Preliminary Analysis of the Anti-biofilm Efficacy of Manuka Honey on Extended Spectrum Β-lactamase Producing Escherichia Coli Tem-3 and Klebsiella Pneumoniae Shv18, Associated with Urinary Tract Infections

open access articleUrinary Tract Infections (UTIs) are one of the most common infections in the UK and many other parts of the world. The prevalence of the Extended Spectrum β-Lactamases (ESBLs) producing UTIs, combined with their ability to form a bio film, has significantly risen and is limiting therapeutic options. This study investigated the anti-bio film activity of Manuka honey on two ESBL producing pathogens, Escherichia coli TEM-3 and Klebsiella pneumonia SHV18, commonly found in UTIs. The ESBL production was confirmed by the double disk synergy method used to confirm the ESBL production. The antibacterial activity of Manuka honey was determined using the agar well diffusion method. The Minimum Inhibitory Concentration (MIC) was established using serially diluted honey ranging from 50% to 1.56%. The effect of Manuka honey on the pathogen bio films was analysed using the Tissue Culture Plate method, with an established MIC and under 24h incubation with the honey. The results indicated that K. pneumonia SHV18 is a stronger bio film producer than E. coli TEM 3. 50% (w/v) MIC Manuka honey appears to fully prevent the plank tonic growth of both strains. A significant reduction of 81% of the E. coli TEM3 (p < 0.001) and 52% of the K. pneumonia SHV18 (p = 0.001) bio film biomass was observed. The E. coli bio films were found to be more sensitive to the 50% (w/v) honey dilution than those produced by K. pneumonia. The study indicated the anti-bio film potency of Manuka honey and its potential to become an alternative treatment for the ESBL producing pathogens associated with UTIs

The Efficacy of Antibiotics in the Treatment of Chlamydia Trachomatis Infections during Pregnancy. A Systematic Review and a Meta-analysis.

open access journalChlamydia trachomatis infections encountered during pregnancy can lead to many complications for both mother and neonate if left untreated. A systematic review and a meta-analysis were conducted to analyse the efficacy of antibiotic treatment infection during pregnancy that established the most effective antibiotic to treat Chlamydia trachomatis infections during pregnancy. PubMed and Cochrane Library were searched to the end of 2019. Randomised control trials that aligned with inclusion criteria of relevant antibiotics Azithromycin, Erythromycin, Clindamycin and Amoxicillin during Pregnancy was included. A systematic review and meta-analysis were used to calculate pooled (i) success of treatment for each antibiotic in several trials and (ii) the total adverse side effects for each drug encountered in several trials. Randomised controlled trials (RCTs) selected included 1917 pregnant patients with Chlamydia infections, and 851 have completed follow up assessments. Data from three RCTs observed a higher success of treatment for Azithromycin versus Erythromycin, fixed effect model, odds ratio (OR) = 0.67, 95% confidence interval (CI), 0.32-1.80. Data from three RCTs observed a higher success of treatment for Clindamycin versus Erythromycin OR = 2.43 95%, CI:2.00-2.60. Data from two RCTs showed a higher success of treatment for Amoxicillin versus Erythromycin OR = 3.74, 95%, CI:0.68-13.66. Analysis established that Erythromycin resulted in the most adverse side effects. Azithromycin was determined as the most effective antibiotic to treat C. trachomatis infections during pregnancy due to the high success of treatment. Clindamycin is capable of eradicating C. trachomatis infections in pregnancy and can be considered a secondary antibiotic treatment with fewer incidences of adverse side effects and high levels of successful treatment of the infection, however, due to a lack of studies more analysis will be required to confirm the efficacy of Clindamycin

Building on-line materials for teaching parasitology to health sciences’ students: initial impressions.

Background: It is widely recognised that the use of web-based teaching resources is an increasingly important method for delivering education, and it will be particularly important in the near future due to the progressively increasing number of health science students and the current number of academics in the “European Higher Education Area”. The study of parasitology and infectious diseases is essential to build professionals in the health sector with the key knowledge and skills to face global public health threats such as food-, water- or vector-borne infectious diseases outbreaks. However, the current time dedicated to the teaching of this discipline in all health sciences degrees at De Montfort University (DMU, Leicester, UK) is very little or non-existent depending on the degree/master. Methods: An innovative teaching group at DMU is trying to fill this gap in the currently available teaching offer in line with new trends in global health education, the large number of students enrolled in any health degree and the increasing number of students that would like to study this discipline (but due to different commitments do not have enough time or resources to study on a full time basis). Thus, an innovative teaching group from different EU Universities (DMU and the Spanish universities: University of San Pablo CEU, University of Alcalá, and University Miguel Hernández de Elche) and clinicians (University Hospitals of Leicester, UK) have started to design, create and develop a complete on-line package in Parasitology for undergraduate and postgraduate students that study health sciences. Results: The e-Parasitology package will be accessible through the DMU website (http://parasitology.dmu.ac.uk) in 2017 and will be focused on infection, prevention and treatment of major and emerging parasitological diseases. Conclusions: This teaching resource will aid our undergraduate and postgraduate students to gain a significant knowledge in parasitology by promoting self-learning and internationalization. This poster will explore one of the first mini-modules developed so far related with Toxocara, a helminthiasis with prevalence rates that can reach as high as 40% or more in parts of the world, and the challenges for its development

Systematic review of antiepileptic drugs’ safety and effectiveness in feline epilepsy

Understanding the efficacy and safety profile of antiepileptic drugs (AEDs) in feline epilepsy is a crucial consideration for managing this important brain disease. However, there is a lack of information about the treatment of feline epilepsy and therefore a systematic review was constructed to assess current evidence for the AEDs’ efficacy and tolerability in cats. The methods and materials of our former systematic reviews in canine epilepsy were mostly mirrored for the current systematic review in cats. Databases of PubMed, CAB Direct and Google scholar were searched to detect peer-reviewed studies reporting efficacy and/or adverse effects of AEDs in cats. The studies were assessed with regards to their quality of evidence, i.e. study design, study population, diagnostic criteria and overall risk of bias and the outcome measures reported, i.e. prevalence and 95% confidence interval of the successful and affected population in each study and in total

Percutaneous Pulmonary Valve Implantation in Small Conduits: A Multicenter Experience

BACKGROUND: Guidelines allow percutaneous pulmonary valve implantation (PPVI) in conduits above 16mm diameter. Balloon dilatation of a conduit to a diameter>110% of the original implant size is also not recommended. We analyzed patients undergoing PPVI in such conditions. METHODS AND RESULTS: Nine patients (May 2008-July 2016) from 8 institutions underwent PPVI in conduits 110%. Mean age and weight of the 14 patients was 12.1 (7.7 to 16) years and 44.9 (19 to 83) kg. Median conduit diameter at PPVI was 12 (10 to 17) mm. Median systolic right ventricular pressure was 70 (40 to 94) mmHg. Procedure was successful in all cases. A confined conduit rupture occurred in 7 patients (50%) and was treated with covered stent in 6. One patient experienced dislocation of 2 pulmonary artery stents that were parked distally. The post-implantation median systolic right ventricular pressure was 36 (28 to 51) mmHg. A fistula between right-ventricle outflow and aorta was found in one patient, secondary to undiagnosed conduit rupture. This was closed surgically. After a median follow-up of 20.16 (6.95 to 103.61) months, all the patients are asymptomatic with no significant RVOT stenosis. CONCLUSIONS: PPVI is feasible in small conduits but rate of ruptures is high. Although such ruptures remain contained and can be managed with covered stents in our experience, careful selection of patients and high level of expertise are necessary. More studies are needed to better assess the risk of PPVI in this population.info:eu-repo/semantics/publishedVersio

LuxS Sequence Alignment of Different Gram-Negative Bacteria, is it Conserved?

Background: Quorum sensing is a mechanism by which bacteria can communicate with each other’s using chemical signals termed autoinducers. Each single cell can detect the concentration of those signals in its surrounding. As the concentration of those signals reaches a threshold, the bacterial population is then ready to make their decision. Quorum sensing has been reported to regulate many biological processes like secondary metabolites production, biofilm formation, virulence, symbiosis, and dormancy. Autoinducer-2 (AI-2) is an important quorum sensing signal involved in inter-species communication. It is synthesized by the enzyme LuxS. In this work, we are trying to find out how much is the similarity of LuxS sequence between different Gram-negatives. This might lead to a better understanding of its evolutionary lineage and conservatively Methods: LuxS DNA complete coding sequence from Escherichia coli (K88, O17:K52:H18, O7:K1, K-12, O83:H1, O104:H4), Klebsiella pneumoniae, Salmonella typhimurium, Yersinia pestis, Dickeya dadantii, Citrobacter freundii, Vibrio fischeri, Haemophilus influenza, Shewanella oneidensis and Aeromonas hydrophila was obtained from Genebank. DNA sequence analysis was carried out by means of multiple sequence alignment using EMBOSS Clustal Omega and aligned using default setting at times with all mentioned strains and, at other times only with the E. coli strains Results: Pairwise sequence alignment data showed varied areas of similarity between different strains even within same species. Percent identity matrix showed a high percentage of similarity between two different species (Salmonella typhimurium and E. coli, 86%) and within the same species (E. coli K88 and O17:K52:H18, 99%). Also, it showed a very low percent of similarity (Dickeya dadantii and E. coli O83:H1, 33%) and only 42% between E. coli K-12 and O83:H1. The dendogram showed unexpected evolutionary pathway of LuxS of which this sequence in E. coli K88 and O104:H4 is closer to S. typhimurium much more than to the other E. coli strains Conclusions: LuxS sequence evolved dramatically even within the same species. This might reflect the effect of quorum sensing interaction effect in different environments which might be a case of convergent evolution, and the communication specificity within the same species and also with different species

Gene Expression Analysis of the AI-2-Controlled Genes and Biofilm Formation-Related Genes of the Antibiotic-Resistant Escherichia coli at Different Growth Stages

Gene Expression Analysis of the AI-2-Controlled Genes and Biofilm Formation-Related Genes of the Antibiotic-Resistant Escherichia coli at Different Growth Stages Baho S.1 and Samarasinghe S.2 1,2 De Montfort University/Leicester/LC/United Kingdom Background- Antibiotic resistance along with biofilm formation represents current major issues faced by healthcare institutions worldwide. Autoinducer-2 (AI-2), which is coded by luxS, is one of the quorum sensing signals that acquired great attention recently for its role in many bacterial activities through inducing many genes’ regulation. Most studies focus on seeking for new antibiotics to treat biofilms but very little focus on the bacterial communication signals and gene expression study at specific stages of bacterial growth. In our work, we are trying to analyze gene expression at different biofilm growth stages in antibiotic resistant Escherichia coli CTX-M-15. This might give new information about biofilm formation mechanism and the role of quorum sensing and its relation to antibiotic resistance, which in turn, might give new insights into using new therapeutic alternatives other than the traditional antibiotic. Methods The amount of biofilm formation at different time points (6, 12, 24 & 48 hours) was measured in E. coli CTX-M-15 using Tissue Culture Plate Assay (TCP). 6 wells tissue culture plates were used in this study using AB broth medium, incubated under static conditions at 37°C, and OD570 values for crystal violet absorbance rate represented biofilm amount. RNA extraction was performed for the growing strain in AB broth at the above mentioned periods of incubation plus a 0-time incubation to be the calibrator, followed by cDNA reaction. qPCR analysis was performed for the selected 7 AI-2 controlled genes plus luxS, 21 biofilm formation-related genes and CTX-M-15 resistance gene. Results Biofilm formation amount was the highest at the 6 hr time of incubation, then started to decrease by time. The expression of 2 out of 21 biofilm formation-related genes (fimA and fimC) showed the highest upregulation at 6 hr time of incubation when 0 time was chosen as the calibrator, while 10 genes at 12 hr time of incubation and 6 genes at 24 and 48 hr time of incubation were upregulated when 6 hr time was chosen as the calibrator. All the 7 AI-2 controlled genes showed their highest upregulation at the 24 hr time of incubation along with luxS and CTX-M-15 resistance gene. Conclusion qPCR and TCP analysis demonstrates that biofilm formation-related genes in comparison with biofilm amount measured by TCP can be utilized as a model to study the up- and down- regulation of such genes. Also, it shows that AI-2 has a profound effect during biofilm formation stages, and antibiotic resistance might be controlled by AI-2 as the resistance gene upregulated along with other AI-2 controlled gene

Multiple Sequence Alignment of NDM-1 DNA Sequence of Different Carbapenemases Producing Gram-Negatives

Multiple Sequence Alignment of NDM-1 DNA Sequence of Different Carbapenemase Producing Gram-Negatives Baho S.1 and Samarasinghe S.2 1,2 De Montfort University/Leicester/LC/United Kingdom Background- NDM-1 is an enzyme that confers resistance to bacteria against a broad range of β-lactam antibiotics. It is one of the most effective Carbapenemases in terms of carbapenem hydrolysis and geographical spread. Therefore, learn how its DNA sequence evolved through testing its sequence in some organisms might give an idea about the effect of geographical distribution and other environmental effects. In our work, we are trying to find out how the wide spreading of DNM-1 among different Gram-negatives could affect the conservatively of its DNA sequence i.e. the percentage of similarity and differences among NDM-1 DNA sequence. This might give possible insights into the development of new resistance against current effective antibiotics which in turns might help to the precedent event for a possible treatment. Methods From GenBank, the complete DNA coding sequences of NDM-1 of Acinetobacter (johnsonii NF114, sp. NF111, sp. NF116), Klebsiella pneumoniae (Res2011-182, K.P-UC-13, K.P-UC-14, K.P-UC-15), Citrobacter freundii NF109, Escherichia coli NF113, Providencia vermicola NF115 and Pseudomonas sp. NF117 were obtained (813 bp for each). DNA sequence analysis was carried out by means of multiple sequence alignment using EMBOSS Clustal Omega and aligned using default setting with all mentioned strains. Results Percent identity matrix showed a 100% of similarity between all Klebsiella strains, it also showed a 100% similarity between Acinetobacter strains, C. freundii NF109, E. coli NF113, P.vermicola NF115 and Pseudomonas sp. NF117. But showed only 44.12% of similarity between all the above-mentioned strains with K. pneumoniae strains. The cladogram showed unexpected evolutionary pathway of NDM-1 between Klebsiella strains and A. johnsonii and A. sp. NF111, compared with A. sp. NF116 and other tested strains. Conclusion NDM-1 sequence evolved dramatically when transferred to some species while still conserved with others. The evolution might arise as a result of a mutation in the resistance sequence while being conserved among different species could be attributed to the convergent evolution

Genotypic Identification of ESBL Producing Urinary Tract Infections In Leicestershire Area, UK

Background: Uropathogenic E. coli is one of the highest producers of extended-spectrum β-lactamases (ESBLs), a major public health concern. Many studies have described the epidemiology and prevalence of ESBL-producing E.coli. (1) Plasmids can transfer between different species of bacteria, carrying resistance genes with them, facilitating the spread of resistance. This study aimed to investigate the prevalence of ESBL genes and plasmid types in Leicestershire, UK. Methods: 380 uropathogenic E. coli ESBL-producing isolates were collected from the Leicester Royal Infirmary in Leicestershire, UK. Isolates were confirmed to be ESBL producers using the MAST ESBL detection kit. This study investigated the presence of ESBL-producing genes (blaCTX-M, blaSHV, blaTEM and blaOXA) by multiplex PCR.A second multiplex PCR assay identified blaCTX-M-1, blaCTX-M-2, blaCTX-M-8, blaCTX-M-9 and blaCTX-M-25. To investigate the relationship between plasmid type and ESBLs, a multiplex PCR-based replicon typing assay was designed to detect IncFIA, IncI1, IncL/M, IncN and IncFII. Results: Prevalence of ESBL genes was as follows: blaCTX-M (37%) blaOXA (5%), blaTEM and blaSHV (2.4%). Multiple resistance genes were detected in 10.2% of isolates. In the second multiplex assay, the order of prevalence of CTX-M genes was as follows: blaCTX-M-1 (56%), blaCTX-M-9 (11%), blaCTX-M-25 (6%), blaCTX-M-8 (0.5%) and blaCTX-M-2 (0.2%). Multiple resistance genes were detected in 3 isolates. blaCTX-M was found to be associated with all the replicon genes other than incL/M. Conclusions: To our knowledge, this is the first study to analyse the prevalence of uropathogenic ESBLs in Leicestershire. Our findings are in line with other authors findings in Europe. Bacteria use plasmids to transfer genes, including those that cause antibiotic resistance. It was found that CTX-M producing E.coli are associated with multiple plasmids, which can be linked to its rapid spread across the world. Epidemiology and prevalence studies help to inform policy about antibiotic stewardship, with the aim to reduce resistance levels in the future

Adaptability to Various Growth Conditions of Biofilm Associated Extended-Spectrum-Beta-Lactamases Producing Bacteria

Open access article. The Publisher's final version can be found by following the URI link.Extended-Spectrum β-Lactamase (ESBL) producing bacteria are becoming increasingly prevalent in biofilmassociated infections. Bacteria form biofilms that allow their survival in hostile environments. The amount of formed biofilm is affected by external environmental factors. This study investigates the effect of specific parameters (media type, incubation condition, and growth stage) on the amount of produced biofilm on antibiotic resistant bacterial strains, Escherichia coli (CTX-M-15, TEM-3, and IMP-type) and Klebsiella pneumoniae (OXA-48, SHV-18, NDM-1, and KPC-3). The amount of biofilm formed was measured at different time points (6, 12, 24 and 48 h) of incubations under static and shaking conditions, using three different types of media (nutrient broth, LB broth, and AB broth). Statistical tests showed that there was a significant difference in biofilm level (p<0.01) for 64 out of 80 tests (80%) when grown under different types of media. Growing under different incubation conditions also showed a statistical difference in biofilm level (p<0.05) for 76 out of 120 tests (63%). Stage of growth of the same species also showed statistical difference, 20 out of 24 tests (83%) for E. coli and 24 out of 24 tests (100%) for K. pneumoniae. These findings suggested that biofilm formation is highly affected by incubation conditions, strains’ stage of growth, and media type demonstrating that these conditions may play a role in adaptability of the ESBLs on different environmental conditions and their increased prevalence in biofilm associated infections