3^rd World Congress and Exhibition on Antibiotics and Antibiotic Resistance July 30-31, 2017 Milan, Italy

Biography:

John James Stewart Cadwell received his degree in Pharmacology from the University of Miami in 1981. He spent additional time studying at the University of Nottingham and the National Institute of Medical Research at Mill Hill, outside of London. He served in various capacities in the Biomedical Research fields until 2000 when he became Founder of FiberCell Systems Inc. a company specializing in the research and supply of hollow fiber bioreactors. He has over 10 publications in the field and three patents relating to hollow fiber systems and is considered a world expert in the field

Abstract:

The recent introduction of the hollow fiber infection model represents an important advancement in the field of in vitro toxicology. Many toxicologists believe in vitro testing methods are a useful, time and cost effective tool for drug discovery but it is generally accepted that many of the available tests are not effective for examining the effects of both time and concentration, and do not closely mimic human kinetics. This is because they do not properly take into account pharmacodynamic actions (what a drug does to the body) and pharmacokinetic actions (what a body does to the drug). With the use of hollow fiber bioreactor cartridge this has now changed. The hollow fiber infection model is a useful addition to standard in vitro toxicology methods as it mimics changes in drug concentration over time, as they would occur in humans. The hollow fiber infection can mimic precisely human bioavailability, study meaningful numbers of organisms to reveal emerging resistance, mimic both adsorption and elimination curves, define total kill in a closed, biosafe system. The clinical utility of the HFIM has been demonstrated and is now endorsed by the EMA. An overview of historic pk/pd models is presented and the utility of the system as it relates to antibiotics and other drugs are discussed.

Biography:

Marie-Agnes Sari is Professor of Biochemistry at Paris Descartes University and Head of the Laboratory of Chemistry and Biochemistry, Pharmacological and Toxicological research (UMR 8601 CNRS) and research interests are linked to metalloenzymes, or enzymes and peptides involved in metal transport, especially iron

Abstract:

Urinary tract infections (UTI), affecting 150 million cases per year worldwide, are the main cause of medical antibiotic prescription. However, uropathogen E. coli (UPEC) responsible for UTI are increasingly resistant to antibiotics, indicating urgent need for new therapeutic alternatives. The goal of our project is to explore hepcidin as a new antimicrobial peptide. Hepcidin is a cysteine rich, β defensin like peptide hormone that acts as the key regulator of body iron metabolism. Hepcidin regulates iron homeostasis in human through the binding of the sole iron exporter protein, ferroportin inducing its internalization prior degradation by lysosomes. Besides the liver that is the major source of bloodstream hepcidin, this peptide is expressed in epithelial barriers that are frequently confronted to pathogen infection, including kidneys. However, hepcidin belongs to defensins family and was observed to have an in vitro antimicrobial activity. We hypothesized that hepcidin was involved in host resistance to UPEC in renal epithelia where iron limit may be efficiently regulated. Hepcidin role against urinary tract infection was investigated using HepC-/-mice and synthetic hepcidin. Several recombinant hepcidins were produced as biologically active, ferroportin-binding peptide in E. coli, or yeast and synthetic hepcidin analogs were prepared and compared for their antibacterial activity on E. coli, B. subtilis and S. aureus. Bacteriostatic activity of hepcidin was demonstrated in vitro on CFT073 (a prototypical pyelonephritis-associated E coli strain). Evidence will be presented demonstrate that hepcidin is an effective mediator against UPEC infection and acts to prevent pathogenesis in the kidney in mice.

Figure 1: Structure of the mouse hepcidine analog and evaluation of its antimicrobial activity.

Biography:

Yasuhiro Igarashi studied at the University of Tokyo and received his PhD degree in Organic Chemistry in 1992. After two years of research activity at Fujifilm Corporation, he conducted Postdoctoral Research at Johns Hopkins University for two years. In 1996, he joined the faculty at Toyama Prefectural University, Japan, as Assistant Professor. In 1998, he was appointed Associate Professor and, in 2009, was appointed Professor. He is the author of up to 200 papers and has been the Editorial Board Member of The Journal of Antibiotics since 2011. For the last 20 years, he has focused his efforts on discovering novel bioactive compounds from natural resources.

Abstract:

Free radicals including reactive oxygen species (ROS) show a wide range of antimicrobial activity through oxidative damage to cellular components such as proteins and DNA. Some inorganic and organic compounds are known to generate ROS. Titanium oxide (TiO₂) catalyzes the formation of superoxide and hydroxy radical under the light. Some porphyrins have ability to generate singlet oxygen that kills viruses. Low specificity of ROS toward biomolecules seems uncontrollable, however it becomes an advantage that can avoid and overcome antibiotic resistance. ROS randomly react with multiple biomolecules, which does not allow microorganisms to acquire resistance. Fossil seashell (FS) is a kind of minerals mined in our district, Toyama, Japan. It has been used as a fertilizer or water-cleaner by local farmers. Pathogen protection effects in crops are known for many years but scientific investigations have not been made to date. In this study, we investigated the antimicrobial activity of FS powder. Its water suspension liquid displayed growth inhibition against representative human pathogens Escherichia coli, Staphylococcus aureus, Candida albicans and Trichophyton rubrum. As an active principle, we detected superoxide in the FS suspension. The major component of FS is calcium carbonate (CaCO₃) which is changed to calcium oxide (CaO) by burning to expel carbon dioxide. As this burning process is essential to activity, CaO is likely responsible for superoxide generation. The detailed mechanism is under investigation. Considering the mode of action, there may be very low possibility of occurrence of resistance to superoxide. Broad antimicrobial spectrum of FS against bacteria and fungi is attractive as a sterilizer. FS is stable for years and only water is needed to prepare FS powder suspension. It can be used for sterilization of food and skin to prevent oral infections or food poisoning. Effectiveness of FS in the treatment of skin or oral infectious diseases shall be investigated to explore its potential in medical usage.

Biography:

Nahla Omer Eltai has completed her PhD from Humboldt University, Berlin, Germany. She did Postdoctoral studies from University of the West of England, UK. She is Research Associate at Biomedical Research Centre, Qatar University. She has published many papers in the field of Antibiotic Resistance. Her research interests are multidisciplinary with emphasis on molecular diagnostic approaches, antimicrobial susceptibility & resistance, test of new natural antimicrobial agents. She is adopting the one health system approach by studying antimicrobial resistance in agriculture, environment and human in Qatar

Abstract:

Antimicrobial resistance (AMR) is a growing public health concern worldwide and is one of the top health challenges facing humanity in the 21^st century. AMR among Enterobacteriaceae is rapidly increasing especially to third-generation cephalosporins and carbapenems. Further, strains carrying mobilized colistin resistance (mcr) genes 1 and 2 have been isolated from humans, food-producing animals, and environment. Uncontrolled use of antibiotics in animals in large scale could be one of the major contributing factors to generation and spread of antibiotic resistance. No studies have been done to evaluate antimicrobial resistance in animals in Qatar. This study aimed at establishing a primary baseline data for prevalence of antimicrobial resistance among food animals in Qatar. 172 fecal samples were obtained from two broiler farms and one live bird market in Qatar and 90 Escherichia coli (E. coli) bacteria were isolated and subjected to antimicrobial susceptibility testing using Etest method. 90% (81/90) of the isolates were resistant to at least one of the 16 tested antibiotics. 15.5% (14/90) of the isolates were colistin resistant, 2.2% (2/90) were extended spectrum β lactamase (ESBL) producers and similar percentages were multi-drug resistant (MDR) to four antibiotic classes. ESBL-producing E. coli and colistin resistant isolates were confirmed using double disc susceptibility testing and PCR, respectively. In Summary, Our results indicate high antimicrobial resistance in food producing animals in Qatar, including ESBL and colistin resistance. Such AMR bacteria could be easily transmitted to humans through consumption of undercooked food or noncompliance with hygiene practices, which mandates prompt development and implementation of stewardship program to control and monitor the use of antimicrobial agents in community and agriculture

Biography:

Kang M S has earned his BS to PhD degrees from Department of Biology at Seoul National University, Korea. After his PhD, he did his Postdoctoral Research Fellowship at Department of Microbiology and Molecular Genetics, Harvard Medical School/Brigham and Women’s Hospital. He then joined as a Professor and Faculty at Samsung Medical Center, Center for Future Medicine. Since 2016, he has been a Chief Scientist and Vice Director of Institute of iNtRON Biotechnology. He is interested in development of bacteriophages and phage –derived bio drugs for effectively controlling bacterial infections

Abstract:

Statement of the Problem: Emergence and dispersal of multidrug-resistant Staphylococcus aureus (SA) (MRSA) are rapid and wide, necessitating an invention of novel class of MRSA-controlling effective antimicrobial drug that is not likely to induce a resistance. This presentation describes our 15 year-long experience in development of a bacteriophage-derived endolysin as promising new antibiotics.

Materials & Method: A bacteriophage SAP-1 specifically infecting SA was isolated from sewage water. Whole genome sequencing revealed typical genomic structure of lytic bacteriophage and identified putative peptidoglycan (PG) hydrolase gene encoding SAL-1 endolysin. The wild type SAL-1 with no extraneous amino acids was highly purified in compliance to a good manufacturing practice (GMP) standard and further formulated to N-Rephasin^® SAL200.

Findings: SAL200 showed to have unique modes of action: It potently and specifically killed planktonic and encapsulated SA as well as biofilm-producing SA cells, but not for other species. SAL200 exhibited strong bactericidal activity against more than 425 clinical isolates including 1 vancomycin-intermediate SA and 336 methicillin-resistant SA isolates. It directly bound to and lysed SA as quickly as within ten minutes and apparently did not induce an emergence of any resistant SA strains from 30-times repeated exposure at sub-lethal concentrations. Intravenous injection of SAL200 significantly prolonged survival of mice and reduced viable bacterial counts in MRSA infection mouse model. Further safety evaluation studies for rodents, dogs and monkeys showed neither death nor severe adverse events. Any abnormal findings-if any- were tolerable and transient.

Conclusion: The SAL200 specifically lysed SA strains including antibiotic-resistant SA strains in vitro and in vivo with excellent in vivo safety.

Biography:

Abstract:

Background: Bacterial resistance to the antibiotics is considered as natural phenomenon that occurs naturally over the time due to genetic changes. However, various factors lead to the acceleration of this process and resulting in antibiotics resistance. These factors include the overuse and the misuse of antibiotics, for example, taking an antibiotic for a viral infection like influenza (World Health Organization, 2016). This may be attributed to consumer’s lack of knowledge about the correct use of antibiotics, or it may be due to their wrong habit about using the antimicrobials. A key among the causes of the increased antimicrobial resistance is self-medication  and drugs on one’s own initiative without consulting a qualified medical practitioner (Bennadi, 2014; p.19; WHO 2001, p.21 ). Bacterial resistance to antibiotics is increasing significantly in the UAE (Al-Dhaheri et al., 2009).Antibiotics use without prescriptions has been reported to be high among university‘s students  worldwide (Javed, 2013; Awad andEltayeb, 2007; Sawalha, 2008; Sarahroodi et al., 2010; Sharif and Sharif, 2013; Ehigaiator et al., 2013; Badiger et al., 2012 and  Kumar et al., 2013). There was no extensive studies that explore the reasons for misusing antibiotics among university students worldwide.  Therefore, it was necessary for this study to investigate this type of irrational use of drugs in UAE.

Objective: The purpose of this research is to measure the prevalence of irrational use of Antibiotics by university students in UAE and identifies the reasons for this irrational drug use using mixed methodological approach.

Methods: A mixed methods research design with a sequential explanatory strategy was adopted to conduct this research (Creswell 2011; Cameron, 2009).   The first quantitative cross-sectional analytical study was conducted from January to April 2014 with 2,875 students at three randomly selected UAE universities. The aim was to measure the prevalence and risk factors for misusing antibiotics among university students. A structured and validated questionnaire was used to collect the responses of the students. SPSS version 20 was used to analyze the data. A second interview study investigated first-year healthcare students at high risk of using antibiotics without prescription, with a purposefully selected sample. The aim was to further explore common themes related to their knowledge, awareness, attitude, views, and perceptions as well as possible strategies to decrease their misuse of antibiotics. Data were analysed thematically for the identification of themes and subthemes within the data through the use of coding

Findings: More than one third of users (484 of 1384, 36%) were reported using antibiotics without prescription during the 90 days prior to the present study. Of those medical students (n=492), 36.8% reported using antibiotics. Also, of those non-medical students (n=856), 36% of them were using the antibiotics, but the difference was statistically not significant (p-value= 0.608). Nine risk factors were identified for using antibiotics.  Being Expatriates (OR = 0.471, 95% CI: 0.326-0.681, p<0.001). Participants who were influenced by the cost of Oral Non-Prescription Drugs (ONPD) (OR = 1.716, 95% CI: 1.175-2.508, p<0.005). Participant believing that ONPD are moderately effective (OR = 0.332, 95% CI:0 .135-0.815, p<0.02) or ineffective (OR =0 .400, 95% CI: 0.161-0.994, p<0.05) had significantly lower odds of using antibiotics in SMP compared to those who believed that ONPD are effective.  Participants with good medication knowledge (OR = 0.619, 95% CI: 0.443-0.866, p < 0.005). Also, participants in their fourth year of study (OR = 0.310, 95%, CI: 0.141-0.681, p < 0.004), fifth year of study (OR = 0.243, 95%, CI: 0.088-0.666, p < 0.01), or sixth year of study (OR = 0.101, 95%, CI: 0.015-0.678, p < 0.02)   had lower odds of using antibiotics in SMP compared to participants in their first year of study. Being a medical student was a risk factor for using antibiotics in SMP (OR = 1.465, 95%, CI: 1.012-2.120, p < 0.05). Overall the second interview study identified four main themes with multiple subthemes related to the use of antibiotics without a physician’s prescription by first-year healthcare students. The thematic analysis of the interviews revealed four main themes:

• Medication habits and practices
• Reasons for self-medication
• Access to antibiotics without a prescription
• Gaps in students’ knowledge regarding antibiotic resistance

Conclusion: The prevalence of using antibiotics is high among university students in UAE. There is a need for an educational intervention to motivate students to be use antibiotics wisely.

Biography:

Tula Krishna Gupta has completed his MD in General Practice and Emergency Medicine from B P Koirala Institute of Health Sciences, Dharan in 2016 September. He is a Senior Resident in General Practice and Emergency Medicine Department of B P Koirala Institute of Health Sciences, Dharan, Nepal.

Abstract:

Introduction: Recently, UTI became more complicated and difficult to treat because of appearance of pathogen with increasing resistance to antimicrobial agents. It is useful to obtain the local sensitivity pattern in the hospital setting so as to guide empirical prescribing.

Methods: This is a prospective hospital based study to identify the organisms causing UTI and their antibiotic susceptibility. A total of 400 urine samples were collected from the patient attending general OPD of BPKIHS, Dharan, Nepal. Consecutive patients presenting with symptoms of UTI had their clean catch midstream urine analyzed. Quantitative urine culture was performed by using calibrated loop direct streaking method of 4 mm diameter nichrome loop delivering 0.001 ml of uncentrifused urine specimen which was inoculated onto cysteine lactose deoxycolate agar medium and incubated at 37 degree centigrade for 24 hours.

Results: Out of 400 patients, 163 (40.7%) showed bacterial growth. Escherichia coli was most common (68.7%), followed by Klebsiella spp. (7.4%). There was a female dominance of 2.79:1 compared to males. Infections were most common in young adults (20-29 years). The most effective antibiotic was Nitrofurantoin (89.7%).

Conclusion: The most common causative organism for UTI was Escherichia coli, and the best first line antibiotic was Nitrofurantoin. This study provides useful information for health practitioners of all levels in the eastern region of Nepal. As the data was collected in the general outpatient department it represents mostly community acquired, rather than hospital acquired urinary infection and may be used as an aid to choice of antibiotic in the community setting.

Biography:

Tie T K has completed his BPharm in 2007 from Curtin University of Technology in Western Australia. He has a Postgraduate Master‘s Degree in Infectious Diseases from the University of Western Australia in 2014. He is currently practicing as an Antimicrobial Stewardship Pharmacist at Changi General Hospital. His research interest is in infectious diseases especially related to Clostridium difficile.

Abstract:

Background: Ertapenem use has been associated with central nervous system (CNS) related adverse effects (AEs) which include delirium, hallucinations, altered mental status (AMS), stroke like symptoms and seizures. We described baseline characteristics and clinical features in 4 patients who developed CNS related AEs while receiving ertapenem during a period of six consecutive months in 2016.

Methods: We retrospectively reviewed patients on ertapenem and identified the ones who developed CNS related AEs attributed to ertapenem use from April till October 2016.

Results: Patient 1 (no renal impairment) was prescribed ertapenem for complicated intra-abdominal infection. He was noted to be drowsy on day 12 of ertapenem. Over the next 1-2 days he developed auditory hallucinations, jerky limb movements and fluctuating level of consciousness. He was switched to aztreonam. Patient 2 (no renal impairment) has developed confusion and hallucinations (visual, auditory and tactile) on day 25 of ertapenem which was being administered for tubo-ovarian abscess. Ertapenem was substituted to meropenem. Patient 3 (known end stage renal failure) developed seizure while on day 28 of ertapenem which was prescribed for bilateral renal abscess. No further episode of seizure was observed after ertapenem was switched to piperacillin/tazobactam. Patient 4 (known chronic kidney disease) was receiving ertapenem for complicated urinary tract infection. He was noted to be delirious on day 9 of ertapenem. Patient was switched to meropenem. In these patients, average age was 65 years. Male to female ratio was 1:1. None of the patients had previous history of seizure or CNS disorders. Two patients had normal renal functions. Toxic and metabolic causes were excluded. Computed tomography (CT) of brain was normal in all cases. Dose was renally adjusted in 2 patients. Ertapenem therapeutic drug monitoring (TDM) was not locally available. Symptom improvement was noted 3-4 days after ertapenem discontinuation in all patients.

Conclusions: Patients who developed these AEs can deteriorate rather unexpectedly which leads to extensive investigations and increase in health care cost. The cost can be minimized if physicians are acquainted to consider ertapenem as an offender agent. However appropriate workup to eliminate alternate etiology also needs to be considered at the same time as dictated by the clinical scenario. Ertapenem TDM might give information on CNS AEs in future studies.

Biography:

Andrea Barbarossa has his expertise in many aspects of Veterinary Pharmacology. These include pharmacokinetics/pharmacodynamics and residues studies, but also antimicrobial resistance. In this context, he took part in a project of the University of Copenhagen focused on identifying synergies between antimicrobial and non-antibiotic drugs commonly used in veterinary medicine as a possible strategy to restore antimicrobial susceptibility in MRSP. In parallel, he took part in a 3 years project promoted by the Emilia Romagna Regional Veterinary Service aimed at understanding how antimicrobials are used in veterinary medicine and at developing specific guidelines for their correct use in pets, swine and bovine. It has already been demonstrated how these tools can be helpful at containing antimicrobial resistance

Abstract:

Statement of the Problem: Antimicrobial resistance (AMR) is considered a major public health issue and there is an increase in its prevalence in both humans and animals. Lately, many studies focused on the role of pets in AMR diffusion, highlighting the lack of data on prescribing patterns and the need for guidelines for the correct use of antimicrobials in these species. Studies in literature demonstrated that the adoption of guidelines helped improving prescribing patterns and reducing antimicrobial use. The purpose of this study was to gather data on antimicrobial prescribing patterns for pets in Emilia-Romagna Region and to use these results while drawing up guidelines for small animal practitioners.

Methodology & Theoretical Orientation: In the context of a regional project on antimicrobial resistance, small animal practitioners were invited by the Veterinary Professional Association of Emilia Romagna to voluntarily complete an online questionnaire, aimed to understand their prescribing habits and how they apply biosecurity measures. A group of experts then analyzed the collected data and, based on recent regulations, scientific data and personal experience, issued practical guidelines. These include specific chapters, tackling the different aspects that could be of interest for the everyday practice of small animal veterinarians.

Findings: The most commonly prescribed antimicrobials resulted penicillins, fluoroquinolones and cephalosporins; critically important antibiotics were employed mainly to treat urinary tract infections. A general awareness of the AMR issue and of the importance of applying biosecurity measures were encouraging outcomes.

Conclusions & Significance: This overview of the prescribing patterns of antibiotics represented a useful tool for issuing guidelines on the correct use of these drugs in small animal practice. Guidelines will be soon available for all veterinarians, hoping that the provided practical information will be a useful for containing antimicrobial resistance.

Biography:

Georg Daeschlein received the MD degree from Christian-Albrechts-Universität Kiel, Kiel, Germany, in 1985 and specializations in Laboratory Medicine, in Microbiology and Epidemiology, in Hygiene and Environmental Medicine and in Dermatology from University Medicine Greifswald, Germany, in 1993, 1995, 2005 and 2014, respectively. In 2011, he acquired his Habilitation in Hygiene and Environmental Medicine. Since 2006, he has been the Registrar and Head of the Dermatologic Laboratory in the Department of Dermatology of the University Medicine Greifswald and conducts the research group plasma-medicine and antimicrobial procedures.

Abstract:

Background: Cold atmospheric plasma (CAP) could be considered as an alternative against multi-drug resistant (MDR) pathogens. Susceptibility to CAP of clinical isolates of enterococci depending on antibiotic resistance was investigated in pure culture regarding the most important resistance patterns.

Patients & Methods: 6 groups of enterococci (39 isolates) were treated on agar using dielectric barrier discharge plasma (DBD). These included enterococci with combined vancomycin- and high-level gentamicin resistance, with high-level resistance to gentamicin (HLGR) only, with vancomycin resistance alone (VRE), and enterococci susceptible to both. Isolates showing high-level resistance to gentamicin were placed together in a separate group, as well as isolates with vancomycin resistance. Susceptibility was evaluated based on the zones of inhibition (ZOI) and examined in terms of the enterococcal group and the degree of drug resistance.

Results: CAP-treatment killed all groups. Significantly smaller ZOI were obtained with MDR-groups: VRE-isolates showed significantly smaller ZOI than non-VRE-isolates and HLGR-isolates showed smaller ZOI than non-HLGR-isolates. The mean of all isolates, irrespective of belonging to groups, showed smaller ZOI with increasing drug resistance.

Conclusions: CAP-treatment killed all isolates of enterococci, but its efficacy depended on the degree of drug resistance and on membership in special resistance groups with particular clinical-outbreak importance.

Figure 1: Mean (SD) ZOI (mm²) of all enterococci by respective resistance group (RG).

Biography:

Fadya Hamadani has acquired her BSc in Pharmacy and MSc in Clinical Pharmacy from College of Pharmacy, Baghdad University (1992, 1998 respectively). She is Assistant Professor in Clinical Pharmacy and since that period, she has built a variety of experience in research, evaluation, teaching in higher education institutions and providing hospital training. Currently she is a full time PhD student at the School of Pharmacy and Pharmaceutical Sciences, Cardiff University. Her PhD project involves identification of issues around safe and effective administration and management of medicines in care homes. Her research interest includes the rational use of antibiotics in care homes.

Abstract:

Statement of the Problem: Residents of care homes are noted for the complexity of their care needs, multiple co-morbidities and the incidence of polypharmacy. This makes them particularly susceptible to medicines related harm. Antibiotic prescribing in care homes is particularly prevalent and their inappropriate administration risks the development of antimicrobial resistance. The aim of this present study was to identify the prevalence and types of prescribing and administration errors related to antibiotics in Care Homes.

Methodology & Theoretical Orientation: A retrospective study was conducted in 12 Care Homes in South Wales over an eight month period (Feb-Oct 2015). Anonymised real-time data was collected from an electronic medicines management system and the prevalence of antibiotic determined. The data was also analyzed for errors related to the prescribing and administration of antibiotics. The type and frequency of medication errors were recorded in excel and selected data was transferred into SPSS for further analysis.

Findings:  A total of 142 (43.16%) residents received at least one antibiotic over the study period. The most commonly prescribed antibiotic was trimethoprim (24.6%).  Some 9499 administrations were analyzed against pre-defined medication error categories with 2247 medication errors identified, 79.6% of residents were exposed to at least one error. The most frequent administration error type was duration error (44.5%) where the antibiotic was administered either for a period shorter or longer than the prescribed duration, followed by dose omissions errors (38.9%). The most common prescribing error type was similarly duration errors (74.3%).

Conclusion & Significance: This evaluation of the management of antibiotics in care homes in South Wales has revealed that the prevalence of antibiotic prescribing along with the incidence of administration and prescribing errors related to antibiotics is significant. This may result in antimicrobial resistance in a particularly vulnerable patient cohort.

Table.1: Prescribing and administration errors for antibiotics in care homes

Biography:

Al-Mehdi Edowik Y is interested in Bacterial Antibiotic Resistance. After earning his Bachelor’s degree in Biomedical Science from Bangor University, he went through a one-year training period in the Public Health Wales Microbiology Department at Ysbyty Gwynedd Hospital, Bangor, where he gained experience in routine microbiology. Subsequently, he conducted an MSc (Master of Science) degree in Molecular Biology and Genetics at Bangor University. Currently he is undertaking a PhD in Molecular Microbiology at Bangor University. The research synopsis of his research is epidemiological gene carriage of ESBL-producing bacteria in UTI patients in North Wales.

Abstract:

The antibiotic nitrofurantoin is indicated for the treatment of acute or recurrent lower urinary tract infections (UTI) which is caused by strains of Escherichia coli, Enterococci and Staphylococci, Citrobacter sp, Klebsiella sp or Enterobactersp. This study analyzed ESBL-producing bacteria isolated from 300 urine samples of UTI patients from three referral hospitals in North Wales, UK. Multiplex PCR amplification of β-lactamase genes of the blaCTX-M groups 1, 2, 9 and 8/25, followed by DNA sequencing revealed one new beta-lactamase gene which is closely related to CTXM‑14 at the protein levels. Recombinant expression of this new bla gene renders Escherichia coli highly resistant to nitrofurantoin. To characterize this new beta-lactamase, the enzyme was purified from the periplasmic space of E. coli or from the growth medium in a highly pure form, utilizing a C-terminal strep-tag. Isoelectric focusing and phostag electrophoresis revealed a high degree of phosphorylation and the presence of at least two distinct protein species. Since this new gene was isolated only from one of the three hospitals, it is quite possible that the new beta-lactamase is responsible for a local increase in nitrofurantoin resistance in UTI patients.

Figure 1: Induction of the novel CTX-M-14 related beta-lactamase renders cells resistant to Nitrofurantoin (E-Test Assay, + =induction of the recombinat enzyme; -= no induction.

Biography:

Álvarez-Martínez F J is a Molecular Biology PhD student at the Universidad Miguel Hernández of Elche. The main theme of his thesis is the identification of natural compounds with antimicrobial capacity to act as antibiotics or in conjunction with those already existing in the treatment of microbial diseases, especially those resistant to traditional drugs. To reach these objectives, computational approaches and in vitro studies using reference microbial strains are used in addition to clinical isolates collected at the General University Hospital of Alicante.

Abstract:

At present, the rise of multi-drug resistant microorganisms represents a serious threat to human health worldwide. The objective of the present work is the discovery of individual natural compounds or mixtures of them that have a remarkable antimicrobial capacity to be used as therapy against microorganisms resistant to antibiotics or as a complement to existing treatments. To carry out the tests of microbial susceptibility, we used ten natural agents with potential antimicrobial activity, including pure compounds and complex mixtures of them. The method chosen was disk-plate inhibition in Mueller Hinton (MH) medium, with positive results being those where the compounds produced an inhibition halo around the disc. The microbial species used in this study were Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Enterobacter spp, Escherichia coli, Serratia marcescens, Salmonella spp, Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Acinetobacter baumanii. All the strains were obtained from clinical isolates in the General University Hospital of Alicante. The most active formulations against microorganisms were two plant extracts and a pure compound, the major component of one of the previous extracts. The microbial species most affected by these natural agents were Staphylococcus aureus (between 62.1% and 72.4% of susceptible strains), Klebsiella pneumoniae (between 12.5% ​​and 25%), Enterobacter spp. 25%) and Stenotrophomonas maltophilia (between 37.5% and 62.5%). Three of the natural formulations tested exhibit a remarkable antimicrobial capacity against species of great clinical importance causing serious nosocomial infections. Given the results obtained, these naturally occurring formulations could be used in conjunction with known antibiotics to increase their efficacy and circumvent microbial resistance mechanisms. The next step is to characterize the effective concentrations of the natural formulations found in the present screening and possible synergy between them. The next tests will be performed using the p96 microdilution plate method.

Figure 1: Antimicrobial assay in p96 plate using ten different concentrations of natural agent against eight different isolates of Staphylococcus aureus.

Biography:

Elisa Rampacci received her degree in Veterinary Medicine from the University of Perugia. Currently, she is a PhD student in Health and Experimental Veterinary Sciences. She is making her experience in the field of in vitro and in vivo studies of antibiotic interactions, efficacy and toxicity of antimicrobial compounds. Particularly, her attention is focused on the treatment of zoonotic infectious diseases. Additionally, her interest is addressed towards pharmaceutical formulations potentially suited for not standard delivery routes associated with minor adverse effects.

Abstract:

Statement of the Problem: Rhodococcus equi is the main infectious agent of pneumonia in foals. This bacterium has recently gained attention as a zoonotic pathogen in immuno depressed people. Two-thirds of cases of R. equi infection in humans involved HIV-infected patients. However, 10%–15% of infections occur in immunocompetent hosts. In human medicine, rhodococcosis may be underdiagnosed because of frequent misidentification for Mycobacterium tuberculosis. Combination treatment with two or more drugs is recommended both in equine and human infection in order to contrast the antibiotic resistance, especially including antibiotics with intracellular activity. The purpose of this study is to investigate the potential synergistic effect against R. equi of different azithromycin/rifampicin combinations formulated as dry powders and to assess their possible cytotoxicity.

Methodology & Theoretical Orientation: A broth microdilution test was performed to determine the minimum inhibitory concentration against R. equi of spray-dried azithromycin and rifampicin prepared independently and binary combinations of azithromycin and rifampicin developed as dry powders according to the ratios 1:1, 2:1 and 1:2 using mini spray-drying technology. The fractional inhibitory concentration indexes (FICIs) were calculated to describe drug interactions. Additionally, the minimum bactericidal concentration (MBC) was established. The cytotoxicity on BEAS-2B cells of azithromycin and rifampicin individually and in combination was assessed performing the MTT colorimetric assay.

Findings: FICIs calculation has confirmed the favorable additive effect of each antimicrobial combo. Moreover, MBC determination demonstrated the more powerful killing activity of combinations than single compounds. The MTT assay provided encouraging data about the combination application. Indeed, reduction of cellular viability below 50% was not observed neither at the highest concentration tested (100 µg/mL).

Conclusion & Significance: The additive interaction between azithromycin and rifampicin and the more powerful bactericidal activity of spray-dried combinations compared to single compounds may result in an improved therapeutic dosage and prevention of drug-resistant bacterial strains.