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Workshop

Dr Helieh Oz has a DVM, MS (U. IL); PhD (U. MN) and clinical translational research certificate (U.KY). Dr Oz is an active member of American Association of Gastroenterology (AGA) and AGA Fellow (AGAF) and associate in Rome Foundation (Functional Gastrointestinal Diseases). Dr Oz is a Microbiologist with expertise in infectious and inflammatory diseases, drugs discovery, pathogenesis, innate/mucosal Immunity, molecular biology, and micronutrient. Dr. Oz has over 90 publications in the areas of chronic inflammatory disorders, microbial and infectious diseases. Dr Oz has served as Lead Editor for special issues including Gut inflammatory, infectious diseases and nutrition 2017 (Mediators of Inflammation); gastrointestinal inflammation, repair: role of microbiome, infection, nutrition (Gastroenterology Research Practice), J. Nutrient and guest Editor for J. Pediatric Infectious Disease. Dr Oz is a member of different editorial board and an avid reviewer for journals. 

Abstract:

Pathogens cause dysregulated immune and inflammatory response. Microbiome in the gastrointestinal tract (GI) acts as a protector of health from disease. Microbiota helps absorption of nutrients and guards against invasive pathogens to dwell in GI. While, antibiotics are required to fight pathogens and infectious diseases, overuse and abuse as well as unintentional consumption of food contaminated with antibiotics affect GI to alter the composition of microbiome. Further, antibiotics shift equilibrium from health into disease status as in infections with Clostridium spp. Infectious protozoan and microbial diseases are transmitted from animals and cause GI inflammation and diarrhea in man. The common preventive practice for infectious diseases and growth promoter in farm animals are continued use (overuse) of antibiotics which enter blood circulation and contaminate eggs, milk, and meat products. Antibiotics are entered food chain and consumed by humans with possible allergic, antibiotic resistance, and other enigmatic side effects. It is estimated that over 80% of antimicrobials are used for prevention and growth promotion in swine, cattle and chickens compared to only 20% used in human therapies. This presentation will aim to discuss unintentional consumption of antibiotic residues in contaminated food products with possible side effects. Association between overuse and abuse of antibiotics in food animal industry will be discussed with outbreaks of major infectious foodborne diseases, altered gut microbiota and dysbiosis with serious complications. In addition, different preventive measures will be discussed including possible applications of new agents as surrogates to substitute antibiotics in food animals.

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Antoine Abou Fayad has completed his PhD at the age of 26 years from Univeristy of St Andrews, UK, and postdoctoral studies from Helmholtz Institute for Pharmaceutical Research in Saarland, Germany. He is currently a junior group leader and an instructor at the department of experimental pathology, immunology and microbiology at the faculty of medicine in the American University of Beirut. He has published more than 12 papers in the field of AMR in reputed journals and has 2 pending patent applications. 

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Antimicrobial resistance (AMR) is a life threatening urgent global health problem. There is increasing concern about the alarming emergence of multidrug-resistant superbugs. Infections resulting from such superbugs are barely responsive to treatment with few if any currently available antibiotics, reviving memories of the pre-antibiotic era and raising concerns regarding a post-antibiotic era. Given the complexity of the AMR challenge and concerns about issues at the interface of human health, animal health, and the environment, it seems particularly important to emphasize the role of a One Health approach in addressing the problem. Priorities should not only consist of strengthened human and animal health surveillance and monitoring for resistant organisms, antimicrobial stewardship programs, infection-control courses, and development of rapid diagnostic tests, but also should include, development and validation of new antimicrobial agents. As noticed, since combination therapeutic strategies will reach a dead-end very soon, novel antimicrobials with unexploited targets are highly needed in a timely manner. Soil microorganisms produce natural products as a significant number of drugs in clinical use are derived from these metabolites. Actinomycetes and Myxobacteria are mainly soil dwelling microorganisms that produce secondary metabolites to be screened for antibacterial activity. More than 80% of clinically utilized antibiotics are either natural products or natural product-derived molecules such as vancomycin, teicoplanin, daptomycin, tetracycline and many others. Our aim is to demonstrate the progress on isolating and identifying novel antimicrobials from  new Actinomycetes and Myxobacteria from Lebanon. 

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Prof. Hak-Ryul Kim has completed his PhD from Auburn University in Auburn, Alabama and postdoctoral studies from University of Maryland, School of Pharmacy in Baltimore, Maryland. He is a professor and Chairman of School of Food Science and Biotechnology, Kyungpook Nartional University in Daegu Korea. He has published more than 65 papers in reputed journals and has been serving as a board member of the International Society of Biocatalysis and Agricultureal Biotechnology. 

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Structural modification of natural lipids by biocatalysis can change their properties or even create novel functionalities. Hydroxy fatty acid, one of oxylipins, can be produced from the microbial bioconversion of natural vegetable oils. Recently 7,10-dihydroxy-8(E)-octadecenoic acid (DOD) was produced with high yield from olive oil containing oleic acid by bacterial strain Pseudomonas aeruginosa PR3, and further study confirmed that DOD contained strong antimicrobial activities against broad range of microorganisms. In this study we tried to modify DOD molecules by physical reaction to create new functionality or to enhance the antimicrobial activity of DOD. After the harsh heat-treatment, a novel furan fatty acid (EODA) was produced from DOD. We confirmed that EODA presented strong antibacterial activity against multidrug-resistant Staphylococcus aureus and also EODA showed a recuperative effect of the beta-lactam antibiotics activity against methicillin-resistant Staphylococcus aureus.

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Temedie-Asogwa Tarilate is a 29 year old pharmacist. She completed her B-Pharm degree at the age of 21 years from Niger Delta University, Wilberforce Island, Nigeria. She holds a masters degree in Clinical Pharmacy from Nnamdi Azikiwe University, Awka, Nigeria. She has an eight (8) years cumulative work experience as a Pharmacist. She has for 5 years now worked as a clinical Pharmacist at the University of Port Harcourt Teaching Hospital.and currently head the geriatrics pharmacy unit of the hospital. She has published two papers in reputable journals and also functions as a preceptor and co-supervise undergraduate pharmacy students.

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Inappropriate use of medicines is widespread in teaching hospitals. The study aims to assess antibiotic use pattern among hospitalised patients in University of Port Harcourt Teaching Hospital (UPTH), Nigeria. A cross-sectional survey was conducted in medical, surgical, paediatrics, and obstetrics and gynaecology wards from October 2015 to March 2016 using seventeen (17) core antimicrobial drug use indicators developed under the Rational Pharmaceutical Management Plus (RPM-Plus) Program of Management Sciences for Health and revised under the Strengthening Pharmaceutical Systems (SPS) Program. A total of 2356 patients’ folders were reviewed retrospectively and data obtained statistically analysed. Additionally, sixteen inpatient physicians were interviewed to understand the reason behind the antibiotic prescribing practice observed. Antibiotics were prescribed in 68.8 percent of hospitalizations. Only 84.2 percent appeared on the essential drug list, 60.4 percent were injections and 39.9 percent were prescribed in generics. Each patient was prescribed 2.74 antibiotics at ₦13,632.00 cost averagely per hospitalization. Cephalosporin antibiotics were most commonly prescribed. The percentage of prescribed antibiotics actually administered was 87.4 percent, while 88.5% were empirically prescribed. About 35.0 percent of antibiotics were prescribed for Respiratory Tract Infections. Only 62.2 percent of key antibiotics were available and there was no systems regulating antibiotics use at UPTH. Physicians reported certain factors influenced the antibiotic use pattern observed. Antibiotics use in UPTH did not conform to acceptable standards. Establishment of local protocols and systems regulating antibiotics use including an antibiotic stewardship programme could improve rational antibiotic use; curtail cost and result in substantial savings.

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Zajeba Tabashsum is a second year Graduate student at the Department of Animal and Avian Sciences in University of Maryland- College Park. Her research area is focused on natural antimicrobials and their implications. She has already published 4 papers in reputed journals and submitted one book chapter. Recently one of her abstract has got accepted for oral presentation and has presented posters in different national/international coferences. 

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With growing trend of back yard, mixed crop-livestock or pasture/organic farming system, cross-contamination of produce specifically leafy-vegetables with enteric pathogens including Salmonella, Listeria and Escherichia coli is a major concern. Again produce-borne infections in the US are rising faster than ever causing thousands of hospitalizations and deaths yearly. It is also huge economic burden and major reason to recall produce products. So post-harvest processing is necessary to decrease contamination by enteric pathogens for safer products. As organic-farmers cannot use synthetic chemicals/antibiotic, they are desperately needed a natural-alternative to eliminate contaminants from produces.  As an alternative approach, we verified the efficacy of berry pomace extract (BPE) which is high in phenolic compounds including flavan, flavanone, glucuronides, glucosides, quinolones, catechol, tannins, quercetin, chlorogenic acid, gallic acid, and xanthoxic acid as sanitizer in controlling enteric pathogen Salmonella from fresh produces. For this purpose spinach and celery were decontaminated for any natural contamination and spiked with Salmonella enterica serovar Typhimurium   (ATCC® 14028™) and dipped into BPE or water for same time period and reduction in bacterial load and alteration in virulent-gene expression level was determined. There was significant reduction in bacterial number from 0.5-3.5 log with BPE treatments when compared to dipping into water also reduced bacterial number but not as efficiently. The expression of virulent-genes were also altered significantly by BPE treatments. Our findings indicated that BPE can be a potential natural alternative to replace synthetic chemical sanitizer to eliminate/reduce major enteric bacterial pathogens particularly Salmonella in produces and limit salmonellosis in human.

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Nataliya Roth has completed her MSc in Academy of Veterinary Medicine in Lviv, Ukraine and MSc at the University of Natural Resources and Life Sciences in Vienna, Austria. Last 10 years she was working in R&D of feed additives producer Biomin Holding GmbH. Last four years she was working on her PhD about antibiotic use, resistance and strategies to overcome resistance on University of Natural Resources and Life Sciences in Vienna.

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The application of antibiotics for the treatment of disease, disease prevention and growth promotion in food-producing animals provides favorable conditions for the selection, persistence and spread of antibiotic-resistant bacteria and their resistance determinants at the farm level. Increasing antibiotic resistance is a major public health concern. Fluoroquinolones are used to treat and prevent poultry diseases worldwide. Fluoroquinolone resistance rates are high in their countries of use. The aim of this study was to evaluate the effect of an acid-based feed additive, as well as fluoroquinolone antibiotics, on the prevalence of antibiotic-resistant E. coli. To study the emergence of antibiotic resistance in Gram-negative bacteria, E. coli are widely accepted as indicator bacteria. A total of 480 broiler chickens (Ross 308) were randomly assigned to three treatments: a control group receiving a basal diet; a group receiving a feed additive (FA) based on formic acid, acetic acid and propionic acid; and an antibiotic enrofloxacin (AB) group given the same diet, but supplemented with enrofloxacin in water. A pooled fecal sample of one-day-old chicks was collected upon arrival at the experimental farm. On day 17 and day 38 of the trial, cecal samples from each of the eight pens were taken, and the count of E. coli and antibiotic-resistant E.coli was determined.

The results of the study show a high prevalence of antibiotic-resistant E. coli in one-day-old chicks. Supplementation of the diet with FA and treatment of broilers with AB did not have a significant influence on the total number of E. coli in the cecal content on day 17 and day 38 of the trial. Supplementation with FA contributed to better growth performance and to a significant decrease (P ≤ 0.05) in E. coli resistant to ampicillin and tetracycline compared to the control and AB groups, as well as to a decrease (P ≤ 0.05) in sulfamethoxazole and ciprofloxacin-resistant E. coli compared to the AB group. Treatment with AB increased (P ≤ 0.05) the average daily weight compared to the control group and increased (P ≤ 0.05) the number of E. coli resistant to ciprofloxacin, streptomycin, sulfamethoxazole and tetracycline; it also decreased (P ≤ 0.05) the number of E. coli resistant to cefotaxime and extended spectrum beta-lactamase-(ESBL)-producing E. coli in the ceca of broilers. 

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Dr. Bernard C. Silvala is the current Department Head of the Medical Laboratory Technology Program at the University of Tabuk. He is an  Assistant Professor in Microbiology. He has published numerous works in Microbiological assays, educational manuals and scientific articles. Also a member of editorial board reviewer of reputable journals.

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Panton-valentine leukocidin (PV-luk) toxin is a very potent toxin responsible for severe and fatal form of Staphylococcal infections. This study aimed to determine the frequency of PV-luk producing methicillin resistant S. aureus (MRSA) and compare the conventional against real time PCR for identification of MRSA. A total number of 47 MRSA isolates were enrolled in the study. All are identified and typed using VITEK system. DNA was extracted and subject to conventional and real time PCR (RT-PCR) for the detection of mecA, luk-PV, orfX, mecA/mecC SCC/orfX junction genes. Forty-five isolates (95.7%) confirmed as MRSA and 2 isolated showed a mixed infection of Methicillin sensitive S. aureus (MSSA) and Methicillin resistant coagulase negative Staphylococci (MRCoNS).  Fifteen out of forty-seven (31.9%) have Panton – Valentine Leukocidin gene (luk-PV) gene.  PV-luk was detected in one third of tested MRSA emerging the possibility to have a potent superbug. In addition to, diagnosis of MRSA is highly accurate when using multiplex real time PCR rather than conventional PCR.

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Richard Mbi Beteck has completed his PhD at the age of 28 years from North-West University, Potchefstroom campus and is currently a postdoctoral fellow at Rhodes University.

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Trichomoniasis is a neglected venereal infection affecting almost 43 million people in Africa and almost 4 million people in the United states of America. It is a very common disease with high prevalence of up to 40 % in some regions in Africa. It is caused by Trichomonas vaginalis, a parasitic protozoan which is transmitted from person to persons during sex. It has been established that infection with T. Vaginalis increases the chances of acquiring and transmitting HIV. It also amplifies the progression of other sexually transmitted diseases. Treatment of Trichomoniasis rely solely on metronidazole and tinidazole, with metronidazole being the mainstay therapy. These agents are effective and safe at low doses required to treat susceptible strains of the parasite. However, serious side effects arose when large doses are used to manage metronidazole tolerant and resistant strains of T. Vaginalis.  With metronidazole tolerant and resistant strains of T. Vaginalis well documented, it is imperative to develop new trichomonacidal agents. Methodology & Theoretical Orientation: Our approach is to develop novel agents that can potentially probe new targets in the parasite and can still be reduced in the parasite hydrogenosome to generate reactive radicals (as does metronidazole). To this effect, we conceptualised and synthesise target compounds containing a metronidazole (MTZ) unit flagged by Schiff bases of varied lipophilicities and hydrogen bonding properties. Findings: Target compounds were screened in vitro against drug susceptible strains of the parasite for preliminary investigation and were established to be potent, with almost 80 % of target compounds exhibiting 100 % parasite inhibition at 10 µM. Conclusion & Significance: There is currently no treatment for metronidazole resistant trichomoniasis. Our approach has delivered very potent agents having the potential to probe additional targets to that of MTZ. They are promising templates to overcome the issue of resistance.
 

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Mengfei is at his last stage of PhD study in the University of Maryland, and he will finish his PhD in May 2018. His research area is focusing on applying modern techniques of microbiology in improving human gut microbiome as well as preventing foodborne enteric diseases. So far he has published 3 book chapters and 10 papers in reputed journals, and he has presented 3 oral talks and more than 10 posters in top-level conferences.

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Microbial horizontal gene transfer is a continuous process and it shapes especially bacterial genomes. However, the niches of microbial ecology as well as synthetic antibiotics or natural antimicrobials in the influence on the evolution of this biological networks are not fully understood yet. So far, very little attention has been paid on how dosages of antibiotics utilized in farm animal production contributes to the overall problem of antibiotic resistance. In this study, we investigated the mechanisms of sub-therapeutic and therapeutic dosages of synthetic antibiotics that foster the exchange of genetic materials in the microbial community in various components of farm animal production system using marked conventional and organic farm isolated Salmonella enterica serovar Typhimurium strains. The relatedness among the S. Typhimurium isolates from this study was investigated based on genome-wide SNP comparisons and the draft genome contigs have been submitted to NCBI. The antibiotic resistant S. Typhimurium isolates from conventional farm cultured in both conventional and organic soils failed to lose their tetracycline resistance, whereas highly sensitive S. Typhimurium strains isolated from organic farm gradually acquired higher tetracycline MICs over time under both types of soils. None of the isolated S. Typhimurium strains developed or lost sulfamethoxazole/trimethoprim resistance in soil environment. The qRT-PCR analysis indicated that differential expression of genes involved in antibiotic resistance in S. Typhimurium isolates was artificially inoculated under antibiotic pressure. The findings suggest some basic patterns in mechanism of S. Typhimurium antibiotic-resistance development in farm-related microbial ecosystems.

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       Dr Rahul Lokhande  , MD in Pulmonary Medicine. Working as Associate Professor in Chest department of BJ Govt Medical college Pune  India. Member of Drug Resistant Tuberculosis Board of Aundh Chest Hospital . He is one of Fogarty Scholar with collaboration with John Hopkins University , Baltimore  USA. Working on  many studies and trial  conducted in BJ government College  along with CTU unit , in field of HIV , Tuberculosis and diabeties , upcoming trials on vaccines for tuberculosis .   Presently working on “Factors for Unfavorable Outcome of Drug Resistant Tuberculosis  “.   

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              Antibiotic resistance is an important concern for the public health authorities at global level. Community based researches in developing countries have shown increase in burden of antimicrobial resistance.  In 1993 World Health Organization (WHO) declared Tuberculosis as a global emergency and promoted directly observed treatment short course (DOTS) in 1995. Millions of lives have been saved compare with the pre DOTS era, high cure rates have been achieved in most of countries worldwide. Global incidence of tuberculosis has been in a slow decline since the early 2000’s. However, the emergence and spread of Multidrug resistant (MDR) tuberculosis, extensively drug resistant ( XDR ) tuberculosis pose a threat to global tuberculosis control .

  Genotypic  and Phenotypic changes in Mycobacterium Tuberculosis ,with lack of adequate laboratory facilities in most of tuberculosis endemic countries leads to missing the diagnosis. Multidrug resistant tuberculosis and XDR tuberculosis greatly complicate patient management within resource poor national tuberculosis program , reducing  treatment  efficacy  and increasing  the cost of treatment .

         Despite nearly 20 years  of WHO promoting activity and >12 years of MDR tuberculosis specific activity , has the country response  to the drug resistant tuberculosis  epidemic been so in effectual ? The current dilemmas, challenges and priority needs for global drug resistance screening and surveillance, improving treatment regimens, and management of outcomes and prevention of drug resistance will be discussed

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Samir Jaoua, PhD from the University of Technology of Compiegne (UTC, France), is currently Professor at the Department of Biological and Environmental Sciences (CAS, QU). He is also Qualified Professor of France Universities “Section 64: Molecular Biology and Biochemistry” and Professor at the University of Sfax (Tunisia) and having been previously Director of Laboratory at the Center of Biotechnology of Sfax (University of Sfax- Tunisia). He is molecular and microbial geneticist, having more than 30 years of experience in the fields of molecular and microbial Biotechnology in different countries: France, Switzerland, Belgium, Germany, Tunisia and Qatar. He is the author of 118 publications with impact factors; H-index: 30 and served as member, Chair, and General Chair of a number of conferences. 

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Bacillus thuringiensis strain BUPM4 is known for its ability to produce a bacteriocin, called Bacthuricin F4 (BF4), which inhibits the growth of several Gram-positive bacteria and particularly Bacillaceae. This study aimed to use the insertional transposon mutagenesis approach for disrupting and thus identifying genes associated with BF4 synthesis. Here, the mini-Tn10 transposon was used to generate a library of B. thuringiensis mutants. Twenty thousand clones were screened for the search of mutants with affected bacteriocin synthesis. Bymolecular hybridization, it was demonstrated that the mini-Tn10 transposition occurred in different sites. Clone MB1, containing a mini-Tn10 single-copy insertion, lost the BF4 synthesis, but maintained its immunity to BF4. The flanking sequences surrounding the mini-Tn10 insertion were cloned and sequenced. The bacteriocinogenic plasmid pIBF4 from Bacillus thuringiensis responsible of Bacthuricin F4 synthesis was isolated and characterized. It has a molecular weight of 19.1 kb. Ninety-five percent of cells retained the pIBF4 plasmid after 200 generations, demonstrating its high stability. pIBF4 was successfully transferred to Bacillus thuringiensis HD1CryB strain with a transfer frequency of 10^-8 transconjugants per donor cell. The study of the recipient host range revealed that pIBF4 is specifically transferable to Bacillus thuringiensis strains with variable transfer frequencies depending on the recipient host strain. 

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Richard Mbi Beteck has completed his PhD from North-West University, Potchefstroom Campus and is currently a Postdoctoral Fellow at Rhodes University.

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Yeon-Jung Lee is a graduate student in School of Food Science and Biotechnology, Kyungpook National University in Daegu Korea.

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Ji-Sun Moon is a graduate student in School of Food Science and Biotechnology, Kyungpook National University in Daegu Korea..

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