dr. Fakhr moved to Tulsa, where he currently runs an active research program in the area of Molecular Typing and Detection of Foodborne Bacterial Pathogens alongside exploring the mechanisms by which these foodborne pathogens develop resistance to antimicrobials
Staphylococcus spp. is a known cause of food poisoning in humans. The prevalence and virulence of staphylococci in retail mushrooms is understudied despite their proposed role as a mecA gene reservoir. One of the objectives of this study was to determine the prevalence of staphylococci in both conventional and organic retail mushrooms sold in the Tulsa, Oklahoma area. Characterizing the isolated strains for their possession of toxin and mecA genes was also aimed at. Molecular typing of the isolated strains to determine their origin and genetic diversity was also performed. A total of 420 samples of retail mushrooms were purchased from retail stores including Asian markets across the Tulsa area. Staphylococcus spp. was isolated from 14 different domestic and imported brands of fresh mushrooms. Prospective colonies of Staphylococcus aureus were identified by PCR and the presence of 18 toxin genes and the mecA gene were screened for by PCR. A total of 297/420 mushroom samples (70.71%) was positive for the presence of Staphylococcus spp. The prevalence of S. aureus in the tested mushroom samples was only 1.6%. The mecA gene was detected in 64/297 (21.55%) of the positive samples. A total of 552 isolated staphylococcal strains were also tested for the presence of 18 toxin genes. The prevalence of enterotoxins ranged from 0.1% to 2.1%. The see, sej and sei genes were the most detected enterotoxins among all isolates and the exfoliative toxin (eta) gene was detected in only one isolate. The genes of toxic shock syndrome toxin 1, leucocidins (lukE/lukD and lukM), and Panton-Valentine leucocidin (PVL) were not detected in any of the staphylococcal isolates screened. A subset of 120 staphylococcal isolates was subjected to 16S rDNA gene sequencing and was molecularly identified. A total of 10 different Staphylococcus species was detected including S. aureus, S. fleurettii, S. saprophyticus, S. vitulinus, S. sciuri, S. xylosus, S. succinus, S. pasteuri, S. warneri, and S. haemolyticus. More than half of the screened S. fleurettii strains carried the mecA gene. Genetic diversity among the identified staphylococci strains showed higher diversity reflecting the mushroom compost environment. Genetic diversity in the mecA gene among a selected number of isolated staphylococci in this study showed a higher degree of DNA homology among the isolates, indicating that these isolates can serve as a reservoir of this important gene in the environment. Molecular typing using PFGE for the identified staphylococci showed a high degree of diversity that varied according to the mushroom brand. PFGE, MLST and spa typing of the isolated S. aureus strains showed two distinct strains of a human origin, supporting that contamination with S. aureus was as a result of human handling of the mushrooms during packaging. In conclusion, the prevalence of Staphylococcus spp. in fresh mushrooms is high and a subset of the strains was shown to harbor enterotoxin genes, which might lead to foodborne poisoning. The mecA screening and genetic diversity results support the hypothesis that those staphylococci other than S. aureus can serve as reservoirs for this methicillin resistance gene.
- Track 1: Bacterial Morphology and Metabolism
Track 3: Bacterial Identification Method
Track 6: Plant Microbe Pathology
Track 7: Veterinary Bacteriology
Drexel University, USA
University of Tulsa, USA
Center for Disease Control and Prevention, USA
Brunilís Burgos-Rivera completed her PhD in Genetics from the University of Georgia in 2012. Currently, she is a microbiologist at the Pertussis and Diphtheria Laboratory at the Centers for Disease Control and Prevention (CDC) working as a contractor. She has served as the Laboratory Coordinator for the Latin American Pertussis Project, a collaboration between CDC, Sabin Vaccine Institute, Pan American Health Organization, and the Ministries of Health in select Latin American Countries to strengthen pertussis surveillance and diagnostics in the Region. More recently, she has been selected as a Fellow to the CDC Laboratory Leadership Service, Class of 2016
The Pertussis and Diphtheria Laboratory at the Centers for Disease Control and Prevention (CDC) developed a real-time PCR (rtPCR) assay for the detection of three Bordetella species. This assay is routinely used at the CDC, public health laboratories in the USA and internationally for pertussis diagnostics. It relies on one approved and validated PCR master mix, Applied Biosystems Taqman Gene Expression Master Mix (GE). In recent years PCR master mixes have been engineered to be resistant to PCR inhibitors, thus potentially providing more sensitive alternative master mixes to be used in this assay. Three commercially available alternative master mixes were identified: Quanta PerfeCTa SuperMix (QS), Quanta PerfeCTa ToughMix (QT) and Quanta PerfeCTa ToughMix with UNG (QTU). The performance of these master mixes was evaluated by comparing target-specific detection levels and consistency among three rtPCR instruments previously validated and currently used for routine pertussis diagnosis. The analytical sensitivity and specificity of these alternative PCR master mixes was assessed by the Bordetella species rtPCR multi-target assay. We report that QS produced Ct values comparable to GE, while QT and QTU produced lower Ct values (≥2 Ct value difference) and thus exhibited higher analytic sensitivity across all assays and instruments. Therefore, before selecting an alternative master mix or rtPCR platform, laboratories should validate their own diagnostic assay to ensure it is producing results with clinically relevant cut-offs, as those previously published.
Essam Badawy has completed his PhD Minia University, Egypt and Postdoctoral studies from Cairo University School of Medicine. He is the Director of Emergency Department, Hera General Hospital, JCI-Accredited Governmental Hospital, MOH, KSA. He is a Senior Consultant Internal Medicine & Professor of Internal Medicine & Immunology, Faculty of Medicine, Minia University. He has published more than 24 papers in reputed journals and has been serving as an Editorial Board Member of repute.
The recent discovery of Human metapneumovirus (hMPV) as a major respiratory pathogen has been made possible by means of RT-PCR. Studies thus far published have mostly been conducted using the molecular approach; clarification of epidemiological, clinical features and using molecular biological techniques for diagnosis of hMPV. 189 patients with suspected viral respiratory tract infections were included and respiratory specimens were analyzed for hMPV by Seeplex respiratory virus detection kit. Detection techniques that were used included virus detection by RT-PCR, DFA-staining and the rapid culture technique known as shell vial amplification using Mabs of nasal wash or aspirate fluid. The study determined that 61 (32.3%) respiratory viruses out of 189 respiratory samples and showed presence of hMPV in 8 (13.1%) of 61 samples and epidemiological data showed that hMPV had variable seasonal activity. Sex patients with positive hMPV (75%) had preexisting serious disorders. By using shell vial cultures with monoclonal antibodies (MAbs), the related isolated virus of the patient with Non-Hodgkin Lymphoma (NHL), showed a plaque of infected cells with small syncytial formations, while that of other seven patients showed single infected cells. All samples with hMPV positive patients by RT-PCR were correlated with whatever DFA staining or shell vial cultures by MAbs. hMPV is a significant pathogen in immunocompromised patients with a risk of high morbidity and mortality. Using combination of diagnostic work up may be useful to confirm detection of hMPV.