High Frequency Of Resistance Research Articles

Influence of the stress level on the execution of the Grooved Pegboard Test.

The Grooved Pegboard Test (GPT) is a widely adopted test to evaluate manual dexterity. A factor that could influence the cognitive process is physical and mental stress, which could be controlled by respiration. Stress can be monitored through heart variability. Consequently, the present study's objective was to investigate the association between the breath relaxation technique and the GPT and correlate the findings with stress parameters. One hundred fourteen participants were recruited. Stress level test, GPT, and breathing exercise for relaxation were performed. Spearman test was adopted to evaluate the correlation, while the Friedman Test and the Dunn Test and the Mann-Whitney Test were performed to detect statistical differences. Correlation existed between GPT and age (r=-0.02), height (r=0.22), weight (r=0.21), and handgrip (r=0.21). The Friedman and Dunn's post-test resulted in significant differences in group 1 and 2 between GPT baseline vs. GPT rest (P<0.01) and GPT baseline vs. GPT relax in group 1 (P<0.01) and group 2 (P<0.0001). A breath relaxation technique does not influence the GPT, and even not significantly, physical stress increases the time to complete the GPT. Instead, high mental stress, stress resistance, and heart frequency decrease the time to complete the GPT.

Sepsis: emerging pathogens and antimicrobial resistance in Ethiopian referral hospitals

BackgroundSepsis due to multidrug resistant (MDR) bacteria is a growing public health problem mainly in low-income countries.MethodsA multicenter study was conducted between October 2019 and September 2020 at four hospitals located in central (Tikur Anbessa and Yekatit 12), southern (Hawassa) and northern (Dessie) parts of Ethiopia. A total of 1416 patients clinically investigated for sepsis were enrolled. The number of patients from Tikur Anbessa, Yekatit 12, Dessie and Hawassa hospital was 501, 298, 301 and 316, respectively. At each study site, blood culture was performed from all patients and positive cultures were characterized by their colony characteristics, gram stain and conventional biochemical tests. Each bacterial species was confirmed using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI TOF). Antimicrobial resistance pattern of bacteria was determined by disc diffusion. Logistic regression analysis was used to assess associations of dependent and independent variables. A p-value < 0.05 was considered as statistically significant. The data was analyzed using SPSS version 25.ResultsAmong 1416 blood cultures performed, 40.6% yielded growth. Among these, 27.2%, 0.3% and 13.1%, were positive for pathogenic bacteria, yeast cells and possible contaminants respectively. Klebsiella pneumoniae (26.1%), Klebsiella variicola (18.1%) and E. coli (12.4%) were the most frequent. Most K. variicola were detected at Dessie (61%) and Hawassa (36.4%). Almost all Pantoea dispersa (95.2%) were isolated at Dessie. Rare isolates (0.5% or 0.2% each) included Leclercia adecarboxylata, Raoultella ornithinolytica, Stenotrophomonas maltophilia, Achromobacter xylosoxidans, Burkholderia cepacia, Kosakonia cowanii and Lelliottia amnigena. Enterobacteriaceae most often showed resistance to ampicillin (96.2%), ceftriaxone (78.3%), cefotaxime (78%), cefuroxime (78%) and ceftazidime (76.4%). MDR frequency of Enterobacteriaceae at Hawassa, Tikur Anbessa, Yekatit 12 and Dessie hospital was 95.1%, 93.2%, 87.3% and 67.7%, respectively. Carbapenem resistance was detected in 17.1% of K. pneumoniae (n = 111), 27.7% of E. cloacae (n = 22) and 58.8% of Acinetobacter baumannii (n = 34).ConclusionDiverse and emerging gram-negative bacterial etiologies of sepsis were identified. High multidrug resistance frequency was detected. Both on sepsis etiology types and MDR frequencies, substantial variation between hospitals was determined. Strategies to control MDR should be adapted to specific hospitals. Standard bacteriological services capable of monitoring emerging drug-resistant sepsis etiologies are essential for effective antimicrobial stewardship.

High Osmotic Stress Increases OmpK36 Expression through the Regulation of KbvR to Decrease the Antimicrobial Resistance of Klebsiella pneumoniae.

ABSTRACTKlebsiella pneumoniae is a pathogen known for its high frequency of antimicrobial resistance. Responses to various environmental stresses during its life can influence the resistance to antibiotics. Here, we demonstrate the role and mechanism of KbvR regulator in the response to environmental osmotic stress and in the effect of osmotic stress on antimicrobial resistance. The kbvR mutant strain exhibited increasing tolerance to high osmotic stress and certain antibiotics, including β-lactams. The expression levels of KbvR and outer membrane porin OmpK36 were upregulated in response to high osmotic stress in the wild type (WT), and the deletion of kbvR decreased the expression level of ompK36. The membrane permeability of the kbvR mutant strain was decreased, which was partly restored through the upregulated expression of OmpK36. The DNA affinity purification sequencing (DAP-seq) and microscale thermophoresis (MST) assay disclosed the binding of KbvR to the promoter of the ompK36 gene, indicating that KbvR directly and positively regulated the expression of OmpK36. The high osmotic stress increased the susceptibility to β-lactams and the expression of ompK36 in the WT strain. However, the increased ompK36 expression and the susceptibility to β-lactams in the kbvR mutant strain under high osmotic stress were lower than those of WT. In conclusion, our study has identified that high osmotic stress in the environment influenced the resistance of K. pneumoniae to antibiotics and that the regulation of KbvR with OmpR on the expression of OmpK36 was involved in countering high osmotic stress to change the antimicrobial resistance.IMPORTANCE Klebsiella pneumoniae is considered a global threat because of the rising prevalence of multidrug-resistant strains and their optimal adaptation to clinical environments and the human host. The sensing and adaption abilities of bacteria to the environmental osmotic stress can change the expression of their outer membrane porins, membrane permeability, and resistance to antibiotics. This study reports that KbvR is a newly found regulator that can be upregulated under high osmotic stress and directly regulate the expression of OmpK36 to change the resistance of K. pneumoniae to β-lactam antibiotics. The results demonstrate how adaptation to high osmotic stress changes the sensitivity of K. pneumoniae to antibiotics. The mechanism can be used to sensitize bacteria to antibiotics and highlight new potential strategies for exploiting shared constraints in governing adaptation to diverse environmental challenges.

Prevalence of Extended-Spectrum β-Lactamase Genes and Antibiotic Resistance Pattern in Clinical Isolates of Acinetobacter baumannii from Patients Hospitalized in Mashhad, Iran

Background: Carbapenem-resistant Acinetobacter baumannii strains are one of the most severe factors in hospital infection worldwide, in which the beta-lactamase enzyme is one of the main resistance mechanisms. Objectives: This study aimed to evaluate the presence of carbapenem-resistant beta-lactamase genes and determine antibiotic resistance patterns in the clinical isolates of A. baumannii from patients hospitalized in the Shahid Kamyab Hospital, Mashhad, Iran. Methods: Out of 286 collected isolates from patients hospitalized in Shahid Kamyab Hospital (from March 2017 to June 2017), 31 isolates were confirmed to be A. baumannii using biochemical tests. Antibiotic susceptibility testing was conducted using the disc diffusion method according to the CLSI standard protocols. The presence of beta-lactamase genes, namely blaVEB, blaPER, blaAmpC, blaVIM, blaIMP, blaSHV, and blaTEM, was detected using polymerase chain reaction. Results: In this study, 31 isolates were identified as A. baumannii, all of which revealed high resistance to ceftazidime, cefixime, ceftriaxone, meropenem, imipenem, cefotaxime and cephalexin. In this case, the lowest resistance (19.35%) was observed against polymixin B. Moreover, blaAmpC, blaTEM, blaSHV, blaPER, and blaVIM were observed in 93.54% (29), 51.61% (16), 48.38% (15), 41.93% (13), and 77% (24) of the isolates, respectively. However, blaVEB and blaIMP were observed in none of the isolates. Conclusions: The results showed high carbapenem resistance and high frequency of beta-lactamase resistance genes among the clinical isolates of A. baumannii.

An innovative cardiac rehabilitation based on the power-force-velocity profile to further improve cardiorespiratory capacities in coronary artery disease patients

Abstract Funding Acknowledgements Type of funding sources: Other. Main funding source(s): Doctoral contract 2018-2022 Background Several studies have shown the importance of the relationship between the power-force-velocity profile (PFVP) and sport performance in elite athletes through optimised exercise training.[1] Optimising the training programme is constantly sought in rehabilitation among patients always younger with coronary artery disease. Nowadays, it is well established that intermittent training should be offered to coronary patients during the rehabilitation cycle.[2] In this continuity, the assessment of the PFVP at the beginning of the cardiac rehabilitation (CR) would allow to better adapt the CR training programme for each patient. Purpose The aim of this study was to compare the effects of two exercise training programmes: a traditional CR versus a new CR relied on patient’s PFVP on cardiorespiratory, functional and autonomic systems in coronary patients. Methods This prospective randomized controlled trial was conducted from May 2020 to July 2021 in an university hospital. A total of 89 patients were randomly assigned (1:1) to test or control group. Before starting CR, participants performed two sprints of 8 s on a cycle ergometer to define the PFVP. The PFVP was analysed to determine whether the participant had a force or velocity deficit. Patients included in test group followed a specific cycling training programme based on their weak point (i.e., specific force training with high resistance and low pedalling frequency on the cycle if the PFVP was oriented in velocity and reversely). While control patients attended a conventional CR programme. The 3-week training intervention consisted of 40 min of cycling, 30 min of walking on treadmill and 20 min of strength training (4/week). Cardiopulmonary exercise test (VO2 at the first ventilatory threshold, SV1 and VO2 peak in ml/min/kg), functional assessments (distance of 6-min walk test, handgrip strength, 10 sit-to-stand repetitions, cholesterol levels, LDL-C and quality of life) and autonomic nervous system (heart rate variability and sensitivity baroreflex) were performed at the baseline and after CR. A two-way ANOVA with one repeated measure (pre vs. post) and one independent factor (test vs. control) was realized. Results The mean age was 61.0 ± 9.6 years, 18% were women. A significant difference was observed in VO2 peak (test: +22.0 ± 19.1% vs. control: +10.2 ± 15.8%, p=0.003) and VO2 SV1 (test: +35.9 ± 33.9% vs. control: +11.9 ± 34.4%, p&amp;lt;0.001), LDL-C (p=0.016) and quality of life (p&amp;lt;0.001). No significant change between groups in other functional tests and autonomic activity occurred after CR programme. Conclusion Cardiopulmonary activity, cholesterol and quality of life were improved after 3-week exercise programme. The novel CR depending on initial PFVP showed greater cardiorespiratory benefits than a conventional CR. Therefore, the PFVP can be used in CR to adapt specifically the content of training sessions.

Molecular characterization of diarrheagenic Escherichia coli isolates from children with diarrhea: A cross-sectional study in four provinces of Mozambique: Diarrheagenic Escherichia coli in Mozambique

Analyze the frequency of diarrheagenic Escherichia coli (DEC) pathotypes and their antimicrobial resistance profiles among children aged <15 years with diarrhea in four Mozambican provinces. A cross-sectional hospital-based surveillance program of diarrhea was implemented in Maputo, Sofala, Zambézia, and Nampula. A single stool sample was collected from each child from May 2014 to May 2017. Culture methods and biochemical characterization were performed to detect E. coli strains. DEC pathotypes were determined by conventional polymerase chain reaction targeting specific virulence genes. Antimicrobial susceptibility was assessed by the Kirby-Bauer method. From 723 specimens analyzed by culture, 262 were positive for E. coli. A total of 208 samples were tested by polymerase chain reaction for DEC identification, of which 101 (48.6%) were positive for a DEC pathotype. The predominant pathotypes were enteroaggregative (66.3%, 67/101), enteropathogenic (15.8%, 16/101), enterotoxigenic (13.9%, 14/101), and enteroinvasive E. coli (4.0%, 4/101). No Shiga toxin-producing E. coli was identified. Regardless of the province, the most frequent pathotype was enteroaggregative E. coli. Isolated DEC presented high frequency of resistance to ampicillin (97.8%), tetracycline (68.3%), chloramphenicol (28.4%), nalidixic acid (19.5%), and gentamicin (14.4%). Children with diarrhea in Mozambique had DEC and higher resistance to ampicillin and tetracycline.

Impedance spectroscopy study of defect/ion mediated electric field and its effect on the photovoltaic performance of perovskite solar cells based on different active layers

Effect of defects mitigation on the photovoltaic (PV) performance of three different types of perovskite solar cells (PSCs), namely, single halide (methylammonium lead triiodide, MAPbI 3 ), mixed halide (MAPbI x Cl 3-x ) and phenyl-C71-butyric acid methyl ester (PC 71 BM) based bulk heterojunction (MAPbI 3 -PC 71 BM) PSCs is studied using impedance and transient spectroscopy as a function of dc voltage bias. Parameters extracted from impedance spectra and transient signals are quantitatively analysed to understand the operation mechanism of each PSC. Among the PSCs, the low frequency capacitance (C lf ) values obtained near to the open circuit voltage (V OC ), where charge carriers are restricted inside the device, are found to be minimum for MAPbI 3 based PSCs. Whereas, bulk capacitance (C hf ), high frequency resistance (R hf ) and low frequency resistance (R lf ) values are obtained to be highest for the MAPbI 3 based device which exhibited the best power conversion efficiency (PCE) of 16%. The extracted impedance parameters suggest lessened defect or ion accumulation, improved charge storage capacity/charge life-time, reduced recombination losses and ion transport for the MAPbI 3 device, favoring performance enhancement. The characteristic time constants for the high frequency and low frequency processes also specify longer charge carrier lifetime and faster interfacial charge transfer process near to V OC in MAPbI 3 based PSCs. In addition, transient photocurrent and photovoltage analysis confirms the existence and interplay of ion induced electric field, built-in and applied external field. C lf , C hf , R lf and R hf values as well as transient responses are observed to be greatly dependent on the chemical composition, grain size, grain boundaries and pinholes formed in the perovskite active layer. Based on the results from impedance measurements and transient spectroscopy, the working mechanism of each PSC is proposed in view of the interplay among ion induced electric field, built-in-field and applied electric field. • Operation of three different perovskite solar cells during J-V characterisation. • Defects/ions ion movement and its consequence on the photovoltaic parameters. • The electric field imbalance due to ionic movement in perovskite solar cells.

Pervasive fungicide resistance in Botrytis from strawberry in Norway: Identification of the grey mould pathogen and mutations

AbstractControl of grey mould, caused by Botrytis spp., is a major challenge in open field strawberry production. Botrytis was isolated from plant parts collected from 19 perennial strawberry fields with suspected fungicide resistance in the Agder region of Norway in 2016. Resistance to boscalid, pyraclostrobin and fenhexamid was high and found in 89.1%, 86.0% and 65.4% of conidia samples, respectively. Multiple fungicide resistance was common; 69.6% of conidia samples exhibited resistance to three or more fungicides. Botrytis group S and B. cinerea sensu stricto isolates were obtained from 19 and 16 fields, respectively. The sdhB, cytb, erg27 and mrr1 genes of a selection of isolates were examined for the presence of mutations known to confer fungicide resistance to boscalid, pyraclostrobin, fenhexamid and pyrimethanil plus fludioxonil, respectively. Allele‐specific PCR assays were developed for efficient detection of resistance‐conferring mutations in cytb. Among B. cinerea isolates, 84.7%, 86.3% and 61.3% had resistance‐conferring mutations in sdhB, cytb and erg27, respectively. A triplet deletion in mrr1, resulting in ΔL497, commonly associated with the multidrug resistance phenotype MDR1h, was detected in 29.2% of Botrytis group S isolates. High frequencies of resistance to several fungicides were also detected in Botrytis from both imported and domestically produced strawberry transplants. Fungicide resistance frequencies were not different among fields grouped by level of grey mould problem assessed by growers, indicating factors other than fungicide resistance contributed to control failure, a fact that has important implications for future management of grey mould.

Evaluation of the Speed, Accuracy and Precision of the QuickMIC Rapid Antibiotic Susceptibility Testing Assay With Gram-Negative Bacteria in a Clinical Setting.

The rapidly changing landscape of antimicrobial resistance poses a challenge for empirical antibiotic therapy in severely ill patients and highlights the need for fast antibiotic susceptibility diagnostics to guide treatment. Traditional methods for antibiotic susceptibility testing (AST) of bacteria such as broth microdilution (BMD) or the disc diffusion method (DDM) are comparatively slow and show high variability. Rapid AST methods under development often trade speed for resolution, sometimes only measuring responses at a single antibiotic concentration. QuickMIC is a recently developed lab-on-a-chip system for rapid AST. Here we evaluate the performance of the QuickMIC method with regard to speed, precision and accuracy in comparison to traditional diagnostic methods. 151 blood cultures of clinical Gram-negative isolates with a high frequency of drug resistance were tested using the QuickMIC system and compared with BMD for 12 antibiotics. To investigate sample turnaround time and method functionality in a clinical setting, another 41 clinical blood culture samples were acquired from the Uppsala University Hospital and analyzed on site in the clinical laboratory with the QuickMIC system, and compared with DDM for 8 antibiotics routinely used in the clinical laboratory. The overall essential agreement between MIC values obtained by QuickMIC and BMD was 83.4%, with an average time to result of 3 h 2 min (SD: 24.8 min) for the QuickMIC method. For the clinical dataset, the categorical agreement between QuickMIC and DDM was 96.8%, whereas essential and categorical agreement against BMD was 91.0% and 96.7%, respectively, and the total turnaround time as compared to routine diagnostics was shown to be reduced by 40% (33 h vs. 55 h). Interexperiment variability was low (average SD: 44.6% from target MIC) compared to the acceptable standard of ±1 log2 unit (i.e. -50% to +100% deviation from target MIC) in BMD. We conclude that the QuickMIC method can provide rapid and accurate AST, and may be especially valuable in settings with high resistance rates, and for antibiotics where wildtype and antibiotic-resistant bacteria have MIC distributions that are close or overlapping.

Association of Antibiotic Resistance Traits in Uropathogenic Escherichia coli (UPEC) Isolates.

Background Antimicrobial resistance (AMR) is a global health problem which is constantly evolving and varies spatially and temporally. Resistance to a particular antibiotic may serve as a selection and coselection marker for the same or different antibiotic classes. Therefore, this cross-sectional study was conducted to predict the association of phenotypic and genotypic resistance traits in uropathogenic Escherichia coli (UPEC). Method A total of 42 UPEC from 83 urine samples were investigated for the prevalence and association of phenotypic and genotypic AMR traits. Antibiogram profiling was carried out by Kirby–Bauer's disc diffusion method and AMR genes (ARGs) were detected by PCR. Result UPECs were isolated from 50.60% (42/83) of the samples examined. Of these, 80.95% of cases were derived from females, and 38.10% of cases were found in the age group of 21–30 years. The isolates were shown to have a high frequency of resistance to tetracycline (92.86%), followed by sulfonamide (71.43%), ampicillin (52.38%), trimethoprim-sulfamethoxazole (47.62%), and 28.57% each to streptomycin, chloramphenicol, and erythromycin. The most prevalent antimicrobial resistance genes (ARGs) in these isolates were tet(A) (78.57%), tet(B) (76.19%), sul1 (61.91%), dfrA1 (35.71%), blaSHV (26.19%), cmlA (19.05%), and CITM, qnrA, and catA1 each at 11.91%. According to statistical analysis, ampicillin, sulfonamide, trimethoprim-sulfamethoxazole, and ciprofloxacin resistance were strongly correlated with the presence of blaSHV, sul1, dfrA1, and qnrA, respectively. Nonsignificant associations were observed between ciprofloxacin-tetracycline, sulfonamide-erythromycin pairs as well as between tet(A) and tet(B) genes. Besides, coselection was also assumed in the case of chloramphenicol resistance genes, namely, catA1 and cmlA. Conclusion Both the phenotypic and genetic resistance traits were found in the UPEC isolates. Statistical association and coselection phenomena among AMR phenotypes and genotypes were also observed but required to be validated in a broad-scale study. However, these findings might have important implications for the development of an AMR prediction model to tackle future AMR outbreaks.

Antimicrobial Resistance of Shigella flexneri in Pakistani Pediatric Population Reveals an Increased Trend of Third-Generation Cephalosporin Resistance.

The rapid emergence of resistance to third-generation cephalosporins in Shigella flexneri is crucial in pediatric shigellosis management. Limited studies have been conducted on molecular pattern of antibiotic resistance of S. flexneri in diarrhea endemic areas of Pakistan. The aim of the study was to analyze the antimicrobial resistance of S. flexneriisolated from pediatric diarrheal patients in Peshawar, Pakistan. A total of 199 S. flexneri isolates (clinical, n = 1 55 and non-clinical, n = 44) were investigated for drug resistance and mutational analysis of selected drug resistance genes. All isolates were found to be highly resistant to amoxicillin/clavulanic acid (88%), followed by trimethoprim-sulfamethoxazole (77%), chloramphenicol (43%), and quinolones (41.6%). About 34.5% S. flexneri isolates were found to be resistant to third-generation cephalosporin. None of the isolates was resistant to imipenem, piperacillin-tazobactam, and amikacin. Interestingly high frequency of third-generation cephalosporin resistance was observed in S. flexneri isolated from non-clinical samples (49%) when compared to clinical samples (30.5%). Furthermore, the most prevalent phenotypic-resistant patterns among third-generation cephalosporin-resistant isolates were AMC,CAZ,CPD,CFM,CRO,SXT (13%) followed by OFX,AMC,CAZ,CPD,CFM,CRO,SXT,NA,CIP (10%). The most frequently detected resistance genes were trimethoprim-sulfamethoxazole (sul2 = 84%), beta-lactamase genes (blaOXA = 87%), quinolones (qnrS = 77%), and chloramphenicol (cat = 64%). No mutation was detected in any drug-resistant genes. We are reporting for the first time the sequence of the blaTEM gene in S. flexneri. Furthermore, high third-generation cephalosporin resistance was observed in the patients who practiced self-medication as compared to those who took medication according to physician prescription. This study shows the high emergence of third-generation cephalosporin-resistant S. flexneri isolates, which is a potential threat to the community in the country. This finding will be helpful to develop a suitable antibiotic prescription regime to treat shigellosis.

Fungicide resistance frequencies of Botrytis cinerea greenhouse isolates and molecular detection of a novel SDHI resistance mutation

Sensitivity of B. cinerea to commonly used fungicides against Gray mold with emphasis to the newer quinone outside inhibitor (QoIs), and succinate dehydrogenase inhibitors (SDHIs) was assessed during a monitoring survey from vegetable greenhouses in four representative regions of Crete. 42% from a total of 168 isolates were simultaneously resistant to boscalid, fluopyram, pyraclostrobin and fenhexamid but not to fludioxonil making this phenylpyrrole fungicide an excellent anti-resistance antifungal agent. Isolates with double resistance to SDHIs and QoIs were found in very high frequencies indicating a selection towards double resistance due to the use of pyraclostrobin-boscalid mixtures. A number of sdhB resistance mutations (H272R, N230I and P225F/H) were found in isolates also carrying the G143A cytb resistance mutation in the above isolates. A novel sdhB point mutation (I274V) was identified for the first time in B. cinerea isolates collected from greenhouses with a fluopyram spray history with specific resistance to SDHIs. A PCR-RFLP diagnostic assay was developed for the detection of this mutation in the sdhB gene. Mutations P225F/H and I274V were found to be associated with fitness penalties in terms of mycelial growth, sporulation or pathogenicity. Results suggest that, in order to retain effective control of gray mold in Crete, appropriate anti-resistance strategies should be implemented taking into account the high double SDHI and QoI resistance frequencies. Additional studies for monitoring the already known and the new SDHI-resistance mutations, are necessary in order to hinder the further spread and establishment of single or double resistant isolates of B. cinerea detected in greenhouses in Crete.

Antimicrobial Resistance of E. coli and Salmonella Isolated from Wild Birds in a Rehabilitation Center in Turkey.

Wildlife plays a critical role as a reservoir for zoonosis especially pathogenic enteric bacteria. In this study we evaluated the presence of E. coli and Salmonella isolates from wild birds and determined their antimicrobial resistance. Intestine and fecal samples from 82 dead wild birds obtained from rehabilitation centre, were examined by microbiological analysis, antibiotic susceptibilities against of 18 antimicrobials and presence of tetracycline resistance genes by multiplex and singleplex PCR were investigated. A total of 51 E. coli were identified as well as Salmonella Kentucky and Salmonella Bisberg. A majority of the E. coli isolates were resistant to lincomysin (100%), penicilline (96.1%), kanamycin (80.4%), tetracycline (68.6%), and oxytetracycline (64.7%). All Salmonella serotypes were resistant to lincomycin, nalidixic acid and penicilline.In addition, 58.82% of E. coli isolates had phenotypic resistance to at least three or more antimicrobials. Our results indicated that the high frequency of tetracycline resistance (68.62%) due to the tet (A), tet (B), and tet (D) genes. This is the first report isolating S. Bisberg and determining antibiotic susceptibility of E.coli and Salmonella isolates from wild birds in Turkey. These results will help providing better understand of the dissemination of antibiotic resistancy in the environment, which can be used to potentially decrease spread through bird migration. Moreover, these results help assess the risk of spread of resistance from wild birds to humans.

Individual and Combined Application of Nematophagous Fungi as Biological Control Agents against Gastrointestinal Nematodes in Domestic Animals.

Gastrointestinal nematodes (GINs) are a group of parasites that threaten livestock yields, and the consequent economic losses have led to major concern in the agricultural industry worldwide. The high frequency of anthelmintic resistance amongst GINs has prompted the search for sustainable alternatives. Recently, a substantial number of both in vitro and in vivo experiments have shown that biological controls based on predatory fungi and ovicidal fungi are the most promising alternatives to chemical controls. In this respect, the morphological characteristics of the most representative species of these two large groups of fungi, their nematicidal activity and mechanisms of action against GINs, have been increasingly studied. Given the limitation of the independent use of a single nematophagous fungus (NF), combined applications which combine multiple fungi, or fungi and chemical controls, have become increasingly popular, although these new strategies still have antagonistic effects on the candidates. In this review, we summarize both the advantages and disadvantages of the individual fungi and the combined applications identified to date to minimize recurring infections or to disrupt the life cycle of GINs. The need to discover novel and high-efficiency nematicidal isolates and the application of our understanding to the appropriate selection of associated applications are discussed.

Isolation and antimicrobial resistance of coagulase-negative staphylococci recovered from healthy tortoises in Minas Gerais, Brazil

ABSTRACT: In the last few decades, there has been a global increase in the adoption of reptiles as companion animals, mainly turtles and tortoises. Considering the popularity of reptiles as pets in Brazil, and a notable lack of data about potentially pathogenic staphylococci in these animals, this study isolated and evaluate the antimicrobial susceptibility of staphylococcal species from healthy tortoises (Chelonoidis carbonaria) in Brazil. During a 12-month period (February 2019 to February 2020), cloacal swabs from 66 healthy tortoises were collected at the Wild Animals Screening Center in Belo Horizonte, Minas Gerais, Brazil. The swabs were plated onto mannitol salt agar for staphylococci isolation, and species identification was performed using MALDI-TOF MS. Antimicrobial susceptibility was investigated using the disk diffusion method, and the presence of the mecA gene was investigated by PCR to detect methicillin resistance. Of the tested animals, 72.7% were positive for staphylococcal isolation. All isolates were coagulase-negative staphylococci (CoNS), and Staphylococcus sciuri (81.3%), and S. xylosus (12.5%) were the most frequently isolated species. The majority of the isolates (56%) were resistant to at least one antimicrobial agent. A high frequency of resistance was observed for penicillin (35.5%) and tetracycline (29.1 %). All strains were susceptible to cefoxitin, chloramphenicol, ciprofloxacin, erythromycin, and gentamicin. All isolates were negative for the mecA gene. The present work suggests that healthy tortoises are mainly colonized by CoNS, especially S. sciuri. Half of the isolates were resistant to at least one antimicrobial, raising questions regarding the possible role of these animals as reservoirs of antimicrobial resistance genes.

High frequency of antimicrobial resistance and virulence gene in Shigella species isolated from pediatric patients in an Iranian Referral Hospital.

Background:Shigella is a main cause of gastroenteritis and it is responsible for 5 to 10 % of diarrhea through the world. The aims of this study were to assess the antibiotic susceptibility pattern and the presence of 3 common virulence genes (sigA, virF, invE) of Shigella strains isolated from patients with gastroenteritis in Children’s Medical Center Hospital, Tehran, Iran.Methods:Over a period of 15 months, all Shigella species collected from the patients with gastroenteritis were entered to the study. Susceptibility testing of all isolates towards different antibiotics was performed using the disk diffusion method and the prevalence of virulence genes was detected by polymerase chain reaction (PCR) technology.Results:Among a total of 183 Shigella strains, 128 Shigella sonnei (70%) and 55 S. flexneri (30%) were isolated. The resistance rate to the antibiotics in S. sonnei strains was higher than S. flexneri. The most sensitive antibiotics for S. flexneri strains were gentamicin (98%), amikacin (85%) and ciprofloxacin (82%), while high resistance rate to trimethoprim-sulfamethoxazole (96%), ampicillin (96%), nalidixic acid (64%) and cefotaxime (60%) was observed. The frequency of invE, virF and sigA gene in S. flexneri strains was 89 %, 93 % and 56 %, respectively; whereas they found in 93 %, 96 %, and 100 % of S. sonnei strains, respectively. SigA gene was identified significantly higher in the S. sonnei strains (100%). There was no significant difference between the presence of virF and invE genes among Shigella strains.Conclusion:The high presence of sigA gene in S. sonnei strains plays an important role in its pathogenesis, and the high frequency of invE and virF genes showed that this classical pathway regulating the expression of Shigella virulence factor genes could play a key role in the pathogenesis of this bacterium. (www.actabiomedica.it)

Frequency of resistance to first-line antiretroviral therapy observed among HIV patients.

Objectives:This study aimed to assess the frequency of first-line antiretroviral therapy (ART) resistance among HIV patients and to identify the factors affecting the drug resistance.Methods:A cross-sectional study was conducted over a sample of 162 HIV-positive patients attending the Medicine Department of Lady Reading Hospital, Peshawar-Pakistan, from July 2020 to January 2021. Blood samples were collected for phylogenic profiles to determine first-line antiretroviral therapy resistance.Results:The frequency of ART resistance was detected in 64.8% of the enrolled HIV patients. Factors such as patient age, gender, comorbidities, and smoking status had no significant impact on drug resistance. While only body mass index (BMI) significantly affected ART resistance among HIV patients. The drug resistance mutations M184V and K103N were detected in the nucleoside reverse transcriptase inhibitors (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs), respectively, whereas the mutations G73SC and I47VA were detected in the protease inhibitors (PIs).Conclusion:There is a high frequency of resistance to first-line antiretroviral therapy among HIV patients presenting to the selected healthcare facility in Peshawar. Furthermore, we found no significant factors impacting ART resistance among HIV patients other than BMI.

PON2 blockade overcomes dexamethasone resistance in acute lymphoblastic leukemia

ABSTRACT Objectives The high frequency of chemotherapy resistance is ultimately responsible for clinical relapse in acute lymphoblastic leukemia (ALL). Nevertheless, the molecular mechanism relevant to glucocorticoid (GC) resistance remains ambiguous. Methods Quantitative real-time polymerase chain reaction and Western blot were performed to detect the expressions of paraoxonase 2 (PON2), Bcl-2 and Bax. shRNA was used to knockdown PON2 expression in SUP-B15 and REH cell. CCK-8 and flow cytometry assay were conducted to monitor the changes of proliferation and apoptosis in ALL cells. The growth of ALL REH cells in vivo was determined using transplanted tumor model. Results This study was designed to identify GC resistance-associated genes by means of the transcriptome chip from the public Gene Expression Omnibus database, and preliminarily investigation of dexamethasone (DEX)-resistance mechanism in ALL. We disclosed that PON2 expression was elevated in ALL patients and especially higher in DEX-resistance ALL patients. Then, cell apoptosis assay suggested that silencing of PON2 dramatically promoted in DEX-resistant ALL cells apoptosis and the activity of Caspase 3 induced by DEX administration. In xenograft tumor model, PON2 knockdown significantly reduced DEX-resistant ALL cells growth in immunodeficient mice. Conclusions Collectively, inhibition of PON2 may represent a novel method to restore the sensitivity of treatment-resistant ALL to GC-induced cell death.

Epidemiological, clinical and laboratory differences between male urethral infections due to Haemophilus spp. and those due to Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium and Ureaplasma urealyticum: A descriptive study.

To describe the epidemiological, clinical and laboratory characteristics of male patients diagnosed with Haemophilus spp. urethral infection and to compare them with the characteristics of male patients diagnosed with N. gonorrhoeae, C. trachomatis, M. genitalium and U. urealyticum urethral infection. Over the past 2 years, an increase in urethral infections due to Haemophilus spp. was observed. All male patients who attended our Department of Sexually Transmitted Infections between January 2018 and February 2019 were retrospectively studied; they underwent conventional bacteriological and multiplex PCR studies in the urethra at the same time. Of the 86 patients studied, a unique microorganism was detected in 76 cases, N. gonorrhoeae in 24, Haemophilus spp. in 21 (16 H. parainfluenzae and 5 H. influenzae), C. trachomatis in 19, M. genitalium in 8 and U. urealyticum in 4; 10 cases presented more than one microorganism. In case of multiple aetiological agents, sexual partnership was multiple. In the Haemophilus group, 81% reported only unprotected oral insertive sex; symptoms lasted for more than one week in 62% of the patients. Haemophilus is an aetiological agent of non-gonococcal urethritis whose incidence is clearly increasing; the main route of transmission is oral sex. The most common reason for consultation is dysuria and testicular pain, while urethral discharge was predominant for the other causes of urethral infection. Due to the high frequency of antibiotic resistance in the Haemophilus group, it is necessary to confirm eradication by performing a test of cure.

Cellular and Extracellular Vesicle RNA Analysis in the Global Threat Fungus Candida auris.

ABSTRACTEmerging and reemerging pathogens are a worldwide concern, and it is predicted that these microbes will cause severe outbreaks. Candida auris affects people with weakened immune systems, particularly those who are hospitalized or are in health care facilities. Extracellular vesicles (EVs) are lipid bilayer structures released by organisms from all domains of life. EVs can deliver functional molecules to target cells, including proteins and nucleic acids, especially RNA molecules. EVs from several pathogenic fungi species play diverse biological roles related to cell-cell communication and pathogen-host interaction. In this study, we describe a data set which we produced by sequencing the RNA content of EVs from C. auris under normal growth conditions and in the presence of the antifungal caspofungin, a first-line drug to treat this fungus. To generate a more complete data set for future comparative studies, we also sequenced the RNA cellular content of EVs under the same conditions. This data set addresses a previously unexplored area of fungal biology regarding cellular small RNA and EV RNA. Our data will provide a molecular basis for the study of the aspects associated with antifungal treatment, gene expression response, and EV composition in C. auris. These data will also allow the exploration of small RNA content in the fungal kingdom and might serve as an informative basis for studies on the mechanisms by which molecules are directed to fungal EVs.IMPORTANCE Candida auris, a relevant emerging human-pathogenic yeast, is the first fungus to be called a global public health threat by the WHO. This is because of its rapid spread on all inhabited continents, together with its extremely high frequency of drug and multidrug resistance. In our study, we generated a large data set for 3 distinct strains of C. auris and obtained cellular small RNA fraction as well as extracellular vesicle RNA (EV-RNA) during normal growth conditions and after treatment with caspofungin, the first-line drug used to treat C. auris infection.