Day :
- Track 1: Bacterial Pathogenesis, Virulence and Counter measures
Track 2: News in Harmful Bacteria
Track 3: Immune Defence against Bacterial Pathogens: Innate and Adaptive Immunity
Track 4: Plant-Microbe Symbiosis and Pathology
Location: Melia Meeting 1 & 2
Chair
Leif A. Kirsebom
Uppsala University, Sweden
Co-Chair
Kei Amemiya
US Army Medical Research Institute for Infectious Diseases, USA
Session Introduction
Agnieszka Grabowiecka
Wrocław University of Technology, Poland
Title: Novel urease inhibitors for the control of ureolytic microbial pathogens
Time : 11:20-11:40
Biography:
Agnieszka Grabowiecka presently as Faculty of Chemistry, Department of Bioorganic Chemistry, Wrocław University of Technology, Poland
Abstract:
Ureolytic activity of bacterial pathogens is a main causative factor of severe clinical conditions concerning the human gastric and urinary tract. The damaging effect to host organism results from alkalization of pathogens environment due to ammonia release from urea molecules. The growing bacterial resistance to antimicrobial agents has stimulated studies on urease inhibition as a way to control colonization with Helicobacter pylori and Proteus mirabilis. Within research of our Bioorganic Chemistry Group a new class of urease inhibitors was proposed using available enzyme crystal structures. Investigated aminophosphonates and aminophosphinates are extended transition state analogs of urease reaction. Their inhibition potency against urease purified from H. pylori and Proteus mirabilis as well as assayed in whole-cell system exceeds clinically approved acetohydroxamate (dissociation constants within micromolar range). Novel inhibitors were verified as agents for maintaining ionic balance and preventing Proteus – induced struvite precipitation in artificial urine. In the presence of the studied compounds maximum urine pH was 7.3 compared to 9.4 in untreated cultures. Precipitate formation was significantly reduced as shown in 31P-NMR analysis. In microscopic examinations only single coffin-like crystals characteristic of early struvite formation were present. In H. pylori reference strains repressed ureolysis reduced ability to survive incubations under strongly acidic conditions. Besides urease inhibition, several structures bearing long aliphatic chains (C5, C6) additionally exhibited moderate bacteriostatic effect. Minimal Inhibitory Concentrations were estimated using broth micro dilution method and metabolic activity assay using tetrazolium salts (MTT). Reduced viability in the presence of urease inhibitors was validated by fluorescence staining using LIVE/DEAD BacLight Kit by Molecular Probes.
János Tamás Padra
University of Gothenburg, Sweden
Title: Aeromonas salmonicida proliferation and quorum sensing in response to mucins isolated from Atlantic salmon skin and intestine
Time : 11:40-12:00
Biography:
János Tamás Padra has completed his PhD at the University of Debrecen, Hungary in 2012. Since then he is a Post-Doctoral member of Mucins in Infection and Cancer team lead by Dr. Sara K. Lindén at Sahlgrenska Academy, University of Gothenburg, Sweden.
Abstract:
Aquaculture is a growing industry increasing the need for understanding host-pathogen interactions in fish. The skin and mucosal surfaces, covered by a mucus layer, is the first point of contact between fish and pathogens. However, knowledge on fish mucin-pathogen interactions is limited. A. salmonicida, the causative agent of furunculosis, is a major infectious threat to aquaculture. We previously demonstrated that the binding of A. salmonicida to Atlantic salmon mucins differ between body sites and is dependent on the presence of the sialic acid: N-acetylneuraminic acid on mucins. Here, we cultured A. salmonicida in the presence of mucins purified from skin, pyloric caeca, proximal and distal intestine from five healthy Atlantic salmons, and analyzed growth rate and bacterial communication through quorum sensing molecules. Intestinal mucins enhanced A. salmonicida growth, whereas skin mucins had no effect. The increase in growth was positively affected by longer glycan chains of mucins and higher ratio of sialic acid. Enzymatic desialylation of mucins enhanced proliferation further. Mucins from all sites decreased the production of autoinducer-II (AI-II) signal molecules. Desialylation organ specifically altered the level of AI-II molecules produced by A. salmonicida. Thus, it appears that although mucins of the intestinal tract stimulated A. salmonicida growth, presumably reflecting that the pathogen senses the right target niche, the sialylated mucin glycans seem to act as a defense mechanism and limit the growth response. Mucin inhibition of QS may be an additional host defense mechanism influenced by the level of sialylation on salmon mucins.
Svitich O.A
Pirogov Russian National Research Medical University, Russia
Title: The role of Candida albicans in the activation of TLR-mediated mechanisms of innate immunity
Time : 12:00-12:20
Biography:
Svitich O.A from mechnikov Scientific Research Institute of Vaccines and Serums, Moscow, Russian Federation.
Abstract:
Introduction. Candidiasis of infants and immuno compromised people is the one of the main problems of medicine. Candida albicans and products of its metabolism play an important role in the pathogenesis of candidiasis. Previously it was found that Toll-like receptor (TLRs) system of innate immunity is directly involved in the process of recognition of fungal agents, such as Candida albicans, Aspergillus fumigatus, Cryptococcus neoformans and some others. Information about mechanisms of interaction of fungi (C. albicans) and system of innate immunity is extremely important, because it allowed to develop new methods of therapy. The aim of this work was to investigate the levels of TLRs and cytokines gene expression in vitro after exposure to C. albicans antigens and in the epithelial cells of the mucosa of cervical canal of pregnant women with candidiasis. Materials and methods. The experiments were carryed out on the Vero cells and mononuclear cells (MNCs). Candida cells were grown on minimal medium with 1% glucose. Antigens were presented after biomass was inactivated by heating. MNCs from peripheral blood of healthy donors were isolated by density gradient of ficoll pack. Selected mononuclear cells (at a concentration of 1*106 cells/ml) or 48-hour monolayer of Vero cells were cultured with lysates of C. albicans in 96-well plates (Costar, USA) at 37°C in the atmosphere of 5% CO2. The study of gene expression of TNF, TLR1, TLR2, TLR3, TLR9 and TLR6 was performed using real-time PCR using the intercalating dye SYBR Green I and fluorescent probes (Synthol, RF). The number of copies of cDNA genes were standartisated by expression of b-actin gene. Vero cells were incubated with C.albicans antigen for 3 and 24 h, MNCs culture - for 12 and 24 hours. In this study we also examined expression levels in the epithelial cells of the cervical canal of 20 pregnant women (27-30 weeks): 10 healthy pregnant women and 10 pregnant patients with candidiasis. Results and discussion. During experiments it was established that in Vero cells after 3 hours of antigenic exposure levels of TLR1, TLR2, TNF genes expression significantly increased and the level of TLR6 expression was unchanged. After 24 hours, levels of expression of innate immunity genes were not statistically different from control values. In the culture of MNCs levels of TLRs genes increased after 12h of antigenic exposure, the level of TNF gene expression - after 24 h. So, the level of TNF gene expression in the experiment corresponded to 3.38±0,34 lg(number of copies of cDNA), relatively 2,56±0,28 lg (number of copies of cDNA in the sample). It can be assumed that in response to C. albicans infection in cultures of Vero cells and mononuclear cells the expression of TLR2 and TNFα increase. Studies have shown that the level of expression of TLR1 gene in epithelial cells of the cervical canal of pregnant women with urogenital infection compared with healthy pregnant women increases 6 times or more, and TLR2 – more, then 10 times. The level of expression of TLR6 in cells of patients with candidiasis has not changed or has been lower than in the cells of healthy pregnant women. Significant correlation was shown between levels of expression of TLR2 and TLR1 (r=0,8921), which is an indirect evidence for the dimerization of these receptors. Thus, the antifungal immune responses, apparently, specified through TLR2/TLR2 and TLR1/TLR2. Further research of TLR-dependent mechanisms of the detection of antifungal immunity will allow to create the new approaches in treatment of fungal infections, based on TLRs ligands.
Josiah Ademola Onaolapo
Ahmadu Bello University, Nigeria
Title: Incidence and antibiotic susceptibility profile of staphylococcus aureus on door handles in Ahamadu Bello University, Zaria
Time : 12:20-12:40
Biography:
Josiah Ademola Onaolapo from Department Of Pharmaceutics and Pharmaceutical Microbiology, Ahmadu Bello University, Zaria, Nigeria
Abstract:
Pathogen microorganisms implicated in most diseases are transferable through contact with infected persons or objects. In this study, door handle in the Faculty of Pharmaceutical Science and Amina female hostels in Ahmadu Bello University, Zaria were evaluated for the presence of Staph. aureus and their antibiotics susceptibility profile using standard microbiological methods. The result showed that out of the 143 door handles sampled (Amina female hostel=89, Pharmacy main block=40, Pharmacy old block=14), the incidence of Staph. aureus was 23.8% (34) with highest occurrence in Amina female hostel (16.8%), followed by Pharmacy main block (4.2%) and Pharmacy old block (2.8%). The antibiotic susceptibility profiles of the isolated Staph. aureus showed that the isolates were 100% susceptible to Ciprofloxacin, Erythromycin and Tetracycline, 97% susceptible to Mupirocin and Cotrimoxazole, 92% to Pefloxacin and Oxacillin, while 9% susceptible to Cefotaxime. Their levels of resistance to the selected antibiotics were very low (3% resistant to Mupirocin and Cotrimoxazole, 8% to Pefloxacin and Oxacillin) except to Cefotaxime of 91% resistance. The result showed that the selected antibiotics are still effective against Staph. aureus isolated from door handles in Ahmadu Bello University (A.B.U), Zaria. The high incidence of Staphylococcus aureus in this study might be attributed to poor hygiene among students and the possibility of transferring pathogenic Staph. aureus through door handles in densely populated environ during disease outbreak is suspected. To curb the spread of pathogenic and resistant Staphylococcus aureus, this study suggest that door handles in A.B.U, Zaria should be replaced with metallic copper surfaces with antimicrobial properties and frequent use of disinfectant/hand sanitizer is recommended. Also proper periodic antibiotic surveillance should be encouraged to have referable documentaries in disease outbreak.
Vanessa Jackson
Cape Peninsula University of Technology, South Africa
Title: Isolation and Identification of potential PAH degrading Bacterial Species from the Diep- and Plankenburg Rivers, Western Cape, South Africa
Time : 12:40-13:00
Biography:
Vanessa Jackson completed her PhD at the age of 32 years from the Cape Peninsula University of Technology, Cape Town, South Africa. She is currently a senior lecturer in the Biotechnology Programme in the Department of Biotechnology and Consumer Science and is co-pi of the Bioresource Engineering Research Group (BioERG) at the Cape Peninsula University of Technology. She has published 12 peer-reviewed accredited scientific publications as both main and co-author and is supervising several Masters and PhD students.
Abstract:
Polycyclic Aromatic Hydrocarbons have been shown to be carcinogenic, teratogenic and mutagenic. PAHs can enter environmental matrices via natural and anthropogenic sources from agricultural and industrial activities. The aim of this study was to isolate and identify potential PAH-degrading microorganisms from the Diep- and Plankenburg Rivers, Western Cape, South Africa. Water and sediment samples were collected monthly over a six month period at six sampling points (three sampling points along each of the Rivers) and kept on ice at 4ºC during transport. For phenotypic identification, standard cultivation on various general and selective media, including Nutrient and Aeromonas isolation agars, as well as Pseudomonas-isolation Agar Base, amongst others, were employed. Each isolate was screened for PAH degradation (acenaphthene, fluorene) using shaking flasks prior to isolation and identification. After screening and cultivation, nineteen isolates were selected and molecularly identified using 16S rRNA extraction followed by primer-specific PCR amplification. The predominant species identified included Raoutella ornithinolytica, Serratia marcescens, Bacillus megaterium and Aeromonas hydrophila. Raoutella ornithinolytica was isolated from most of the media utilised, including Pseudomonas Agar Base supplemented with CFC and Pseudomonas Agar Base supplemented with CN, while Aeromonas hydrophila grew on all culture media except Pseudomonas Agar Base. Raoutella ornithinolytica is a member of the Enterobacteriaceae while Aeromonas sp. closely resembles Enterobacteriaceae, which could account for its growth on most media. Bacterial species that display tolerance to these compounds could have great beneficial effects in the area of bioremediation of PAH-contaminated sites.
Mouna Jrad
University of Sfax, Tunisia
Title: Antibacterial properties of new bacterium isolated from date palm leaves affected by the brittle leaf disease
Time : 13:00-13:20
Biography:
Mouna Jrad from Laboratory of Plant Biotechnology Applied to Crop Improvement, Faculty of Science of Sfax. University of Sfax, Sfax 3038, Tunisia
Abstract:
An actinomycete strain was isolated from date palm leaves affected by brittle leaf disease. The strain was classified as a new bacterium, an endophytic strain. Identification was based primarily on the morphological characteristics. This strain was grown on a medium rich tryptic soy broth (TSB) with K2HPO4 (1 mM) and MgSO4, 7H2O (2 mM) at 28 °C and neutral pH. It is a Gram-positive filamentous bacterium with broad spectrum antimicrobial activity. The optimum pH for the product was 7 to 7.4. The highest product yield was with glucose and tryptone at 1 % (w/v) as carbon and nitrogen sources, respectively. Incubation between 25 °C and 30 °C for 24 hours was optimal for the bioactive metabolites production. The product remained stable up to 50 hours. The bioactive metabolites produced by endophytic bacteria was partially purified and studied for antibacterial characteristics using the wells technique. Analysis by bio-autography and gas chromatography/mass spectroscopy (GC/MS) of the supernatant from precipitation with 100% ammonium sulphate indicated that the strain produced a high percentage of fatty acid. Additionally, a cyclic compound could be involved in the antibacterial activity. After silica plate scraping, bioactivity disappeared at 70 °C. Bioactive molecules may be involved in the brittle leaf disease because its product by a strain isolated from date palm leaves affected by the brittle leaf disease enough. In addition, this endophytic has been a best lipolytic activity and produces a lipidic biomolecules, this results explain the implication of metabolites in the brittle leaf disease. Finally, determination of MIC and MBC of bioactive molecules against Strains (Bt,Bs).
Likou Zou
Sichuan Agricultural University, China
Title: Antibiotic and disinfectant resistance of Salmonella from retail meats in china
Time : 14:05-14:25
Biography:
Likou Zou has completed his PhD at the age of 28 years from Sichuan Agricultural University and Postdoctoral studies from Sichuan University School of Life Science. He is the Director of Lab of Microbiology, Dujiangyan Campus of Sichuan Agricultural University. He has published more than 20 papers in reputed journals and has been serving as an Editorial Board Member of Journal of Sichuan Agricultural University.
Abstract:
Salmonella is a leading cause of food borne illnesses and deaths worldwide. The frequent use of antibiotic and disinfectant in food production and processing may have imposed a selective pressure and contributed to the emergence of resistant microbes. However, little information is available regarding the antibiotic and disinfectant resistance of Salmonella isolates from retail meats in China. Salmonella isolates were recovered from chicken, beef and pork purchased from selected supermarket. Antibiotic susceptibility was tested according to the standard disk diffusion method. The MICs of disinfectants were determined using the agar dilution method. All isolates were screened for the presence of disinfectant resistance genes and further analyzed for genetic relatedness by PFGE. The predominant serotype was Salmonella Derby (34.8%) followed by S. Enteritidis (12.3%) and S. Rissen (9.0%). Overall, 82.8% isolates were resistant to OTC, 66.3% to TMP, 28.8% to AMX, 23.9% to AMP, 20.3% to LEV, 11.7% to GEN, 11.7% to EFT, 15.3% to CIP and 3.7% to AMC respectively. The MICs of the disinfectants cetyltrimethylammonium bromide and cetylpyridinium chloride were 8-128 mg/L and 8-256 mg/L. The qac and sugE(p) gene was found less prevalent (0.0% to 14.7%). Up to 71 distinct PFGE types were identified among the 163 Salmonella isolates. PFGE revealed that the resistant isolates were associated with the sampling supermarkets or groceries. Salmonella can serve as a critical vector in spreading disinfectant and antibiotic resistance. The use of disinfectant in food processing environments may have played a role in the emergence of antibiotic and disinfectant resistant bacteria.
Arnelia N Paulse
Cape Peninsula University of Technology, South Africa
Title: Microbial pollutants in stagnant water in an informal settlement in the Western Cape, South Africa
Time : 14:25-14:45
Biography:
Arnelia Paulse obtained her PhD from the Cape Peninsula University of Technology, Cape Town, South Africa in 2009. She is currently a lecturer in the Department of Environmental and Occupational Studies and is one of the co-pi of the Bioresource Engineering Research Group (BioERG) at the Cape Peninsula University of Technology. She has published 9 peer-reviewed accredited scientific publications as both main and co-author and is supervising several Masters students. Dr. Paulse has successfully graduated two Masters Students.
Abstract:
Inadequate sanitation and poor drainage in informal settlements result in greywater being stagnant at the base of communal taps. This water could potentially cause health problems to those who come in contact with it. This study is aimed at determining microbial pollutant levels in stagnant greywater in the RR Section of Khayelitsha, Western Cape, South Africa. Sampling of stagnant greywater was conducted twice monthly (January to May 2013) from the base of six communal taps. Enumeration techniques employed included the Most Probable Number (MPN), Heterotrophic Plate Count (HPC) and Flow Cytometric (FCM) techniques. Identification techniques utilised included the API 20E, RapID™ ONE and BBL Crystal™ Gram Positive (GP) Identification (ID) systems. The highest MPN, faecal coliform and E. coli counts were 1.6 x 108 microorganisms/100mâ„“ (Sites A, B; weeks 3 and 5), 4.7 x 106 microorganisms/100mâ„“ (Site B; week 5) and 1.8 x 106 microorganisms/100mâ„“ [Site A (week 5); Site F (week 5)], respectively. The highest total FCM count was 3.4 x 107 microorganisms/mâ„“ (Site A, week 5). The FCM technique showed significantly (p < 0.05) higher counts than both the MPN and HPC techniques. The most common microorganisms isolated included Escherichia coli, Klebsiella pneumonia, Klebsiella oxytoca, Acinetobacter baumannii/calcoaceticus, Enterobacter cloacae, Corynebacterium species and Bacillus cereus. The presence of these microorganisms in addition to significantly high microbial counts obtained, raises major health concerns as these organisms present in high numbers may lead to serious waterborne diseases, especially in informal settlements where population densities are extremely high.
Zaki A Siddiqui
Aligarh Muslim University, India
Title: Interactions of plant pathogenic bacteria and plant growth promoting rhizobacteria with plant parasitic nematodes
Time : 15:05-15:25
Biography:
Zaki Anwar Siddiqui is Professor, Department of Botany, Aligarh Muslim University Aligarh, India. He did his Graduation, Post-Graduation, MPhil and PhD from Aligarh Muslim University, Aligarh. He was Visiting Professor in Kyoto University, Japan from 2007-2008. He has published more than 115 research papers and review articles and edited 2 Books published from Springer, Netherlands. He has worked as Principal investigator of two major research projects on the Biological control of plant diseases and he is also a reviewer of more than 20 journals of his field. He has delivered invited talks in several national and international conferences.
Abstract:
Interactions of plant parasitic nematodes with plant pathogenic bacteria and plant growth promoting rhizobacteria (PGPR) are quite different. Interaction of nematodes with plant pathogenic bacteria results in disease complexes. Each disease complex is generally distinct from another and largely dependent on the type of nematode parasitism involved. Nematodes interaction increase development of plant disease caused by bacteria in different ways such as predisposing agent, modifying physiology of host tissues, breaking host resistance to bacterial pathogens, acting as vectors of bacterial pathogens and changing the rhizosphere microflora. Prior occupancy of host tissues by nematode or bacterium has different influence on the disease severity. Sometimes, the presence of both the nematode and the bacterial pathogen is necessary for production of certain types of symptoms. Interaction of PGPR with plant parasitic nematodes on the other hand, reduced disease severity caused by nematodes. Inoculation of plants with PGPR result in multiple effects on plant growth that includes improved seed germination, root development, water uptake, plant growth promotion solubilization of insoluble phosphorus, production of siderophores, phytohormones, lowering of ethylene concentration, production of antibiotics and induced systemic resistance. Significant control of plant parasitic nematodes has been demonstrated by PGPR. Recent progress in our understanding of their diversity, colonizing ability and mechanisms of action, formulation and their application may facilitate their development as reliable biocontrol agents against plant parasitic nematodes.
Mohammed Shukur
University of Nottingham, United Kingdom
Title: Immune responses to Mycobacterium avium infection and the role of MAPK pathway in regulation of signal transduction during the infection
Time : 15:25-15:45
Biography:
Mohammed Shukur is currently a PhD student in School of Veterinary Medicine and Science, University of Nottingham, UK. He has completed his MSc from School of Veterinary Medicine University of Duhok.
Abstract:
Mycobacterial infection is a major problem in both human and animal health. Several cell signaling pathways are involved in the mediation of expression of a number of cytokines induced by mycobacterial infection. The mitogen activated protein kinase (MAPK) pathway plays a crucial role in the pathogenesis of mycobacterial infection. In this study we aimed to evaluate the role of ERK and p38 MAPK in cellular regulation by Mycobacterium avium infection in human and chicken macrophages-like cells. THP-1 and HD11 cells were infected with eight clinical isolates of Mycobacterium avium. The cells were pre-treated with highly specific inhibiters of the ERK (PD98059 or U0126) and p38 (SB203580) pathways 30 min prior to infection and the levels of cytokine production at 6 and 24 hour post infection were assessed using ELISA and RT-qPCR. M. avium infection resulted in differential expression of cytokines and chemokines in THP-1 and HD11 cells. Treatment of the cells with PD98059 or U0126 inhibited production of cytokines in THP-1 and completely blocked their expression in HD11 cells. In addition, p38 inhibition differentially modulated cytokine production in THP-1 cells compared to non-inhibited M. avium-infected cells. It inhibited release of IL-6 while the level of IL-1β and TNF-α showed an increase in THP-1 cells in response to M. avium infection following treatment of the cells with SB203580. The results suggest that signalling events are significantly different in avian and human cells following M. avium infection. Both p38 and ERK are involved in regulation of IL-6 production while p38 negatively regulates IL-1β and TNF-α production in response to M. avium in THP-1 cells. In HD11 cells, it is suggested that multiple signal pathways simultaneously participate in regulation of cytokine signal transduction during M. avium infection.
Quesada-Gomez C
Universidad de Costa Rica, Costa Rica
Title: Recombination in the substrate recognition domain of TcdB confers a variant glucosyl transferase activity and a distinctive cytopathic effect to a Clostridium difficile strain from the hypervirulent clade II
Time : 16:00-16:20
Biography:
Carlos Quesada-Gómez is a lecturer on Anaerobic Bacteriology, School of Microbiology, University of Costa Rica. He has a M.Sc. in Medical Bacteriology and is currently a Ph.D. Candidate working with Clostridium difficile. His research group is working on the molecular characterization of different isolates by PFGE, toxinotyping, MLST and whole genome sequence methods which have made possible the description of the hipervirulent NAP1/027 and other novel genotypes in Costa Rica and Latin America. C. difficile pathogenesis is another area of study in which the level of virulence is being associated with the genotypes and its toxins using cellular and animal models
Abstract:
Clostridium difficile NAP1 strains are responsible for nosocomial outbreaks worldwide. The increased pathogenic potential of these strains has been attributed to over production of toxins and fluoroquinolone resistance. During a C. difficile nosocomial outbreak in Costa Rica, a strain was found to induce a distinct cytopathic effect (CPE) different from the canonical arborizing CPE. The strain was further identified as a NAP1 isolate (NAP1V) of the hypervirulent clade II that harbored a silent mutation in the gyrA gene. After toxin purification, cells treated with TcdB of the NAP1V strain (TcdBV), displayed a rounded and surface detached phenotype resembling that induced by C. difficile TcdB toxin A-negative strains. The effect mediated by the TcdBV was due to differences in substrate preferences that resulted in different glucosylation patterns of the various GTPases. Whereas TcdB from classical NAP1/027 strains glucosylated RhoA, Rac and Cdc42, TcdBV did not used RhoA as substrate and displayed less affinity for Cdc42. Sequence comparison of the functional domains of TcdBv with other C. difficile strains along with comparative genomic analysis revealed that TcdBv is a NAP1 toxin but with modifications within the enzymatic domain. The enzymatic domain is identical to that of a NAP9/017 strains (A-B+). We also provide evidence that the NAP1 strains glucosylate a broader spectrum of GTPases in vitro as both toxins glucosylated Rap and R-Ras. These findings provide insight into the role of the glucosyl tranferase activity in the pathogenesis of C. difficile variant TcdB strains and the hyper virulent NAP1 strains.
Malem Flores
Velasquez St. University of the Philippines, Philippines
Title: Diversity, bioactivity, and secondary metabolite profiles of bacteria isolated from different tissue of the mollusk gastropod Truncatella species
Time : 16:20-16:40
Biography:
Malem Flores works at the Marine Science Institute, Velasquez St. University of the Philippines, Philippines.
Abstract:
The emergence of drug resistance for many diseases has been for decades the driving force behind the pursuit of naturally-occurring bioactive compounds. Among the candidate drug leads, marine sources are one of the most promising owing to the vast biodiversity of marine organisms. However, marine gastropods with shells are an overlooked source of bioactive secondary metabolites because they are well-protected by their shell covering and some produce venoms. In this study, several bacteria were isolated from different tissues of Truncatella sp., a small gastropod mollusk that is found on land very close to seawater (splash zone). Identification of each isolate using 16S sequence analysis revealed a diversity of Gram-negative and Gram-positive bacteria, with Actinobacteria as the dominating group. Furthermore, secondary metabolites produced by Truncatella-associated bacteria showed promising antimicrobial and anti-cancer properties. Some of these bioactive metabolites manifested narrow spectrum antimicrobial activity against Staphylococcus aureus or Pseudomonas aeruginosa while having weak inhibitory effect on lactic acid bacteria commensals. Likewise, there were extracts that exhibited selective cytotoxic activity on the lung cancer cell line, A549 or colorectal cancer cell, HCT 116 with moderate or no cytotoxicity on the normal kidney cell line, MDCK. Chemical profiles of secondary metabolites determined by reverse-phase high performance liquid chromatography (RP-HPLC) demonstrated common and unique patterns of metabolites for each isolate. This suggests that Truncatella is a potential source of bioactive metabolite-producing bacteria which can be valuable for drug discovery and that secondary metabolite profiles may present distinct phenotypic traits of the microorganisms. Importantly, this is the first study to report on the genetic and chemical profile diversity of bacteria associated with Truncatella and their corresponding antimicrobial and anticancer properties.
Médea Padra
University of Gothenburg, Sweden
Title: Host-Helicobacter interactions in the mucus niche
Time : 16:40-17:00
Biography:
Médea Padra completed PhD student in the Mucins in Infection and Cancer Team at the Department of Medical Biochemistry and Cell biology, University of Gothenburg, Sweden
Abstract:
Helicobacter pylori chronically infects half of the world’s human population and is the main etiological agent causing gastric ulcers and cancer. Helicobacter suis colonizes the gastric mucosa and in pigs it is associated with gastritis, decreased daily weight gain and possibly gastric ulcer disease. H. suis is also the most prevalent non-H. pylori Helicobacter species in the human stomach and is associated with peptic ulcer disease, gastric mucosa-associated lymphoid tissue lymphoma and chronic gastritis. Helicobacters are prone to developing antibiotic resistance, and recurrence frequently occurs. H. pylori-infected rhesus monkeys and human children secreting mucins with less H. pylori binding capacity develop higher H. pylori density infections and gastritis, supporting that the ability of secreted mucins to bind to H. pylori protects the gastric epithelium. Gastric mucin turnover is impaired during H. pylori infection, which in turn creates a more stable environment for H. pylori to colonize long term. H. pylori and H. suis bound to human and pig mucins via the mucin glycans, with distinctly different specificites: the most notable difference being that, H. suis bound to neutral glycans with an acidic pH optimum. This unusual binding mode may be the reason for that H. suis in addition to colonizing the mucus layer also colonizes the acid producing parietal cells. We found over 300 glycan structures on mucins from 16 humans, and each individual carried 60-120 structures, whereas we found around 250 structures in four pigs. Mucins differently regulated Helicobacter proliferation, gene expression and downstream interactions with host cells. This regulation appeared to occur via at least three glycan response pathways. Both in primate and pig models of infection, host glycosylation was altered during infection, which affected the ability of these pathogens to interact with their hosts, and the outcome of the interaction. In conclusion, when a Helicobacter encounters the mucins that builds up the mucus layer, its behavior changes in a host specific manner in response to the mucin glycans. This “changed” pathogen then interacts with the host epithelial cells, which responds by changing its mucins and mucin glycans, which in turn changes the pathogen again, in a constant host-pathogen adaptation and response process.
Enayatollah Kalantar
Alborz University of Medical Sciences, Iran
Title: Extreme antibiotic resistant Acinetobacter baumannii related pneumonia in a regional hospital
Time : 17:00-17:20
Biography:
Enayatollah Kalantar is a Faculty Member of School of Medicine, currently is the Research Director of Alborz University of Medical Sciences, Editor In Chief for International Journal of Enteric pathogens. He has been awarded as the Best Researcher several times.
Abstract:
Background: A. baumanniiis emerging as an important pathogen which has received a great interest during the last two decades. Furthermore, such pathogens are challenging to treat because of extensive antimicrobial drug resistance. Aim: Purpose of this study is to report our experience with five cases of A. baumannii related pneumonia infections, seen in a regional hospital in Karaj, Iran. Method: Retrospective analysis of eight cases of Acinetobacter baumannii related pneumonia infections presenting a regional hospital in Karaj, Iran during ????? 2014 were done. Results: All the cases were admitted to the emergency department with loss of consciousness and then transferred to Intensive Care Unit. Sputum/pulmonary secretions culture revealed A. baumannii. During the treatment, various antibiotics like meropenem, vancomycin, colistinand amikacin was prescribed but all the eight cases died because of Acinetobacter baumannii related pneumonia infections. Conclusion: Our cases bring forth the burden of Acinetobacter baumannii related pneumonia which has been associated with significant mortality and as illustrated in our cases infection with A. baumannii can be rapidly fatal, thus stressing the importance of reporting A.baumannii isolates in cases of pneumonia. Physicians should be aware of the remarkable virulence and drug resistance.
Oumar Traore
University of Ouagadougou, Burkina Faso
Title: Occurrence of Vibrio cholerae in fish and water from a reservoir and a neighboring channel in Ouagadougou, Burkina Faso
Time : 17:20-17:40
Biography:
Oumar Traore is doing his PhD at the age of 35 years at the University of Ouagadougou and is an Office Worker as Microbiologist at National Laboratory of Public Health and work on the field of water quality. He has one publication and one submitted in reputed journals and still writing manuscripts for submission. He participated in different international meeting. He supervised master students and trainee from the National School of Public Health.
Abstract:
Introduction: Vibrio cholerae is a human pathogen and natural inhabitant of aquatic environments. In this study we surveyed the occurrence of V. cholerae in fish harvested from a reservoir Barrage de Tanghin in Ouagadougou, water of which is increasingly used for fishing and for irrigation of vegetables. The water is of poor quality, because several channels convey municipality’s discharges directly to the reservoir. Methodology: A total of 238 fish (Oreochromis niloticus), 25 samples of reservoir water and 55 samples of channel water were analyzed for the presence of V. cholerae. Results: Altogether, 13 V. cholerae strains were isolated from the fish (from 5% of the samples), 1 (4%) from the reservoir water and 4 (7%) from the channel water. All the strains were identified as non-O1/non-O139 V. cholerae and they did not possess the Cholera Toxin–encoding ctxA gene. The antimicrobial susceptibility testing showed that the strains were in general susceptible but approximately half of them were resistant to ampicillin. Conclusion: Although no strains of epidemic V. cholerae serotypes were encountered, it would be important to monitor the microbiological quality of this extensively used water resource and its products for vibrios as well as for other enteropathogenic bacteria.
- Track 5: Clinical Aspects of Bacterial Infections
Track 6: Epidemiology, Emerging Infectious Diseases and Public Response Planning
Track 7: Medical Microbiology - From Benchtop to Clinic
Location: Melia Meeting 1 & 2
Chair
Ashraf M. Ahmed
King Saud University, Saudi Arabia
Co-Chair
Antonio Fasanella
Anthrax Reference Centre of Italy, Italy
Session Introduction
Ashraf M. Ahmed
King Saud University, Saudi Arabia
Title: Bacterial challenge in the control measures of mosquito-borne diseases: The battle against mosquito vectors
Time : 10:30-10:50
Biography:
Prof. Ahmed has completed his PhD from Keele University, UK, on 2002 and has ongoing Research Fellowship at Keele from 2004 until now, and Fellow of the Roial Society, UK, in 2004. He is a Professor of Medical Entomology at Zoology Department at El-Minia University, Egypt, (currently at King Saud University, Saudi Arabia). His current research interest is “mosquito immunity and biocontrol”, aiming at utilizing the immune response of mosquito vectors against mosquito-borne disease agents, and biological agents in the biocontrol measures against mosquito vectors. He has successfully achieved several grants for research projects, and got two currently ongoing granted projected (1.6 million Saudi Riyals each). One grant is for isolating native mosquito Bacillus thuringiensis bacteria with enhanced larvicidal activities for use in the battle against mosquito vectors in Saudi Arabia. The other grant is for isolating immune peptides from honey-bees for use as natural antibiotic against the American Foal Broad disease (AFB) that threatens the global Apiary industry. He had successfully monitored undergraduate, postgraduate students and research scholars. His academic output consists of more than 26 papers in reputed journals, membership of several Scientific societies, attended several local, national & international conferences and invited for main talks in many international conferences.
Abstract:
Malaria, kills several millions annually worldwide and is transmitted by anopheline mosquitoes. The recently evolved problem in the human battle against malaria is the pesticide resistance of the vector and the rapid spread of multiple anti-malarial drug-resistance of malaria parasite. Great efforts are currently being done to utilize the vector immune system (refractoriness) in this battle. This strategy is being seriously tested regardless of the concomitant reproductive cost, the price of immune induction that must be paid by the vector. This, in fact, may reduce the importance of this immune-based control strategy in the battle against such mosquito-borne diseases. In spite of the research emphasis on the use of transgenic pathogen-refractory mosquitoes, insecticides are still the main method for controlling mosquito vectors, although being environmental pollutants and facing vector resistance that now threatening the effective life of these chemical compounds. Thus, recent studies have refocused interest on mosquitocidal bacteria as useful eco-friend alternatives to conventional insecticides, suggesting them as bio-control candidates in the human battle against mosquito-borne diseases. Yet, Bacillus thuringiensis (Bt) faced no resistance from insect host due to both the interactions among its multiple toxins and their respective receptors in mosquito midgut, and it is likely that these multiple intermolecular interactions are the major reasons for the absence of passive resistance to Bt in mosquitoes. This talk will discuss these scenarios in terms of the costs of vector immune-control strategy, Bt mosquitocidal mechanisms and new Bt isolates with enhanced mosquitocidal toxicity.
Emanuele Campese & Antonio Fasanella
Anthrax Reference Centre of Italy, Italy
Title: Bacillus anthracis strains circulating in Albania
Time : 10:50-11:10
Biography:
Emanuele Campese was born on 19th May 1985. In 2012 he got a Master’s degree in Environmental Biology at the Faculty of Biology of the University of Bari. He participated in several courses and workshops. In December 2013 he won a grant at the Department of the National Reference Center for Anthrax at the Istituto Zooprofilattico of Puglia and Basilicata. Main fields of his research activities are the development and improvement of techniques for the isolation of Bacillus anthracis from soil and molecular epidemiology of anthrax in developing countries. He is author and co-author of scientific papers published on international journals and conference communications.
Abstract:
Anthrax, whose causative agent is Bacillus anthracis, is a non-contagious infectious disease that affects several animal species, the human one included. Domestic and wild ruminants represent the most susceptible categories. The bacterial agent has the characteristic of producing spores that can survive in the environment for several decades. Anthrax in Albania is an endemic disease characterized by few outbreaks involving a very low number of animals. Nineteen samples of soil coming from burial sites and 11 strains of Bacillus spp isolated from died animals of different districts of Albania were examined. The analysis of soil samples revealed that 11 of them were contaminated with anthrax spores, while only 8 strains were confirmed as Bacillus anthracis. The analysis of CanSNPs showed that all isolates belong to lineage A major subgroup A.Br.008/009 (Trans-Eurasian or TEA strains). The MLVA test at 15 loci showed three different genotypes: Albania GT/1, Albania GT/2 and Albania GT/3. Two distinct genotypes (Albania GT/2 and GT/3) were found in the same burial site in the district of Kukës. All the genotypes are genetically very similar to each other, which confirms the hypothesis that all of them are the results of the evolution of a local common ancestral strain. However it is not excluded that in the course of further investigation, strains belonging to other lineages can be found, as it has been observed in Italy, where, in addition to the dominant genotype TEA, there are ecological niches of B.Br.CNEVA and A.Br.005/006 in the regions Northern Italy.
Arwa Alumran
University of Dammam, Saudi Arabia
Title: The Parental Use of Antibiotics in Children in Saudi Arabia
Time : 11:10-11:30
Biography:
Arwa has completed her PhD at the age of 28 years from Queensland University of Technology, Australia. She is currently holding a position as an assistant professor at the University of Dammam, Saudi Arabia. She has published 5 papers in reputed journals and has been serving as an editorial board member of repute.
Abstract:
The overuse of antibiotics especially in children is becoming one of the most important public health issues worldwide. This study aims to assess the factors underlying the parental use of antibiotics for children in Saudi Arabia. This is a cross-sectional study design. 1104 Parents (52% were mothers) of children younger than 12 years old were recruited from schools parental meetings in the Eastern Province of Saudi Arabia. Ordinal logistic regression was used to identify the factors influencing the parental use of antibiotics in children. Risk factors considered included parents’ demographic information, child health-related information, along with the PAPA scales, a validated instrument that measures parents’ knowledge and beliefs, behaviours, adherence, seeking information, and awareness about antibiotic resistance. There is a high association between the frequency of cold episodes and the number of antibiotics used for the youngest child in the family during the previous year. Three parent-related psychosocial aspects appeared to be significantly associated with the parents’ tendency to use an antibiotic in their children: Knowledge and beliefs, behaviors, and seeking health-related information. Also, parent’s geographical background is associated with their use of antibiotics. The strong association between the number of cold incidence and the amount of antibiotics used suggests an evidence for antibiotic misuse in Saudi Arabia. In addition, to stronger regulations relating to dispensing antibiotics, the Saudi government should implement public health interventions aimed at advocating parents to appropriate use of antibiotics, and the potential dangers in their misuse.
Liaqat Ali Chaudhry
SBAH-CITY Rehablitation Hospital & Medical Center Riyadh, K.S. Arabia
Title: Prevalence of Diabetes type-2 & Pulmonary Tuberculosis among Filipinos and Treatment outcomes: A surveillance study in Eastern Saudi Arabia
Time : 11:30-11:50
Biography:
MBBS- King Edward Medical College university Lahore1981. Post Graduation: 1-DTCD —Diploma in Tuberculosis & Chest Diseases -Punjab University Lahore-Pakistan. 2-MCPS--- College of Physicians and Surgeons Pakistan. 3-MRCP,FRCP---The Royal College of Physicians Dublin-Ireland. 4-FACCP (USA) Worked in Pakistan: House physician, Registrar, Assistant professor at King Edward Medical College Lahore,1982-1987. Worked K.Saudi Arabia. Joined MOH of Saudi Arabia worked as, a- Specialist Physicians chest specialist on Feb-1988. b- Consultant Pulmonologist and Chief of Tuberculosis Center, Deptt.of Internal Medicine & Chest diseases Dammam medical complex and Honorary Associate & Professor Dammam Medical University, Eastern Province K.Saudi Arabia – 2011. c- Chairman Internal Medicine & Consultant Invasive
Abstract:
Objective: To study prevalence of diabetes type-2 and pulmonary tuberculosis among Filipino patients and treatment out comes. Tuberculosis centre of Dammam medical complex (MOH) is a referral centre for the Eastern Saudi Arabia where patients from all government and private hospitals having open pulmonary tuberculosis are admitted for isolation till they are rendered noninfectious. All patients are treated for 6 months under DOTS strategy with 4 drugs (2HRZE) for 2 months as initial intensive phase and 2 drugs (HR) for 4 months as continuation phase. Methods and materials: We retrospectively reviewed clinical records of 1388 patients admitted with open pulmonary tuberculosis between Jan- 2003 and June-2010. Results: Among 1388 patients, 39% (n = 542) were Saudis and 61% (n = 846) were non-Saudis. Among these 12.39% (n = 172) were Filipinos,153 males and 19 females respectively. Out of 1388 patients, 114 (7.17%) were found to have diabetes type-2. Among these diabetics, majority n = 91 (79.82%) were Filipinos. Sputum conversion was late in diabetic patients resulting in relatively longer hospital stay compared to fellow patients having only tuberculosis. Conclusion: Our study has shown that one possible risk factor for tuberculosis is diabetes. Majority of TB patients having diabetes type-2, n = 91 (79.82%) were Filipinos. Their sputum conversion was relatively late and their hospital stay was longer than their fellow patients having only tuberculosis. Our findings are in agreement with the current literature on the correlation of diabetes and tuberculosis.
Mahasin Wadi
Dar ALuloom University, Saudi Arabia
Title: Antibacterial Activity of Sudanese Bee Honey: Laboratory and Clinical Evaluation
Time : 11:50-12:10
Biography:
Mahasen Wadi completed the PhD from AL Neelain University , Medical Microbiology. Faculty of Medical Laboratory Sciences 2010 Khratoum Sudan. MSc. University of Khartoum, Sudan. Medical Microbiology & Pharmacology 1987. She joined work at Dar ALUloom University, College of Medicine, Riyadh , Saudi Arabia August 2014, She Worked at the central research laboratory Khartoum , Sudan. She joined work at King Saud University, Saudi Arabia, Riyadh 1988, Department of Clinical Laboratory Science, and Medical Microbiology. She worked in the area of Bee Honey as a natural antimicrobial product. She published a numbers of papers in reputed journals and Participated in many Internationals and national conferences. She issued a patent research about the antimicrobial activity of Sudanese bee honey. She attended many workshops and seminars. Awarded certificate of prestigious Author for the journal of Bacteriology &Parasitology 2011. Awarded a medal on participating in workshop at King Saud University Saudi Arabia 2011. Awarded many appreciation and thanks certificate in scientific activities. Member of many international associations: German Apitherapy Society, American Apitherapy Society, International Bee Research Association, European Society of Clinical Microbiology and Infectious Disease ESCMID and Sudanese Veterinary Association. She served as reviewer of various journals.
Abstract:
Honey is recognized as an effective topical treatment of burns and wounds. In many cases it is being used with success on infections not responding to standard antibiotic and antiseptic therapy. Fifteen samples of bee honey from different localities in Sudan were tested against five standard bacterial strains; Bacillus subtilis, Staphylococcus aureus, representing Gram positive bacteria, and Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeurginosa representing Gram–negative bacteria. All honey samples exerted inhibitory effects on both Gram-positive and Gram negative organisms. The clinical isolates obtained from infected wounds of twelve hospitalized patients at Omudrman Teaching Hospital, showed seven Pseudomonas and five Staphylococcus according to the cultural, microscopically and biochemical characteristics. All honey samples exerted inhibitory effects against the clinical isolates. Daily applications of honey on septic wounds, chronic wounds, ulcers and pyogenic abscess of the twelve hospitalized patients, gave favorable results, typified by promotion of granulation tissue and epithelization of the infected wounds.
Ahmad Homoud Al-Hazmi
Al Jouf University, Saudi Arabia
Title: Knowledge, attitudes and practice of primary healthcare physicians concerning the occupational risks of hepatitis B virus in Al Jouf Province, Saudi Arabia
Time : 12:10-12:30
Biography:
Ahmad Homoud Al-Hazmi is an Assistant Professor, Family Physician Consultant at Department of Family Medicine, College of Medicine, AlJouf University, Saudi Arabia
Abstract:
Background: Hepatitis B virus (HBV) infection is a well-recognized occupational risk for all healthcare workers (HCWs) worldwide. Aim: This study aimed to assess the knowledge, attitudes and practices of primary healthcare (PHC) physicians regarding the occupational risks of HBV. Method: In this cross-sectional study, a questionnaire survey was administered to 145 physicians of primary care centers in the Al Jouf Province of Saudi Arabia. The questionnaire contained questions concerning the knowledge base, attitudes and practices of physicians regarding the occupational risks of HBV. Result: The response rate of 82.8% yielded 120 questionnaires for analysis. The majority of physicians surveyed 99 (82.5%) felt at high risk of contracting and spreading HBV. The vast majority, 115 (95.6%) considered the HBV vaccine safe for all ages. Of the total 101 (84.2%) were vaccinated. Only 44 (36.7%) physicians recognized that HBV is resistant to alcohol and to some detergents. During surgical procedures, only a minority of the physicians uses double gloves and wears glasses. Almost all physicians were willing to subscribe to regular training programs concerning HBV. Conclusions: A lack of knowledge was determined and the practices of our physicians concerning the occupational risks of HBV appeared inappropriate. More education focusing on HBV is recommended.
Abdullah Abdulrahman Bin Salamah
King Saud University college of Medicine, Saudi Arabia
Title: Profile of viral hepatitis in Saudi Arabia
Time : 12:30-12:50
Biography:
Abdullah Bin Salamah, a senior medical student at King Saud University, Riyadh, Saudi Arabia.
Abstract:
The study was conducted to investigate the profile of hepatitis in Kingdom of Saudi Arabia, and to determine which age group hepatitis viruses most commonly infect. The epidemiology of viral hepatitis in Saudi Arabia has undergone major changes, concurrent with major socio-economicdevelopments over the last two to three decades.This disease represents a major public health problem in Saudi Arabia resulting in the need for considerable healthcare re-sources.A retrospective cross sectional analysis of the reported cases of viral hepatitis was conducted based on the reports of The Ministry of Health in Saudi Arabia about Hepatitis A, B and C infections in all regions from the period of January 2006 to December 2010. The study demonstrated that incidence of viral Hepatitis is decreasing, except for Hepatitis B that showed minimal increase. Of hepatitis A, B, and C, Hepatitis B virus (HBV) was the most predominant type, accounting for (53%) of the cases, followed by Hepatitis C virus (HCV) (30%) and HAV (17%).HAV infection predominates in children (5–14 years) with 60% of viral hepatitis cases, HBV in young adults (15–44 years) with 69% of viral hepatitis cases, and HCV in older adults (>45 years) with 59% of viral hepatitis cases.Despite significant changes in the prevalence of viral hepatitis A, B and C, it remains a major public health problem in Saudi Arabia, however it showed a significant decline in the last two decades that could be attributed to the vaccination programs and the improved health facilities. Further researches are needed to identify the risk factors making a specific age group or a specific region in Saudi Arabia targeted for a specific type of hepatitis viruses.
Shekari M
Hormozgan University of Medical Science, Iran
Title: Association of genetic polymorphism of the DNA base excision repair gene (APE-1 Asp/148 Glu) and HPV type (16/18) with the risk of cervix cancer in north Indian population
Time : 12:50-13:10
Biography:
Shekari M works as an Associated Professor at Department of Medical Genetics, Hormozgan University of Medical Sciences, Bandar Abbas -Iran
Abstract:
Cervical cancer is one of the most common neoplastic diseases affecting women with a combined worldwide incidence of almost half a million new cases. Reduced DNA repair capacity (DRC) can render a high risk of developing many types of cancer; including cervical cancer. Polymorphisms in DNA repair genes may contribute the genetic instability and carcinogenesis. Smoking experience and use of oral contraceptives have been confirmed to be risk factors for cervical cancer. The purpose of the present study was therefore to investigate APE-1 genotypes (Asp/Asp, Asp/Glu, Glu/Glu) with different histological subtypes in cases compared with controls. It has been observed that Asp/Glu with Glu/Glu genotypes that combined we observed statistically significant with protective effect for developing of cervix cancer (OR-0.51, 95% CI 0.31-0.83, p-0.006). The combined Asp/Glu with Glu/Glu genotypes who were using oral contraceptives was shown to be statistically significant with reduced risk of cervical cancer (OR-0.22 95% CI- 0.11-0.47, p-0.0002). It has been suggested that significantly correlation between HPV 16 and users of oral contraceptives in certain APE-1 genotypes with reduced risk in developing cervix cancer. In conclusion we observed statistical significant association with reduced risk of cervix cancer in APE-1 with different genotypes, though on the other hand, in association between HPV type 18 and those having SCC highly increased the risk of cervical cancer was observed.
Rashid Ramazanzadeh
Kurdistan University of Medical Sciences, Iran
Title: Effect of ciprofloxacin and levofloxacin alone or in combination with N-acetylcysteine on biofilm-producing Enterococcus faecalis: Contaminated root canals of extracted human teeth
Time : 14:45-15:05
Biography:
Rashid Ramazanzadeh is a Faculty of Medicine at Cellular & Molecular Research Center and Microbiology Department, Kurdistan University of Medical Science, Iran
Abstract:
Introduction: Biofilm producing-Enterococcus faecalis remains a major challenge in the disinfection of an infected root canal system. The aim of this in vitro study was to investigate the antibacterial effect of ciprofloxacin (CIP), levofloxacin (LEV) and their combination with N-acetylcysteine (NAC) in root canals infected with E. faecalis biofilms. Methods: Ninety one single root teeth were prepared. The antimicrobial effectiveness of seven-day intra canal medication with increasing concentrations (MIC to 1000MIC) of calcium hydroxide (CaOH2), CIP, LEV, CIP plus NAC (8 mg/ml) and LEV plus NAC was evaluated using the roots infected with E. faecalis for 30 days. Colony-forming unit (CFU) counting was performed. Biofilm formation and structural changes were monitored using scanning electron microscopy (SEM). Results: All the chemotherapeutic agents were significantly more effective than CaOH2 (P<0.05). There was no significant difference between combined treatment with NAC and single antibiotic treatments (P>0.05) except for LEV plus NAC which showed significantly higher reduction in CFU/ml than LEV at concentrations of 50 MIC and above (P<0.05). The effectiveness of the antibiotics was significantly positively correlated with their concentrations (r>0.8; P=0.000). SEM examination confirmed the formation of mature biofilms and a complete accordance between reductions in CFU/ml and destruction of biofilm structures was found. Conclusions: The local application of CIP and LEV and in vitro synergism of LEV and NAC led to more antimicrobial efficacy than CaOH2 against E. faecalis biofilms in root canals of extracted human teeth.
Liaqat Ali Chaudhry
SBAH-CITY Rehablitation Hospital & Medical Center Riyadh, K.S. Arabia
Title: Fascinating case reports published
Time : 13:55-14:15
Biography:
Liaqat Ali Chaudhry completed MBBS from King Edward Medical College university Lahore1981. He completed his post-graduation in Diploma in Tuberculosis & Chest Diseases -Punjab University Lahore-Pakistan, MCPS from College of Physicians and Surgeons Pakistan and MRCP, FRCP from The Royal College of Physicians Dublin-Ireland. He worked as house physician, Registrar, Assistant professor at King Edward Medical College Lahore, 1982-1987. Later he joined MOH of Saudi Arabia worked as Specialist Physicians chest specialist on Feb-1988, Consultant Pulmonologist and Chief of Tuberculosis Center, Dept. of Internal Medicine & Chest diseases Dammam medical complex and Honorary Associate & Professor Dammam Medical University, Eastern Province K. Saudi Arabia – 2011 and Chairman Internal Medicine & Consultant Invasive.
Abstract:
1- Case Report A 35-year-old immuno-competent male with open pulmonary tuberculosis associated with extra-ordinary extensive extra-pulmonary tuberculosis. 2- Milliary Tuberculosis with Unusual Paradoxical Response at 3 Weeks of Antituberculous Treatment. 7071/CR/10 Q3/22/01 Journal of the College of Physicians and Surgeons Pakistan 2012, Vol. 22 (01): 0 3- Paraplegia is not a diagnosis: Spinal tuberculosis deserves a place on the clinical radar screen: Awakening call to clinicians. 4- Clinical consequences of non-compliance with directly observed therapy short course (DOTS): Story of a recurrent defaulter.
Akeni O Augustine
University of Benin, Nigeria
Title: Modeling the Nigerian public health competencies in developing responsive public health care systems in early filovirus (Ebola) outbreaks in developing Africa
Time : 14:15-14:35
Biography:
Augustine Omena Akeni born 8 June, 1985 obtained MpH from the Public Health Management department of the University of Benin and is presently a PhD research candidate of the University. He has industry experience in Public Health administrative structural design and management. Proficient in Public Health Facility Administration. He assisted in formulation and initial implantation of the reviewed Edo State Primary Health Care Policy program. Assisted in design and implantation of the Edo State ICT program on data collation of public responsiveness to the State free Mother and Child Health Care Program. He has writing a number of academic papers on public health care administration.
Abstract:
Recent re-emergence of the filovirus Ebola outbreak in West Africa has stare up a narratives of a poor health and economies infrastructures of the developing countries of these West African states. The narratives hold that infectious diseases pose a huge public health challenges in the spaces where contacts with largely unknown reservoir occurs and were poor early warning systems as well as preparedness are handicapped by slow mobilization of responsive public health care resources. Complicated by fear of the viruses becoming endemic; the work understudies the recent Ebola virus in West Africa and the Nigerian responsive case study. The work focuses on development of the Nigeria model of a responsive systemic public health engagement in improving potentials for containment of infectious disease outbreaks at the national level. We argued that as the Nigerian case; that there was inherent capacity for development of quick responsive system model after the resource capacity of any national health care system which ultimately determined spreads of infectious disease as it defines the difference between stemmed case of outbreak or full epidemic as success depends on how swiftly resources can quickly be made available in stemming new outbreaks.
Sajjad Yazdansetad
Science and Research Branch University, Iran
Title: Biosynthesis of silver nanoparticles using Pseudomonas fulva as a novel silver nanoparticles producing isolate
Time : 14:35-14:55
Biography:
Sajjad Yazdansetad is a PhD candidate in Microbiology at Department of Microbiology, Science and Research Branch Univesity, Tehran, Iran. His main research fields are molecular microbiology and biotechnology. He has presented over 10 papers in conferences. Now, he studies silver nanoparticles as a novel approach in treatment of bacterial infections and biomedical applications.
Abstract:
Silver nanoparticles (Ag NPs) are the most interesting nanoparticles due to their wide variety of applications in various fields especially biomedical. In contrast, a promising usage of Ag NPs as antimicrobial agent is well known and has already found applications in antimicrobial paint coatings, textiles, water treatment, wound healing, medical devices and HIV prevention as well as treatment. In this study, we have reported the successful synthesis of silver nanoparticles by Pseudomonas fulva as a novel silver nanoparticles producing isolate. Pseudomonas fulva, a silver nanoparticles producing organism was isolated from vinegar. The isolate was confirmed by standard biochemical tests and 16S rDNA typing method. The ability of the organism was evaluated by adding 1 M solution of silver nitrate (AgNo3) to the microorganism culture medium. Production of the silver nanoparticles was investigated by X-ray diffraction (XRD), spectrophotometry, transmission electron microscopy (TEM) and fourier transform infrared spectroscopy (FTIR) analysis. The biosynthesis of silver nanoparticles was observed with the color change of the microorganism culture medium from yellow to dark brown. The sterile culture medium (without microorganism) supplemented with AgNO3 was used as a negative control. XRD indicated the silver nanoparticles diffraction peaks which were ranged from 2θ=30° to 2θ=80°. A peak at 430 nm for silver nanoparticles was seen by spectrophotometer. TEM micrograph revealed the fairly uniform size and spherical shapes of silver nanoparticles in the range of 50 to 100 nm. FTIR absorption spectra with range of 500 to 3500 cm-1 were also observed for silver nanoparticles. Biosynthesis of nanoparticles by microorganism is a green and eco-friendly technology. In this study, we introduced the novel silver nanoparticles producing isolate (Pseudomonas fulva), for the first time. Our study indicated that the isolate has robust potential in production of silver nanoparticles. Further study is required to explore the silver nanoparticles production of P. fulva.
- Track 8: Animal Bacterial Diseases
Track 9: Advances in Antimicrobials, Vaccines and Therapeutics
Track 10: Diagnostic Pathology of Infectious Disease
Location: Melia Meeting 1 & 2
Chair
B Y Chin
International Medical University, Malaysia
Co-Chair
Tony Velkov
Monash University, Australia
Session Introduction
B Y Chin
International Medical University, Malaysia
Title: Carbon monoxide: A poison to microbes
Time : 10:00-10:20
Biography:
B Y Chin received her degree 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, US, Department of energy in Richland, Washington. She 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 Sciences and Associate Dean of 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 hemo-proteins in order to restore physiological balance and sustain life. In this presentation we will review and discuss recent reports that have propelled and possible establish the paradoxical use of CO, once viewed as a toxic molecule, now as a host defense molecule agent against pathogens.
Tony Velkov
Monash University, Australia
Title: Development of novel antimicrobial lipopeptides against MDR Gram-negative ‘superbugs’
Time : 09:20-09:40
Biography:
Dr Tony Velkov completed PhD in 2000 from Monash University. His anti-infective discovery research is at the leading edge globally. He was awarded a NHMRC Research Fellowships in 2006, 2011 and 2014. The quality and impact of his independent research was recognized by the NHMRC with an Excellence Award in 2011. He has published over 50 papers in high-caliber journals, 3 book chapters and 15 conference presentations. The dynamic team he leads consists of 3 postdocs, 3 RAs and 9 PhD students. Over the last 6 years, he has obtained >$9M funding from the NIH, NHMRC and foundations.
Abstract:
The octapeptins are a family of naturally occurring cyclic lipopeptide antibiotics with a broad antimicrobial spectrum against multidrug-resistant (MDR) Gram-negative and Gram-positive ‘superbugs’. Octapeptins were discovered over 30 years ago, consequently there is limited information on their chemical biology and structure-activity-relationships (SAR). Accordingly, the broad aim of our study is to explore the chemical biology of the octapeptins through a SAR approach. Synthetic chemistry will be employed to generate a large series of novel analogs that will be evaluated against the biological systems the octapeptins are known to modulate. The best approach to achieve this is through the substitution of existing amino acids with novel derivatives rather than chemical modification of the side-chain functionalities. This can only be achieved through a ‘total synthesis’ approach, which is a particularly challenging task considering the complex chemical structure of the octapeptins. To the best of our knowledge, we are the first group to have developed an efficient routine ‘total synthesis’ of octapeptins on a scale that would allow for mechanistic investigations of antibacterial activity and resistance. For the first time our novel approach will interface the chemistry and biology of these important antibiotic compounds to investigate the underlying SAR with the purpose of creating new functions. More specifically the ultimate aim is to med-chem out liabilities such as nephrotoxicity whilst concomitantly improving the antimicrobial activity and spectrum, thereby creating the foundations for a new generation of safer and more efficacious antibiotics.
C. Diaz
Fundación MEDINA, Spain
Title: New Antibacterial Agents against Gram-negative Pathogens from a Fundacion MEDINA’s Microbial Natural Products Collection Screening
Time : 09:40-10:00
Biography:
Caridad Díaz completed his degree in Biochemist and Biology at the Granada University and she has worked as researcher in the screening and target validation department from Fundación MEDINA since 2010. She has published more than 5 papers in reputed journals and has presented more than 10 communications in International congress.
Abstract:
New antibacterial agents for treating Gram-negative bacteria are one of the main unmet medical needs because bacterial resistance represents a major issue for all classes of antibiotics. In this regard and considering that the majority of the antibiotics in the marketplace or clinical development derive from screening approaches with natural products, we developed a high throughput discovery program with the Fundación MEDINA’s microbial natural products collection. This collection contains more than 131,000 extracts and covers an uncommonly unexplored broad chemical space resulting from a variety of fermentation products. From that screening, several new compounds have been identified with activity against Gram-negative pathogens possessing a previously unreported chemical scaffold. They may represent a starting point for the development of a novel treatment of infections. These new compounds (MDN-057, MDN-0114, MDN-0116 and MDN-0119), with unprecedented structures, with molecular weights ranging from 282 to 544 Da and with MICs (Minimal Inhibitory Concentration) in the 40-128 µg/ml range against Acinetobacter baumannii and other resistant clinical strains, are described in this work
Promod Mehta
Maharshi Dayanand University, India
Title: Comparing several gene targets of Mycobacterium tuberculosis for designing multiplex-PCR: It’s implications for diagnosis of extra pulmonary tuberculosis
Time : 09:00-09:20
Biography:
Promod Mehta presently as Director at Centre for Biotechnology, Maharshi Dayanand University, India
Abstract:
Diagnosis of extra pulmonary tuberculosis (EPTB) exhibits true challenges due to paucibacillary nature of specimens. An early diagnosis of EPTB is needed to initiate anti-tubercular therapy to avoid unnecessary morbidity and mortality. During the last decade, Polymerase Chain Reaction (PCR) assays have emerged important tools in diagnosing EPTB specimens. Many studies have shown the inability of a single gene target to detect M. tuberculosis with certainty in biological specimens. A careful selection of gene targets is imperative for designing a Multiplex-Polymerase Chain Reaction (M-PCR) assay. We compared various gene targets of Mycobacterium tuberculosis that is IS6110, genes encoding MPB-64 (mpb64; Rv1980c), 38 kDa (pstS1; Rv0934), 65 kDa (hsp65; Rv0440), 30 kDa (fbpB; Rv1886c), ESAT-6 (esat6; Rv3875) and CFP-10 (cfp10; Rv3874) proteins and devR (Rv3133c) by PCR assays on the same 105 EPTB specimens. The mpb64 showed the highest sensitivity followed by IS6110, hsp65, 38 kDa, 30 kDa, esat6, cfp10 and devR. This study showed the authenticity of mpb64+IS6110 in designing an M-PCR assay as high sensitivity (96% in confirmed EPTB cases and 89% in clinically suspected EPTB cases) and specificity (100%) was observed using clinical diagnosis as the gold standard. These results along with the clinical findings and histopathological/cytological observations may help for an early diagnosis of EPTB. This simple and cost-effective test may also be a better alternative than rpoB (encoding RNA β polymerase subunit) based Xpert assay especially in resource-poor settings.
Gamal Wareth
Friedrich-Loeffler-Institut, Germany
Title: Investigation and molecular typing of Brucella isolated from dairy cattle herd immunized with Brucella abortus RB51 vaccine in Egypt
Time : 10:20-10:40
Biography:
Gamal Wareth has obtained his Bachelor degree (BVSc) from faculty of Veterinary Medicine, Assuit University, Egypt in 2002. He completed his Master degree in Veterinary Pathology in 2009 at Benha University, Egypt. Then he enrolled for DVM in Free University of Berlin in collaboration with Friedrich-Loeffler-Institut (FLI), Germany. He is currently works in NRL of brucellosis at Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses in Jena, Germany. The main areas of his interest are development of diagnostic products; proteomics, epidemiology, molecular epidemiology and microbiology of pathogen level 3 agents (Brucella spp., Francisella tularensis, Burkholderia mallei/pseudomallei, Bacillus anthracis, Yersinia pestis).
Abstract:
Brucellosis is still a common in capacitating zoonoses that affecting wide range of domesticated and wild animals as well as humans. Brucella was firstly described by Sir David Bruce in 1887 in human. Fifty two years later (1939), the disease was reported in Egypt for the first time in a scientific report, however the doubt still exist if, whether the disease is endemic in Egypt since thousands of years. Since then the disease remained endemic nationwide and affecting all livestock animals. In Egypt, brucellosis control programs for bovines are based on a test and slaughter policy as well as vaccination. Recently, Mass vaccination with attenuated B. abortus RB51 has been approved nationwide; however some may lead to abortion in pregnant females. The present study was carried out in supposed Brucella-free dairy cow farm in Egypt to investigate a numerous occurrence of abortions occurred after immunization with B. abortus strain RB51. Among pregnant vaccinated cows, 10% aborted three months post vaccinations. All aborted cows show positive reaction with Rose Bengal Test (RBT) and Complement Fixation Test (CFT). B. abortus biovar one was isolated from four aborted fetuses. Conventional biochemical analysis revealed two B. abortus smooth type and two rough types. The use of the Bruce ladder PCR provided clear differentiation to the field strain and excludes the RB51 vaccine strain. Genotyping analysis of isolates by Multiple Loci VNTR Analysis (MLVA-16) cleared that there are two different isolates in the herd with low genetic diversity. Isolated B. abortus are located in a different cluster from the RB51 vaccinal strains which used in immunization of the herd. In Conclusion: B. abortus bv1 is the source of outbreak but not the vaccine. RB51 vaccine would only reduce the number of abortions caused by field strains but offer no protection from infection. The most likely source of infection appears to be the uncontrolled introduction of the agent via vectors or latent carrier. Eradication of brucellosis is a multidirectional process required combination between ordinary control program and high biosecurity level in the farms.
Memon Z
Liaquat University of Medical & Health Sciences, Pakistan
Title: Serum trace elements in active pulmonary tuberculosis by atomic absorption spectrophotometer
Time : 10:55-11:15
Biography:
Zainab Manzoor Memon Ph.D. Research Scholar in Biochemistry. Presently she is working at Liaquat University of Medical & Health Sciences, Jamshoro, Pakistan. She has eight international research publications and presented her research work at national and international conferences. She has attended many academic workshops, symposium and has three years research experience. She has expertise on bio analytical techniques such as AAS, Spectrophotometer, AAA, Ultra Centrifuge Machine, HPLC, Microlab 300 & Chemistry Analyzer. She also has command on statistical software including SPSS and Minitab.
Abstract:
Background: Tuberculosis (TB) is a curable disease though it is still remains a major public health problem worldwide especially in developing countries. TB ranks as the second leading cause of death throughout the world. Pakistan ranked as fourth in the midst of 22 elevated challenging tuberculosis nations. Trace elements have involved in many biochemical and physiological functions. Poor concentration of trace elements has been resulting clinical outcome. Objectives: The objective of study was to find out the alteration in serum trace elements levels in active pulmonary tuberculosis patients and was compared it with normal healthy volunteers with no sign and symptoms of TB. Method: A total of 248 active pulmonary tuberculosis patients were selected from Liaquat University of Medical & Health Sciences, Jamshoro, Sindh, Pakistan, Liaquat University Hospital, Hyderabad, Sindh, Pakistan, Rajputana Hospital, Hyderabad, Sindh, Pakistan and Institute of Chest Diseases, kotri, Sindh, Pakistan. The subjects were recruited from both genders with same age group 20-70 of years. Blood samples of active pulmonary tuberculosis patients with sputum smear positive were collected and were compared with healthy control subjects without symptoms of pulmonary tuberculosis. Approval was taken from all hospitals. Informed consent was also taken from each patients and healthy participants. Serum trace element analysis was carried out by flame atomic absorption spectrophotometer (FAAS). Results: Among 248 active pulmonary tuberculosis patients, 104 (42%) was male patients while 144 (58%) was female subjects respectively. The interpretation of outcome was made on the basis of reference ranges. In our study we were found that the mean serum zinc, iron and magnesium were significantly low in active pulmonary tuberculosis patients (58.1 mg/dl), (34.2 mg/dl) and (0.6 mg/dl). While mean serum calcium was elevated compare to normal healthy control subjects.
Majed A Halwani
King Abdulaziz city for Science and Technology, Saudi Arabia
Title: Comparison for encapsulated gentamicin efficiency within different neutral and negative charged liposomal formulations for treating pathogenic lung bacteria
Time : 11:15-11:35
Biography:
Majed Halwani currently working as an Associate Research Scientist at Antimicrobial and Vaccine Development unit, King Saud bin Abdulaziz University for health sciences, Kingdom of Saudi Arabia - Riyadh
Abstract:
Background: Associated pneumonia with antimicrobial resistant pathogens is a worldwide concern. New antimicrobial agents are sparse and such infections will become a major challenge. Liposomal formulations are of proven superiority in delivering drugs to end organs and have the ability to protect drugs from metabolites and other host's harsh materials and at the same time reduce toxicity. The goal of this study is to evaluate different neutral and negative liposomal-gentamicin formulations for their encapsulation efficiency, stability, and antibacterial activities. Method: Neutral liposomal-gentamicin (NLG), negative charged liposomal-gentamicin-1 (NELG-1) & negative charged liposomal-gentamicin-2 (NELG-2) were prepared by dehydration-rehydration method. The particles sizes in the formulations were measured by light scattering technique. Loaded gentamicin within different liposomal nanoparticles was measured by microbiological assay. Minimum inhibitory concentration (MIC), minimum bactericidal concentrations (MBC), and killing time curve were studied on several bacterial gram negative / positive strains. Liposomal formulations’ stability were studied within different biological conditions include plasma, sputum, and BAL at 37 áµ’C. Results: The encapsulation efficiency of gentamicin within NLG, NELG-1, and NELG-2 were 1.8%, 37.2% and .43.6% respectively. Formulations stability of each of NLG, NELG-1, and NELG-2 in presence of some of the biological fluids such as bronchial alveolar lavage (BAL), human plasma, and sputum were studied at 37áµ’C for 48 hours. NLG, NELG-1, and NELG-2 were kept interestingly amount of gentamicin among those biological fluids, but when exposed to human plasma their content of gentamicin were mild dropped down. Bacterial eradication of several planktonic pathogenic ATCC strains by NLG was 2-fold lower than free gentamicin, whereas NELG-1 and NELG-2 eradicated them as same as free gentamicin. The killing time curve values at 1, 2 and 4 time MIC for our formulations against P. aeruginosa were better than free gentamicin. Interestingly, NELG-2 reduced biofilm formation by 2-fold on day 2 better than NLG and NELG-1 compared to free gentamicin. Conclusion: All formulations exhibited a bactericidal effect able to eradicate bacterial strains at low concentration when compared to free gentamicin. On the other hand, these formulations exhibited improved killing time and anti-biofilm forming. Our endeavors are ongoing to examine theses formulations toxicity profile.
Alessandro Pini
University of Siena, Italy
Title: A novel synthetic antimicrobial peptide for the cure of gram-negative infections: Mechanism of action, efficacy in vivo, toxicity, bio-distribution and resistance selection
Time : 11:35-11:55
Biography:
Alessandro Pini is a Professor of Biochemistry at the University of Siena, Italy. He is Founder and President of SetLance SRL, a start-up company in Siena with a special focus in the identification and early development of peptide based-drug. He is author of dozens of publications and inventor in 10 patents.
Abstract:
A synthetic antimicrobial peptide was identified some years ago as possible candidate for the development of a new antibacterial drug. The peptide showed a MIC 90 below 1.5 µM for Pseudomonas aeruginosa and Klebsiella pneumonia. In models in vivo of P. aeruginosa lethal infections the peptide and its pegylated form, allowed a survival percentage ranging between 65-80% in sepsis and lung infections when injected IV, and completely resolved skin infections when administered topically. Plasma clearance demonstrated different kinetics for both peptides, with a higher persistence for the pegilated one after two hours from injection. Bio-distribution in organs did not show significant uptake differences between the two peptides. Contrary to colistin, the molecule here described did not select resistant mutants in bacterial cultures. Moreover it resulted non-genotoxic and with an in vivo toxicity comparable to antimicrobial peptides already used in clinic. The characterizations here reported are part of a preclinical development plan that should bring the molecule to clinical trials in the next years.
Onaolapo J A
Ahmadu Bello University, Nigeria
Title: Antibiotic susceptibility profile of methicillin resistant Staphylococci aureus in poultry farm, in Zaria, Nigeria
Time : 11:55-12:15
Biography:
Josiah Ademola Onaolapo from Department Of Pharmaceutics and Pharmaceutical Microbiology, Ahmadu Bello University, Zaria, Nigeria
Abstract:
Methicillin antibiotic is not commonly used in veterinary practice in the hospitals due to its toxicity, but the wide spread of its gene (MecA) calls for concern in livestock.The epidemiological and antibiotic susceptibility of Staph. aureus in Zaria, Nigeria was carried out in this study due to the increasing resistance associated with Staph. aureus in poultry birds. In this study, 250 samples of chicken droplets were collected from five different poultry farms (50 samples from each farm) within Zaria metropolis. Eighty eight (88) isolates of Staph. aureus were confirmed using standard microbiological methods. The antibiotic susceptibility pattern showed that the isolates where 90.8% susceptible to Ciprofloxacin, 76.2% to Vancomycin, 72.2% to Pefloxacin, 65.6% to Gentamicin, 58.8% to Methicillin, 57.6% to Oxacillin, 49.6% to Ampicillin and 25.3% to Tetracycline. Their percentage resistance varied from 9.2, 23.9, 27.8, 34.4 and 42.4 for Ciprofloxacin, Vancomycin, Pefloxacin, Gentamycin and Oxacillin, respectively. The isolates showed high resistance 74.7% and 50.4% to Tetracycline and Ampicillin respectively while 41.2% of the isolates were resistant to Methicillin and produces β-lactamase enzyme. Seventy five percents (75%) of the isolates had MIC value of ≥64µg/ml while 25% had MIC ≤2 µg/ml. The MARI result showed that 40% of the isolates had MAR index of ≤0.3 while 60% had MARI of ≥0.4; indicating that the Staph. aureus tested were pre-exposed to the antibiotics used in this study. Further study on the 42.4% isolates that were resistant to Oxacillin showed that 60.5% and 64.8% were still resistant on mannitol salt agar impregnated with 4 µg/ml of Vancomycin and 67.6% and 70% of the same isolates grew on brain heart infusion agar impregnated with 6 µg/ml of Vancomycin after 24 and 48 hours incubation at 37o C respectively. This study showed high incidence of Staph. aureus and antibiotics resistance among poultry birds in Zaria, Nigeria and calls for antibiotic surveillance and education of the poultry farm workers to curb the wide spread of resistance gene which could be transferred in zoonotic diseases.
Sanjib Bhakta
University of London, Uk
Title: Common non-steroidal anti-inflammatory drugs (NSAIDs) in the fight against tuberculosis (TB)
Time : 12:15-12:35
Biography:
Following a BSc (Hons), an MSc and a PhD in Molecular Biology & Biochemistry from world class Universities & Research Institutions in India, Dr Sanjib Bhakta joined the Oxford University, Department of Pharmacology as an ISIS innovation Senior Research Scholar and shortly after he was awarded with a Wellcome Trust International Travelling Fellowship. He graduated from The Queen’s College, University of Oxford completing a second doctoral degree (DPhil) and received a “Sir William Paton Prize†in Pharmacology. He attained his first academic appointment at Birkbeck as a University Lecturer to lead his interdisciplinary research to tackle infectious diseases like TB.
Abstract:
Tuberculosis (TB) remains a serious healthcare issue, more than two decades on from the first time it was declared as a global health emergency. Control of the disease has become increasingly difficult because of the alarming rise of antibiotic resistance in Mycobacterium tuberculosis, the etiological agent of TB. Development of new and effective drugs with novel mechanisms of action is thus of paramount importance to tackle antibiotic resistance. Novel chemical entities require at least a decade to be commercially available and repurposing drugs offers a solution to circumvent the investment of time and other resources (1). Certain common non-steroidal anti-inflammatory drugs (NSAIDs) have proven to be selectively bactericidal against replicating, non-replicating and multi-drug-resistant clinical isolates of M. tuberculosis (2, 3). Our primary focus is to repurpose carprofen and investigate their novel mechanisms of action in M. tuberculosis to help design more potent inhibitors in the future. To this effect we have followed both target-based and whole-cell approaches. Whole-cell transcriptomic analyses have revealed the effects of the drugs on a selected set of genes involved in key metabolic pathways that also play essential roles in antimicrobial resistance. Furthermore, the NSAIDs showed influence on the expression levels of proteins involved in cell-wall homeostasis and dormancy mechanisms. The most active NSAID, Carprofen was found to be a bactericidal drug that also inhibited the formation of mycobacterial biofilms and exhibited strong efflux pump inhibitory properties thus demonstrating their great potential in reversing antibiotic resistance.
Arun G Ingale
North Maharashtra University, India
Title: DENV3 envelope protein of dengue virus: A potential target for the vaccine designing
Time : 12:35-12:55
Biography:
Arun G Ingale has obtained his PhD in Biotechnology from Sant Gadge Baba Amravati University, India. He was the Founder, Head, Department of Biotechnology, Dr Babasaheb Ambedkar Marathwada University, India. He is President of Society for Biotechnology and Bioinformatics, India. He is also the Editor in-Chief of the Journal of Biotechnology and Bioinformatics (JBB) and International Journal of Modern Biotechnology. His primary field is immunology with research emphasis on CD antigens and the structure-function prediction using Bioinformatics approach. He has recently entered the developing field of Lectin Biosensor and Glyco-Nanobiotechnology research. In Genomics research area he has been working on constructions of transgenic Okra against pest (Lipidopteron) and pigeon pea against bollworm. In proteomics research area he is working on proteomics of lectin and other plant and microbial proteins. In bioinformatics, he has developed a database of CD markers and Toxin database is on completion. He has published several research papers in national and International journal of repute. He has submitted protein and nucleotide sequences on NCBI and viral protein models are being submitted in PDB database. Currently, he is holding Major research projects as a Principal Investigator and Head Department of Biotechnology, North Maharashtra University.
Abstract:
Dengue is arthropod-borne viral disease mainly found in Southeast Asia, the Pacific and the Americas. There are four antigenically discrete serotypes of dengue viruses (DENV1-4) each of which is competent of causing dengue fever, hemorrhagic fever and dengue shock syndrome and currently circulating in these areas. An estimated 50 million dengue infection cases occur globally with around 500,000 cases of hemorrhagic dengue and 20,000 deaths per year. In India, numbers of dengue fever cases are increasing every year. Currently, vaccines and antiviral drugs are under investigation could also make vital contribution to dengue control in the future. Hence, there is dire aspiration to develop new vaccine or drugs that are more targeted fewer lethal and more successful against this virus. Here we analyzed structural components of DENV3 envelope protein for development of homology model and prediction of their antigenic determinants. This work may contribute to effectively target and cultivate operational epitope vaccine to defend the host from the virus.
Bincy Joseph
Kerala Veterinary and Animal Sciences University, India
Title: Effective protection by high efficiency bicistronic DNA vaccine against anthrax expressing protective antigen and catalytically inactivated lethal factor
Time : 13:40-14:00
Biography:
Bincy Joseph has completed her Graduation in Veterinary Sciences (BVSc & AH) from Kerala Agricultural University with KAU merit scholarship. She has completed Post graduation in Veterinary Bacteriology from Indian Veterinary Research Institute with ICAR Junior Research Fellowship and completed Doctorate in Veterinary Bacteriology from Indian veterinary research institute with Senior Research Fellowship. She has published 8 research articles in various reputed journals and participated in more than 10 national and international conferences and presented research papers.
Abstract:
Limitations of currently available anthrax spore vaccine necessitate the development of an improved vaccine for animals. In the present study lethal factor gene of Bacillus anthracis was made catalytically inactive by primer based site directed mutagenesis. To explore whether immunization with plasmid encoding this mutated lethal factor (mLF) and protective antigen (PA) can provide protection against anthrax, a bicistronic DNA vaccine encoding PA and mLF was then made along with mono-cistronic constructs encoding PA/ mLF. The ability of the constructs to express the encoded genes was verified by transfection in MDBK cells followed by indirect immunofluorescence analysis. To investigate the immunogenic potential of the made constructs immunization trials were conducted in mice. After primary immunization with these DNA vaccines the mice were boosted with DNA vaccines, recombinant proteins or formalin inactivated spores (FIS) on 14thand 28th days post vaccination. Subsequently, indirect ELISA, toxin neutralization assays (TNA) and monitoring of cytokines (IL-4, IL-2 and IFN-γ) were done to monitor the immune response. The direct challenge test of immunized mice was done using 1000 LD50 of virulent B. anthracis IVRI strain. The results showed that the heterologous prime boost regimen involving priming with bicistronic DNA construct encoding PA and mLF or mono-cistronic DNA construct encoding PA and boosting with recombinant proteins can provide better protection (66.66%) compared to other groups. At the same time immunization with bicistronic DNA construct encoding PA and mLF and boosting with recombinant proteins could provide higher antibody titer, toxin neutralization titer and Th1 and Th2 response (p<0.0001) compared to all other groups illustrating that DNA vaccine encoding PA and mLF conferred a broader spectrum of immune reaction than PA alone. The increase in serum concentration of IL-2, 1L-4 and IFN-γ indicated that both humoral and cell mediated immune response were elicited by DNA vaccination. Thus, the results of the present study indicatedthe feasibility of DNA prime protein boost based immunization strategy based on PA and mLF being developed into nontoxic, effective and stable anti-anthrax vaccine.
Whika Febria Dewatisari
Universitas Terbuka, Indonesia
Title: Antimicrobial activity of the saponin extract of Sansevieria trifasciata var. Golden Hahnii
Time : 14:00-14:20
Biography:
Whika Febria Dewatisari is an Academic Staff at Department of Mathematic and Natural Science in Open University (Universitas Terbuka) Bandar Lampung-a public university in Indonesia. She is a Biology Lecturer and started working at 23 years old. She teaches courses in Plant Embriology, Microbiology and Ecology
Abstract:
Sansevieria trifasciata used as an ornamental plant. It also used as a traditional medicine for influenza, cough and inflammation of the respiratory tract. The roots and leaves of S. trifasciata contains many secondary metabolites such as saponins that efficacious as a cough remedy to treat sprains, injuries hit, venomous snake bites, ulcers, cough, inflammation of the respiratory tract and hair growth. Microbes are used to seeing the antibacterial saponins activities of S. trifasciata are Escherichia coli and Staphylococcus aureus. The main reason for used these microbes because E. coli is a bacterium that causes diarrhea and S. aureus is one of the bacteria that cause cough in humans. Plants were used S. trifasciata var. Golden Hahnii. Based on research by Dewatisari (2008), states that the variety had the highest saponin content of S. trifasciata is Golden Hahnii among other varieties and the parts that had the highest saponin were in its roots. This article aims to explain quantitatively the effectiveness of saponin compounds from the roots of S. trifasciata var. Golden Hahnii as anti-microbials in inhibiting the growth of bacteria S. aureus and E. coli. Methods of data collection were sample preparation, a preliminary test, the extraction of saponins, antibacterial activity test and isolate compound separation by TLC (Thin layer Chromatography). This study includes the extraction of the active compounds in the roots of S. trifasciata by maceration with methanol 90%. Separation of the active compound was conducted by TLC. Eluent used was chloroform; methanol; water with various concentration (13: 4: 1), (65; 50: 10), (20: 60: 4), 20: 60: 10). Antibacterial test conducted by the disc diffusion method against S. aureus and E. coli. Identification of test compounds triterpenoid saponins using foam and Liebermann-Burchard color test (LB). The results showed that the extract of the roots has potential as anti-bacterial. The extract was able to inhibit the growth of E. coli and S. aureus. At the optimum concentration of 200 ppm produced inhibition zone was 26.5 mm to 20.2 mm for S. aureus and E. coli. Best eluent to separate the triterpenoid saponins in root extracts of S. tifasciata was chloroform; methanol; water at a concentration (20: 60: 4) with 3 separate visible stains are: 0.135; 0.85; 0.815. Mechanism root extract as an anti-bacterial S. trifasciata was synergistic. It was seen from the inhibition zone, for E. coli isolates I=5.52 mm and isolates II=2.50 mm, for S. aureus isolates I=1.52 mm and isolates II=0.58 mm while the isolates III is not effective as an anti-bacterial.
Yanmin Hu
University of London, UK
Title: Antimicrobial peptide novicidin synergises with rifampicin, ceftriaxone and ceftazidime against antibiotic-resistant gram-negative Enterobacteriaceae in vitro
Time : 14:20-14:40
Biography:
Yanmin Hu is a Senior Research Fellow at St George’s, University of London. Her main research interests are in tuberculosis and antibiotic discovery. The scientific and intellectual imperatives for her research include new drugs and better drug regimen for tuberculosis and other important infectious diseases; improved chemotherapy to eradicate persistent bacteria; molecular approaches to understand the processes of infection and pathogenesis of M. tuberculosis and other pathogens.
Abstract:
Infections caused by gram-negative bacteria species such as those in the Enterobactericeae family are responsible for high rates of morbidity and mortality. The spread of antibiotic resistance amongst gram-negative pathogens is a serious clinical threat requiring urgent attention. Traditional novel drug development inevitably leads to emergence of new resistant bacterial strains, rendering the new drugs ineffective. Therefore reviving the therapeutic potentials of existing antibiotics represents an attractive novel strategy. Novicidin, a novel cationic antimicrobial peptide is effective against gram-negative bacteria. The actions of novicidin in combination with rifampicin, ceftriaxone and ceftazidime were investigated. We performed in vitro investigations to test against 94 antibiotic resistant clinical Gram-negative isolates and 7 strains containing New Delhi metallo-β-lactamase-1 (NDM-1). The effects of combining novicidin with rifampicin, ceftriaxone and ceftazidime were examined using the chequerboard method and time kill curves. A fluorescence assay was used to investigate the depolarisation of the bacterial cell membrane by novicidin. The post antibiotic effect was measured. The cytotoxicity and haemolysis of novicidin were examined using neutral red uptake in the L929 fibroblast cell line and lysis of human blood. Novicidin combined with rifampicin showed synergy with over 70% of the tested gram-negative clinical isolates (n=94) and NDM-1 strains (n=7) reducing the MIC significantly. The combination of novicidin with ceftriaxone and ceftazidime showed synergistic effects with more than 89.7% of ceftriaxone-resistant strains and 94.1% of ceftazidime-resistant strains. These synergistic combinations were also demonstrated using time kill studies with multiple strains. We also demonstrated that novicidin altered the cytoplasmic membrane potential by membrane depolarisation against both Escherichia coli and an isolate from the Klebsiella-Enterobacter-Serratia (KES) group. Furthermore, novicidin was shown to increase the post-antibiotic effect (PAE) when combined with rifampicin or ceftriaxone. Novicidin showed low haemolytic activity and conservation of cell viability in the cell culture post treatment. We demonstrated that novicidin strongly rejuvenates the therapeutic potencies of ceftriaxone or ceftazidime against ceftriaxone or ceftazidime resistant gram-negative bacteria in vitro. In addition, novicidin boosted the activity of rifampicin. This strategy can have major clinical implications in our fight against antibiotic resistance bacterial infections.
Hosseini Seyed Davood
Razi Vaccine and Serum Research Institute, Iran
Title: Amplification, cloning and expression of Brucella melitensis bp26 gene (OMP28) isolated from Markazi province (Iran) and purification of Bp26 protein
Time : 14:40-15:00
Biography:
Hosseini Seyed Davood is currently as Dean for Razi Vaccine & Serum Research Institute Central Area branch(Arak), Iran
Abstract:
Brucellosis is a debilitative disease that imposes costs on both economy and society. It is shown that although the vaccine can prevent abortion, it does not provide complete protection against infection. In Iran, Brucella melitensis is a common causative agent for brucellosis and BP26 protein of this bacterium having a good antigenesity and an important vaccine candidate. In this study B. melitensis bp26 gene was cloned first in to PTZ57R/T vector and accessed on the PET28a vector and sequenced. Recombinant vector transformed and expressed in to E. coli BL21 (DE3) and then recombinant protein was purified with Ni-NTA column of chromatography against His tag. Obtained rOmp28 could be used as a research experimental tool to find its potential as a detection kit and vaccine candidate.
Collins Njie Ateba
North West University, South Africa
Title: Genetic characterization of antibiotic resistant Enterococcus faecalis isolated from groundwater in North West Province, South Africa
Time : 15:00-15:20
Biography:
Collins Njie Ateba has completed his PhD from the North West University - South Africa. He also received professional training in the Centre for Medical Genetics, Yerevan State University, Yerevan – Armenia in 2006 in the Department of Microbiology- Tartu University Tartu – Estonia in 2007 and the Lethbridge Research Station – Lethbridge Alberta, Canada in 2014. He is currently an Associate Professor in the Department of Biological Sciences, Microbiology Division, North West University –Mafikeng Campus and is head of the Water, Food Safety and Phage Therapy/Biocontrol Research Laboratory. He is actively involved in research training and lecturing at both undergraduate and postgraduate levels. He has been serving as a host mentor for the DST/NRF internship program from 2011 till date. He has published more than 30 papers in reputed journals and serving as an Editorial Board Member of repute. He has presented research papers in a number of conferences locally and internationally.
Abstract:
Enterococcus species are commercial organism usually inhabit the gastrointestinal tract of humans and animals. They are the indicators of faecal contamination in water and food and mostly enter into host through faecal-oral route. The presence of some pathogenic strains of Enterococci specifically vancomycin resistant Enterococcus faecalis cause nosocomial infection which become more prevalent in recent years and make situation more worse. In South Africa, mostly people in rural areas have no access to portable water and they depend on untreated water source. Therefore, they are at high risk of exposure to pathogenic microbes. Keeping this in view, present study was conducted to determine the presence of vancomycin resistant Enterococcus faecalis from the underground water from North West Province, South Africa. Enterococci were isolated from 60 borehole tap water samples on Bile Esculin agar. The presumptive isolates were analysed for the characteristics of Enterococcus faecalis using preliminary (Gram staining, catalase test, growth in 6.5% NaCl broth and haemolysis) and confirmatory (species specific 16S rRNA and ddlE. faecalis gene specific PCR) tests. The screened isolates were further subjected to antimicrobial susceptibility for 13 different antibiotics by standard disk diffusion method. Also, these isolates were screened for the presence of vancomycin resistant genes (vanA and vanB) and transposable elements (Tn1546 and Tn1547). A total of 253 presumptive isolates were obtained on Bile Esculin Agar and analysed for characteristics of Enterococcus faecalis using preliminary and confirmatory tests. A molecular typing confirmed 122 isolates as Enterococcus faecalis which were further screened for antimicrobial susceptibility. A large proportion of the isolates were multiple antibiotic resistant and showed resistance to vancomycin, penicillin, amoxicillin, ampicillin, tetracycline and erythromycin. The presence of vanB gene was observed in 83 isolates followed by 63 isolates harbouring vanA gene. Only 27 isolates which harbour vanA showed the presence of Tn1546 whereas none of the vanB positive isolates showed the presence of Tn1547. Conclusions: This study showed the presence of vancomycin resistant E. faecalis (multiple antibiotic resistant) from the underground tap water which is a cause of concern. Therefore, there should be a regular surveillance to monitor antibiotic resistant bacteria to prevent its spreading in nature. Further, detailed molecular study is required to know the deep insight into the genetic jugglery in these bacteria as in the present study a high level incompatibility between phenotypes and genotypes were observed.