Terms Of Antibiotic Resistance Research Articles

Differences in Biofilm Formation by Methicillin-Resistant and Methicillin-Susceptible Staphylococcus aureus Strains.

Staphylococcus aureus (S. aureus) is a common pathogen involved in community- and hospital-acquired infections. Its biofilm formation ability predisposes it to device-related infections. Methicillin-resistant S. aureus (MRSA) strains are associated with more serious infections and higher mortality rates and are more complex in terms of antibiotic resistance. It is still controversial whether MRSA are indeed more virulent than methicillin-susceptible S. aureus (MSSA) strains. A difference in biofilm formation by both types of bacteria has been suggested, but how only the presence of the SCCmec cassette or mecA influences this phenotype remains unclear. In this review, we have searched for literature studying the difference in biofilm formation by MRSA and MSSA. We highlighted the relevance of the icaADBC operon in the PIA-dependent biofilms generated by MSSA under osmotic stress conditions, and the role of extracellular DNA and surface proteins in the PIA-independent biofilms generated by MRSA. We described the prominent role of surface proteins with the LPXTG motif and hydrolases for the release of extracellular DNA in the MRSA biofilm formation. Finally, we explained the main regulatory systems in S. aureus involved in virulence and biofilm formation, such as the SarA and Agr systems. As most of the studies were in vitro using inert surfaces, it will be necessary in the future to focus on biofilm formation on extracellular matrix components and its relevance in the pathogenesis of infection by both types of strains using in vivo animal models.

Benzalkonium chloride forces selective evolution of resistance towards antibiotics in Salmonella enterica serovar Typhimurium

BackgroundAlthough food-grade disinfectants are extensively used worldwide, it has been reported that the long-term exposure of bacteria to these compounds may represent a selective force inducing evolution including the emergence of antibiotic resistance. However, the mechanism underlying this correlation has not been elucidated. This study aims to investigate the genomic evolution caused by long-term disinfectant exposure in terms of antibiotic resistance in Salmonella enterica Typhimurium. MethodsS. Typhimurium isolates were exposed to increasing concentrations of benzalkonium chloride (BAC) and variations of their antibiotic susceptibilities were monitored. Strains that survived BAC exposure were analyzed at whole genome perspective using comparative genomics, and Sanger sequencing-confirmed mutations in ramR gene were identified. Next, the efflux activity in ramR-mutated strains shown as bisbenzimide accumulation and expression of genes involved in AcrAB-TolC efflux pump using quantitative reverse transcriptase PCR were determined. ResultsMutation rates of evolved strains varied from 5.82 × 10−9 to 5.56 × 10-8, with fold increase from 18.55 to 1.20 when compared with strains evolved without BAC. Mutations in ramR gene were found in evolved strains. Upregulated expression and increased activity of AcrAB-TolC was observed in evolved strains, which may contribute to their increased resistance to clinically relevant antibiotics. In addition, several indels and point mutations in ramR were identified, including L158P, A37V, G42E, F45L, and R46H which have not yet been linked to antimicrobial resistance. Resistance and mutations were stable after seven consecutive cultivations without BAC exposure. These results suggest that strains with sequence type (ST) ST34 were the most prone to mutations in ramR among the three STs tested (ST34, ST19, ST36). ConclusionsThis work demonstrated that disinfectants, specifically BAC forces S. Typhimurium to enter a specific evolutionary trajectory towards antibiotic resistance illustrating the side effects of long-term exposure to BAC and probably also to other disinfectants. Most significantly, this study provides new insights in understanding the emergence of antibiotic resistance in modern society.

Cupuassu juice fermentation by Lactiplantibacillus plantarum Lp62 produces an antioxidant enriched probiotic beverage.

The present work aimed to produce a cupuassu juice (Theobroma grandiflorum) fermented by the probiotic bacterium Lactiplantibacillus plantarum Lp62 and to analyze its antioxidant potential, antimicrobial activity, and resistance to biological barriers. The fermented beverage showed an increase in the content of phenolics, flavonoids, and antioxidant potential. The culture showed antagonistic activity against pathogens, but this result was not observed when the juice was tested. The probiotic strain remained viable under refrigeration, even in an acidified environment, and survived simulated gastrointestinal transit in vitro. L. plantarum Lp62 showed 30% adherence to HT-29 intestinal cells and proved to be safe in terms of antibiotic resistance and production of virulence factors. Fermentation increased the functional characteristics of cupuassu juice. This drink proved to be a good vehicle for the delivery of the probiotic bacteria L. plantarum Lp62.

ДИНАМИКА АНТИБИОТИКОРЕЗИСТЕНТНОСТИ КЛИНИЧЕСКИХ ИЗОЛЯТОВ KLEBSIELLA PNEUMONIAE, ВЫДЕЛЕННЫХ ОТ ПАЦИЕНТОВ С ВТОРИЧНОЙ БАКТЕРИАЛЬНОЙ ИНФЕКЦИЕЙ ПРИ COVID-19 ЗА ПЕРИОД 2019–2021 гг.

Gram-negative bacteria (GAB) continue to lead in terms of antibiotic resistance among the causative agents of opportunistic infections. K.pneumoniae occupies a special place among GOBs, causing secondary bacterial infections with a high frequency in patients with COVID-19. Objective. To study the dynamics of sensitivity/resistance to antibiotics of K.pneumoniae clinical isolates isolated from biological material in patients with secondary bacterial infection with COVID-19. Materials and methods. In terms of sensitivity/resistance, 535 clinical isolates of K.pneumoniae isolated from patients with COVID-19 in various regions of the Republic of Belarus in the period 2019–2021 were studied. Results. The proportion of clinical isolates of K.pneumoniae resistant to antibiotics against the background increased in 2021 compared to 2019 and was for carbapenems in 2019 ertapenem – 46,4±3,35%, imipenem – 35,8±3,19% and meropenem – 39,6±3,28% and in 2021 ertapenem 66,7±4,74% (p=0,000603), imipenem 54,4±4,66% (p=0,001253) and meropenem 54,5±4,49% (p=0,008182). K.pneumoniae showed a high frequency of resistance to colistin: against the background of COVID-19 in 2019 – 95,0±4,87%, in 2020 – 100,0±0,0% (p=0,311608), in 2021 – 55,3±7,25% (p=0,000046). Conclusion. Among clinical isolates of K.pneumoniae, widespread resistance to most antibiotics was observed, as well as an increase in resistance to carbapenems.

Implications of bacteriological study in complicated and uncomplicated acute appendicitis.

Bacteriological sample in the presence of intraabdominal free fluid is necessary to adapt the antibiotherapy and to prevent the development of resistance. The aim was to evaluate the differences between uncomplicated (UAA) and complicated acute appendicitis (CAA) in terms of bacterial culture results and antibiotic resistance, and to evaluate the factors linked with CAA. We performed a single-center, retrospective observational study of all consecutive patients who presented with appendicular peritonitis and underwent emergent surgery in a tertiary referral hospital in Brussels, Belgium, between January 2013 and December 2020. The medical history, parameters at admission, bacterial culture, antibiotic resistance, and postoperative outcomes of 268 patients were analyzed. UAA was considered catarrhal or phlegmonous inflammation of the appendix. CAA was considered gangrenous or perforated appendicitis. Positive microbiological cultures were significantly higher in the CAA group (68.2% vs. 53.4%). The most frequently isolated bacteria in UAA and CAA cultures were Escherichia coli (37.9% and 48.6%). Most observed resistances were against ampicillin (28.9% and 21.7%) and amoxicillin/clavulanic acid (16.4% and 10.5%) in UAA and CAA, respectively. A higher Charlson comorbidity index, an elevated white blood cell count, an open procedure, and the need for drainage were linked to CAA. Culture results, group of bacterial isolation, and most common isolated bacteria were not related to CAA. CAA presented a higher rate of positive cultures with increased identification of gram-negative bacteria. Bacterial culture from the peritoneal liquid does not reveal relevant differences in terms of antibiotic resistance.

Acinetobacter baylyi Strain BD413 Can Acquire an Antibiotic Resistance Gene by Natural Transformation on Lettuce Phylloplane and Enter the Endosphere.

Antibiotic resistance spread must be considered in a holistic framework which comprises the agri-food ecosystems, where plants can be considered a bridge connecting water and soil habitats with the human microbiome. However, the study of horizontal gene transfer events within the plant microbiome is still overlooked. Here, the environmental strain Acinetobacter baylyi BD413 was used to study the acquisition of extracellular DNA (exDNA) carrying an antibiotic resistance gene (ARG) on lettuce phylloplane, performing experiments at conditions (i.e., plasmid quantities) mimicking those that can be found in a water reuse scenario. Moreover, we assessed how the presence of a surfactant, a co-formulant widely used in agriculture, affected exDNA entry in bacteria and plant tissues, besides the penetration and survival of bacteria into the leaf endosphere. Natural transformation frequency in planta was comparable to that occurring under optimal conditions (i.e., temperature, nutrient provision, and absence of microbial competitors), representing an entrance pathway of ARGs into an epiphytic bacterium able to penetrate the endosphere of a leafy vegetable. The presence of the surfactant determined a higher presence of culturable transformant cells in the leaf tissues but did not significantly increase exDNA entry in A. baylyi BD413 cells and lettuce leaves. More research on HGT (Horizontal Gene Transfer) mechanisms in planta should be performed to obtain experimental data on produce safety in terms of antibiotic resistance.

Antibiotic prophylaxis at the time of dental implant placement: a cost-effectiveness analysis

BackgroundAntibiotic prophylaxis during implant placement may improve implant short term survival. Nevertheless, use of antibiotics carries risks of adverse effects and antibiotic resistance. The aim of the present study is to compare the use of antibiotics in dental implant procedures in terms of costs and effectiveness.MethodsA decision-tree model was developed using TreeAge Pro Healthcare software. Two strategies were compared: Antibiotics and No antibiotics in implant placement procedures. The costs were calculated considering direct costs for implant placement, antibiotic costs, and costs for implant replacement in case of failure. Effectiveness was defined in terms of General Oral Health Assessment Index. Outcomes were evaluated as Incremental Cost Effectiveness Ratio (ICER). One-way sensitivity analysis and Probabilistic Sensitivity Analysis were performed for the most influential variables to test parameter uncertainty. Patient and healthcare perspectives were considered.ResultsAntibiotic prophylaxis resulted to be cost-effective compared to no use of antibiotics (ICER = 14,692,64 and ICER = 3841,18, respectively for patient’s and healthcare perspective). The cost of antibiotics, cost of implant replacement in case of failure and probability of adverse effects significantly influenced the results.ConclusionsFrom an individual patient perspective, antibiotic strategy can be considered cost-effective, even when the cost of antibiotic therapy increases. We can conclude that the administration of antibiotics in association with implant placement is recommended in clinical practice, as it increases the success rate and makes the treatment more effective. However, attention should be placed when healthcare perspective is considered, particularly in terms of antibiotic resistance that may impact public health and associated costs.

The Exploration of Complement-Resistance Mechanisms of Pathogenic Gram-Negative Bacteria to Support the Development of Novel Therapeutics.

Resistance to antibiotics in Bacteria is one of the biggest threats to human health. After decades of attempting to isolate or design antibiotics with novel mechanisms of action against bacterial pathogens, few approaches have been successful. Antibacterial drug discovery is now moving towards targeting bacterial virulence factors, especially immune evasion factors. Gram-negative bacteria present some of the most significant challenges in terms of antibiotic resistance. However, they are also able to be eliminated by the component of the innate immune system known as the complement system. In response, Gram-negative bacteria have evolved a variety of mechanisms by which they are able to evade complement and cause infection. Complement resistance mechanisms present some of the best novel therapeutic targets for defending against highly antibiotic-resistant pathogenic bacterial infections.

Antibiotic susceptibility of Bacillus velezensis.

We evaluated the antibiotic minimum inhibitory concentrations (MICs) of 123 Bacillus velezensis strains predominantly isolated from fermented soybean foods from Korea. When the 2018 European Food Safety Authority breakpoint values for Bacillus spp. were applied, all the strains were sensitive to chloramphenicol, clindamycin, erythromycin, gentamicin, kanamycin, tetracycline and vancomycin, and eight strains (6.5%) were resistant to streptomycin. The population distribution in MIC tests with streptomycin was continuous and the profile was clearly different from that expected for acquired antibiotic resistance. As of 25 October 2021, there were 181 complete published genomes of B. velezensis strains; 175 (96.7%) and 136 (75.2%) of these strains, respectively, possess potential tetracycline and streptomycin resistance genes tetL and ant(6) in the chromosome. In Bacillus licheniformis, SpeG confers resistance to clindamycin and there is an 'speG' gene annotated in the genomes of 180 B. velezensis strains; however, the gene products exhibit≤26.6% amino acid identity with that from B. licheniformis DSM 13T. All the potential antibiotic resistance genes in the 181 B. velezensis strains were intrinsic, and traits of lateral gene transfer were not found. In this context, B. velezensis may not present a high risk in terms of antibiotic resistance in food fermentation or human use.

Carbapenem Resistance in Acinetobacter nosocomialis and Acinetobacter junii Conferred by Acquisition of blaOXA-24/40 and Genetic Characterization of the Transmission Mechanism between Acinetobacter Genomic Species.

ABSTRACTCarbapenem resistance is increasing among Gram-negative bacteria, including the genus Acinetobacter. This study aimed to characterize, for the first time, the development of carbapenem resistance in clinical isolates of Acinetobacter junii and Acinetobacter nosocomialis conferred by the acquisition of a plasmid-borne blaOXA-24/40 gene and also to characterize the dissemination of this gene between species of Acinetobacter. Carbapenem-resistant A. nosocomialis HUAV-AN66 and A. junii HUAV-AJ77 strains were isolated in the Arnau de Vilanova Hospital (Spain). The genomes were sequenced, and in silico analysis were performed to characterize the genetic environment and the OXA-24/40 transmission mechanism. Antibiotic MICs were determined, and horizontal transfer assays were conducted to evaluate interspecies transmission of OXA-24/40. Carbapenems MICs obtained were ≥64 mg/L for HUAV-AN66 and HUAV-AJ77. Genome analysis revealed the presence in both strains of a new plasmid, designated pHUAV/OXA-24/40, harboring the carbapenem-resistance gene blaOXA-24/40 and flanked by sequences XerC/XerD. pHUAV/OXA-24/40 was successfully transferred from A. nosocomialis and A. junii to a carbapenem-susceptible A. baumannii strain, thus conferring carbapenem resistance. A second plasmid (pHUAV/AMG-R) was identified in both clinical isolates for the successful horizontal transfer of pHUAV/OXA-24/40. blaOXA-24/40-carrying plasmids of the GR12 group and showing high identity with pHUAV/OXA-24/40 were identified in at least 8 Acinetobacter species. In conclusion the carbapenemase OXA-24/40 is described for the first time in A. nosocomialis and A. junii. In both isolates the blaOXA-24/40 gene was located in the GR12 pHUAV/OXA-24/40 plasmid. GR12 plasmids are implicated in the dissemination and spread of carbapenem resistance among Acinetobacter species.IMPORTANCE Acinetobacter baumannii is one of the most relevant pathogens in terms of antibiotic resistance. The main resistance mechanisms are the carbapenem-hydrolyzing class D β-lactamases (CHDLs), especially OXA-23 and OXA-24/40. In addition to A. baumannii, there are other species within the genus Acinetobacter, which in general exhibit much lower resistance rates. In this work we characterize for the first time two clinical isolates of Acinetobacter nosocomialis and Acinetobacter junii, isolated in the same hospital, carrying the carbapenemase OXA-24/40 and displaying high resistance rates to carbapenems. By means of bioinformatics analysis we have also been able to characterize the mechanism by which this carbapenemase is horizontally transferred interspecies of Acinetobacter spp. The dissemination of carbapenemase OXA-24/40 between non-baumannii Acinetobacter species is concerning since it prevents the use of most β-lactam antibiotics in the fight against these resistant isolates.

Screening Chemical Composition and Bioactive Properties of Mentha x piperita L. Essential Oil and Extract

In recent years, medicinal and aromatic herbs, which contain variety of antimicrobial compounds and have no risk to human health in terms of antibiotic resistance, are increasingly used as alternatives to antibiotics. In the study, chemical composition, total phenolic content, antioxidant and antimicrobial activity of essential oil and extract of Mentha x piperita L. growing wild in Eskisehir were investigated. Carvone (55.8%), limonene (12.8%), 1.8-cineole (8.7%) and trans-dihydrocarvone (6.4%) were determined as the main constituents of the essential oil, while the major components of the extract were carvone (56.4%), 1.8-cineole (14.1%) and trans-dihydrocarvone (8.4%). The total phenolic contents of essential oil (2204.33 µg GAE/g) were significantly higher than of extract (744 µg GAE/g). The essential oil and the extract were inhibited 84.08% and 42.59% by the DPPH radical, 91.87% and 48.40% by the ABTS radical, respectively. The highest inhibition zone was observed against S. aureus (23 mm). However, minimum inhibition concentration (MIC) values of the essential oil were ranged between 2.5% and 10% (v/v), while MIC values of the extract were determined as 10% and >10% (v/v). The present study demonstrated that M. piperita could be used in pharmaceutical and food applications as a natural antioxidant and antimicrobial substance.

Effect of Promoter Region Sequence Variations in Relation to Antibiotic Resistance of Methicillin- Resistant Staphylococcus Aureus

Multidrug resistant bacterial strains are one of the concerns of healthcare sector. Among these, Methicillin- resistant Staphylococcus aureus (MRSA) is one of the major hospitals acquired multidrug resistant strains. MRSA shows it is resistant against methicillin and other penicillin like antibiotics by producing PBP2a (penicillin binding protein 2a) which is encoded by its mecA gene. In this process mecR1 and mecI gene act as regulator of mecA gene. The aim of this study is to identify sequence variance located in mecA, mecR1 and mecI promoter region and the effect of those changes in relation to pathogenicity of these MRSA strains. In this current research, work wound infections samples were collected from 90 patients who were infected during post-surgical mamagement. Samples were collected from Rajshahi Medical college hospital, Bangladesh. In antibiotic sensitivity test, MRSA was found 100% sensitive against only Choramphenicol and Bacitracin. It also shows partial resistance against Amikacin, Impenem, Doxycycline, Gentamycin and Neomycin. In HRM, analysis six types of genotypes (TT, AA, CC, GG, GA and CT) were identified in mecA, mecR1 and mecI genes. Two types of mutation (T>C and G>A) were found in current study. This HRM analysis was further correlate with antibiotic sensitivity test in terms of antibiotic resistance and sequence variance.

Antibiotic Resistance and Pathogenomics of Staphylococci Circulating in Novosibirsk, Russia.

A total of 394 strains of staphylococci found in humans and pets in Novosibirsk, Siberian Russia, were characterized in terms of antibiotic resistance and corresponding genes. Two coagulase-positive and 17 coagulase-negative species were identified. The majority of isolates, with the exception of S. haemolyticus and hospital S. epidermidis isolates, were sensitive to most of the tested antibiotics, and isolates from pets displayed the lowest level of resistance. Nevertheless, methicillin-resistant (MRS) and/or multidrug-resistant (MDR) isolates were found in all prevailed species, including coagulase-negative. A set of genes corresponding to the detected resistance was identified: mecA (beta-lactam resistance), aac(6′)-Ie-aph(2″)-Ia, aph(3′)-IIIa, ant(4′)-Ia (aminoglycoside-modifying enzymes), ermA/ermC, and msrA (macrolide resistance). Complete genome analysis for ten MDR S. epidermidis and five MDR S. haemolyticus isolates revealed additional antibiotic resistance genes mphC, qacA/qacB, norA, dfrC/dfrG, lnuA, BseSR, and fosB. NorA, dfrC, and fosB were present in all S. epidermidis genomes, whereas mphC and msrA were identified in all S. haemolyticus ones. All investigated MDR S. epidermidis and four of five S. haemolyticus strains were moderate or strong biofilm producers, whereas multiple genes responsible for this function and for virulence and pathogenicity were identified mostly in S. epidermidis, but were less frequently represented in S. haemolyticus.

Comparative study of Staphylococcus aureus from burn patients and healthcare workers in aburn center, Yazd, Iran.

Healthcare workers (HCWs) are proposed as the potential source of transmission of Staphylococcus aureus to hospitalized patients, especially in burn units. This study aimed to investigate S.aureus from burn wound infections and those from the nose of HCWs in terms of antibiotic resistance, the presence of Panton-Valentine leucocidin-encoding gene (pvl) and the arginine catabolic mobile element (ACME), and the ability for biofilm formation. Also, the genetic diversity of isolates was assessed using staphylococcal proteinA (spa) typing and staphylococcal cassette chromosome mec (SCCmec) typing. Overall, regarding the studied factors, significant differences were found neither between isolates from patients and HCWs nor between methicillin-resistant and methicillin-susceptible isolates (except for multidrug resistance which was significantly higher in MRSA). The most frequent SCCmec types were typeI andIII. ACME-arcA was only detected in isolates from patients and similarly the presence of ACME-opp3 was the most prevalent in this group. The presence of common clonal complexes among patient isolates and more importantly between isolates from patients and HCWs is warning. The high prevalence of virulence factors, both in MRSA and MSSA, emphasizes the importance of MSSA in burn centers. Finding no significant difference in the presence of virulence-associated factors between isolates from patients and HCWs demonstrates the need to take HCWs into account as important reservoirs.

Microbial communities of biofilms developed in a chlorinated drinking water distribution system: A field study of antibiotic resistance and biodiversity

Antibiotic resistance and biodiversity were investigated in microbial communities attached to inner surfaces of water supply fittings in a chlorinated drinking water distribution system (DWDS) supplied by two independent water treatment plants (WTPs) drawing the same source water. The investigation of the effect of the season, the applied water treatment technology, and type, material, and age of water supply fittings on both antibiotic resistance and biodiversity in biofilms involved collection of tubercles during summer and winter seasons throughout the DWDS. A total of 16 samples were collected (8 per season) from areas supplied by two independent WTPs. Culturable aerobic antibiotic resistant bacteria (ARB) proved more prevalent in summer. Various antibiotic resistance genes (ARGs) were detected, confirming the role of biofilms as ARGs reservoirs, but the abundances of quantified genes (sulI, ermB, qacEΔ1, intI1) were low (a range of <LOQ to 2313 gene copies/mg dry mass of tubercles) throughout the DWDS. In terms of microbial community composition, Proteobacteria were dominant in each sample (51.51–97.13%), and the most abundant genus was Desulfovibrio (0.01–66.69%) belonging to sulphate-reducing bacteria. Biodiversity of microbial communities was shaped by many coexisting factors, including season, water supply fitting material, and sampling site location. Spatial distribution analysis revealed that although only samples collected at the same sampling sites were similar to each other in terms of antibiotic resistance, some samples collected in the close proximity were similar in terms of biodiversity. This suggests that antibiotic resistance spreads only locally over small distances in drinking water biofilms. Although actual drinking water biofilms have been previously investigated in terms of microbial biodiversity, this is the first study that characterised both antibiotic resistance and biodiversity of microbial communities attached to inner surfaces of a real DWDS functioning for decades.

Direct identification of bacterial and human proteins from infected wounds in living 3D skin models

Trauma is one of the leading causes of death in people under the age of 49 and complications due to wound infection are the primary cause of death in the first few days after injury. The ESKAPE pathogens are a group of bacteria that are a leading cause of hospital-acquired infections and a major concern in terms of antibiotic resistance. Here, we demonstrate a novel and highly accurate approach for the rapid identification of ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.) directly from infected wounds in 3D in vitro skin models. Wounded skin models were inoculated with bacteria and left to incubate. Bacterial proteins were identified within minutes, directly from the wound, by liquid extraction surface analysis mass spectrometry. This approach was able to distinguish closely related strains and, unlike genomic approaches, can be modified to provide dynamic information about pathogen behaviour at the wound site. In addition, since human skin proteins were also identified, this method offers the opportunity to analyse both host and pathogen biomarkers during wound infection in near real-time.

Evaluation of Antibiotic Resistance Trend among Cystic Fibrosis Patients: A Single Center Study from 2014-2019

Background: Antibiotic resistance in Cystic Fibrosis (CF) patients has been a serious issue in their survival. The objective of this study was to investigate the antibiogram trend in serial admissions among CF patients. Materials and Methods: In this retrospective descriptive-analytical study, from 2014 to 2019, 95 known cases of CF patients with repeated admissions at Masih Daneshvari Hospital, Tehran, Iran, were evaluated. Demographic data and para-clinical parameters were evaluated. Fourteen antibiotic disks were used to determine antibiotic resistance. Resistance trend in 2014 and 2018 was compared. Results: Out of 95 patients, 48 patients (52.2%) were female. The mean age of patients was 19.43 ± 6.7 years. Pseudomonas positive culture in the first admission was 68.4%; while it was 45.3% in the second admission and 28.4% in the third admission (p> 0.05). Staph positive culture was 20%; 18.9%, and 16.8%, respectively (p> 0.05). Pseudomonas aeruginosa was the most sensitive to vancomycin (93.8%), followed by colistin (93.3%) and ceftazidime (65.2%). There was no correlation between patients' age and multidrug resistance (MDR). FEV1 was significantly lower in both the patient with positive pseudomonas (p-value: 0.01), and culture and MDR (p= 0.023). Furthermore, in terms of antibiotic resistance over time, resistance to colistin statistically decreased from 25% in 2014 to 2% in 2018 (p= 0.02). Conclusion: Vancomycin, Colistin, Ceftazidim, Imipenem, Amikacin, and Gentamycin had the highest drug sensitivity; while Cefotaxime, Clindamycin, and Chloramphenicol antibiotics had a low sensitivity. From 2014 to 2019, resistance to Colistin dramatically decreased.

Identification and Antibiotic Resistance Assessment of Ensifer adhaerens YX1, a Vitamin B12 -Producing Strain Used as a Food and Feed Additive.

This study provides phenotypic and molecular analyses of the antibiotic resistance of Ensifer adhaerens strain YX1 (CICC 11008s), a strain that was identified using a polyphasic taxonomy approach. The antibiotic resistance profile of E. adhaerens YX1 was assessed using the Clinical & Laboratory Standards Inst. (CLSI) method. The strain was susceptible to ciprofloxacin, levofloxacin, norfloxacin, ofloxacin, gentamicin, tobramycin, chloramphenicol, tetracycline, imipenem, and ceftazidime, and resistant to kanamycin, streptomycin, fosfomycin, and nitrofurantoin. The antibiotic resistance genes nsfA, nsfB, fosA, aph, and aadA1 were not detected in E. adhaerens YX1 via PCR using gene-specific primers. Subsequently, the genome sequence of E. adhaerens was screened for antibiotic genes. Although no antibiotic resistance genes were identified using the ResFinder database, five genes copies of one resistance gene, adeF, were detected using the Comprehensive Antibiotic Resistance Database (CARD). The results of this study will be useful for understanding the phenotypic and genotypic aspects of E. adhaerens antibiotic resistance. No safety issues were identified for E. adhaerens YX1 in terms of antibiotic resistance. Performing similar studies will be conducive to the safety assessment and control of the use of E. adhaerens in the food and feed industry. PRACTICAL APPLICATION: Few relevant reports are currently available regarding antibiotic resistance assessments or other safety evaluations for Ensifer adhaerens. Because of a lack of relevant information on the safety of this bacterium, including the genetic basis of antibiotic resistance in the production strain, it has not been recommended for use in the "qualified presumption of safety" (QPS) list and subsequent updated lists. The current study shows no safety issue of E. adhaerens YX1 in terms of its antibiotic resistance. These results are important as they provide an initial basis for an understanding of the antibiotic resistance/susceptibility of E. adhaerens YX1 (CICC 11008s), which produces vitamin B12 and is widely used in the food and feed industry.

Efficacy of Short Course Antibiotic Prophylaxis in Surgical Treatment of Closed Fracture

Introduction: Unnecessary and rampant use of antibiotic for perioperative prophylaxis are not only increasing cost of treatment but also are hazardous in terms of increasing antibiotic resistance, super infection with resistant pathogens and antibiotic toxicity.&#x0D; Objective: Our primary objective is to assess the efficacy of short course antibiotic prophylaxis in preventing surgical site infection (SSI) in surgery of closed orthopedic fracture.&#x0D; Methodology: Patients fulfilling all the inclusion criteria and willing and able to give informed consent were included in the study. All participants were treated by standard surgical technique following standard surgical protocol. Short course antibiotic prophylaxis was administered in peri-operative period to all patients in given format: Three divided dose of intravenous 1.5 gram cefuroxime during peri-operative period followed by no intravenous or oral antibiotics; 1st dose half an hour before incision; 2nd and 3rd dose repeated at 12 hours interval.&#x0D; Results: Of total 100 patient included, three developed surgical site infection till one year follow-up. All three patients with surgical site infection were treated successfully with intravenous antibiotic based on the culture sensitivity report. The average cost of antibiotic for short course antibiotic prophylaxis was 1500 NPR for each patient.&#x0D; Conclusions: There was no significant increase in rate of surgical site infection with short course antibiotic prophylaxis, rather it was less expensive and may have less adverse effect, in terms of antibiotic resistance and antibiotic side effects on long run.

Biofilm Forming Antibiotic Resistant Gram-Positive Pathogens Isolated From Surfaces on the International Space Station.

The International Space Station (ISS) is a closed habitat in a uniquely extreme and hostile environment. Due to these special conditions, the human microflora can undergo unusual changes and may represent health risks for the crew. To address this problem, we investigated the antimicrobial activity of AGXX®, a novel surface coating consisting of micro-galvanic elements of silver and ruthenium along with examining the activity of a conventional silver coating. The antimicrobial materials were exposed on the ISS for 6, 12, and 19 months each at a place frequently visited by the crew. Bacteria that survived on the antimicrobial coatings [AGXX® and silver (Ag)] or the uncoated stainless steel carrier (V2A, control material) were recovered, phylogenetically affiliated and characterized in terms of antibiotic resistance (phenotype and genotype), plasmid content, biofilm formation capacity and antibiotic resistance transferability. On all three materials, surviving bacteria were dominated by Gram-positive bacteria and among those by Staphylococcus, Bacillus and Enterococcus spp. The novel antimicrobial surface coating proved to be highly effective. The conventional Ag coating showed only little antimicrobial activity. Microbial diversity increased with increasing exposure time on all three materials. The number of recovered bacteria decreased significantly from V2A to V2A-Ag to AGXX®. After 6 months exposure on the ISS no bacteria were recovered from AGXX®, after 12 months nine and after 19 months three isolates were obtained. Most Gram-positive pathogenic isolates were multidrug resistant (resistant to more than three antibiotics). Sulfamethoxazole, erythromycin and ampicillin resistance were most prevalent. An Enterococcus faecalis strain recovered from V2A steel after 12 months exposure exhibited the highest number of resistances (n = 9). The most prevalent resistance genes were ermC (erythromycin resistance) and tetK (tetracycline resistance). Average transfer frequency of erythromycin, tetracycline and gentamicin resistance from selected ISS isolates was 10−5 transconjugants/recipient. Most importantly, no serious human pathogens such as methicillin resistant Staphylococcus aureus (MRSA) or vancomycin-resistant Enterococci (VRE) were found on any surface. Thus, the infection risk for the crew is low, especially when antimicrobial surfaces such as AGXX® are applied to surfaces prone to microbial contamination.