Day 3 :
- 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
B Y Chin
International Medical University, Malaysia
Monash University, Australia
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.
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.
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
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.
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
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
Promod Mehta presently as Director at Centre for Biotechnology, Maharshi Dayanand University, India
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.
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
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).
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.
Liaquat University of Medical & Health Sciences, Pakistan
Time : 10:55-11:15
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.
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.
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
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
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 ᵒC. 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 37ᵒC 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.
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
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.
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.
Ahmadu Bello University, Nigeria
Title: Antibiotic susceptibility profile of methicillin resistant Staphylococci aureus in poultry farm, in Zaria, Nigeria
Time : 11:55-12:15
Josiah Ademola Onaolapo from Department Of Pharmaceutics and Pharmaceutical Microbiology, Ahmadu Bello University, Zaria, Nigeria
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.