Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 132

Search results for: Klebsiella pneumoniae

132 Investigation of Biofilm Formation in Clinical Strains of Klebsiella pneumoniae and Klebsiella rhinoscleromatis

Authors: Gulcan Sahal, Nermin Hande Avcioglu, Isil Seyis Bilkay

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Klebsiella species which are natural colonizers of human upper respiratory and human gastrointestinal tracts are also responsible for every reoccurring nosocomial infections by means of having ability to form slimy layers known as biofilm on many surfaces. Therefore, in this study, investigation of biofilm formation in K. pneumoniae and K. rhinoscleromatis and examination of each Klebsiella strains’ clinical information in the light of their biofilm formation results were aimed. In this respect, biofilm formation of Klebsiella strains was analyzed via crystal violet binding assay. According to our results, biofilm formation levels of K. pneumoniae and K. rhinoscleromatis strains were different from each other. Additionally, in comparison to K. rhinoscleromatis strains, K. pneumoniae was observed to include higher amounts of strong biofilm forming strains. Besides, it was also seen that clinical information of patients from which strong biofilm forming Klebsiella strains were isolated were similar to each other. Our results indicate that there should be more precautions against K. pneumoniae which includes higher amount of strong biofilm forming strains.

Keywords: biofilm formation, Klebsiella pneumoniae, Klebsiella rhinoscleromatis, biosystems engineering

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131 Determination of Klebsiella Pneumoniae Susceptibility to Antibiotics Using Infrared Spectroscopy and Machine Learning Algorithms

Authors: Manal Suleiman, George Abu-Aqil, Uraib Sharaha, Klaris Riesenberg, Itshak Lapidot, Ahmad Salman, Mahmoud Huleihel

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Klebsiella pneumoniae is one of the most aggressive multidrug-resistant bacteria associated with human infections resulting in high mortality and morbidity. Thus, for an effective treatment, it is important to diagnose both the species of infecting bacteria and their susceptibility to antibiotics. Current used methods for diagnosing the bacterial susceptibility to antibiotics are time-consuming (about 24h following the first culture). Thus, there is a clear need for rapid methods to determine the bacterial susceptibility to antibiotics. Infrared spectroscopy is a well-known method that is known as sensitive and simple which is able to detect minor biomolecular changes in biological samples associated with developing abnormalities. The main goal of this study is to evaluate the potential of infrared spectroscopy in tandem with Random Forest and XGBoost machine learning algorithms to diagnose the susceptibility of Klebsiella pneumoniae to antibiotics within approximately 20 minutes following the first culture. In this study, 1190 Klebsiella pneumoniae isolates were obtained from different patients with urinary tract infections. The isolates were measured by the infrared spectrometer, and the spectra were analyzed by machine learning algorithms Random Forest and XGBoost to determine their susceptibility regarding nine specific antibiotics. Our results confirm that it was possible to classify the isolates into sensitive and resistant to specific antibiotics with a success rate range of 80%-85% for the different tested antibiotics. These results prove the promising potential of infrared spectroscopy as a powerful diagnostic method for determining the Klebsiella pneumoniae susceptibility to antibiotics.

Keywords: urinary tract infection (UTI), Klebsiella pneumoniae, bacterial susceptibility, infrared spectroscopy, machine learning

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130 Nanotechnology-Based Treatment of Klebsiella pneumoniae Infections

Authors: Lucian Mocan, Teodora Mocan, Matea Cristian, Cornel Iancu

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We present method of nanoparticle enhanced laser thermal ablation of Klebsiella pneumoniae infections, using gold nanoparticles combined with a specific growth factor and demonstrate its selective therapeutic efficacy. Ab (antibody solution) bound to GNPs (gold nanoparticles) was administered in vitro and determined the specific delivery of the nano-bioconjugate into the microorganism. The extent of necrosis was considerable following laser therapy, and at the same time, normal cells were not seriously affected. The selective photothermal ablation of the infected tissue was obtained after the selective accumulation of Ab bound to GNPs into bacteria following perfusion. These results may represent a major step in antibiotherapy treatment using nanolocalized thermal ablation by laser heating.

Keywords: gold nanoparticles, Klebsiella pneumoniae, nanoparticle functionalization, laser irradiation, antibody

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129 In silico Subtractive Genomics Approach for Identification of Strain-Specific Putative Drug Targets among Hypothetical Proteins of Drug-Resistant Klebsiella pneumoniae Strain 825795-1

Authors: Umairah Natasya Binti Mohd Omeershffudin, Suresh Kumar

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Klebsiella pneumoniae, a Gram-negative enteric bacterium that causes nosocomial and urinary tract infections. Particular concern is the global emergence of multidrug-resistant (MDR) strains of Klebsiella pneumoniae. Characterization of antibiotic resistance determinants at the genomic level plays a critical role in understanding, and potentially controlling, the spread of multidrug-resistant (MDR) pathogens. In this study, drug-resistant Klebsiella pneumoniae strain 825795-1 was investigated with extensive computational approaches aimed at identifying novel drug targets among hypothetical proteins. We have analyzed 1099 hypothetical proteins available in genome. We have used in-silico genome subtraction methodology to design potential and pathogen-specific drug targets against Klebsiella pneumoniae. We employed bioinformatics tools to subtract the strain-specific paralogous and host-specific homologous sequences from the bacterial proteome. The sorted 645 proteins were further refined to identify the essential genes in the pathogenic bacterium using the database of essential genes (DEG). We found 135 unique essential proteins in the target proteome that could be utilized as novel targets to design newer drugs. Further, we identified 49 cytoplasmic protein as potential drug targets through sub-cellular localization prediction. Further, we investigated these proteins in the DrugBank databases, and 11 of the unique essential proteins showed druggability according to the FDA approved drug bank databases with diverse broad-spectrum property. The results of this study will facilitate discovery of new drugs against Klebsiella pneumoniae.

Keywords: pneumonia, drug target, hypothetical protein, subtractive genomics

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128 Phylogenetic Analysis of Klebsiella Species from Clinical Specimens from Nelson Mandela Academic Hospital in Mthatha, South Africa

Authors: Sandeep Vasaikar, Lary Obi

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Rapid and discriminative genotyping methods are useful for determining the clonality of the isolates in nosocomial or household outbreaks. Multilocus sequence typing (MLST) is a nucleotide sequence-based approach for characterising bacterial isolates. The genetic diversity and the clinical relevance of the drug-resistant Klebsiella isolates from Mthatha are largely unknown. For this reason, prospective, experimental study of the molecular epidemiology of Klebsiella isolates from patients being treated in Mthatha over a three-year period was analysed. Methodology: PCR amplification and sequencing of the drug-resistance-associated genes, and multilocus sequence typing (MLST) using 7 housekeeping genes mdh, pgi, infB, FusAR, phoE, gapA and rpoB were conducted. A total of 32 isolates were analysed. Results: The percentages of multidrug-resistant (MDR), extensively drug-resistance (XDR) and pandrug-resistant (PDR) isolates were; MDR 65.6 % (21) and XDR and PDR with 0 % each. In this study, K. pneumoniae was 19/32 (59.4 %). MLST results showed 22 sequence types (STs) were identified, which were further separated by Maximum Parsimony into 10 clonal complexes and 12 singletons. The most dominant group was Klebsiella pneumoniae with 23/32 (71.8 %) isolates, Klebsiella oxytoca as a second group with 2/32 (6.25 %) isolates, and a single (3.1 %) K. varricola as a third group while 6 isolates were of unknown sequences. Conclusions/significance: A phylogenetic analysis of the concatenated sequences of the 7 housekeeping genes showed that strains of K. pneumoniae form a distinct lineage within the genus Klebsiella, with K. oxytoca and K. varricola its nearest phylogenetic neighbours. With the analysis of 7 genes were determined 1 K. variicola, which was mistakenly identified as K. pneumoniae by phenotypic methods. Two misidentifications of K. oxytoca were found when phenotypic methods were used. No significant differences were observed between ESBL blaCTX-M, blaTEM and blaSHV groups in the distribution of Sequence types (STs) or Clonal complexes (CCs).

Keywords: phylogenetic analysis, phylogeny, klebsiella phylogenetic, klebsiella

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127 Antibacterial Activity of Green Synthesis Silver Nanoparticles from Moringa Oleifera

Authors: Ali Fadhel Ahmed, Tuqa Abdulkareem Hameed

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Moringa oleifera (leaves and seeds) ethanolic and aqueous extracts were tested for antibacterial activity. The effect of plant extracts on three types of bacterial species: Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae, was investigated. Using the agar well diffusion method, ethanolic extracts of Moringa oleifera demonstrated a significant antibacterial effect on the forty tested bacterial strains. Seed-induced inhibition zones (ethanolic extracts)were ranged from16 to 24 mm in diameter against S. aureus, respectively, whileE. coli and K. pneumonia had no effect. Gram-positive and Gram-negative bacteria were not affected by alcoholic and aqueous plant leaf extracts. The purpose of this present study was to look at the cytotoxic effects of M.Oleifera plant (alcoholic extracts).

Keywords: moringa oleifera, escherichia coli, klebsiella pneumoniae, staphylococcus aureus

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126 High Frequency of Chlamydophila Pneumoniae in Children with Asthma Exacerbations

Authors: Katherine Madero Valencia, Carlos Jaramillo, Elida Dueñas, Carlos Torres, María Del Pilar Delgado

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Asthma, described as a chronic inflammatory condition of the airways, courses accompanied by episodes known as exacerbations, characterized by a worsening of symptoms. Among the triggers, some allergen-irritative and infectious agents are found, including Chlamydophila pneumoniae which seems to play an increasingly important role. In this paper a PCR was used to detect C. pneumoniae in order to estimate the frequency of infections caused by this agent in pediatric patients with asthma exacerbations. C. pneumoniae distribution throughout the study period was also evaluated. 175 nasopharyngeal aspirates from children with asthma exacerbations were analyzed by PCR and sequencing. A global prevalence of C. pneumoniae of 53.71% was obtained. This study highlights a high circulation of C. pneumoniae during the study period, in children of all ages and especially in children under 5 years old. Molecular tests applied permit a rapid detection and improved our knowledge about these infections in children with asthma.

Keywords: Chlamydophila pneumoniae, detection, molecular techniques, pediatric asthma

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125 Isolation and Molecular Characterization of Lytic Bacteriophage against Carbapenem Resistant Klebsiella pneumoniae

Authors: Guna Raj Dhungana, Roshan Nepal, Apshara Parajuli, , Archana Maharjan, Shyam K. Mishra, Pramod Aryal, Rajani Malla

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Introduction: Klebsiella pneumoniae is a well-known opportunistic human pathogen, primarily causing healthcare-associated infections. The global emergence of carbapenemase-producing K. pneumoniaeis a major public health burden, which is often extensively multidrug resistant.Thus, because of the difficulty to treat these ‘superbug’ and menace and some term as ‘apocalypse’ of post antibiotics era, an alternative approach to controlling this pathogen is prudent and one of the approaches is phage mediated control and/or treatment. Objective: In this study, we aimed to isolate novel bacteriophage against carbapenemase-producing K. pneumoniaeand characterize for potential use inphage therapy. Material and Methods: Twenty lytic phages were isolated from river water using double layer agar assay and purified. Biological features, physiochemical characters, burst size, host specificity and activity spectrum of phages were determined. One most potent phage: Phage TU_Kle10O was selected and characterized by electron microscopy. Whole genome sequences of the phage were analyzed for presence/absence of virulent factors, and other lysin genes. Results: Novel phage TU_Kle10O showed multiple host range within own genus and did not induce any BIM up to 5th generation of host’s life cycle. Electron microscopy confirmed that the phage was tailed and belonged to Caudovirales family. Next generation sequencing revealed its genome to be 166.2 Kb. bioinformatical analysis further confirmed that the phage genome ‘did not’ contain any ‘bacterial genes’ within phage genome, which ruled out the concern for transfer of virulent genes. Specific 'lysin’ enzyme was identified phages which could be used as 'antibiotics'. Conclusion: Extensively multidrug resistant bacteria like carbapenemase-producing K. pneumoniaecould be treated efficiently by phages.Absence of ‘virulent’ genes of bacterial origin and presence of lysin proteins within phage genome makes phages an excellent candidate for therapeutics.

Keywords: bacteriophage, Klebsiella pneumoniae, MDR, phage therapy, carbapenemase,

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124 Activity of Malate Dehydrogenase in Cell Free Extracts from S. proteamaculans, A. hydrophila, and K. pneumoniae

Authors: Mohamed M. Bumadian, D. James Gilmour

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Three bacterial species were isolated from the River Wye (Derbyshire, England) and identified using 16S rRNA gene sequencing as Serratia proteamaculans, Aeromonas hydrophila and Klebsiella pneumoniae. Respiration rates of the strains were measured in order to determine the metabolic activity under salt stress. The highest respiration rates of all three strains were found at 0.17 M and 0.5 M NaCl and then the respiration rate decreased with increasing concentrations of NaCl. In addition, the effect of increasing concentrations of NaCl on malate dehydrogenase activity was determined using cell-free extracts of the three strains. Malate dehydrogenase activity was stimulated at NaCl concentrations up to 0.5 M, and a small level of activity remained even at 3.5 M NaCl. The pH optimum of the malate dehydrogenase in cell-free extracts of all strains was higher than pH 7.5.

Keywords: fresh water, halotolerant pathogenic bacteria, 16S rRNA gene, cell-free extracts, respiration rates, malate dehydrogenase

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123 Bacteriological Spectrum and Resistance Patterns of Common Clinical Isolates from Infections in Cancer Patients

Authors: Vivek Bhat, Rohini Kelkar, Sanjay Biswas

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Introduction: Cancer patients are at increased risk of bacterial infections. This may due to the disease process itself, the effect of chemotherapeutic drugs or invasive procedures such as catheterization. A wide variety of bacteria including some emerging pathogens are increasingly being reported from these patients. The incidence of multidrug-resistant organisms particularly in the Gram negative group is also increasing, with higher resistance rates seen to cephalosporins, β-lactam/β-lactam inhibitor combinations, and the carbapenems. This study documents the bacteriological spectrum of infections and their resistance patterns in cancer patients. Methods: This study includes all bacterial isolates recovered from infections cancer patients over a period of 18 months. Samples included Blood cultures, Pus/wound swabs, urine, tissue biopsies, body fluids, catheter tips and respiratory specimens such as sputum and bronchoalveolar lavage (BAL). All samples were processed in the microbiology laboratory as per standard laboratory protocols. Organisms were identified to species level and antimicrobial susceptibility testing was performed manually by the disc diffusion technique or in the Vitek-2 (Biomereux, France) instrument. Interpretations were as per Clinical laboratory Standards Institute (CLSI) guidelines. Results: A total of 1150 bacterial isolates were cultured from 884 test samples during the study period. Of these 227 were Gram-positive and 923 were Gram-negative organisms. Staphylococcus aureus (99 isolates) was the commonest Gram-positive isolate followed by Enterococcus (79) and Gr A Streptococcus (30). Among the Gram negatives, E. coli (304), Pseudomonas aeruginosa (201) and Klebsiella pneumoniae (190) were the most common. Of the Staphylococcus aureus isolates 27.2% were methicillin resistant. Only 5.06% enterococci were vancomycin resistant. High rates of resistance to cefotaxime and ciprofloxacin were seen amongst E. coli (84.8% & 83.55%) and Klebsiella pneumoniae (71 & 62.1%) respectively. Resistance to carbapenems (meropenem) was high at 70% in Acinetobacter spp.; however all isolates were sensitive to colistin. Among the aminoglycosides, amikacin retained good efficacy against Escherichia coli (82.9%) and Pseudomonas aeruginosa (78.1%). Occasional isolates of emerging pathogens such as Chryseobacterium indologens, Roseomonas, and Achromobacter xyloxidans were also recovered. Conclusion: The common infections in cancer patients include respiratory, wound, tract infections and sepsis. The commonest isolates include Staphylococcus aureus, Enterococci, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. There is a high level of resistance to the commonly used antibiotics among Gram-negative organisms.

Keywords: bacteria, resistance, infection, cancer

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122 Antibacterial Potentials of the Leaf Extracts of Siam Weed (Chromolaena odorata) on Wound Isolates

Authors: M. E. Abalaka, O. A. Falusi, M. Galadima, D. Damisa

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The antimicrobial activity of aqueous, ethanolic and methanolic extracts of Chromolaena odorata (Siam weed) was evaluated against four wound isolates: Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae at the concentrations of 200mg/ml, 100mg/ml, 50mg/ml and 25mg/ml respectively. S. aureus and E. coli showed high susceptibility to the various extracts than the other test isolates. The aqueous extract showed activity against Staphylococcus aureus with a mean diameter of zone of inhibition of 16 ± 3.00 at concentration of 200mg/ml and as low as 8 ± 0.00 at concentration of 25mg/ml; E. coli showed susceptibility with a mean diameter of zone of inhibition of 18 ± 2.00 and 10 ± 0.00 at a concentration of 200mg/ml and 25mg/ml respectively. Pseudomonas aeruginosa and Klebsiella pneumoniae were resistant to the aqueous extract. Methanol extract showed activity against Staphylococcus aureus with a mean diameter of zone of inhibition at 28 ± 4.00 and 12 ± 2.30 at a concentration of 200mg/ml and 25mg/ml respectively; while E. coli was susceptible with mean diameter of zone of inhibition of 18 ± 2.00 and as low as 12 ± 0.00 at a concentration of 200mg/ml and 50mg/ml respectively, Pseudomonas aeruginosa showed considerable susceptibility with mean diameter of zone of inhibition of 13 ± 1.00 and 12 ± 0.00 at a concentration of 200mg/ml and 100mg/ml respectively. The ethanol extract showed activity against S. aureus with a mean diameter zone of inhibition of 15 ± 2.00 and 9 ± 0.00 at a concentration of 200mg/ml and 25mg/ml respectively: E. coli showed susceptibility with a mean diameter zone of inhibition of 20 ± 4.00 and 13 ± 2.00 at a concentration of 200mg/ml and 25mg/ml respectively. Pseudomonas aeruginosa showed considerable susceptibility with a mean diameter zone of inhibition of 13 ± 1.00 and 9 ± 0.00 at a concentration of 200mg/ml and 100mg/ml respectively. The results above indicate the efficacy and potency of the crude extracts of Chromolaena odorata leaf on the tested wound isolates.

Keywords: antibacterial, Chromolaena odorata, leaf extracts, test isolates

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121 Extra-Pulmonary Mycoplasma Pneumoniae Infection in a Healthy 25-Year-Old Female: A Case Report

Authors: Minna Chang

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Introduction: M. pneumoniae is a respiratory pathogen, which commonly causes upper and lower respiratory infections. It primarily affects children and young adults. Respiratory symptoms are well recognized, but extrapulmonary involvement is also common. Other systems that have been implicated in the disease include: skin, mucus membranes, central, peripheral nervous systems, cardiovascular, haematological, renal, and musculoskeletal systems. Here, we report a case of an otherwise healthy, young female with M. pneumonia, who presented with right upper quadrant abdominal pain. Case presentation: a healthy 25-year-old female was referred to A&E by her general practitioner, after presenting with fever, malaise, and right upper quadrant pain. M. pneumoniae was confirmed retrospectively by serology. The patient made a full recovery after a six-day course of doxycycline 100mg. Conclusion: M. pneumonia is a well-established cause of respiratory infections in children and young adults. Febrile illness with multisystem involvement, even in the absence of respiratory symptoms, should raise suspicion of M. pneumoniae infection in healthy, young adults. Our case illustrates the multi-system involvement of M. pneumoniae, which was initially missed, due to paucity of respiratory symptoms at presentation.

Keywords: infectious diseases, mycoplasma pneumoniae, respiratory infections, extra-pulmonary manifestations

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120 Characterization of Mycoplasma Pneumoniae Causing Exacerbation of Asthma: A Prototypical Finding from Sri Lanka

Authors: Lakmini Wijesooriya, Vicki Chalker, Jessica Day, Priyantha Perera, N. P. Sunil-Chandra

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M. pneumoniae has been identified as an etiology for exacerbation of asthma (EQA), although viruses play a major role in EOA. M. pneumoniae infection is treated empirically with macrolides, and its antibiotic sensitivity is not detected routinely. Characterization of the organism by genotyping and determination of macrolide resistance is important epidemiologically as it guides the empiric antibiotic treatment. To date, there is no such characterization of M. pneumoniae performed in Sri Lanka. The present study describes the characterization of M. pneumoniae detected from a child with EOA following a screening of 100 children with EOA. Of the hundred children with EOA, M. pneumoniae was identified only in one child by Real-Time polymerase chain reaction (PCR) test for identifying the community-acquired respiratory distress syndrome (CARDS) toxin nucleotide sequences. The M. pneumoniae identified from this patient underwent detection of macrolide resistance via conventional PCR, amplifying and sequencing the region of the 23S rDNA gene that contains single nucleotide polymorphisms that confer resistance. Genotyping of the isolate was performed via nested Multilocus Sequence Typing (MLST) in which eight (8) housekeeping genes (ppa, pgm, gyrB, gmk, glyA, atpA, arcC, and adk) were amplified via nested PCR followed by gene sequencing and analysis. As per MLST analysis, the M. pneumoniae was identified as sequence type 14 (ST14), and no mutations that confer resistance were detected. Resistance to macrolides in M. pneumoniae is an increasing problem globally. Establishing surveillance systems is the key to informing local prescriptions. In the absence of local surveillance data, antibiotics are started empirically. If the relevant microbiological samples are not obtained before antibiotic therapy, as in most occasions in children, the course of antibiotic is completed without a microbiological diagnosis. This happens more frequently in therapy for M. pneumoniae which is treated with a macrolide in most patients. Hence, it is important to understand the macrolide sensitivity of M. pneumoniae in the setting. The M. pneumoniae detected in the present study was macrolide sensitive. Further studies are needed to examine a larger dataset in Sri Lanka to determine macrolide resistance levels to inform the use of macrolides in children with EOA. The MLST type varies in different geographical settings, and it also provides a clue to the existence of macrolide resistance. The present study enhances the database of the global distribution of different genotypes of M. pneumoniae as this is the first such characterization performed with the increased number of samples to determine macrolide resistance level in Sri Lanka. M. pneumoniae detected from a child with exacerbation of asthma in Sri Lanka was characterized as ST14 by MLST and no mutations that confer resistance were detected.

Keywords: mycoplasma pneumoniae, Sri Lanka, characterization, macrolide resistance

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119 Chemical Composition and Antibacterial Activity of Ceratonia siliqua L. Growing in Boumerdes, Algeria

Authors: N. Meziou-Chebouti, A. Merabet, Y. Chebouti N. Behidj

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This work is a contribution to the knowledge of physicochemical characteristics of mature carob followed by evaluation of the activity, antimicrobial phenolics leaves and green pods of Ceratonia siliqua L. physicochemical study shows that mature carob it has a considerable content of sugar (50.90%), but poor in proteins (7%), fat (8%) and also has a high mineral content. The results obtained from phenolic extracts of leaves and green pods of Ceratonia siliqua L. show a wealth leaf phenolic extract especially flavonoids (0,545 mg EqQ/g) relative to the extract of green pods (0,226 mgEqQ/g). Polyphenols leaves have a slightly inhibitory effect on the growth of strains: Staphylococcus aureus, Escherichia coli, Klebsiella pneumoiae, Streptococcus sp and Sanmonella enteritidis, a strong inhibitory effect on the growth of Pseudomonas strain aerogenosa. Moreover, polyphenols pod have a slightly inhibitory effect on the growth of Streptococcus sp strains, Pseudomonas and aerogenosa Sanmonella enteritidis, a slightly inhibitory effect on the growth of Klebsiella pneumoniae strains, E. coli and Staphylococcus aureus.

Keywords: antimicrobial activity, bacteria, clove, Ceratonia siliqua, polyphenols

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118 Antibacterial Activity of Northern Algerian Honey

Authors: Messaouda Belaid, Salima Kebbouche-Gana, Djamila Benaziza

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Our study focuses on determining the antibacterial activity of some honeys from northern Algeria. To test this activity, the agar well diffusion methods was employed. The bacterial strains tested were Staphylococcus aureus, Bacillus subtilis, Streptococcus faecalis, Klebsiella pneumoniae, Escherichia coli, and Pseudomonas aeroginosae. The results showed that all the microbes tested were inhibited by all honey used in this study but Those bacteria that appear to be more sensitive to all honey tested are Staphylococcus aureus and Pseudomonas aeroginosae.

Keywords: honey, antibacterial activity, Northern Algeria, Staphylococcus aureus

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117 Analysis of Non-Coding Genome in Streptococcus pneumoniae for Molecular Epidemiology Typing

Authors: Martynova Alina, Lyubov Buzoleva

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Streptococcus pneumoniae is the causative agent of pneumonias and meningitids throught all the world. Having high genetic diversity, this microorganism can cause different clinical forms of pneumococcal infections and microbiologically it is really difficult diagnosed by routine methods. Also, epidemiological surveillance requires more developed methods of molecular typing because the recent method of serotyping doesn't allow to distinguish invasive and non-invasive isolates properly. Non-coding genome of bacteria seems to be the interesting source for seeking of highly distinguishable markers to discriminate the subspecies of such a variable bacteria as Streptococcus pneumoniae. Technically, we proposed scheme of discrimination of S.pneumoniae strains with amplification of non-coding region (SP_1932) with the following restriction with 2 types of enzymes of Alu1 and Mn1. Aim: This research aimed to compare different methods of typing and their application for molecular epidemiology purposes. Methods: we analyzed population of 100 strains of S.pneumoniae isolated from different patients by different molecular epidemiology methods such as pulse-field gel electophoresis (PFGE), restriction polymorphism analysis (RFLP) and multilolocus sequence typing (MLST), and all of them were compared with classic typing method as serotyping. The discriminative power was estimated with Simpson Index (SI). Results: We revealed that the most discriminative typing method is RFLP (SI=0,97, there were distinguished 42 genotypes).PFGE was slightly less discriminative (SI=0,95, we identified 35 genotypes). MLST is still the best reference method (SI=1.0). Classic method of serotyping showed quite weak discriminative power (SI=0,93, 24 genotypes). In addition, sensivity of RFLP was 100%, specificity was 97,09%. Conclusion: the most appropriate method for routine epidemiology surveillance is RFLP with non-coding region of Streptococcsu pneumoniae, then PFGE, though in some cases these results should be obligatory confirmed by MLST.

Keywords: molecular epidemiology typing, non-coding genome, Streptococcus pneumoniae, MLST

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116 Conversion of Glycerol to 3-Hydroxypropanoic Acid by Genetically Engineered Bacillus subtilis

Authors: Aida Kalantari, Boyang Ji, Tao Chen, Ivan Mijakovic

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3-hydroxypropanoic acid (3-HP) is one of the most important biomass-derivable platform chemicals that can be converted into a number of industrially important compounds. There have been several attempts at production of 3-HP from renewable sources in cell factories, focusing mainly on Escherichia coli, Klebsiella pneumoniae, and Saccharomyces cerevisiae. Despite the significant progress made in this field, commercially exploitable large-scale production of 3-HP in microbial strains has still not been achieved. In this study, we investigated the potential of Bacillus subtilis to be used as a microbial platform for bioconversion of glycerol into 3-HP. Our recombinant B. subtilis strains overexpress the two-step heterologous pathway containing glycerol dehydratase and aldehyde dehydrogenase from various backgrounds. The recombinant strains harboring the codon-optimized synthetic pathway from K. pneumoniae produced low levels of 3-HP. Since the enzymes in the heterologous pathway are sensitive to oxygen, we had to perform our experiments in micro-aerobic conditions. Under these conditions, the cell produces lactate in order to regenerate NAD+, and we found the lactate production to be in competition with the production of 3-HP. Therefore, based on the in silico predictions, we knocked out the glycerol kinase (glpk), which in combination with growth on glucose, resulted in improving the 3-HP titer to 1 g/L and the removal of lactate. Cultivation of the same strain in an enriched medium improved the 3-HP titer up to 7.6 g/L. Our findings provide the first report of successful introduction of the biosynthetic pathway for conversion of glycerol into 3-HP in B. subtilis.

Keywords: bacillus subtilis, glycerol, 3-hydroxypropanoic acid, metabolic engineering

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115 Antimicrobial Activity of Ethnobotanically Selected Medicinal Plants Used in the Treatment of Sexually Transmitted Diseases

Authors: Thilivhali Emmanuel Tshikalange, Phiwokuhle Mamba

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Ten medicinal plants used traditionally in the treatment of sexually transmitted diseases (STDs) and urinary tract infections (UTIs) were selected from an ethnobotanical database developed in Mpumalanga. The plants were investigated for their antimicrobial activity against five bacterial strains (Escherichia coli, Klebsiella oxytoca, Klebsiella pneumoniae, Neisseria gonorrhoeae and Staphylococcus aureus) and one fungal strain (Candida albicans). Eight of the plants inhibited the growth of all microorganisms at a concentration range of 0.4 mg/ml to 12.5 mg/ml. Acacia karroo showed the most promising antimicrobial activity, with a minimum inhibitory concentration (MIC) of 0.4 mg/ml on Staphylococcus aureus and 0.8 mg/ml on Neisseria gonorrhoeae. All ten plants were further investigated for their antioxidant activities using the DPPH scavenging method. Acacia karroo and Rhoicissus tridentata subsp. cuneifolia showed good antioxidant activity with IC50 values of 0.83 mg/ml and 0.06 mg/ml, respectively. The toxicity of plants was determined using the XTT reduction method against Vero cells. None of the ten plants showed toxicity on the cells. The obtained results confirmed that Acacia karroo and possibly Rhoicissus tridentata subsp. cuneifolia have the potential of being used as antimicrobial agents in the treatment of STDs and UTIs. These results support and validate traditional use of medicinal plants studied.

Keywords: antimicrobial, antioxidant, Neisseria gonorrhoeae, sexually transmitted diseases

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114 Antimicrobial Activity of Oil Extracted from the Almonds of the Fruits of Argania spinosa in the West of Algeria (Mostaganem)

Authors: Nassima Behidj-Benyounes, Nadjiba Chebouti, Thoraya Dahmane, Amina Henni

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This work examines the study of the antimicrobrial effect of oil extracted from the seeds of Argania spinosa L. (Sapotaceae) in the area of Stida (Mostaganem). This natural substance is extracted by using the Soxhlet. The antimicrobial activity of this oil is evaluated on several microorganisms. It has been tested on five bacterial strains; Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Bacillus subtilis and Staphylococcus aureus. The extract has been studied by using Candida albicans. It should be noted that these agents are characterized by a high frequency of contamination and pathogenicity. Through this study, we note that these microorganisms are moderately sensitive to the argan oil.

Keywords: Argania spinosa, oil, several microorganisms, almonds, antimicrobial activity

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113 Septic Pulmonary Emboli as a Complication of Peripheral Venous Cannula Insertion

Authors: Ankita Baidya, Vanishri Ganakumar, Ranveer S. Jadon, Piyush Ranjan, Rita Sood

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Septic embolism can have varied presentations and clinical considerations. Infected central venous catheters are commonly associated with septic emboli but peripheral vascular catheters are rarely implicated. We describe a rare case of septic pulmonary emboli related to infected peripheral venous cannulation caused by an unusual etiological agent. A young male presented with complaints of fever, productive cough, sudden onset shortness of breath and cellulitis in both the upper limbs. He was recently hospitalised for dengue fever and administered intravenous fluids through peripheral venous line. The patient was febrile, tachypneic and in respiratory distress, there were multiple pus filled bullae in left hand alongwith swelling and erythema involving right forearm that started at the site of cannulation. Chest examination showed active accessory muscles of respiration, stony dull percussion at the base of right lung and decreased breath sounds at right infrascapular, infraaxillary and mammary area. Other system examination was within normal limits. Chest X-ray revealed bilateral multiple patchy heterogenous peripheral opacities and infiltrates with right-sided pleural effusion. Contrast-enhanced computed tomography (CECT) chest showed feeding vessel sign confirming the diagnosis as septic emboli. Venous Doppler and 2D-echocardiogarm were normal. Laboratory findings showed marked leucocytosis (22000/mm3). Pus aspirate, blood sample, and sputum sample were sent for microbiological testing. The patient was started empirically on ceftriaxone, vancomycin, and clindamycin. The Pus culture and sputum culture showed Klebsiella pneumoniae sensitive to cefoperazone-sulbactum, piperacillin-tazobactum, meropenem and amikacin. The antibiotics were modified accordingly to antimicrobial sensitivity profile to Cefoperazone-sulbactum. Bronchoalveolar lavage (BAL) was done and sent for microbiological investigations. BAL culture showed Klebsiella pneumoniae with same antimicrobial resistance profile. On day 6 of starting cefoperazone-sulbactum, he became afebrile. The skin lesions improved significantly. He was administered 2 weeks of cefoperazone–sulbactum and discharged on oral faropenem for 4 weeks. At the time of discharge, TLC was 11200/mm3 with marked radiological resolution of infection and healed skin lesions. He was kept in regular follow up. Chest X-ray and skin lesions showed complete resolution after 8 weeks. Till date, only couple of case reports of septic emboli through peripheral intravenous line have been reported in English literature. This case highlights that a simple procedure of peripheral intravenous cannulation can lead to catastrophic complication of septic pulmonary emboli and widespread cellulitis if not done with proper care and precautions. Also, the usual pathogens in such clinical settings are gram positive bacteria, but with the history of recent hospitalization, empirical therapy should also cover drug resistant gram negative microorganisms. It also emphasise the importance of appropriate healthcare practices to be taken care during all procedures.

Keywords: antibiotics, cannula, Klebsiella pneumoniae, septic emboli

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112 Chemical Compositon and Antimicrobial Activity of Daucus aristidis Coss. Essential Oil in Pre-Flowering Stage from Algeria

Authors: M. Lamamra, H. Laouer, A. Adjaoud, Sahli Farida

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Essential oils can have significant antimicrobial activities and can successfully replace antibiotics that show their ineffectiveness against resistant germs. The chemical composition of the essential oil obtained by hydrodistillation from the aerial part of Daucus aristidis (Apiaceae) at the pre-flowering stage was investigated for the first time, by GC and GC-MS and evaluated for in vitro antimicrobial activity by the disk diffusion method. The Main components of D. aristidis oil were α-pinene (20.13%), cedrol (20.11%), and E- asarone (18.53%). The oil exhibited an antibacterial activity against almost strains tested except for Klebsiella pneumoniae ATCC 700603 K6 and Enterococcus faecalis ATCC 49452, the oil of D. aristidis had no activity against all fungi tested.

Keywords: α-pinene, antimicrobial activity, Daucus aridtidis, essential oil

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111 Peculiarities of Microflora of Odontogenic Inflammatory Processes in the Central Kazakhstan Region

Authors: Aliya Tokbergenova, Maida Tusupbekova, Daulet Dzhangaliyev, Alena Lavrinenko

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Background: Odontogenic phlegmons are ranked the first among pyoinflammatory processes in the frequency of hospitalization in maxillofacial surgery in the post-Soviet countries. The main role in etiology is played by obligate anaerobes and aerobes. According to numerous data, the structure of aerobic pathogens is dominated by staphylococci and gram-negative bacteria. Aim: The research aim is to study the microflora of the purulent discharge odontogenic inflammatory processes. Materials and methods: A total of 220 patients have been examined, of which 120 patients aged 25-59 years have been included in the research who did not have comorbidity hospitalized in the maxillofacial hospital in Karaganda (Kazakhstan) from January 2016 to July 2017. The bacteriological research has been carried out on the basis of the multiaccess laboratory of the KSMU, through the Matrix Assisted Laser Desorption/Ionization (MALDI) apparatus. The material sample was pus from the inflammation focus, taken during the operating period. Results: According to the research among 120 patients (100%), 15 patients (12.5%) have had microorganisms not grown. From 105 (87.5%) bacteriological results, it has been revealed the following 1) Streptococcus: 51 (42.5%): Streptococcus beta-haemolytic: 17 (14.2%), Streptococcus pneumoniae: 12 (10%), Streptococcus anginosus: 8 (6.6%), Streptococcus oralis: 8 (6.6%), Streptococcus constellatus: 6 (5.0%); 2) Staphylococci: 27 (22.5%): Staphylococci aureus: 14 (11.7%) and Staphylococci epidermidis: 13 (10.8%); 3) Pseudomonas aeruginosa: 12 (10%); 4) Neisseria: 11 (9.1%): Neisseria mucosa: 5 (4.1%) and Neisseria macacae: 6 (5.0%); 5) Klebsiella pneumoniae: 2 (1.7%); 6) Stenotrophomonas maltophilia: 2 (1.7%). 15 patients (12.5%) experienced complications in the form of 1) The dissemination of the process in 10 patients (8.4%). 2) Osteomyelitis in 3 (2.5%). 3) Mediastinitis in 1 (0.8%). 4) Sinusitis in 1 (0.8%). 15 patients (100%) were carried out repeated bacteriological examination, the following was revealed: 1) Streptococcus: 10 (66.7%): Streptococcus beta-haemolytic: 4 (26.7%), Streptococcus pneumoniae: 2 (13.3%), Streptococcus аnginosus: 2 (13.3%), Streptococcus oralis: 1 (6.7%), Streptococcus constellatus: 1 (6.7%); 2) Staphylococci: 4 (26.7%): Staphylococci aureus: 3 (20%) and Staphylococci epidermidis: 1 (6.7%); 3) Pseudomonas aeruginosa: 1 (6.7%). Conclusions: Thus, according to our research data, streptococci predominate in the odontogenic processes microflora in aerobic flora in the central Kazakhstan region, which refutes the leading role of staphylococci in the development of odontogenic inflammatory processes, thus creating prerequisites for studying new treatment approaches.

Keywords: maxillofacial surgery, microflora, odontogenic phlegmons, pyo-inflammatory

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110 Qualitative Phytochemical Screening and Antibacterial Evaluation of Sohphlang: Flemingia Vestita

Authors: J. K. D. M. P. Madara, R. B. L. Dharmawickreme, Linu John, Ivee Boiss

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Flemingia vestita, commonly known as ‘Sohphlang’ is an important medicinal plant found in the North-Eastern region of India, which is traditionally recognized for its anthelmintic properties. This study was aimed to evaluate the phytochemical constituents and antibacterial activity of the tuber skin extracts of the plant species. Methanol, acetone, and water were used to obtain the solvent extractions of the skin peel extracts. Concentrated extracts of skin peel were tested using previously established qualitative phytochemical assays. The antibacterial efficacy of methanol tuber skin extract was tested against Gram-negative and positive microorganisms, namely, Klebsiella pneumonia, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Mycobacterium tuberculosis strains. Agar well diffusion method was employed to determine the zone of inhibition of the plant extracts. Obtained data were statistically analyzed. Methanol extracts of Flemingia vestita were found to be effective against Bacillus subtilis and Mycobacterium tuberculosis at concentrations of 0.5 mg/ml. The reported zone of inhibition for the two strains was 13.3mm ± 0.57 and 16.3mm ± 4.9, respectively. However Klebsiella pneumoniae, Pseudomonas aeruginosa and Escherichia coli were resistant to the plant extracts with no zone of inhibition. Alkaloids, glycosides, and phenols were found to be present in aqueous, methanol, and acetone extracts of the plant in qualitative phytochemical analysis.

Keywords: flemingia vestita, antibacterial activity, phytochemical screening, well diffusion method

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109 Retrospective Study of Positive Blood Cultures Carried out in the Microbiology Department of General Hospital of Ioannina in 2017

Authors: M. Gerasimou, S. Mantzoukis, P. Christodoulou, N. Varsamis, G. Kolliopoulou, N. Zotos

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Purpose: Microbial infection of the blood is a serious condition where bacteria invade the bloodstream and cause systemic disease. In such cases, blood cultures are performed. Blood cultures are a key diagnostic test for intensive care unit (ICU) patients. Material and method: The BacT/Alert system, which measures the production of carbon dioxide with metabolic organisms, is used. The positive result in the BacT/Alert system is followed by culture in the following selective media: Blood, Mac Conkey No 2, Chocolate, Mueller Hinton, Chapman and Sabaureaud agar. Gram staining method was used to differentiate bacterial species. The microorganisms were identified by biochemical techniques in the automated Microscan (Siemens) system and followed by a sensitivity test on the same system using the minimum inhibitory concentration MIC technique. The sensitivity test is verified by a Kirby Bauer-based test. Results: In 2017 the Laboratory of Microbiology received 3347 blood cultures. Of these, 170 came from the ICU. 116 found positive. Of these S. epidermidis was identified in 42, A. baumannii in 27, K. pneumoniae in 12 (4 of these KPC ‘Klebsiella pneumoniae carbapenemase’), S. hominis in 8, E. faecium in 7, E. faecalis in 5, P. aeruginosa in 3, C. albicans in 3, S. capitis in 2, K. oxytoca in 2, P. mirabilis in 2, E. coli in 1, S. intermidius in 1 and S. lugdunensis in 1. Conclusions: The study of epidemiological data and microbial resistance phenotypes is essential for the choice of therapeutic regimen for the early treatment and limitation of multivalent strains, while it is a crucial factor to solve diagnostic problems.

Keywords: blood culture, bloodstream, infection, intensive care unit

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108 Microbiological Profile of UTI along with Their Antibiotic Sensitivity Pattern with Special Reference to Nitrofurantoin

Authors: Rupinder Bakshi, Geeta Walia, Anita Gupta

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Introduction: Urinary tract infections (UTI) are considered to be one of the most common bacterial infections with an estimated annual global incidence of 150 million. Antimicrobial drug resistance is one of the major threats due to widespread usage of uncontrolled antibiotics. Materials and Methods: A total number of 9149 urine samples were collected from R.H Patiala and processed in the Department of Microbiology G.M.C Patiala. Urine samples were inoculated on MacConkey’s and blood agar plates by using calibrated loop delivering 0.001 ml of sample and incubated at 37 °C for 24 hrs. The organisms were identified by colony characters, gram’s staining and biochemical reactions. Antimicrobial susceptibility of the isolates was determined against various antimicrobial agents (Hi – Media Mumbai India) by Kirby-Bauer disk diffusion method on Muller Hinton agar plates. Results: Maximum patients were in the age group of 21-30 yrs followed by 31-40 yrs. Males (34%) are less prone to urinary tract infections than females (66%). Out of 9149 urine sample, the culture was positive in 25% (2290) samples. Esch. coli was the most common isolate 60.3% (n = 1378) followed by Klebsiella pneumoniae 13.5% (n = 310), Proteus spp. 9% (n = 209), Staphylococcus aureus 7.6 % (n = 173), Pseudomonas aeruginosa 3.7% (n = 84), Citrobacter spp. 3.1 % (70), Staphylococcus saprophyticus 1.8 % (n = 142), Enterococcus faecalis 0.8%(n=19) and Acinetobacter spp. 0.2%(n=5). Gram negative isolates showed higher sensitivity towards, Piperacillin +Tazobactum (67%), Amikacin (80%), Nitrofurantoin (82%), Aztreonam (100%), Imipenem (100%) and Meropenam (100%) while gram positive showed good response towards Netilmicin (69%), Nitrofurantoin (79%), Linezolid (98%), Vancomycin (100%) and Teicoplanin (100%). 465 (23%) isolates were resistant to Penicillins, 1st generation and 2nd generation Cehalosporins which were further tested by double disk approximation test and combined disk method for ESBL production. Out of 465 isolates, 375 were ESBLs consisting of n 264 (70.6%) Esch.coli and 111 (29.4%) Klebsiella pneumoniae. Susceptibility of ESBL producers to Imipenem, Nitrofurantoin and Amikacin were found to be 100%, 76%, and 75% respectively. Conclusion: Uropathogens are increasingly showing resistance to many antibiotics making empiric management of outpatients UTIs challenging. Ampicillin, Cotrimoxazole, and Ciprofloxacin should not be used in empiric treatment. Nitrofurantoin could be used in lower urinary tract infection. Knowledge of uropathogens and their antimicrobial susceptibility pattern in a geographical region will help inappropriate and judicious antibiotic usage in a health care setup.

Keywords: Urinary Tract Infection, UTI, antibiotic susceptibility pattern, ESBL

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107 Antibacterial Activity of Methanol Extract of Punica Granatum Linn. (Punnicaceae) Fruit Peel Against Selected Bacterial Species

Authors: Afzan Mahmad, Santibuana Abd Rahman, Gouri Kumar Dash, Mohd. Syafiq Bin Abdullah

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Antibacterial activity of the methanol extract of fruit peel of Punica granatum Linn (Family: Punicaceae) was evaluated against two Gram positive and two Gram negative bacteria. The Gram positive bacteria included Staphylococcus aureus, Streptococcus pneumoniae and the Gram negative organisms included Escherichia coli and Pseudomonas aeruginosa respectively. The culture media used for antibacterial assay was Mueller Hinton agar for the growth of S. aureus, E. coli, and P. aeruginosa. The media used for the growth of S. pneumoniae was Mueller Hinton blood agar. The antibacterial assay was performed through Disc diffusion technique. The methanol extract was tested at three different concentrations (50, 100 and 200 mg/ml). Standard antibiotic discs containing vancomycin (30 μg) for S. pneumoniae, penicillin (10 units) for S. aureus, ceftriaxone (30 μg) for E. coli and ciprofloxacin (5 μg) for P. aeruginosa were used for the activity comparison. The results of the study revealed that the extract possesses antibacterial activity against S. aureus, S. pneumoniae and P. aeruginosa at all tested concentrations. The maximum zone of inhibition of 19 mm of the extract at 200 mg/ml was observed against S. pneumoniae. However, no zone of inhibition was observed against E. coli at the tested concentrations of the extract. Based on the results obtained in this study, it may be concluded that the fruit peel of P. granatum possess broad spectrum of antibacterial activity against a number bacteria.

Keywords: Punica granatum Linn., methanol extract, antibacterial, zone of inhibition

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106 A Retrospective Cross Sectional Study of Blood Culture Results in a Tertiary Hospital, Ekiti, Nigeria

Authors: S. I. Nwadioha, M. S. Odimayo, J. A. Omotayo, A. Olu Taiwo, O. E. Olabiyi

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The current study was conducted to determine the epidemiology and antibiotic sensitivity pattern of bacteria isolated from blood of septicemic patients for improved antibiotic therapy. A three-year descriptive study has been carried out at Microbiology Laboratory, Ekiti State University Teaching Hospital, Ado Ekiti, from April 2012 to April 2015. Information compiled from patients’ records includes age, sex, isolated organisms and antibiotic susceptibility patterns. Three hundred and thirteen blood cultures were collected from neonatology and pediatrics wards, Out Patients’ Department (OPD) and from other adult patients. Forty-one cultures yielded mono microbial growth (no polymicrobial growth), giving an incidence of 13.1% positive blood culture (N=41/313). There were 58.4% Gram-negative bacilli and 41.6% Gram-positive cocci in the microbial growth. Bacteria isolated were Staphylococcus aureus 34%(14/41), Klebsiella species22% (9/41), Enterococci 17%(7/41), Proteus species12%(5/41), Escherichia coli 7%(3/41) and Streptococcal pneumoniae 7%(3/41). There was a (35%) higher occurrence of septicemia in neonates than in any other age groups in the hospital. Bacterial sensitivity to 13 antibiotic agents was determined by antibiotics disc diffusion using modified Kirby Bauer’s method. Gram-positive organisms showed a higher antibiotic sensitivity ranging from 14- 100% than the Gram-negative bacteria (11-80%). Staphylococcus aureus and Klebsiella species are the most prevalent organisms. The third generation Cephalosporins (Ceftriaxone) and Floroquinolone(Levofloxacin, Ofloxacin) have proved reliable for management of these blood infections.

Keywords: blood cultures, septicemia, antibiogram, Nigeria

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105 Infrared Spectroscopy in Tandem with Machine Learning for Simultaneous Rapid Identification of Bacteria Isolated Directly from Patients' Urine Samples and Determination of Their Susceptibility to Antibiotics

Authors: Mahmoud Huleihel, George Abu-Aqil, Manal Suleiman, Klaris Riesenberg, Itshak Lapidot, Ahmad Salman

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Urinary tract infections (UTIs) are considered to be the most common bacterial infections worldwide, which are caused mainly by Escherichia (E.) coli (about 80%). Klebsiella pneumoniae (about 10%) and Pseudomonas aeruginosa (about 6%). Although antibiotics are considered as the most effective treatment for bacterial infectious diseases, unfortunately, most of the bacteria already have developed resistance to the majority of the commonly available antibiotics. Therefore, it is crucial to identify the infecting bacteria and to determine its susceptibility to antibiotics for prescribing effective treatment. Classical methods are time consuming, require ~48 hours for determining bacterial susceptibility. Thus, it is highly urgent to develop a new method that can significantly reduce the time required for determining both infecting bacterium at the species level and diagnose its susceptibility to antibiotics. Fourier-Transform Infrared (FTIR) spectroscopy is well known as a sensitive and rapid method, which can detect minor molecular changes in bacterial genome associated with the development of resistance to antibiotics. The main goal of this study is to examine the potential of FTIR spectroscopy, in tandem with machine learning algorithms, to identify the infected bacteria at the species level and to determine E. coli susceptibility to different antibiotics directly from patients' urine in about 30minutes. For this goal, 1600 different E. coli isolates were isolated for different patients' urine sample, measured by FTIR, and analyzed using different machine learning algorithm like Random Forest, XGBoost, and CNN. We achieved 98% success in isolate level identification and 89% accuracy in susceptibility determination.

Keywords: urinary tract infections (UTIs), E. coli, Klebsiella pneumonia, Pseudomonas aeruginosa, bacterial, susceptibility to antibiotics, infrared microscopy, machine learning

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104 Evaluation of Antibiotic Resistance and Extended-Spectrum β-Lactamases Production Rates of Gram Negative Rods in a University Research and Practice Hospital, 2012-2015

Authors: Recep Kesli, Cengiz Demir, Onur Turkyilmaz, Hayriye Tokay

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Objective: Gram-negative rods are a large group of bacteria, and include many families, genera, and species. Most clinical isolates belong to the family Enterobacteriaceae. Resistance due to the production of extended-spectrum β-lactamases (ESBLs) is a difficulty in the handling of Enterobacteriaceae infections, but other mechanisms of resistance are also emerging, leading to multidrug resistance and threatening to create panresistant species. We aimed in this study to evaluate resistance rates of Gram-negative rods bacteria isolated from clinical specimens in Microbiology Laboratory, Afyon Kocatepe University, ANS Research and Practice Hospital, between October 2012 and September 2015. Methods: The Gram-negative rods strains were identified by conventional methods and VITEK 2 automated identification system (bio-Mérieux, Marcy l’etoile, France). Antibiotic resistance tests were performed by both the Kirby-Bauer disk-diffusion and automated Antimicrobial Susceptibility Testing (AST, bio-Mérieux, Marcy l’etoile, France) methods. Disk diffusion results were evaluated according to the standards of Clinical and Laboratory Standards Institute (CLSI). Results: Of the totally isolated 1.701 Enterobacteriaceae strains 1434 (84,3%) were Klebsiella pneumoniae, 171 (10%) were Enterobacter spp., 96 (5.6%) were Proteus spp., and 639 Nonfermenting gram negatives, 477 (74.6%) were identified as Pseudomonas aeruginosa, 135 (21.1%) were Acinetobacter baumannii and 27 (4.3%) were Stenotrophomonas maltophilia. The ESBL positivity rate of the totally studied Enterobacteriaceae group were 30.4%. Antibiotic resistance rates for Klebsiella pneumoniae were as follows: amikacin 30.4%, gentamicin 40.1%, ampicillin-sulbactam 64.5%, cefepime 56.7%, cefoxitin 35.3%, ceftazidime 66.8%, ciprofloxacin 65.2%, ertapenem 22.8%, imipenem 20.5%, meropenem 20.5 %, and trimethoprim-sulfamethoxazole 50.1%, and for 114 Enterobacter spp were detected as; amikacin 26.3%, gentamicin 31.5%, cefepime 26.3%, ceftazidime 61.4%, ciprofloxacin 8.7%, ertapenem 8.7%, imipenem 12.2%, meropenem 12.2%, and trimethoprim-sulfamethoxazole 19.2 %. Resistance rates for Proteus spp. were: 24,3% meropenem, 26.2% imipenem, 20.2% amikacin 10.5% cefepim, 33.3% ciprofloxacin and levofloxacine, 31.6% ceftazidime, 20% ceftriaxone, 15.2% gentamicin, 26.6% amoxicillin-clavulanate, and 26.2% trimethoprim-sulfamethoxale. Resistance rates of P. aeruginosa was found as follows: Amikacin 32%, gentamicin 42 %, imipenem 43%, merpenem 43%, ciprofloxacin 50%, levofloxacin 52%, cefepim 38%, ceftazidim 63%, piperacillin/tacobactam 85%, for Acinetobacter baumannii; Amikacin 53.3%, gentamicin 56.6 %, imipenem 83%, merpenem 86%, ciprofloxacin 100%, ceftazidim 100%, piperacillin/tacobactam 85 %, colisitn 0 %, and for S. malthophilia; levofloxacin 66.6 % and trimethoprim/sulfamethoxozole 0 %. Conclusions: This study showed that resistance in Gram-negative rods was a serious clinical problem in our hospital and suggested the need to perform typification of the isolated bacteria with susceptibility testing regularly in the routine laboratory procedures. This application guided to empirical antibiotic treatment choices truly, as a consequence of the reality that each hospital shows different resistance profiles.

Keywords: antibiotic resistance, gram negative rods, ESBL, VITEK 2

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103 Antimicrobial Activity of Nauclea lotifolia (African Peach) Crude Extracts against Some Pathogenic Microorganism

Authors: Muhammad Isah Legbo

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The phytochemical screening and antimicrobial activity of Nauclea lotifolia fruit, leaf and stem-bark extracts at various concentration of (20.0,10.0, 5.0, and 2.5 mg/ml) were evaluated against some pathogenic microorganisms such as Escherichia coli, Klebsiella pneumoniae, Salmonella typhi, Staphylococcus aureus, Aspergillus niger and Candida albicans. The antimicrobial activity was assayed using agar well diffusion method. The result obtained show appreciable inhibitory effort of acetone, aqueous and methanolic extracts of Nauclea lotifolia. However, result obtained was less active compared to that of the control antibiotic (Ciprofloxacillin). The minimum inhibitory concentration (MIC) was determined using serial doubling dilution method and ranged from 5.0-10.0mg/ml, while the minimum bactericidal concentration (MBC) was determined by plating various dilution of extracts without turbidity and the result ranged from 5.0-7.5mg/ml. The phytochemical screening revealed the presence of alkaloid, anthraquinones, flavonoids, resin, steroid and saponin. The activities of the plant extract therefore justify their utilization in the treatment of various ailments associated with the test organism.

Keywords: Nauclea, lotifolia, antimicrobial, pathogens, saponin, extract

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