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:

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.