Comparison of multidrug-resistant extraintestinal pathogenic Escherichia coli from human, foods and animals to investigate the possible chains of transmission

Background Globally, antimicrobial drug resistant Escherichia coli is the most common etiological agent of invasive disease in humans. In Europe, increasing proportions of infections due to third generation cephalosporins (3GCs) and/or fluoroquinolone resistant extraintestinal pathogenic E. coli (ExPEC) strains are reported. It has been shown that multidrug resistant (MDR) E. coli can be transmitted from animals to humans and based on existing evidence, poultry is the food animal source most closely linked to human E. coli. However, lack of reliable data makes it difficult to assess the attributable risk of different food sources and their impact on human health. Objectives In the present study, our objective was to investigate the antimicrobial resistance profile, phylogenetic background and virulence factors of E. coli isolates from broiler chicken meat sold at retail in Palermo, Italy and to compare with the geographically and temporally matched collection of isolates from humans with infections due to this bacterium. Materials and Methods Isolation of MDR E. coli was performed during April 2013-December 2014 on a total of 250 food samples including 180 raw broiler chicken meat , 17 pork meat , 14 cow meat 19 cheese and 20 ready to eat foods samples and a total of 200 de-identified FQ-R and ESBL-producing E. coli isolates were collected from Ospedale Civico in Palermo , Italy. Susceptibility to a panel of nine antimicrobial agents was determined and the isolates resistant to at least three classes of antibacterial drugs were defined as MDR. PCR assays were carried out to detect extended spectrum β-lactamase (ESBL), plasmid-mediated AmpC β-lactamase and plasmid-mediated quinolone resistance (PMQR) genes, phylogenetic group and ExPEC-associated traits. A single nucleotide polymorphism (SNP) PCR was done to detect E. coli sequence type (ST)131. Enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) and Raman spectroscopy were performed to analyze the relatedness among ExPEC isolates. Results E. coli were isolated from all of the chicken samples. However, multi-drug resistant (MDR) E. coli were isolated from 152 (84.4%) of the 180 chicken samples. No MDR E. coli isolates were found in other types of food samples. B1 was the most prevalent (114 isolates, 48.1%) followed by groups D (80 isolates, 33.7%), A (30 isolates, 12.7%) and B2 (13 isolates, 5.5%); phylogenetic group B2 was the most prevalent phylogroup among clinical isolates (136 isolates, 68.5%) followed by groups D (33 isolates, 16.8%), A (20 isolates, 10.6%) and B2 (9 isolates, 4.6%), respectively. ESBLs and AmpC β-lactamases were detected by PCR in 233 (98.3%) and 29 (12.2%) of chicken meat isolates and 194 (97.5%) and 34 (17.2%) of clinical isolates, respectively. PMQR determinants were detected in 215 (90.7%) of chicken isolates and 183 (91.9%) of clinical isolates, respectively. Based on the molecular definition of ExPEC, 58 and 156 isolates from chicken meat and clinical samples were attributed with the status of ExPEC . SNP-PCR results confirmed that a total of 91.9% (125/136) and 69.2% (9/13) of the clinical and chicken meat isolates of E. coli of B2 phylogroup population were identified as ST131, respectively. Both ERIC-PCR and Raman Spectroscopy analysis showed a large heterogeneity among isolates. Conlusion The results of this study show an alarmingly high prevalence of MDR E. coli and especially ExPEC isolates from broiler chicken meat in our geographic area. The ongoing use of antimicrobial drugs in livestock should be urgently revised and stopped, particularly in the poultry sector

Extended-spectrum ß-lactamase, AmpC-producing, and fluoroquinolone-resistant Escherichia coli in retail broiler chicken meat, Italy

Background: Globally, antimicrobial drug-resistant Escherichia coli is among the most common etiological agents of invasive disease in humans. In Europe, increasing proportions of infections due to third-generation cephalosporins and/or fluoroquinolone-resistant extraintestinal pathogenic E. coli (ExPEC) strains are reported. E. coli from poultry are those more closely linked to human E. coli, but lack of reliable data makes it difficult to assess the attributable risk of different food sources. In the present study, our objective was to investigate the antimicrobial resistance profile, phylogenetic background, and virulence factors of E. coli isolates from broiler chicken meat sold at retail in Palermo, Italy. Materials and Methods: Isolation of multidrug resistant (MDR) E. coli was performed during April-December 2013 on a total of 163 chicken meat samples. Susceptibility to a panel of nine antimicrobial agents was determined. PCR assays were carried out to detect extended-spectrum β-lactamase (ESBL), plasmid-mediated AmpC β-lactamase, and plasmid-mediated quinolone resistance (PMQR) genes, phylogenetic group, and ExPEC-associated traits. A single nucleotide polymorphism (SNP) PCR was done to detect E. coli sequence type (ST)131. Results: One hundred thirty-four isolates from 109 meat samples were MDR. B1 was the most prevalent phylogenetic group (47.8%), followed by groups D (25.4%), A (22.3%), and B2 (4.5%). ESBLs and AmpC β-lactamases were detected by PCR in 132 (98.5%) and 15 (11.2%) isolates. PMQR determinants were detected in 122 (91%) isolates. Twenty-two MDR isolates met the molecular definition of ExPEC. SNP-PCR results confirmed that four B2 isolates were ST131. Enterobacterial Repetitive Intergenic Consensus sequence-PCR analysis showed a large heterogeneity with 55 unique profiles and 31 clusters including 2-4 isolates. Conclusions: An alarmingly high prevalence of MDR E. coli from broiler chicken meat is evident in our geographic area. The ongoing use of antimicrobial drugs in livestock should be urgently restricted, particularly in the poultry sector

Comparing Neonatal Intensive Care Unit Nursing Support in Mothers With Newborn Abstinence Syndrome (NAS) and Mothers of Healthy Neonates

Background: The experience of having neonates in the Neonatal Intensive Care Unit (NICU) is a psychological crisis. It might cause many emotional problems for parents. Entire parental support is among the duties of the healthcare team. Therefore, this study aimed to compare the nursing support received by the mothers with Newborn Abstinence Syndrome (NAS) and the mothers of other neonates admitted to the NICU.Methods: The present cross-sectional descriptive-analytic study was conducted in the selected hospitals in Kerman Province, Iran. In total, 62 mothers with NAS and 61 non-addicted mothers with neonates admitted to the NICU were selected through convenience sampling method. The inclusion criteria were neonates under the care of parents, neonate admitted to the NICU for at least 24 hours, opiate dependence in the case group mothers, and no substance dependence in the control group mothers. The amount of nursing support for mothers having neonates with NAS was compared with that of the control mothers. The study groups were homogenized in terms of the study variables (neonate age, gender, and the duration of hospitalization). The required data were collected by the Nurse-Parent Support Tool (NPST) and analyzed in SPSS.Results: The study results revealed that among the neonates of 123 mothers, 75 (60.97%) were boys, and 58(39.02%) were girls. The majority of neonates in both groups were breastfed. The Mean±SD age of the mothers in the case and control group were 31.93±7.25 and 28.99±4.36 years, respectively. The nursing support level was desirable in both groups, and no significant difference was found in this regard (P>0.05). Furthermore, the level of nursing support in emotional, information-communication, self-esteem, and quality caregiving support dimensions was desirable in both groups.Conclusion: The obtained results revealed that nurses’ support was desirable in both groups. The prevalence of maternal addiction and the impact of this social harm on neonates who were admitted are essential. Furthermore, families having neonates with NAS need more support from the healthcare staff and nurses, in comparison with healthy parents; thus, the importance of this issue should be addressed in training and briefing courses for nurses

Drug resistance patterns of bacteria isolated from patients with nosocomial pneumonia at Tehran hospitals during 2009-2011

INTRODUCTION: Nosocomial pneumonia remains an important cause of mortality and morbidity worldwide. Surveillance programs play an important role in the identification of common etiologic agents and local patterns of antimicrobial resistance. METHODOLOGY: In this study we determined the frequency and antimicrobial susceptibility of pathogens isolated from patients with nosocomial pneumonia during 2009 to 2011. RESULTS: A total of 642 bacteria were isolated from 516 suspected samples. Acinetobacter baumannii (21.1%, n = 136), was the commonest isolated pathogen followed by Pseudomonas aeruginosa (17.4%, n = 112), Staphylococcus aureus (15.8%, n = 102) and enterococci (8.4% n = 54). The most effective therapeutic agents against A. baumannii were polymyxin B (95.5% susceptible), ceftriaxone/tazobactam (72% susceptible) and levofloxacin (52.9% susceptible). Polymixin B (89.2% susceptible), ceftriaxone/tazobactam (89.2% susceptible) and piperacillin-tazobactam (80.3% susceptible) were found to be the most active agents against P. aeruginosa. Extended-spectrum beta-lactamases were detected among isolates of K. pneumoniae (45.4%) and E. coli (20.3%). Overall, the prevalence of methicillin-resistant S. aureus and vancomycin resistant enterococci were 80.4% and 40.7% respectively. Linezolid was found to be the most active antibiotic against these pathogens. The etiology of 50% of the nosocomial infection cases was polymicrobial. CONCLUSIONS: The combination of ceftriaxone/tazobactam seems to be beneficial agent against multidrug-resistant Gram-negative bacilli isolated form respiratory tract infections. The results of our study can be used for guiding appropriate empiric therapy in this geographic region

Medication Errors Experiences of Nurses who Working in Hospitals of Isfahan at 1391

Introduction: Patient safety has long been a major concern for healthcare professionals. Today, hospitals should provide patient safety in order to gain accreditation. Among patient safety issues, medication error rate has been considered as a major indicator of healthcare quality. The different aspects of medication error are not clearly known and the nurses are the main source of knowledge in this field. Therefore, the researcher decided to study the experiences of nurses who have been faced with a medication error in order to clarify the various aspects of this phenomenon. It is through the exact identification of this phenomenon that it can be managed. Method: In the present study, a phenomenological qualitative approach was used. Participants in this study were nurses working in hospitals of Isfahan and had experienced medication error. Purposive sampling was adopted in the study and sample size was dependent on the data saturation. A total of 10 participants were chosen. Data collection was carried out through unstructured interviews and analysis through Colaizzi’s Method. Results: From the findings of this study, six main themes expressing participants’ experiences emerged. The six main themes include: The context of error, the error range, negligence, the consequence of error, concealment of error, and reporting of error. Conclusion: Nurses report their errors when they feel safe and do not see harmful results for them. Therefore, hospitals should review their policies on error reporting to ensure they actively encourage nurses to report medication errors, and to support a blame-free culture in the organization and a systembased approach to deal with the error. Keywords: Nurses’ medication errors, Nurses’ experiences, Patient safety, Qualitative researc

Extraintestinal pathogenic Escherichia coli sequence type 131 H30-R and H30-Rx subclones in retail chicken meat, Italy

Extraintestinal pathogenic Escherichia coli sequence type 131 (ST131), typically fluoroquinolone-resistant (FQ-R) and/or extended-spectrum β-lactamase (ESBL)-producing, has emerged globally. Among clinical isolates, ST131, primarily its H30-R and H30-Rx subclones, accounts for most antimicrobial-resistant E. coli and is the dominant E. coli strain worldwide. We assessed its prevalence and characteristics among raw chicken meat samples on sale in Palermo, Italy. A collection of 237 fluoroquinolone resistant and ESBL/AmpC producing E. coli isolates, which had been isolated from processed retail chicken meat in the period May 2013-April 2015, was analyzed. Established polymerase chain reaction methods were used to define ST131 and its H30 subclones, ESBL, AmpC, and plasmid-mediated quinolone resistance (PMQR) determinants. Amplified Fragment Length Polymorphism (AFLP) was performed to assess the relatedness among ST131 isolates. Out of the 237 E. coli isolates, 12 isolates belonged to the phylogenetic group B2. Based on the molecular definition of ExPEC, all isolates were attributed with the status of ExPEC. SNP-PCR results confirmed that nine isolates were ST131. SNP-PCR for H30-R and H30-Rx subclones showed that six and three ExPEC ST131 were positive for H30-R and H30-Rx, respectively. The results of AFLP showed that, except for four isolates grouped into two clusters which proved to be indistinguishable, the isolates under study were genetically heterogeneous. To the best of our knowledge, this is the first report of H30-R and H30-Rx subclones in animal food samples. Our findings appear to support the role of food chain in their transmission to humans

Ancient and recent differences in the intrinsic susceptibility of Mycobacterium tuberculosis complex to pretomanid.

OBJECTIVES: To develop a robust phenotypic antimicrobial susceptibility testing (AST) method with a correctly set breakpoint for pretomanid (Pa), the most recently approved anti-tuberculosis drug. METHODS: The Becton Dickinson Mycobacterial Growth Indicator Tube™ (MGIT) system was used at six laboratories to determine the MICs of a phylogenetically diverse collection of 356 Mycobacterium tuberculosis complex (MTBC) strains to establish the epidemiological cut-off value for pretomanid. MICs were correlated with WGS data to study the genetic basis of differences in the susceptibility to pretomanid. RESULTS: We observed ancient differences in the susceptibility to pretomanid among various members of MTBC. Most notably, lineage 1 of M. tuberculosis, which is estimated to account for 28% of tuberculosis cases globally, was less susceptible than lineages 2, 3, 4 and 7 of M. tuberculosis, resulting in a 99th percentile of 2 mg/L for lineage 1 compared with 0.5 mg/L for the remaining M. tuberculosis lineages. Moreover, we observed that higher MICs (≥8 mg/L), which probably confer resistance, had recently evolved independently in six different M. tuberculosis strains. Unlike the aforementioned ancient differences in susceptibility, these recent differences were likely caused by mutations in the known pretomanid resistance genes. CONCLUSIONS: In light of these findings, the provisional critical concentration of 1 mg/L for MGIT set by EMA must be re-evaluated. More broadly, these findings underline the importance of considering the global diversity of MTBC during clinical development of drugs and when defining breakpoints for AST

The 2021 WHO catalogue of Mycobacterium tuberculosis complex mutations associated with drug resistance: a genotypic analysis.

Background: Molecular diagnostics are considered the most promising route to achievement of rapid, universal drug susceptibility testing for Mycobacterium tuberculosis complex (MTBC). We aimed to generate a WHO-endorsed catalogue of mutations to serve as a global standard for interpreting molecular information for drug resistance prediction. Methods: In this systematic analysis, we used a candidate gene approach to identify mutations associated with resistance or consistent with susceptibility for 13 WHO-endorsed antituberculosis drugs. We collected existing worldwide MTBC whole-genome sequencing data and phenotypic data from academic groups and consortia, reference laboratories, public health organisations, and published literature. We categorised phenotypes as follows: methods and critical concentrations currently endorsed by WHO (category 1); critical concentrations previously endorsed by WHO for those methods (category 2); methods or critical concentrations not currently endorsed by WHO (category 3). For each mutation, we used a contingency table of binary phenotypes and presence or absence of the mutation to compute positive predictive value, and we used Fisher's exact tests to generate odds ratios and Benjamini-Hochberg corrected p values. Mutations were graded as associated with resistance if present in at least five isolates, if the odds ratio was more than 1 with a statistically significant corrected p value, and if the lower bound of the 95% CI on the positive predictive value for phenotypic resistance was greater than 25%. A series of expert rules were applied for final confidence grading of each mutation. Findings: We analysed 41 137 MTBC isolates with phenotypic and whole-genome sequencing data from 45 countries. 38 215 MTBC isolates passed quality control steps and were included in the final analysis. 15 667 associations were computed for 13 211 unique mutations linked to one or more drugs. 1149 (7·3%) of 15 667 mutations were classified as associated with phenotypic resistance and 107 (0·7%) were deemed consistent with susceptibility. For rifampicin, isoniazid, ethambutol, fluoroquinolones, and streptomycin, the mutations' pooled sensitivity was more than 80%. Specificity was over 95% for all drugs except ethionamide (91·4%), moxifloxacin (91·6%) and ethambutol (93·3%). Only two resistance mutations were identified for bedaquiline, delamanid, clofazimine, and linezolid as prevalence of phenotypic resistance was low for these drugs. Interpretation: We present the first WHO-endorsed catalogue of molecular targets for MTBC drug susceptibility testing, which is intended to provide a global standard for resistance interpretation. The existence of this catalogue should encourage the implementation of molecular diagnostics by national tuberculosis programmes. Funding: Unitaid, Wellcome Trust, UK Medical Research Council, and Bill and Melinda Gates Foundation