2^nd World Congress and Exhibition on Antibiotics and Antibiotic Resistance October 13-15, 2016 Manchester, UK

Thanyaluck Siriyong is a PhD candidate in Microbiology from Prince of Songkla University, Thailand. She has completed her BSc and MSc in Thai Traditional Medicine from Prince of Songkla University. She is currently a Visiting Doctoral Researcher at the University of St Andrews, Scotland, UK. Her project is supported by the Thailand Research Fund through the Royal Golden Jubilee PhD Program co-funded by the British Council through Newton Fund

The Pseudomonas aeruginosa MexAB-OprM, MexXY-OprM, MexCD-OprJ and MexEF-OprN are well characterized efflux systems of the resistance-nodulation-cell division (RND) family and are able to export a wide variety of structurally diverse compounds, including many clinically administered antibiotics. The pumps are expressed upon exposure to antibiotics and mutations in local or global regulator genes can occur, leading to overexpression of these efflux pumps and to multidrug resistance (MDR) phenotypes. To overcome MDR, efflux pump inhibitors are a potential solution. Efflux pump inhibitors from natural products have been extensively explored as potential sources because of fewer adverse effects, low cost and traditional medicine. Many plant alkaloids have been previously reported as efflux pump inhibitors such as curcumin, reserpine, capsaicin and piperine. This study has determined the effects of steroidal alkaloids from Holarrhena antidysenterica and its major steroidal alkaloid conessine as efflux pump inhibitors against P. aeruginosa. Steroidal alkaloids and conessine combined with various antibiotics were investigated for synergistic activity against P. aeruginosa PAM1020 (WT), P. aeruginosa PAM1032 (nalB), P. aeruginosa PAM1033 (nfxB), P. aeruginosa PAM1034 (nfxC) and P. aeruginosa PAM1626 (∆mex). Conessine (32 mg/L) reduced MICs of piperacillin, meropenem and levofloxacin in MexEF-OprN overexpressed strain (P. aeruginosa PAM1034). However, synergistic activity obtained from steroidal alkaloids at 1024 mg/L demonstrated less effects than conessine. The results suggested that conessine could be applied as an efflux pump inhibitor to restore antibiotic activity by inhibiting efflux pump system in P. aeruginosa

Baodong Ling has completed his MD from West China Medical University and worked as a Visiting Scholar in University of Zurich in 1996. He is a distinguished Professor in Pharmacology. He is also the Director of Small Molecule Drugs Key Laboratory of Sichuan Province and Institute of Materia Medica. He has published more than 60 papers in reputed journals and has been serving as an Editorial Board Member of Chinese Journal of Antibiotics and World Notes on Antibiotics.

Nosocomial infections caused by quinolone-resistant Acinetobacter baumannii (QRAB) have been increasing in recent years, posing a threat to public health. In this study, we aimed to determine the mechanisms of quinolone resistance in A. baumannii isolates and investigate the occurrence of carbapenem resistance gene blaOXA-23 and aminoglycoside resistance gene armA among these QRAB. Totally, 101 A. baumannii isolates were collected from one university hospital in western China between 2011 and 2012. All isolates were identified and confirmed by sequencing analysis of 16S rRNA internal transcribed spacer and recA. The minimal inhibitory concentrations (MICs) of ciprofloxacin and levofloxacin were determined by agar dilution. Plasmid-mediated quinolone resistance determinants (PMQRs) were performed with PCR assay, whilst gyrA and parC genes of QRAB were amplified and sequenced. The results showed that 77 QRAB harbored mutations of gyrA and parC, whereas PMQRs were not detected. Additionally, 41 isolates had resistance to aminoglycosides and carbapenems due to the expression of a 16S rRNA methylase armA (with high level resistance) and acquisition of OXA-type carbapenemase OXA-23. Obviously, most of sequence types belonged to clonal complex 92 in these 41 isolates, which demonstrated that these isolates had a common origin and caused nosocomial infection. An A. baumannii clone producing OXA-23, armA along with mutations of quinolone resistance-determining regions (QRDRs) has been identified as an emerging and rapidly spreading pathogen. To our knowledge, this is the first report of hospital dissemination of A. baumannii carrying blaOXA-23, armA and mutations of QRDRs in QRAB in western China

Renee received her BS in Microbiology from the University of South Florida in 2013. During her bachelor's degree she performed undergraduate research, first in the laboratory of our collaborator, Dr. Roman Manetsch, in the Department of Chemistry, before joining the Shaw lab in the Summer of 2012. During both research stints she worked on the development on novel antimicrobial agents targeting drug-resistant bacteria. At graduation Renee received the departmental award for Outstanding Undergraduate Researcher. She transitioned to an PhD student in Spring 2014, continuing her work on novel antibiotic development, specifically focusing on the ESKAPE pathogens.

Antibiotic resistance is a major global health issue for the 21st century as an increasing number of bacterial species have become multi-drug resistance. Many species have become resistant to the majority or all of antibiotics available and extremely toxic drugs are being used to cure these infections because there is nothing left to treat them. The increase of antibiotic resistance combined with the decrease of FDA approval for novel therapeutics is driving us rapidly towards a post antibiotic era where antibiotics will be obsolete. If this lack of novel therapeutics continues, resistant antibiotic infections will be the leading cause of death by the year 2050. Mixture based synthetic combinatorial library screening offers a tremendous enhancement for the rate of drug discovery. This is due to the fact that the activity of millions of compounds can be assessed through the testing of exponentially fewer samples. To this end, we utilized the synthetic combinatorial screening to identify anti-resistance agents that cause multi-drug resistant species to be once again susceptible to clinical antibiotics that are no longer effective. From the initial screening we selected a triamine peptide library that decreased the effective concentration of tetracycline towards a clinical multi-drug resistant Pseudomonas aeruginosa by 4-fold without displaying any inhibitory effects alone. Deconvolution of this library was performed using the positional scanning approach to identify the functional groups at each variant position that created the greatest decrease in the effective concentration of tetracycline. Triamine lead agents were shown to decrease the tetracycline 90% effective concentration by an average 7.4-fold. In addition, these compounds displayed efflux inhibition of ethidium bromide of Gram negative and Gram positive organisms. The lead agents were not effective towards bacterial mutant strains lacking a major efflux pump revealing specificity towards efflux inhibition. These results demonstrate that employing synthetic combinatorial libraries to screen for anti-resistance agents can create a fundamental shift away from the traditional screening processes by introducing a rapid approach to discover novel agents that create susceptibility in multi-drug resistant species.

Natascia Bruni has completed her PhD from University of Turin and Postdoctoral studies from high synthesis school of Gargnano, Italy. She is the Director of Research and Development in Candioli Pharma Organization. She has published more than 10 papers in reputed journals.

Otitis externa (OE) is the most common disease of the ear canal in dogs and it has a multifactorial etiology (e.g., atopic dermatitis, food allergy, etc). Microorganisms most frequently isolated are Staphylococcus intermedius group (SIG), Pseudomonas aeruginosa, Proteus vulgaris and Malassezia pachydermatis. Traditionally, OE treatments include topical and or systemic use of antibiotics, antifungals and glucocorticoids. Because of the rising antibiotic-resistance phenomenon, development of alternative antimicrobial agents is encouraged. Bovine lactoferricin is a multi-functional peptide derived from proteolytic cleavage of bovine lactoferrin. It has proved antibacterial, antifungal and immunostimulatory activity. The aim of this study was to evaluate the action of a patent pending mixture (LAS™, filed with number MI2014A 00130 on 24/09/2014) of lactoferricin peptides associated with two plant extracts with anti-inflammatory effects in dogs with erythematous ceruminous otitis externa and bacterial or Malassezia spp., overgrowth. The product was tested in 10 dogs, selected according to the good clinical practice, showing recurrent erythematous and ceruminous OE. Treatment consisted in topical daily administration of the mixture. The patients were submitted to the clinical-instrumental evaluations during the inclusion examination day and at days 7, 14 and 21 of treatment. Each visit included otoscopy and evaluation of the ear condition using a modified CADESI system and cytology of the ear, performed with a sterile swab. The obtained samples were stained by Diff Quick® (Diff-Quik, Medion Diagnostics AG, Dudingen, Switzerland) and examined at 40/HPF for the count of keratinocytes, bacteria and Malassezia spp. The software used was SPSS (version 19; SPPS Inc., Chicago, IL, USA). Data obtained were analyzed with Shapiro-Wilk, Kruskal-Wallis and Friedman tests. A P value<0.05 was considered statistically significant. Results underlined a statistically significant reduction of both ear canal lesions and CADESI score. Moreover, cytological examinations showed a reduction in the number of keratinocytes, bacteria and Malassezia spp. The topical product (LAS™) was effective to treat and control bacterial and or Malassezia overgrowth in dogs with erythematous and ceruminous otitis externa. It showed antibacterial and antifungal activity in addition to ceruminolytic action.

Background: Acute Bacterial Skin and Skin Structure Infections (ABSSSI) are the 4th most frequent cause of in-hospital infections in Portugal and a major driver for resource utilization. Vancomycin and linezolid are widely used in ABSSSI but have been associated with a prolonged length of stay (LOS) of 26.5 and 19.8 days, respectively. Dalbavancin is a new lipoglycopeptide antibacterial agent that is active against gram positive pathogens and demonstrates a long half-life, allowing for a two-dose regimen to treat ABSSSIs (1000 mg IV followed, one week later, by 500 mg IV). This study aimed to evaluate its effect on LOS and disease burden in Portugal. Material & Methods: The natural history of ABSSSI and treatment implications was modeled with the following assumptions: Linezolid as comparator for dalbavancin as 1st line treatment and vancomycin as 2nd line treatment for both; efficacy and safety endpoints as reported in randomized clinical trials; health resource utilization estimates based on observational Portuguese data; assessment of results in the overall microbiologically evaluable population and a separate analysis for methicillin-resistant Staphylococcus aureus (MRSA); therapy failure opportunity exists to 1st line therapy but no subsequent therapy failures and time horizon of up to 19.81 days (1st line therapy) and up to 14.0 days (2nd line therapy). Results: The average LOS with dalbavancin was estimated to be 10.4 days compared to the average LOS for linezolid (19.8 days), this represents an average reduction of 9.4 days in the hospital LOS for ABSSSI patients. Regarding the burden of disease, dalbavancin is estimated to additionally decrease the in-hospital death rate by 1.6% in the overall microbiologically evaluable population and by 1.8% in MRSA infected population when compared to linezolid. Conclusions: Dalbavancin may be an effective pathway to provide IV antibiotic therapy for patients with ABSSSI with a favorable safety profile and a two-dose regimen. Its use as an in-hospital 1st line IV antibiotic therapy to treat ABSSSI has the potential to decrease LOS and burden of disease when compared to 1st line use of linezolid.

Helena Sofia Antão, MD, MSc PharmD graduated in medicine from Nova Medical School, Portugal and earned a master of science from Lisbon Católica University. She works as a medical advisor at the medical department of Angelini Portugal where she coordinates clinical research projects, provides support to pharmacovigilance and ensures scientific support to several therapeutic areas including antibacterials and infectious diseases. She works as a medical doctor at Centro Hospitalar de Lisboa Ocidental emergency room.

Sylvia Adel Daniel Rofael is an Assistant Lecturer in Faculty of Pharmacy, University of Alexandria, Egypt. She has completed her BSc in Pharmaceutical Sciences and her MSc degree in Pharmaceutical Microbiology from University of Alexandria. She is currently pursuing her PhD degree in University College of London, UK through a Newton Mosharafa Scholarship sponsored by the British Council Egypt, British Embassy Egypt and the Egyptian Ministry of Higher Education.

A concern about emergence of new antibiotic resistant strains in health care environment due to extensive exposure of hospital bacteria to sub-inhibitory concentrations of biocides has been expressed. Our aim was to reveal any possible link between adaptation to biocides and resistance to antibiotics in hospitals. A total of 66 clinical and environmental bacterial isolates; isolated from the Main University Hospital in Alexandria were screened for their susceptibility to 22 commonly used broad spectrum antibiotics and six biocides; benzalkonium chloride (BK), cetrimide (CET); chlorhexidine (CHX); povidone-iodine; sodium hypochlorite and dettol®. Then selected hospital isolates in addition to standard strains were adapted to the biocides by passing them in gradually increasing biocide concentrations. The maximum obtained minimum inhibitory concentrations (MICs) were >3200 mg/L for BK and CET; >1000 mg/L for CHX. Cross resistance to antibiotics was then tested in the biocide adapted isolates using Stoke’s method. Screening results revealed a moderate positive correlation between antibiotic resistance and biocide tolerance where Spearman correlation coefficient ranged between 0.376-0.278 (p<0.05). The stable adaptation to BK, CET and CHX resulted in reduced susceptibility towards certain antibiotics; amikacin, gentamicin, imipenem, ciprofloxacin, levofloxacin, chloramphenicol, ceftazidime, doxycycline, tetracycline, cefoperazone, cefotaxime and cefepime. In many cases, the observed cross resistance moved the strains from being “sensitive” to being “intermediately sensitive” or even “resistant” to antibiotics. Therefore, biocides should be handled with care in health care settings avoiding sub-lethal concentrations. Cross resistance between biocides and antibiotics can aggravate the existing problem of antibiotic resistance in the healthcare system.

During a screen of seaweed associated marine microorganisms for their ability to produce antimicrobial metabolites, a bacterial strain which later named (P) demonstrated antimicrobial activity against a variety of gram-positive pathogens. Ribotyping analysis showed the strain had high sequence identity (99%) to Serratia plymuthica. Tests with a type strain of S. plymuthica obtained from the DSMZ strain bank revealed that (P) did not produce a similar spectrum of antimicrobial activity and initial studies of the inhibitory activity of the antimicrobial compound/s produced by (P) demonstrated that it has a low molecular weight, pH and heat resistant and was not affect by enzymes like (Proteinase K). Also, a modified method of transposon mutagenesis (Larsen et al., 2002) has been used to identify gene/s involved in antimicrobial production and 14 mutants defective in antimicrobial production were isolated and partially characterized. Sequence analysis of three of these mutants revealed the presence of transposon insertions in genes encoding proteins similar to polyketide synthases. Aims The aim of this project is to investigate the production of secreted antimicrobial activity in S. plymuthica (P) and the genetic analysis of (P) using transposon mutagenesis and cloning the genomic regions that code for antimicrobial activity to determine the number of genes involved and the structure and position of the identified genes in the bacterial genome. Also, the effect of various growth conditions on the production of secreted anti- microbial activity and the purification and basic characterization of the secreted antimicrobial compound/s.

Ann Lok Yan So received her PhD in chemical biology from The University of Hong Kong in 2012. She is currently working on novel antibiotics development in Hong Kong Polytechnic University as a research fellow. She is particularly interested in developing new drugs against antibiotics resistant bacteria. Potential drug candidates include small organic molecules, antimicrobial peptides and silver nanoparticles.

Emergence of antibiotics resistant bacterial pathogens and the rapid spreading around the world threaten to take us back to the pre-antibiotic era. Concerns are raised to promote personal hygiene to minimize spread of infections and appropriate use of antibiotics to slow down the development of antibiotics resistance in pathogens. However, recent emergence of resistance towards new classes of antibiotics, for example, carbapenem and colistin resistant bacteria has urged the need of new antibiotics. In this study, we synthesized and characterized small monodispersed spherical silver nanoparticles (AgNPs) coated with polyethylene glycol (PEG) with average diameter of 2.56 nm. The stable, water soluble, biocompatible AgNPs were prepared in a simple manner in ethanol under ambient light at 4 ºC with purification to remove unreacted raw materials. The antibacterial activity of the AgNPs towards a range of both antibiotics sensitive and resistant Gram positive and Gram negative bacteria was examined. The tested Gram positive bacterial strains included Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Bacillus subtilis and Streptococcus mutans, while Gram negative strains included Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae and Pseudomonas aeruginosa, covering a wide range of bacterial pathogens responsible for infections in many parts of the human body, for example, pneumonia, infections in bloodstream, urinary tract, bone and joint, wound and surgical site. Moreover, combinational antibacterial activity of the AgNPs with conventional antibiotics was also investigated. The simple preparation of this antibacterial AgNPs may be suitble for the fomulation to combat a broad range of bacterial infections.

Georgios A Delis is a Lecturer in Veterinary Pharmacology in the School of Veterinary Medicine, Aristotle University of Thessaloniki, Greece. He has published research on pharmacokinetics, pharmacodynamics and pharmacokinetic/pharmacodynamic correlation of antimicrobial agents as well as on analytical methods for the determination of drugs in biological fluids.

Understanding the antimicrobials’ pattern of bacterial killing can both ensure therapeutic efficacy and prevent the development of resistance. The scope of this study was to explore the pharmacodynamics of selected bactericidal agents, namely amikacin (AMK), amoxicillin/clavulanic acid (AMC), ceftazidime (CAZ), marbofloxacin (MAR) and enrofloxacin (ENR), against Escherichia coli strains isolated from dogs. Minimum Inhibitory Concentrations (MICs) were determined against clinical and commensal E. coli strains (n=5) by the macro-dilution method. Interpretation of MIC values was performed according to CLSI and EUCAST guidelines. Based on MIC results, kill curves were studied by viable counts determination. The Area Between the Bactericidal and the Control curves (ABBC) was selected as the antimicrobial effect (E) index, whereas maximum effect (Emax) and half maximal effective concentration (EC50) values were obtained by use of the Sigmoidal Emax model. The ratio of EC50 to MIC was also determined. Investigational strains displayed considerable variability in their susceptibility (susceptible, intermediately susceptible and resistant). In terms of Emax, AMK yielded higher ABBC values than all other antimicrobials, altogether with a lower EC50 to MIC ratio, indicating that a substantial bactericidal effect of 50% of Emax can be observed at levels down to 0.34×MIC. MAR required levels higher than 0.6×MIC to yield a half maximal effect. On the other side of the spectrum, CAZ displayed low efficacy. Interestingly, it was the most potent drug (MIC=1 μg/mL, EC50=0.12 μg/mL). Integration of the findings of this study with pharmacokinetic data could provide insight regarding the predicted efficacy by current dosage schemes of the specific antimicrobials in dogs.

Essam J Alyamani has completed his PhD from Northeastern University in Boston, USA and Postdoctoral studies from Ottowa University, Canada and Northeastern University, Chicago USA. He is the Director of Microbial Biotechnology at the National Center for Biotechnology, KACST. He has published more than 20 papers in reputed journals and has been serving as a Grant and journals Reviewer since 2008.

Acinetobacter baumannii is a common opportunistic pathogen that causes major nosocomial infections in hospitals. In this study, we hypothesized a high prevalence of A. baumannii ESBL (extended spectrum betalactamase) among all collected isolates. A. baumannii isolates (n=107) from ICU (Intensive care unit) of local hospitals in Makkah were phenotypically and genotypically characterized. The identity and antibiotic susceptibility of A. baumannii strains were determined using the Vitek-2 system. The identified ESBL producers were further analyzed by PCR and sequencing followed by MLST typing. blaTEM, blaSHV and the blaCTX-M group genes 1, 2, 8, 9 and 25 were investigated. Furthermore, blaOXA51 like and blaOXA23 like genes were also examined in the carbapenem resistant A. baumannii isolates. Our data indicated a high prevalence of A. baumannii ESBL producers among the collected strains. Of the 107 A. baumannii isolates, 94% were found to be resistant to cefepime and ceftazidime and aztreonam using the Vitek 2 system. The genes detected encoded TEM, OXA-51 like and OXA-23 like enzymes and CTX-M group proteins 1, 2, 8, 9 and 25. MLST typing identified eight sequence type (ST) groups. The most dominant STs were ST195 and ST557 and all of them belong to worldwide clonal complex (CC) 2. This study has shown that there is a high prevalence of antimicrobial resistance in A. baumannii. The diversity of STs may suggest that new ESBL strains are constantly emerging. The molecular diversity of the ESBL genes in A. baumannii may have contributed to the increased antimicrobial resistance among all isolates

Hager Ali Saleh is a Lecturer of Health Services Management at Faculty of Public Health and Health Informatics, Umm Al Qura University. She has completed her Master’ degree in Hospital Administration in 2010 and in 2006, she has completed her Bachelor’s degree in Pharmaceutical Sciences, Alexandria University. She is a certified Professional in Healthcare Quality since 2013. She has also worked for 4 years as a Pharmacist, Drug Information Center Coordinator and Patients’ Rights Officer at Alexandria Fever Hospital, Egypt. She has published five research papers and her research areas of interest are Patients' Rights and Safety, Performance Improvement and Pharmacovigilance.

Aim: The aim of the study was assessing knowledge, attitude and current practice of physicians towards antibiotic prescribing in Alexandria Fever Hospital. Methods: Self administering questionnaire published as a Google form in the hospital official Facebook group. The questionnaire was designed after reviewing the literature. Results: Physicians are aware of antibiotic resistance problem in their hospital and in the country although they prefer over prescribing rather than under prescribing of antibiotics. Although 50% indicated that they are confident in the area of antibiotic resistance 45% did not receive a regular training in that area. All physicians agreed that inappropriate use of antimicrobial agents may result in antimicrobial resistance. The most of physicians (75%) agreed that the main cause of inappropriate use of antibiotics is lack of effective hospital polices and (80%) believed that their education will help in controlling of antibiotics resistance. Conclusion: There is a considerable unmet training and education needs for physicians in the area of antimicrobial prescribing therefore, there is a need to increase the teaching and training courses about antibiotics in the hospital and encourage the physicians to attend. The hospital antimicrobial guidelines need revision and to be well published between physicians.

Tarek BENAMEUR completed his PhD in Clinical and Experimental Pharmacology at the University of Angers (France). He joined the University of Liverpool and the MRC-Human Genetic Unit in Edinburgh (U.K) as a post-doctoral research fellow. Currently, he resumed the position of Assistant Professor within the College of Medicine at King Faisal University; actively involved in conducting different research projects and involved in teaching the competency-based curriculum of the University of Groningen. He has a considerable track record of publications in leading international journals. He presented his work in many national and international conferences and was invited speaker in many conferences.

Background: Antibiotics self-prescription is a worldwide public health problem due to irrational use. Medical students have the potential to improve the awareness to limit inappropriate use of antibiotics within their communities. In Saudi Arabia this practice seems to be emerging challenge for the healthcare provider. However, the prevalence and the reasons of this malpractice remains unknown within the students’ populations. Objective: Estimate the prevalence of self-prescription with antibiotics, assess the knowledge and attitudes of medical and non-medical students in the Eastern Region of Saudi Arabia. Methods: A descriptive, cross-sectional study based on pre-tested questionnaire. Convenient sampling involved face-to-face questionnaire of 235 students was adopted. Results: In this study, about 58 % of students are practicing self-prescription. This substantiates high self-prescription prevalence among medical and non-medical students (50 %, 67.3 %), respectively. Nearly, 75 % of medical students knew that antibiotics are used for bacterial infection compared to only 27 % of non-medical students. Confusion about the role of antibiotics in treating infections was the most critical, with more than 72 % of non-medical students failed to identify that antibiotics do not eradicate viral infections which is alarmingly high. Importantly, 87 % of medical students knew that over-use of antibiotics without medical prescription can have adverse effects compared with 58.9 % of non-medical students who agreed with this belief. This statement is affected by the specialty (p<0.001). Despite, the awareness of medical students about antibiotics self-medication is unsafe and mal-practice (79.7 %), the prevalence of antibiotics self-prescription remains relatively high among all respondents. Conclusion: The prevalence of self-medication with antibiotics is noticeably high in all students’ population. We highly recommend improving the health education to better address this malpractice and improve the students’ knowledge, attitudes and awareness towards the antibiotics use and safety.