In vitro susceptibility to fosfomycin in clinical and environmental extended-spectrum beta-lactamase producing and/or ciprofloxacin-non-susceptible Escherichia coli isolates

ABSTRACT Extended-spectrum beta-lactamase producing and ciprofloxacin-non-susceptible Escherichia coli are clinical and environmental issues. We evaluated the susceptibility profile of fosfomycin in non-susceptible E. coli isolated from urine and the environment. We measured the activity of fosfomycin against 319 and 36 E. coli strains from urine and environmental isolates, respectively, collected from rivers. Fosfomycin resistance profiles were investigated using the minimal inhibitory concentration (MIC), according to the Clinical and Laboratory Standards Institute (CLSI) and the European Committee for Antimicrobial Susceptibility Testing (EUCAST) guidelines. Antibiotic susceptibility testing revealed that 5% and 6.6% of urine samples were non-susceptible to fosfomycin according to CLSI and EUCAST guidelines, respectively. The fosfomycin MIC50/90 was 0.5/4 mg/L. Of the 36 E. coli isolates from river water, 11.1% and 13,8% were non-susceptible to fosfomycin according to CLSI and EUCAST, respectively (range ≤0.25 ≥512 mg/L). All the isolates with MIC ≥512 mg/L for fosfomycin showed the fosA3 gene. Fosfomycin resistance was more frequent in the environment than in clinical samples.

Microbiological profile of bloodstream infections and antimicrobial resistance patterns at a tertiary referral hospital in Amazon, Brazil

ABSTRACT Background: Bloodstream infections (BSI) are a global health issue, leading to high mortality and morbidity among hospitalized patients. Methods: A retrospective, observational and descriptive study was conducted by reviewing blood culture records collected from patients with suspected BSI, between January 2017 and December 2019. Results: The most frequent antimicrobial resistant (AMR) pathogens were methicillin-resistantStaphylococcus aureus(MRSA) (40%), methicillin-resistantS. epidermidis (MRSE) (9.5%), and extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae (35.3%). Conclusions: Our findings underscore the importance of continued vigilance and advocate for the rational use of antimicrobial agents.

Extended-spectrum and AmpC ß-lactamases screening and antibiotic resistance profile of Escherichia coli isolated from urine / Análisis de las ß-lactamasas de espectro extendido y tipo AmpC y el perfil de resistencia a antibióticos de Escherichia coli aisladas de orina

Introduction: Antimicrobial resistance is a global concern since infections by resistant pathogens are associated with higher mortality and morbidity. Objective: To assess the prevalence of Escherichia coli isolates producing extended-spectrum and AmpC beta-lactamase (ESBL) in urine samples from patients at the Hospital Metropolitano de Santiago in Dominican Republic. Methods: Pathogen identification and antibiogram were carried out by the automated systems BD Phoenix or Microscan®. General information and past medical history were gathered from patients with a positive urine culture for E. coli. Manual ESBL/AmpC screening was performed with the commercial ESBL+AmpC screen disc kit from Liofilchem Laboratory, Italy. Results: One or both of the studied phenotypes were present in 36% of the analyzed isolates. Among the risk factors for the detection of E. coli producing ESBL and/or AmpC in urine were male gender, advanced age, placement of urinary catheter, arterial hypertension, neoplasms, and coexistence of two or more comorbidities. Apart from cephalosporins resistance, isolates producing ESBL and/or AmpC also showed higher resistance to other antibiotics, such as gentamicin (66.7%), ciprofloxacin and levofloxacin (83.3%), and ampicillin (91.7%). Furthermore, 85% of the ESBL/AmpC producing samples were multidrug resistant (resistant to 1 or more drugs in at least 3 different antibiotic categories). Conclusions: The high prevalence of antimicrobial resistance found in this study highlights the importance of implementing national and global measures to tackle the problem, especially in developing countries such as the Dominican Republic, where resources are scarce.

Introducción: La resistencia antimicrobiana es un grave problema global, pues las infecciones causadas por patógenos resistentes están asociadas con una mayor mortalidad y morbilidad. Objetivos: Analizar la prevalencia de aislados de Escherichia coli productores de β-lactamasas de espectro extendido (BLEE) y tipo AmpC procedentes de muestras de orina de pacientes del Hospital Metropolitano de Santiago en la República Dominicana. Métodos: La identificación del patógeno y el antibiograma fueron llevados a cabo mediante los sistemas automáticos BD Phoenix o Microscan®. Se recolectó información general y la historia médica de pacientes con un cultivo de orina positivo para E. coli. La detección de BLEE/AmpC se realizó de manera manual con el estuche comercial ESBL+AmpC de Liofilchem Laboratory, de Italia. Resultados: Un 36 % de las muestras analizadas mostraron uno o ambos fenotipos estudiados. Como factores de riesgo para la detección en orina de E. coli productoras de BLEE o AmpC se encontraron: sexo masculino, edad avanzada, colocación de un catéter urinario, hipertensión, neoplasmas y coexistencia de comorbilidades. Además de resistencia a las cefalosporinas, los aislados productores de BLEE y AmpC revelaron también elevada resistencia a otros antibióticos como gentamicina (66,7 %), ciprofloxacina y levofloxacina (83,3 %), y ampicilina (91,7 %). Un 85,0 % de las muestras productoras de BLEE/AmpC fueron multidrogorresistentes. Conclusiones: La elevada prevalencia de resistencia antimicrobiana encontrada en este estudio refleja la importancia de tomar medidas nacionales y globales para contener el problema, especialmente en países en desarrollo como República Dominicana, donde los recursos son escasos.

Risk factors for ESBL-positive Escherichia coli urinary tract infections / Factores de riesgo para infección urinaria por Escherichia coli BLEE positiva

Abstract Objective: to determine the association of prior antibiotic use, prior hospitalizations, prior urinary tract infections, age, sex and comorbidities in adult patients hospitalized with urinary tract infections caused by extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli. Materials and methods: a case-control study carried out in the hospital setting of private clinics in Lima. Thirty cases and 30 controls were included, with cases defined as hospitalized patients with an ESBL-producing E. coli urinary tract infection diagnosed by urine culture, and controls defined as hospitalized patients without ESBL-producing E. coli infection. Data were taken from incident cases. A bivariate analysis was performed followed by multivariate logistic regression using the significant variables from the bivariate analysis. Results: the associated factors were: prior antibiotic use OR: 261 (22.5-11,017.4), prior hospitalization OR: 4.6 (1.39-16.1), and prior urinary tract infection OR: 36 (6.9-227.2). After adjusting for potential confounding factors using logistic regression, the main statistically significant associated factor was prior antibiotic use, OR: 97.7 (8.4-1,128.3, p<0.000). Conclusion: evidence was found that prior antibiotic use is a risk factor significantly associated with ESBL E. coli urinary tract infections. (Acta Med Colomb 2022; 47. DOI:https://doi.org/10.36104/amc.2022.2131).

Resumen Objetivo: establecer la asociación del uso previo de antibióticos, hospitalizaciones previas, infección urinaria previa, edad, sexo y comorbilidades en pacientes adultos hospitalizados con infección urinaria por Escherichia coli productora de beta lactamasas de espectro extendido (BLEE). Material y métodos: estudio caso control, realizado en clínicas privadas de Lima en ámbito hospitalario. Se incluyeron 30 casos y 30 controles, definiéndose como caso al paciente hospitalizado que cuente con diagnóstico de infección urinaria por urocultivo de E. coli productora de BLEE y como control al paciente hospitalizado sin infección por E. coli BLEE. Se recolectó la información de casos incidentes. Se realizó un análisis bivariado y regresión logística multivariable con las variables que fueron significativas en el análisis bivariado. Resultados: los factores asociados fueron: uso previo de antibióticos OR: 261 (22.5-11017.4), hospitalización previa OR: 4.6 (1.39-16.1), infección urinaria previa OR: 36 (6.9-227.2). Al ajustar por variables potencialmente confusoras mediante regresión logística, se observó que el principal factor asociado con significación estadística fue el uso previo de antibióticos, OR: 97.7 (8.4-1128.3, p<0.000). Conclusión: se encontró evidencia de que el uso previo de antibióticos es un factor de riesgo asociado significativamente a infección urinaria por E. coli BLEE. (Acta Med Colomb 2022; 47. DOI:https://doi.org/10.36104/amc.2022.2131).

MBL PRODUCING P.PENNERI - A MAJOR THREAT TO THE HOSPITAL

Introduction: Proteus spp. has been the signi?cant cause of wound infections as they commonly colonize the wound. Simultaneously multiple drug resistance mechanisms also pose a therapeutic challenge. P.penneri is commonly misidenti?ed as P.mirabilis which is multidrug resistant. The current study was Aim and objectives: conducted to determine the prevalence of MBLs in P.penneri among wound infections at a tertiary care hospital from Western Maharashtra. Specimens collected from wounds of OPD and IPD patients were examined by standard Material and Methods: bacteriological methods. All Proteus isolates were subjected to Antimicrobial susceptibility and MBL production test as per CLSI guidelines. Total 1826 wound samples were screened over the period of May 2017 to July 2018. Overall Results: prevalence of Proteus spp. was 7.12%, P.mirabilis being the commonest. Among all isolates 53.68% and 37.04% of P.mirabilis and P.vulgaris were ESBL producers, respectively. The rate for MBL production was 11.58% and 0% for P.mirabilis and P.vulgaris, respectively. On the contrary 75% of P.penneri isolates were ESBL producers and 12.5% were MBL producers. Discussion and conclusion: Emergence of ESBL and MBL producers is of special concern as Proteus spp. is intrinsically resistant to tigecycline and colistin. Identi?cation of P.penneri from clinical specimens is necessary, due to its multidrug resistance which makes clinical treatment extremely dif?cult. This will limit its control and eradication especially from wound infections as they are common colonizers. Therefore isolation of such beta lactamases producing P.penneri shall be considered as an alarming sign to control the spread of this superbug.

Extended Spectrum Beta Lactamase producing Gram negative bacteria - An emerging menace in Neonatal Sepsis.

Introduction: Neonatal sepsis caused by extended spectrum beta lactamase (ESBL) producing Gram negative bacteria (GNB) is associated with significantly high mortality and morbidity. Clinical features and risk factors for such neonatal sepsis can help in identifying it early. Objectives: Aim of the study was to estimate the incidence, risk factors, clinical features and antibiotic sensitivity of GNB and outcomes of ESBL GNB in neonatal sepsis. Methodology: A prospective observational conducted at regional tertiary care health center. Statistical analysis was carried out with SPSS version 23.0. Results: A total of 87 cases of Gram negative neonatal sepsis were included in study. Male: female was 1.7:1. Forty nine (56.3%) isolates were ESBL positive strains. The clinical features in order of frequency were shock, lethargy, sclerema, disseminated intravascular coagulation and severe thrombocytopenia. Out born neonates (p=0.03), late onset sepsis (p=0.05) and mechanical ventilation (p=0.002) were the risk factors for ESBL GNB sepsis. Mortality associated with ESBL sepsis was 26.5%. Carbapenems and Piperacillin + Tazobactum were most sensitive antibiotics and high resistant for cephalosporins was observed. Conclusion: ESBL GNB neonatal sepsis is an emerging threat with high mortality in Neonatal Intensive care unit.

In Silico Modeling and Docking Analysis of CTX-M-5, Cefotaxime-Hydrolyzing ?-Lactamase from Human-Associated Salmonella Typhimurium

Objective: To computationally model the CTX-M-5 ?-lactamase and establish its structure, which is exclusively present in human-associated Salmonella. Methods: The CTX-M-5 aminoacid sequence (Uniprot ID:O65975) of Salmonella enterica subsp. enterica serovar typhimurium was retrieved from UniProt database and subjected to homology modeling using MODELLER 9v7. The homology models were duly validated using RAMPAGE tool by generating Ramachandran plots, ERRAT graphs, and ProSA score. DoGSiteScorer server and ConSurf server were used to detect the cavities, pockets, and clefts to identify conserved amino acid sites in the predicted model. Subsequently, the modeled structure was docked using CLC Drug Discovery Workbench against proven drugs and known inhibitors. Results: Obtained high-quality homology model with 91.7% of the residues in favorable regions in Ramachandran plot and qualified in other quality parameters. Docking studies resulted in a higher dock score for PNK (D-benzylpenicilloic acid) molecule when compared to other reported inhibitors. Conclusion: This in silico study suggests that the compound PNK could be an efficient ligand for CTX-M-5 ?-lactamase and serve as a potent inhibitor of CTX-M-5.

Antimicrobial Resistance: The 'Other' Pandemic!

Antimicrobial resistance is projected to kill 10 million people by 2050. The biggest driver of antimicrobial resistance is excessive/unrestricted use of antimicrobials in humans and animals. Antimicrobial resistance is a problem in all types of pathogens including bacteria, mycobacteria, viruses, fungi, and parasites both globally and India and in both adults and children. The areas of greatest concern for India is the epidemic of MDR and XDR tuberculosis and resistance in gram-negative pathogens. The alarming rate of extended spectrum beta lactamase (ESBL) production in Enterobacteriales in both community and health care–associated infections is driving carbapenem use. Rates of carbapenem resistance are now signifcantly high in health care–associated gram negative pathogens with associated high mortality rates. The key solution to this antimicrobial resistance crisis needs participation of all stakeholders and lies in promoting rational antimicrobial therapy

Extended-spectrum beta-lactamases among Klebsiella pneumoniae from Iraqi patients with community-acquired pneumonia

SUMMARY OBJECTIVE: Beta-lactams resistance is a major clinical problem in treating pneumonia. This study aimed to detect the extended-spectrum beta-lactamases (ESBL) genes in Klebsiella pneumoniae among patients with community-acquired pneumonia (CAP) in Al-Najaf City, Iraq. METHODS: A total of 511 sputum samples were obtained from all suspected patients with CAP in Al-Najaf City, Iraq, from March 2020 to September 2020. Sputum samples were subjected to microbiological tests. The disk diffusion method was used to test antibiotic sensitivity. Production of ESBLs was identified using phenotypic and genotypic methods. RESULTS: The total prevalence of K. pneumoniae was 31.9% (163/511). Using CHROM agar, 41 (25.2%) isolates were ESBL producers. The imipenem 0.0% (n=0/41) and norfloxacin 0.0% (n=0/41) were the most effective antibiotics. The multiplex polymerase chain reaction showed that 46.3% (n=19/41) of isolates harbored ESBL genes. Out of 19 ESBL producers, 47.4% and 15.8% harbored blaCTX-M and blaSHV, respectively. While blaCTX-M and blaSHV genes were detected in 7 (36.8%) isolates, simultaneously. CONCLUSIONS: The imipenem and norfloxacin can be used in empirical treatment of K. pneumoniae isolates in Iraq. The emergence of K. pneumoniae strains harboring ESBL resistance genes necessitates the development of a regular surveillance program to prevent the spreading of these isolates more in Iraqi health care systems.

Risk factors for acquisition of ESBL-producing Escherichia coli and Klebsiella pneumoniae on non-ventilator-associated hospital-acquired pneumonia in a tertiary care hospital in Indonesia

Aims@#This study was aimed to identify the risk factors for the acquisition of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae on non-ventilator hospital-acquired pneumonia (NV-HAP) patients in a tertiary care hospital in Indonesia.@*Methodology and results@#A case-control study was performed between March 31, 2018, and August 31, 2019. Twenty-eight ESBL-producing E. coli and K. pneumoniae isolates and 28 susceptible strains of E. coli and K. pneumoniae obtained from NV-HAP patients were included in this study. Phenotypic screening for ESBL production was performed by the Vitek2 system and subsequently confirmed by double-disk synergy tests. The use of 3rd generation cephalosporin as initial antibiotic therapy for more than three days was the significant risk factor for the acquisition of ESBL-producing E. coli and K. pneumoniae among NV-HAP patients (odds ratio [OR] 41.827; p=0.001). The length of stay of patients with NV-HAP acquiring the ESBL strains was longer than 10 days (OR 17.334; p=0.001).@*Conclusion, significance and impact of study@#The use of 3rd generation cephalosporin as the initial antibiotic for NV-HAP should be restricted to prevent the emergence of ESBL-producing strains. Infection prevention measures are required to control the acquisition of ESBL-producing E. coli and K. pneumoniae in NV-HAP patients.

Detección de ß-lactamasas de espectro extendido en Escherichia coli aisladas de ecosistemas dulceacuícolas de La Habana / Detection of extended-spectrum ß;-lactamases in Escherichia coli isolated from freshwater ecosystems in Havana

Introducción: Las cepas de Escherichia coli productoras de β-lactamasas de espectro extendido son patógenos multirresistentes y una de las bacterias que más contribuyen con la resistencia antibiótica bacteriana en la clínica. Sin embargo, se aíslan cada vez con más frecuencia de ambientes naturales, tales como los ecosistemas acuáticos en los cuales se emplea como un indicador de contaminación fecal. Objetivo: Evaluar la susceptibilidad a los antibióticos y la producción de enzimas ß-lactamasas de espectro extendido de aislados de Escherichia coli procedentes de ecosistemas dulceacuícolas de La Habana. Métodos: Se analizaron 43 aislados de E. coli provenientes de los ríos Almendares, Quibú y Luyanó de La Habana. Se determinó la susceptibilidad a 18 antibióticos y la producción fenotípica de ß-lactamasas de espectro extendido según las normas del Instituto de Estándares para el Laboratorio Clínico. La detección molecular de las enzimas se realizó mediante reacción en cadena de la polimerasa. Se calculó el índice de multirresistencia a los antibióticos y los patrones de resistencia de cada aislado de E. coli- ß-lactamasas de espectro extendido. Resultados: El 65 por ciento de los aislados de E. coli fueron resistentes al menos a un antibiótico y el 35 por ciento fueron sensibles a todos los antibióticos. El fenotipo ß-lactamasas de espectro extendido fue detectado en siete aislados; de estos, cuatro fueron portadores del gen bla CTX-M-1 y tres presentaron bla TEM. El 37 por ciento de los aislados de E. coli mostraron valores de índices de multirresistencia a los antibióticos menores que 0,22; el 16 por ciento de 0,22; el 9,3 por ciento mayor que 0,5; y el 5 por ciento mayor que 0,7. Los aislados de E. coli-BLEE mostraron corresistencia a las familias de las tetraciclinas, quinolonas, aminoglucósidos y macrólidos. Conclusiones: La presencia de aislados ambientales multirresistentes de E. coli productores de ß-lactamasas de espectro extendido en ecosistemas dulceacuícolas de La Habana destaca la necesidad de implementar estrategias de control para prevenir la diseminación de estos aislados en los ambientes naturales(AU)

Introduction: Extended-spectrum β-lactamase-producing Escherichia coli strains are multiresistant pathogens and one of the bacteria contributing most greatly to bacterial antibiotic resistance in clinical practice. However, they are increasingly isolated from natural environments, such as aquatic ecosystems, where they are used as fecal pollution indicators. Objective: Evaluate antibiotic susceptibility and extended-spectrum ß-lactamase enzyme production in Escherichia coli isolates from freshwater ecosystems in Havana. Methods: An analysis was conducted of 43 E. coli isolates from the rivers Almendares, Quibú and Luyanó in Havana. Determination was made of susceptibility to 18 antibiotics and phenotypic production of extended-spectrum ß-lactamases according to standards from the Clinical and Laboratory Standards Institute. Molecular detection of the enzymes was performed by polymerase chain reaction. Estimation was carried out of the antibiotic multiresistance index and the resistance patterns of each extended-spectrum E. coli ß-lactamase isolate. Results: Of the E. coli isolates studied, 65 percent were resistant to at least one antibiotic, whereas 35 percent were sensitive to all antibiotics. The extended-spectrum ß-lactamase phenotype was detected in seven isolates, of which four were carriers of the gene bla CTX-M-1 and three contained bla TEM. 37 percent of the E. coli isolates displayed antibiotic multiresistance index values below 0.22, 16 percent of 0.22, 9.3 percent above 0.5 and 5 percent above 0.7. ESBL E. coli isolates displayed co-resistance to the families tetracyclines, quinolones, aminoglycosides and macrolides. Conclusions: The presence of multiresistant extended-spectrum ß-lactamase-producing environmental E. coli isolates in Havana freshwater ecosystems highlights the need to implement control strategies aimed at preventing the spread of these isolates in natural environments(AU)

Detection of the main multiresistant microorganisms in the environment of a teaching veterinary hospital in Brazil / Detecção dos principais microrganismos multirresistentes no ambiente de um hospital veterinário de ensino no Brasil

Contamination of the veterinary hospital environment with multiresistant pathogens endangers not only hospitalized animals, but also the workplace safety of veterinarians and nurses, animal guardians and, when in case of a teaching hospital, veterinary students. The objective of this study was to map the main points of bacterial contamination of a veterinary teaching hospital in Brazil to identify multiresistant microorganisms and their antimicrobial resistance genes. Samples were collected from 39 different locations of a veterinary school hospital which comprised a pool according to each hospital environment. In certain environments, more than one pool has been collected. All samples were collected in quadruplicates for the selective isolation of the main multiresistant microorganisms: methicillin-resistant Staphylococcus (MRS), vancomycin resistant Enterococcus (VRE), cephalosporinases and/or extended-spectrum beta-lactamase-producing Gram-negative bacteria (ESBL) and Carbapenemase-producing (CP). After isolation and identification of isolates, multiplex-PCR reactions were performed to detect the main genes for each microorganism and antimicrobial susceptibility tests with the main antibiotics used for each bacterial group according to CLSI. Of the 39 veterinary teaching hospital sites collected, all (100%) had at least one of the microorganisms surveyed, and 17.95% (n=7) of the sites were able to isolate the four pathogens. From the 94 pools collected, it was possible to isolate MRS in 81.91% (n=77), VRE in 12.77% (n=12), cephalosporinases and/or ESBL in 62.77% (n=59) and CP in 24.47%. (n=23). Regarding MRS, the mecA gene was detected in all isolates. All isolated VREs were identified as Enterococcus faecalis and presented the vanA gene. Regarding cephalosporinases and/or ESBL, 89.83% (n=53) of the isolates presented the blaTEM gene, 57.63% (n=34) the blaOXA-1 gene, 37.29% (n=22) blaCTX-M gene from some group (1, 2, 9 ou 8/25) and 20.34% (n=12) the blaSHV gene. It was possible to identify the main microorganisms responsible for causing nosocomial infections in humans (VRE, MRS, ESBL and CP) in the veterinary hospital environment, suggesting a source of infection for professionals and students of veterinary medicine, placing a high risk for public health.(AU)

A contaminação do ambiente hospitalar veterinário com patógenos multirresistentes coloca em perigo não apenas os animais hospitalizados, mas também a segurança no local de trabalho de veterinários e enfermeiros, responsáveis por animais e, quando se tratar de um hospital de ensino, estudantes de veterinária. O objetivo deste estudo foi mapear os principais pontos de contaminação bacteriana de um hospital veterinário de ensino no Brasil, identificando microorganismos multirresistentes e seus genes de resistência antimicrobiana. As amostras foram coletadas em 39 locais diferentes de um hospital de escola veterinária, que compreendia um pool de acordo com o ambiente de cada hospital. Em certos ambientes, mais de um pool foi coletado. Todas as amostras foram coletadas em quadruplicados para o isolamento seletivo dos principais microorganismos multirresistentes: Staphylococcus resistente à meticilina (MRS), Enterococcus resistente à vancomicina (VRE), bactérias Gram-negativas produtoras de cefalosporinases e/ou beta-lactamase de espectro estendido (ESBL) e produtoras de carbapenemase (PC). Após o isolamento e identificação dos isolados, foram realizadas reações de PCR multiplex para detectar os principais genes de cada microorganismo e testes de susceptibilidade a antimicrobianos com os principais antibióticos utilizados para cada grupo bacteriano de acordo com o CLSI. Dos 39 locais do VCH coletados, todos (100%) possuíam pelo menos um dos microrganismos pesquisados e 17,95% (n=7) dos locais foram capazes de isolar os quatro patógenos. Dos 94 pools coletados, foi possível isolar MRS em 81,91% (n=77), VRE em 12,77% (n=12), ESBL em 62,77% (n=59) e carbapenemases em 24,47% (n=23). Em relação ao MRS, o gene mecA foi detectado em todos os isolados. Todos os VREs isolados foram identificados como Enterococcus faecalis e apresentaram o gene vanA. Em relação às cefalosporinases e/ou ESBL, 89,83% (n=53) dos isolados apresentaram o gene blaTEM, 57,63% (n=34) o gene blaOXA-1, 37,29% (n=22) o gene blaCTX-M de algum grupo e 20,34% (n=12) o gene blaSHV. Foi possível identificar os principais microrganismos responsáveis por causar infecções nosocomiais em humanos (VRE, MRS, ESBL e CP) no ambiente hospitalar veterinário, sugerindo uma fonte de infecção para profissionais e estudantes de medicina veterinária, colocando alto risco para a saúde pública.(AU)

Detection of the main multiresistant microorganisms in the environment of a teaching veterinary hospital in Brazil

ABSTRACT: Contamination of the veterinary hospital environment with multiresistant pathogens endangers not only hospitalized animals, but also the workplace safety of veterinarians and nurses, animal guardians and, when in case of a teaching hospital, veterinary students. The objective of this study was to map the main points of bacterial contamination of a veterinary teaching hospital in Brazil to identify multiresistant microorganisms and their antimicrobial resistance genes. Samples were collected from 39 different locations of a veterinary school hospital which comprised a pool according to each hospital environment. In certain environments, more than one pool has been collected. All samples were collected in quadruplicates for the selective isolation of the main multiresistant microorganisms: methicillin-resistant Staphylococcus (MRS), vancomycin resistant Enterococcus (VRE), cephalosporinases and/or extended-spectrum beta-lactamase-producing Gram-negative bacteria (ESBL) and Carbapenemase-producing (CP). After isolation and identification of isolates, multiplex-PCR reactions were performed to detect the main genes for each microorganism and antimicrobial susceptibility tests with the main antibiotics used for each bacterial group according to CLSI. Of the 39 veterinary teaching hospital sites collected, all (100%) had at least one of the microorganisms surveyed, and 17.95% (n=7) of the sites were able to isolate the four pathogens. From the 94 pools collected, it was possible to isolate MRS in 81.91% (n=77), VRE in 12.77% (n=12), cephalosporinases and/or ESBL in 62.77% (n=59) and CP in 24.47%. (n=23). Regarding MRS, the mecA gene was detected in all isolates. All isolated VREs were identified as Enterococcus faecalis and presented the vanA gene. Regarding cephalosporinases and/or ESBL, 89.83% (n=53) of the isolates presented the blaTEM gene, 57.63% (n=34) the blaOXA-1 gene, 37.29% (n=22) blaCTX-M gene from some group (1, 2, 9 ou 8/25) and 20.34% (n=12) the blaSHV gene. It was possible to identify the main microorganisms responsible for causing nosocomial infections in humans (VRE, MRS, ESBL and CP) in the veterinary hospital environment, suggesting a source of infection for professionals and students of veterinary medicine, placing a high risk for public health.

RESUMO: A contaminação do ambiente hospitalar veterinário com patógenos multirresistentes coloca em perigo não apenas os animais hospitalizados, mas também a segurança no local de trabalho de veterinários e enfermeiros, responsáveis por animais e, quando se tratar de um hospital de ensino, estudantes de veterinária. O objetivo deste estudo foi mapear os principais pontos de contaminação bacteriana de um hospital veterinário de ensino no Brasil, identificando microorganismos multirresistentes e seus genes de resistência antimicrobiana. As amostras foram coletadas em 39 locais diferentes de um hospital de escola veterinária, que compreendia um pool de acordo com o ambiente de cada hospital. Em certos ambientes, mais de um pool foi coletado. Todas as amostras foram coletadas em quadruplicados para o isolamento seletivo dos principais microorganismos multirresistentes: Staphylococcus resistente à meticilina (MRS), Enterococcus resistente à vancomicina (VRE), bactérias Gram-negativas produtoras de cefalosporinases e/ou beta-lactamase de espectro estendido (ESBL) e produtoras de carbapenemase (PC). Após o isolamento e identificação dos isolados, foram realizadas reações de PCR multiplex para detectar os principais genes de cada microorganismo e testes de susceptibilidade a antimicrobianos com os principais antibióticos utilizados para cada grupo bacteriano de acordo com o CLSI. Dos 39 locais do VCH coletados, todos (100%) possuíam pelo menos um dos microrganismos pesquisados e 17,95% (n=7) dos locais foram capazes de isolar os quatro patógenos. Dos 94 pools coletados, foi possível isolar MRS em 81,91% (n=77), VRE em 12,77% (n=12), ESBL em 62,77% (n=59) e carbapenemases em 24,47% (n=23). Em relação ao MRS, o gene mecA foi detectado em todos os isolados. Todos os VREs isolados foram identificados como Enterococcus faecalis e apresentaram o gene vanA. Em relação às cefalosporinases e/ou ESBL, 89,83% (n=53) dos isolados apresentaram o gene blaTEM, 57,63% (n=34) o gene blaOXA-1, 37,29% (n=22) o gene blaCTX-M de algum grupo e 20,34% (n=12) o gene blaSHV. Foi possível identificar os principais microrganismos responsáveis por causar infecções nosocomiais em humanos (VRE, MRS, ESBL e CP) no ambiente hospitalar veterinário, sugerindo uma fonte de infecção para profissionais e estudantes de medicina veterinária, colocando alto risco para a saúde pública.

Sensibilidad in vitro de Escherichia coli a la fosfomicina en urocultivos en un Servicio Asistencial Privado de Asunción / In vitro sensitivity of Escherichia coli to fosfomycin in urine cultures in a private healthcare service in Asunción

La fosfomicina es un antibiótico natural, que actúa sobre la síntesis de la pared celular, con actividad bactericida y de amplio espectro. En este trabajo se evaluó la sensibilidad in vitro de aislados de Escherichia coli (E. coli), incluidos aquellos que producen Beta Lactamasa de Espectro Extendido (BLEE) obtenidos a partir de urocultivos, tomados en diferentes lapsos de colección de datos, en personas de ambos sexos. Fueron incluidos 260 muestras de orina con desarrollo de E. coli provenientes de pacientes que concurrieron al Laboratorio San Roque. El aislamiento e identificación bacteriana se realizó según métodos convencionales y la sensibilidad a los antimicrobianos por el método de difusión de disco. Para la detección de la sensibilidad frente a fosfomicina fueron utilizados discos de 200 µg con el agregado de 50 µg de glucosa 6-fosfato. Se observó frente a los antibióticos evaluados mayor sensibilidad a fosfomicina (98,5%) y nitrofurantoína (97,7%). Ciprofloxacina, trimetoprima y la combinación sulfametoxazol/trimetoprima exhibieron frente a los mismos aislados sensibilidad menor y muy similar entre ellos, con 64,2%, 61,2% y 61,2% respectivamente. En 44 (16,9%) de los aislados de E. coli se detectó la presencia de BLEE y es destacable la alta sensibilidad que mostraron fosfomicina y nitrofurantoína, aún frente a los aislados BLEE positivos, con frecuencias de 90,9% y 93,2%, respectivamente. En resumen, la alta sensibilidad demostrada en el presente estudio por E. coli ante la fosfomicina, abre la posibilidad de considerar a este antibiótico de primera elección en las infecciones urinarias bajas, aún en los casos de gérmenes productores de BLEE, en la población de nuestro país.

Fosfomycin is a natural antibiotic, which acts on the synthesis of the cell wall, with broad spectrum bactericidal activity. In this study, the in vitro sensitivity of Escherichia coli (E. coli) isolates was evaluated, including those that produce Extended Spectrum Beta Lactamase (ESBL) obtained from urine cultures taken at different data collection times, in people of both sexes. Were included 260 urine samples with development of E. coli from patients who attended the San Roque laboratory. Bacterian isolation and identification was carried out according to conventional methods and antimicrobial sensitivity by the disk diffusion method. For detection of sensitivity to fosfomycin, 200 µg discs were used with the addition of 50 µg of glucose 6-phosphate. A greater sensitivity for fosfomycin (98.5%) and nitrofurantoin (97.7%) was observed against the evaluated antibiotics. Ciprofloxacin, trimethoprim and the sulfamethoxazole / trimethoprim combination exhibited in front of the same isolated lower sensitivity and very similar among them, with 64.2%, 61.2% and 61.2% respectively. In 44 (16.9%) of the E. coli isolates the presence of ESBL was detected and the high sensitivity shown by fosfomycin and nitrofurantoin is noteworthy, even compared to the positive ESBL isolates, with frequencies of 90.9% and 93,2%, respectively. In summary, the high sensitivity demonstrated in the present study by E. coli to fosfomycin opens the possibility of considering this first-choice antibiotic in lower urinary infections, even in ESBL-producing germs, in the population of our country.

Antimicrobial-resistant enterobacteria in surface waters with fecal contamination from urban and rural communities

Abstract INTRODUCTION: Inadequate wastewater treatment and fecal contamination have a strong environmental impact on antimicrobial resistance (AMR). This study evaluated the profile of AMR enterobacteria and fecal contamination from four surface waters: Jiquiriça-Brejões River and Cabrito, Tororó, and Abaeté Lagoons. METHODS: We analyzed AMR β-lactamase genes using the polymerase chain reaction method and fecal contamination using Coliscan®. RESULTS: We found high levels of fecal contamination, β-lactamase producers, and AMR genes (blaOXA-48, blaSPM, and blaVIM) in all waterbodies. CONCLUSIONS: Poor sanitation evidenced by fecal contamination and human activities around these surface waters contributed to the distribution and increase in AMR enterobacteria.

Identificación de los mecanismos resistencia enzimáticos en uropatógenos de pacientes ambulatorios de un hospital público de San Lorenzo, Paraguay; 2015-2019 / Identification of enzyme resistance mechanisms in pathogens from an outpatient clinic in a public hospital in San Lorenzo, Paraguay; 2015-2019

Introducción: El propósito del estudio es determinar en los urocultivos, la prevalencia de los mecanismos enzimáticos de resistencia encontrados in vitro. Materiales y métodos: Se realizó un estudio retrospectivo, de ambos sexos, mayores de 18 años, que acudieron al Consultorio Externo de Clínica Médica y Urgencias por síntomas de infección urinaria. Se incluyeron todos los urocultivos en los que se aislaron uropatógenos con recuento ≥ 105 UFC/mL. Se excluyeron los urocultivos polimicrobianos, los que no contaban con antibiograma o aquellos con datos clínicos incompletos. Resultados: Se identificaron 1031 urocultivos que cumplieron con los criterios establecidos para la realización del estudio. El 56% correspondió al sexo femenino y el 43% al masculino. La edad media de las mujeres fue de 52± 20 años y el de los hombres fue de 62±16 años. Los uropatógenos más frecuentes fueron Escherichia coli 553 (52% en promedio) seguida de Klebsiella pneumoniae con 148 (14% en promedio). Urocultivos de varones: El principal mecanismo de resistencia de Escherichia coli fueron las BLEE, 55 aislamientos (91%); seguida de las MBL, 3 aislamientos (5%) y KPC, 2 aislamientos (3%). En Klebsiella pneumoniae en 53 aislamientos se puedo observar: BLEE, 31 aislamientos (58%); seguida de las KPC 13 aislamientos (25%) y MBL, 9 aislamientos en (16%). Urocultivos en mujeres: Las enzimas de Escherichia coli fueron 81 aislamientos, de los cuales fueron BLEE, 79 aislamientos (97%); seguido de las KPC, 1 aislamiento (1%) y las MBL, 1 aislamiento (1%). En Klebsiella pneumoniae se pudo observar los siguientes mecanismos enzimáticos en base a 35 aislamientos; BLEE, 19 aislamientos (54%), seguida de las KPC, 12 aislamientos (34%) y por último, MBL, 4 aislamientos (13%). Conclusión: En las IVU de nuestro estudio, Escherichia coli y Klebsiella pneumoniae fueron las principales bacterias que originan resistencia a los antibióticos y la BLEE fue la enzima más frecuentemente identificada en ambos sexos.

Introduction: The objective of this study was to assess frequency of enzymatic resistance mechanisms isolated from community urinary tract infections (UTI) determined in vitro in urine cultures. Objectives: This is a retrospective study, a total of 1031 urine samples were included from patients with urinary tract infection who had consulted at the Outpatient Clinic and Emergencies Services. The following information was recorded, age, sex, urine sample. All urine cultures in which pathogens with a count of ≥ 105 CFU / mL were included. Were excluded polymicrobial urine cultures, those without an antibiogram or those with incomplete clinical data. Results: A total of 1031 urine samples met inclusion criteria, 56% of patients were female and 43% male. The mean age of the women was 52 ± 20 years and in men was 62 ± 16 years. 553 (52%) E. coli and 148 (14%) Klebsiella strains were isolated from community samples. Male urine cultures: The main resistance mechanism of Escherichia coli was ESBLs, 55 isolates (91%); followed by MBL, 3 isolates (5%) and KPC, 2 isolates (3%). In Klebsiella pneumoniae (53 isolates); ESBL, 31 isolates (58%); followed by KPC 13 isolates (25%) and MBL, 9 isolates in (16%). Female urine culture: Escherichia coli enzymes 81 isolates, of which ESBLs were 79 isolates (97%); followed by KPC, 1 isolate (1%) and MBL, 1 isolate (1%). In Klebsiella pneumoniae the following enzymatic mechanisms could be observed based on 35 isolates; ESBL, 19 isolates (54%), followed by KPCs, 12 isolates (34%) and finally, MBL, 4 isolates (13%). Conclusion: The result of our study showed high prevalence of Escherichia coli and Klebsiella pneumoniae causing resistance to antibiotics in culture bacteria from urine samples of patients with UTI, and ESBL was the main ß-lactamase resistance mechanism in Klebsiella and E. coli isolates in both male and female.

Isolation and Molecular Characterization of Extended Spectrum Beta Lactamase Producing Escherichia coli from Milk

Milk plays a major role as a source of nutrition in the diet but contaminated milk can be a source of harmful bacteria. Escherichia coli is opportunistic pathogen and is responsible for a wide range of infections. The prevalence of pathogenic multi-drug resistant extended-spectrum β-lactamase (ESBL)-producing E. coli is increasing and becoming a global concern. A study was carried out to isolate ESBL producing E. coli from 150 milk samples from Anand and around villages. Total 94(62.66%) samples were found positive as E. coli by isolation on MacConkey and Eosin Methylene Blue agar which were confirmed by primary & biochemical tests including Gram’s staining. Antibiotic sensitivity test (ABST) was performed against 6 antibiotics and isolates found resistant to Aztrionem: 58(61%), Cefoxitin: 20(21%), Ceftriaxone: 56(59%), Ceftazidime: 62(65%), Cefpodoxime: 34(44.73%) & Ceftazidime + Clavulanic acid: 8(8.5%). A total 34(36.17%) ESBL producing E. coli were phenotypically confirmed by ABST and Epsilometer test. Genotypic confirmation of 34 isolates was done by PCR and isolates found positive for bla CTX M-3 gene: 18(52.94%), bla CTX M-9 gene 6(17.64%), bla SHV gene: 5(14.70%) and bla TEM gene: 5(14.70%). In summary, analyzed milk samples were found to have a health risk for consumers due to contamination by ESBL producing E. coli, their pathogenicity and treatment failure as a result of antibiotic resistance

Enterobacteria responsible for urinary infections: a review about pathogenicity, virulence factors and epidemiology

Urinary tract infections (UTIs) are one of the most common types of bacterial infections in humans, bothin the community and in health care settings. UTIs include a range of clinical entities ranging in severityfrom asymptomatic infection to acute cystitis, prostatitis, pyelonephritis, and urethritis. This is one of themost common diseases encountered in medical practice today, affecting people of all ages, from newborn togeriatric age group. The bacteria most commonly implicated as agents responsible for UTIs generally originatein the intestine and include Escherichia coli, Pseudomonas spp., Streptococcus spp., Proteus spp., Klebsiellaspp., Staphylococcus spp., Neisseria gonorrhoeae, Chlamydia trachomatis, and Candida spp. There arevirulence factors of urinary pathogens that promote adhesion to mucosal surfaces and subsequent infections.The multiresistant of these enterobacteria responsible for UTIs is a major public health problem. Antibioticresistance remains a major problem, especially in the developing countries where hygiene conditions are stillprecarious and antibiotic use is often abusive and poorly controlled. The multidrug-resistant Enterobacteriaceaemost implicated in UTIs by extended-spectrum beta-lactamases (ESBL) and carbapenemase production are:Klebsiella pneumoniae, E. coli, and Proteus spp., not to mention Acinetobacter baumanii and Pseudomonasaeruginosa, the most carbapenemase producing. The detection of ESBL and carbapenemase production isbased mainly on phenotypic and genotypic tests.

Antimicrobiana de óleos essenciais e extratos glicólicos de plantas medicinais sobre enterobactérias resistentes a beta-lactâmicos de amplo espectro / Evaluation of the antimicrobial activity of essential oils and glycolic extracts of medicinal plants on wide spectrum beta-lactamic resistant enterobacteria

O avanço da resistência bacteriana tem sido considerado pela OMS uma das maiores ameaças à saúde pública do século XXI. Os patógenos multirresistentes aumentam as taxas de mortalidade e elevam o custo do tratamento antibiótico no mundo todo, tanto em hospitais quanto na comunidade. As opções terapêuticas estão ficando cada vez mais escassas frente a tantos mecanismos de resistência. Dentre eles, as enzimas, como as beta-lactamases de espectro estendido (ESBL), se destacam pelo grande espectro de hidrólise, inativando penicilinas e cefalosporinas. A disseminação da capacidade de produção dessas enzimas é facilitada pela localização dos genes em plasmídeos, e atualmente está presente tanto em ambientes hospitalares quanto na comunidade. Novas alternativas estão sendo pesquisadas, e possíveis fontes de substâncias com atividade antimicrobiana são as plantas, já amplamente utilizadas na medicina popular. Entre elas, podemos citar a arnica (Arnica montana) e a erva-cidreira (Cymbopogon citratus), que tem atividade antimicrobiana descrita na literatura, e apresentaram atividade contra enterobactérias em testes prévios deste grupo de pesquisa. Neste trabalho, foram testadas as atividades in vitro de extratos glicólicos e óleos essenciais destas duas plantas contra enterobactérias produtoras de ESBL, tendo sido testadas 42 cepas, e dentre elas, 12 apresentaram teste positivo para ESBL. Foi realizada a técnica de difusão em ágar, que mostrou que os micro-organismos apresentaram certa sensibilidade aos extratos glicólicos, com um maior destaque para o extrato de arnica. Sendo assim, é sugerido que esses extratos possam ser utilizados como alternativa em produtos saneantes de superfícies hospitalares e de unidades de cuidado à saúde, sendo uma alternativa aos produtos sintéticos potencialmente tóxicos comumente empregados.

The advancement of bacterial resistance has been considered by WHO as one of the greatest threats to public health in the 21st century. Multidrug-resistant pathogens increase mortality rates and raise the cost of antibiotic treatment worldwide, both in hospitals and community. Therapeutic options are becoming increasingly scarce in face of so many resistance mechanisms. Among them, enzymes, such as extended-spectrum beta-lactamases (ESBL), stand out for the large spectrum of hydrolysis, inactivating penicillins and cephalosporins. The dissemination of the production capacity of these enzymes is facilitated by the location of genes in plasmids, and is currently present both in hospital environments and in the community. New alternatives are being researched, and possible sources of substances with antimicrobial activity are plants, which are already widely used in folk medicine. Among them, we can mention arnica (Arnica montana) and lemon balm (Cymbopogon citratus), which has antimicrobial activity described in literature, and showed activity against enterobacteria in previous tests of this same research group. In this work, in vitro activities of glycolic extracts and essential oils of these two plants were tested against ESBL-producing enterobacteria. 42 strains were tested, and among them, 12 tested positive for ESBL. Agar diffusion technique was performed, which showed that the microorganisms showed some sensitivity to glycolic extracts, with greater emphasis on the extract of arnica. Therefore, it is suggested that these extracts can be used as an alternative in sanitizing products on hospital surfaces and health care units, being an alternative to the potentially toxic synthetic products commonly used.

Quality microbiological diagnostics and antimicrobial susceptibility testing, an essential component of antimicrobial resistance surveillance and control efforts in Pacific island nations

Problem@#Emerging bacterial antimicrobial (antibiotic) resistance (AMR) is a global threat to human health. However, a majority of lower income countries do not have microbiological diagnostic testing for prompt, reliable confirmation of bloodstream infection and identification of AMR.@*Context@#Clinicians in Pacific island nations are increasingly challenged by patients who have infection due to antimicrobialresistant bacteria. Treatment of infection remains empirical because of a lack of diagnostic testing capacity and may follow guidelines that were formulated without reference to local measures of AMR prevalence. There is limited understanding among clinicians of microbiology testing and test interpretation.@*Action@#Examine the lessons learnt from pilot laboratory development programmes in two Pacific island nations that focused on establishing standard procedures for micrological diagnostics and antimicrobial susceptibility testing (AST) and on improving the training of clinicians to increase their use of laboratory services.@*Outcome@#The pilot programmes addressed a range of logistical difficulties and evaluated two blood culture systems. They also examined and improved internal QC implementation and evaluated the prevalence of AMR.@*Discussion@#Continued development of microbiological diagnostic capability in the Pacific region is paramount. Pacific Island nations need to develop the capability of at least one central laboratory to culture AMR pathogens and subject them to quality-controlled AST or arrange for suitable referral to a nearby country.