Evaluation of rapid identification methods for foodborne diarrhea Campylobacter and detection of drug resistance / 中华检验医学杂志

Objective:To explore and evaluate a appropriate suitable method for detection of Campylobacter and antibiotic sensitivity test for foodborne diarrhea in clinical laboratories. Methods:Pre-experiment:a total number of 400 fecal samples of patients with foodborne diarrhea were prospectively collected from the intestinal disease clinic of Beijing Tongren Hospital from September 2017 to January 2018. Double-hole filtration culture method and modified cefoperazone charcoal deoxycholate (CCD) agar culture method were used for fecal culture in micro-aerobic environment for 48 hours, and then suspicious colonies were identified by matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. Meanwhile, C. jejuni and C. coli were detected by real-time quantitative polymerase chain reaction(qPCR). Large sample verification: 2 062 fecal samples of patients with foodborne diarrhea in three hospitals of different levels in different areas of Beijing were collected for qPCR detection and culture from April 2018 to March 2019. The antimicrobial sensitivity test (AST) of C. jejuni and C. coli was performed according to the disk diffusion method and agar dilution method recommended by Clinical and Laboratory Standards Institute and National Antimicrobial Resistance Monitoring System for Enteric Bacteria. The results of the three detection methods and the consistency of the two antibiotic sensitivity tests were compared. Results:In the pre-experiment, the positive rates of Campylobacter ( jejuni/coli) detected of qPCR, double-hole filtration culture and modified CCD agar culture were 9.0% (36/400), 5.0% (20/400)and 3.5% (14/400), and the difference was statistically significant ( P<0.01). The samples with negative result of qPCR were negative by both culture methods. The total positive rates of Campylobacter detected by qPCR was 8.1% (168/ 2 062)including 7.0% (144/2 062) for C. jejuni and 1.2% (24/2 062) for C. coli. The samples with positive qPCR results were cultured by double-hole filtration culture method and the positive rate was 61.9%(104/168), among which, the positive rate of C. jejuni and C. coli were 58.3%(84/144) and 83.3%(20/24) respectively, which was not significantly different from the detection rate and culture positive rate in the pre-test ( P>0.1). The resistance rates of C. jejuni and C. coli to ciprofloxacin were 94.0%(94/100) and 100.0%(24/24) and to erythromycin were 6.0%(6/100) and 33.3%(8/24). The results from two antibiotic sensitivity test methods were consistent (Kappa>0.75). Conclusions:qPCR is rapid, sensitive and easy to operate, so it is suitable for routine development in clinical laboratories. The double-hole filtration culture method is beneficial to the acquisition of strains and is essential for the further study of Campylobacter. There was no significant difference between agar dilution method and disk diffusion method in antibiotic sensitivity test. Campylobacter showed a very high resistance rate to quinolones, which was no longer suitable for the treatment of Campylobacter foodborne diarrhea in Beijing area. Macrocyclic lipid antibiotics should be preferred.

Comparación de los perfiles de resistencia antimicrobiana de Escherichia coli uropatógena e incidencia de la producción de betalactamasas de espectro extendido en tres establecimientos privados de salud de Perú / Comparison of Escherichia coli antibiotic-resistance profiles and incidence of beta- lactamase phenotypes in three private health facilities in Perú

Introducción. La aparición de enterobacterias multirresistentes y productoras de betalactamasas de espectro extendido en pacientes ambulatorios con infecciones urinarias representa un problema de salud pública en Perú. Objetivo. Comparar los perfiles de resistencia de Escherichia coli uropatógenas e identificar los fenotipos de cepas productoras de betalactamasas de espectro extendido en tres establecimientos privados de salud localizados en las regiones de la costa, la sierra y la selva de Perú. Materiales y métodos. Se llevó a cabo durante el 2016 un estudio descriptivo de 98 muestras de orina de pacientes con infección urinaria, 35 procedentes de Lima (costa), 38 de Juliaca (sierra) y 25 de Iquitos (selva), en el que se determinó la sensibilidad antimicrobiana utilizando ocho discos antibióticos. Asimismo, se evaluó la producción de betalactamasas de espectro extendido con discos de cefotaxima, de ceftazidima o de su combinación, con ácido clavulánico en agar Mueller-Hinton. Resultados. Se identificaron 18 perfiles de resistencia que incluían desde los sensibles a todos los antibióticos hasta los resistentes simultáneamente a siete antibióticos, con el 18,4 % de aislamientos resistentes a un antibiótico y el 54,0 % de multirresistentes. Se detectó producción de betalactamasas en el 28,6 % de las cepas procedentes de la región de Puno. También, se observó un mayor número de casos en el rango de edad de 31 a 45 años con resistencia a ceftazidima, ceftriaxona, gentamicina y trimetoprim-sulfametoxazol en el establecimiento de salud de Puno. Conclusión. Los perfiles de resistencia variaron según la localización geográfica del establecimiento de salud, observándose mayor resistencia a los antibióticos en la región de la sierra de Perú, con el 28,6 % de cepas productoras de betalactamasas de espectro extendido.

Introduction: The appearance of multidrug-resistant and beta-lactamase producing enterobacteria in outpatient care facilities represent a public health problem in Perú. Objective: To compare the resistance profiles of uropathogenic Escherichia coli and to identify extended-spectrum beta-lactamase-producing phenotypes in three private health facilities located in the Peruvian coast, Andean and jungle regions. Materials and methods: We conducted a descriptive study on 98 urine samples from Lima (coast), Juliaca (Andean region) and Iquitos (jungle region) during 2016. We determined the antimicrobial susceptibility in 35 samples from Lima, 38 from Juliaca and 25 from Iquitos using eight antibiotic disks in samples from patients diagnosed with urinary infection. We also evaluated the production of extended-spectrum beta-lactamases with cefotaxime and ceftazidime disks and a combination of both with clavulanic acid on Mueller-Hinton agar. Results: We identified 18 resistance profiles ranging from those sensitive to others simultaneously resistant to seven antibiotics: 18.4% resistant to one and 54.0% to multiple antibiotics. We detected beta-lactamase production in 28.6% of the strains from the Puno region. Likewise, we observed a greater number of cases with resistance to ceftazidime, ceftriaxone, gentamicin, and trimethoprim-sulfamethoxazole in Puno's health facility in patients within the 31 to 45 year age range. Conclusion: Resistance profiles varied according to the geographical location of the health facilities under study. Resistance to antibiotics was higher in the Andean region with 28.6% of strains producing extended-spectrum beta-lactamases.

Clinical features of children with burkholderia gladioli bloodstream infection and drug susceptibility of burkholderia gladioli / 第二军医大学学报

Objective To explore the clinical features of children with Burkholderia gladioli (B. gladioli) bloodstream infection and the drug susceptibility of B. gladioli. Methods The clinical data of 63 children with B. gladioli bloodstream infection admitted to Wuhan Children’s Hospital, Tongji Medical College of Huazhong University of Science and Technology from Jan. 2015 to Jan. 2016 were retrospectively analyzed, and 81 children with non-bacterial infectious diseases in the same period were enrolled as controls. The C-reactive protein (CRP) level, procalcitonin (PCT) level and white blood cell (WBC) counts of children were compared between the two groups. B. gladioli was isolated from the blood samples of children and cultured for preliminary identifying by Phoenixtm100 automatic microorganism identification instrument and confirming by MALDI-TOP MS mass spectrometer. The in vitro antimicrobial susceptibility testing of B. gladioli were performed by Kirby-Bauer method. Results The children infected with B. gladioli were mainly infants, with 52 cases (82.54%) being three years old or below. All cases had serious underlying diseases, including bronchitis, pneumonia and leukemia. Compared with the control group, the PCT level, CRP level, and WBC counts in the children of the B. gladioli group were significantly increased (all P<0.05). According to the drug susceptibility criteria of Pseudomonas aeruginosa, the isolated B. gladioli was highly sensitive to amikacin, gentamycin, tetracycline, minocycline, cotrimoxazole, ciprofloxacin, levofloxacin, imipenem, meropenem, cefepime, piperacillin, piperacillin/tazobactam and cefoperazone/sulbactam, but had low resistance to chloramphenicol and high resistance to ceftazidime and aztreonam. Conclusion Children infected with B. gladioli are mainly infants aged≤3 years old, with low immunity and poor resistance. Blood culture and CRP level, PCT level and WBC counts can be used as diagnostic indicators of disease outcomes. Piperacillin/tazobactam and cefoperazone/sulbactam should be the first selected drugs for the treatment of children with B. gladioli bloodstream infection.

Toxicological, phytochemical, and antibacterial assessment of Chlorella vulgaris and Spirulina platensis powder in albino rats. A preliminary study

Objectives: To describe acute toxicity, antibacterial activity and phytochemical assessment of Chlorella vulgaris and Spirulina platensis powders. Material and Methods: FeCl3 test, Wagner test, Keller Killiani test, frothing test, alkaline solution and dilute acid; concentrated sulphuric acid were used for phytochemical analysis. Antibacterial screening of the extracts was conducted using the disc gel diffusion method in E. coli, S. aureus and B. cereus clinical strains. In order to evaluate acute toxicity and its effects on haematological and biochemical parameters; 15 albino rats were grouped into five groups: I (powder of aqueous extract of Chlorella vulgaris), II (powder of methanol extract of Chlorella vulgaris), III (powder of aqueous extract of Spirulina platensis), IV (powder of methanol extract of Spirulina platensis) and V (control). The dosage was 25g/day/rat. After six days, haematological and biochemical parameters and gross pathologic changes were evaluated. Results: Alkaloids and flavonoids were detected from the methanol extracts of both Chlorella vulgaris and Spirulina platensis (Arthrospira). Only cardiac glycosides and steroids were detected from Spirulina's extracts. Chlorella vulgaris extracts significantly inhibited B. cereus. Rats fed with Chlorella vulgaris and Spirulina platensis powder showed an increase in white blood cell counts and haemoglobin level compared to negative control rats (p<0.001). Serum glumatic oxalate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT) had normal values but significative differences between groups (p<0.001). Conclusion: This powder is rich in bioactive phytochemicals but only Chlorella's extracts have shown antibacterial effect. Signs of toxicity weren't found in any parameter.

Objetivos: Describir la toxicidad aguda, efecto antibacteriano y análisis fitoquímico de los polvos de Chlorella vulgaris y Spirulina platensis. Materiales y métodos: Se realizaron las pruebas de FeCl3, Keller Killiani, de saponinas, solución alcalina y de concentración de ácido sulfúrico para el análisis fitoquímico. El efecto antibacteriano de los extractos fue evaluado mediante el método de difusión con discos en cepas de E. coli, S. aureus y B. cereus. Para evaluar la toxicidad aguda y los efectos del polvo en parámetros hematológicos y bioquímicos, se agruparon 15 ratas albinas en cinco grupos: I (polvo de extracto acuoso de Chlorella vulgaris); II (polvo de extracto metanólico de Chlorella vulgaris); III (polvo de extracto acuoso de Spirulina platensis); IV (polvo de extracto metanólico de Spirulina platensis), y V (grupo control). La dosis usada fue de 25 g/día/rata. Después de seis días, se evaluaron todos los parámetros y cambios macroscópicos en los órganos. Resultados: Se encontraron alcaloides y flavonoides en los extractos metanólicos de Chlorella vulgaris y Spirulina platensis (Arthrospira). Se detectaron glucósidos cardiacos y esteroides en los extractos de Spirulina platensis. Los extractos de Chlorella vulgaris inhibieron el crecimiento de Bacillus cereus. Las ratas alimentadas con los polvos de Chlorella vulgaris y Spirulina platensis incrementaron el conteo de leucocitos y los valores de hemoglobina comparados con el grupo control (p<0,001). Las transaminasas (SGOT y SGPT) se encontraron en valores normales, pero con diferencias significativas entre los grupos (p<0,001). Conclusiones: Estos polvos son ricos en componentes fitoquímicos activos, pero solo los extractos de Chorella vulgaris mostraron efecto antibacteriano. No se encontraron signos de toxicidad aguda en ningún parámetro.

Resistência antimicrobiana e tipagem molecular de pseudomonas aeruginosa isoladas de feridas crônicas / Antimicrobial resistance and molecular typing of pseudomonas aeruginosa isolated from chronic wounds

Introdução: Para a correta reparação das feridas, as diversas fases do processo de cicatrização devem ocorrer na sequencia correta, numa intensidade ideal. Vários fatores afetam a cicatrização das feridas ao interferir em uma ou mais fases deste processo, tal como, a presença de infecção. Objetivo geral: Analisar as cepas de Pseudomonas aeruginosa encontradas nas feridas crônicas tratadas com gel de carboximetilcelulose a 2% ou com placa de poliuretano. Método: Pesquisa observacional descritiva, com abordagem quantitativa, realizada através da coleta de material biológico de feridas crônicas de pacientes atendidos em serviços ambulatoriais, empregando swabs. A pesquisa foi aprovada pelo Comitê de Ética em Pesquisa (Hospital Universitário Antônio Pedro ­ UFF) com número de parecer 815.353. As cepas de P. aeruginosa foram identificadas por MALDI-TOF MS, submetidas a testes de susceptibilidade aos antimicrobianos, identificação de genes de virulência através de PCR e tipagem molecular através de PFGE. Resultados: Das 43 feridas a partir das quais foramcoletados swabs, em 31 (72,09%) obteve-se isolamento de P. aeruginosa (foram identificadas 48 cepas). Estas feridas têm 3,6 vezes mais chances de desenvolverem infecção quando comparadas àquelas a partir das quais esse microrganismo não foi isolado. As cepas isoladas dos pacientes em uso de gel de carboximetilcelulose a 2% apresentaram maiores taxas de resistência a gentamicina e ciprofloxacino (ambos com 7,89%). Já as cepas isoladas dos pacientes tratados com placa de poliuretano, destacaram-se pela resistência a ciprofloxacino (90%). Foram identificadas três cepas multirresistentes de duas feridas tratadas com placa de poliuretano impregnada com prata. Foram positivas para presença do gene exoS 26 cepas (54,16%), e 13 (27,08%), para o gene exoU. Observou-se mesmo perfil de PFGE entre as cepas coletadas em diferentes momentos de onze pacientes, enquanto que em seis pacientes as cepas coletadas em diferentes momentos foram distintas. Não houve semelhança de padrões de fragmentação de DNA entre cepas derivadas de pacientes diferentes. Conclusão: A maioria das feridas não apresentava sinais clínicos de infecção. Foram identificadas 48 cepas de P. aeruginosa. O isolamento deste microrganismo é fator de risco para desenvolvimento de infecção. As cepas de P. aeruginosa têm baixos índices de resistência antimicrobiana, com apenas três cepas multiresistentes. Os desbridamentos realizados nas feridas crônicas não têm sido efetivos para descolonização de P. aeruginosa, já que um mesmo clone bacteriano foi identificado na ferida em diferentes momentos, na maioria dos casos

Introduction: For proper wound healing, various stages of the healing process must occur in a correct sequence and an ideal intensity. Several factors affect the wounf healing on one or more phases of this process, such as the presence of infection. General objective: To analyze Pseudomonas aeruginosa strains found in chronic wounds treated with 2% carboxymethylcellulose gel or polyurethane plate. Method: Descriptive observational research with a quantitative approach, carried out through the collection of biological material of chronic wounds of patients attended in outpatient services, using swabs. The study was approved by the Research Ethics Committee (Academic Hospital Antonio Pedro - UFF) with number 815.353. P. aeruginosa strains were identified by MALDI-TOF MS, subjected to antimicrobial susceptibility testing, identification of virulence genes by PCR and molecular typing by PFGE. Results: Of the 43 wounds from which swabs were collected, at 31 (72.09%) was obtained isolation of P. aeruginosa (48 strains have been identified). These wounds are 3.6 times more likely to develop infection when compared to those from which this microorganism was not identified. The strains isolated from patients using 2% carboxymethyl cellulose gel showed more resistance rates to gentamicin and ciprofloxacin (both 7.89%). Already the strains isolated from patients treated with polyurethane plate, highlighted by the resistance to ciprofloxacin (90%). Three multiresistant strains were identified from two wounds treated with polyurethane plate impregnated with silver. 26 strains (54.16%) were positive for the presence of exoS gene and 13 (27.08%), for the exoU gene. It was observed even PFGE profile among strains collected at different times of eleven patients, while in six patients, strains collected at different times were different. There was no resemblance DNA fragmentation patterns among strains derived from different patients. Conclusion: Most of the wounds showed no clinical signs of infection. 48 strains of P. aeruginosa have been identified. The isolation of this microorganism is a risk factor for development of infection in chronic wounds. Strains of P. aeruginosa demonstrated low antimicrobial resistance rates and only three multi-resistant strains were identified. The debridement performed in chronic wounds is not effective for removing colonization by P. aeruginosa, because same bacterials clones was identified in the wound swabs collected in same patients at different times in most cases

Study on disk diffusion method for antimicrobial susceptibility test of Candida isolated from patients with lung cancer / 肿瘤研究与临床

Objective To compare the ROSCO disk diffusion method with broth microdilution method (CLSI, M27-A) for antimicrobial susceptibility test of Candida species isolated from patients with lung cancer. Methods Danish ROSCO company disk diffusion testing method and bio Merieux ATB FUNGUR2 were applied to test 5-flucytosine, fluconazole, itraconazole and amphotericin B antimicrobial susceptibility for 78 Candida species strains isolated from patients with lung cancer. Results Through evaluating the susceptibility to 5-flucytosine, amphotericin B, fluconazole and itraconazole by disk diffusion method, the Kappa value was 0.89. The sensitive strains detected by one method did not show resistance in another method. The sensitive rates of 78 strains of Candida species to 5-flucytosine, amphotericin B, fluconazole and itraconazole were 88.20 %, 89.17 %, 56.34 % and 52.12 %. The susceptibility of C.albicans, C.tropicalis, C.glabrata and C.krusei to four kinds of antifungal agents was 90.95 %, 85.71 %, 67.50 % and 41.67 %respectively. Conclusions Results of disk diffusion method coincide well with broth microdilution method. It can be chosen as a clinical routine method for antimicrobial susceptibility test.

Pathogenic organisms analysis of 259 cases of suppurative endophthalmitis / 中国基层医药

ObjectiveToanalyzethepathogensofsuppurativeendophthalmitis,theirdistributionanddrug susceptibility.Provide valuable information for the clinical options in effective use of antimicrobial agents.Methods Collected 259 cases of endophthalmitis from the patients with aqueous humor and vitreous fluid for pathogens and drug sensitivitytestinordertocarryontheanalysisoftheresults.Results 113casesofbacterialculturepositivein259 cases of specimens,with a detection rate of 43.63%.6 cases of multiple bacteria infection,and 2 case of fungal infec-tion.The total separation of pathogens was 120 strains.89 strains were gram positive coccus,accounting for 74.17%, 11 were gram positive bacillus strains, accounting for 9.17%, 13 strains of gram-negative bacilli, accounted for 10.83%,gram-negative in 5strains(4.17%),fungus in 2strains(1.67%).Conclusion Main pathogenic bacteria suppurative endophthalmitis is gram-positive, the pathogen species distribution and antimicrobial susceptibility of change should be timely monitored and the rational using of antibiotics can reduce the generation of drug-resistant strains.

Monitoramento e caracterização molecular de Staphylococcus aureus em lesões cutâneas crônicas tratadas com hidrogel e placa de poliuretano / Monitoring and molecular characterization of Staphylococcus aureus in chronic skin lesions treated with hydrogel and polyurethane plate

Este estudo teve como objetivo analisar a presença de cepas de Staphylococcus aureus nas lesões cutâneas crônicas tratadas com hidrogel a 2% e placa de poliuretano. Para tal, foi realizada pesquisa descritiva, com abordagem quantitativa, tendo como campo de pesquisa o Ambulatório de Reparo de Feridas do Hospital Universitário Antônio Pedro (HUAP/UFF) e a Policlínica Comunitária da Engenhoca (PCE), ambos localizados na cidade de Niterói, RJ. A análise microbiológica foi realizada no Laboratório de Controle Microbiológico da Faculdade de Farmácia da Universidade Federal Fluminense (UFF) e no Instituto de Microbiologia Paulo de Góes da Universidade Federal do Rio de Janeiro (UFRJ). A coleta de dados foi realizada em duas etapas, a primeira relacionada ao exame clínico com a identificação do paciente e descrição clínica das lesões e a segunda relacionada à coleta do espécime clínico utilizando-se como instrumento de coleta o swab. Todas as cepas de S. aureus analisadas foram identificadas por MALDI-TOF e a suscetibilidade a antimicrobianos foi determinada pelo teste de disco-difusão em meio sólido, seguindo as normas recomendadas do CLSI (Clinical and Laboratory Standards Institute). A reação em cadeia da polimerase (PCR) foi empregada na detecção do gene mecA e genes lukF-PV e lukS-PV. A diversidade clonal foi verificada pelo PFGE (pulsed-field gel electrophoresis). O S. aureus foi detectado em 82,9% (29/35) dos pacientes em uso de hidrogel e em 100% (8/8) dos pacientes em uso de placa de poliuretano. Os pacientes em uso de placa de poliuretano com prata apresentaram mais sinais clínicos de infecção quando comparados aos pacientes em uso de hidrogel a 2%, principalmente pela presença do exsudato purulento. A razão de prevalência demonstrou que os pacientes que usaram a placa de poliuretano com prata tiveram pelo menos 3,6 vezes maior chance de apresentarem infecção nas feridas quando comparados com os pacientes em uso de hidrogel. A maioria dos S. aureus identificados, em ambos os campos de pesquisa, apresentou resistência à penicilina, meticilina, eritromicina e clindamicina. Em cinco pacientes (10 cepas- 20%) foi observada amplificação para o gene mecA, demonstrando a colonização por MRSA. Não foi observada amplificação para os genes lukF-PV e lukS-PV. Embora tenha sido detectada grande diversidade genética entre as cepas analisadas, o mesmo padrão se repetiu entre os S. aureus coletados em dois momentos diferentes nos mesmos pacientes. No entanto, as amostras 1 e 32/32* apresentaram o mesmo padrão de fragmentação de DNA pelo PFGE. Através da razão de prevalência, determinou-se que os pacientes com MSSA têm 16 vezes mais chance de ter infecção em feridas quando comparados aos pacientes com MRSA. Assim, o tratamento com hidrogel a 2% ou placa de poliuretano não interferiu na colonização por S. aureus. Além disso, verificamos que o uso de placa de poliuretano com prata não é indicado para feridas infectadas quando o paciente possuir como comorbidades hipertensão arterial e insuficiência venosa crônica

This study aimed to analyze the presence of Staphylococcus aureus in chronic wounds treated with hydrogel 2% and polyurethane plate. To do this, was done a descriptive study with a quantitative approach. The field research was the Wound Repair Clinic at the University Hospital Antônio Pedro (HUAP / UFF) and the Community Polyclinic of the Engenhoca, both located in the city of Niterói, RJ. The microbiological analysis was conducted at the Laboratory of Microbiological Control of the Faculty of Pharmacy of the University Federal Fluminense (UFF) and the Institute of Microbiology Paulo de Góes of the University Federal of the Rio de Janeiro (UFRJ). Data collection was carried out in stages, the first related to the clinical examination with the patient identification and clinical description of the wounds and the other step related to the collection of the clinical specimen using the swab. All strains of S. aureus analyzed were identified by MALDI-TOF and susceptibility to antibiotics was determined by disk diffusion test on solid medium following the standards recommended by CLSI (Clinical and Laboratory Standards Institute). The polymerase chain reaction (PCR) was used in the detection of the mecA gene and lukF-PV and luks-PV genes. The clonal diversity was verified by PFGE (pulsed-field gel electrophoresis). The S. aureus was detected in 82.9% (29/35) of patients using hydrogel and in 100% (8/8) the patients using polyurethane plate. Patients using polyurethane plate with silver presented more clinical signs of infection when compared to patients using hydrogel 2%, mainly by the presence of purulent exudate. The prevalence ratio showed that patients who used polyurethane plate with silver had at least 3.6 times more chance to have infection in the wound when compared to patients using hydrogel. Most S. aureus identified in both research fields presented resistance to penicillin, methicillin, erythromycin and clindamycin. Five patients (10 strains - 20%) were observed amplification for the gen mecA demonstranting colonization by MRSA. There was no amplification for lukF-PV and lukS-PV genes. Although was detected a big genetic diversity among the strains analyzed, the same pattern repeated among the S. aureus collected at two different times from the same patients. However, samples 1 and 32/32* showed the same fragmentation pattern by PFGE. Through the prevalence ratio identified that patients with MSSA had 16 times more likely to have infection in wounds when compared to patients with MRSA. Thus, the treatment with hydrogel 2% or polyurethane board did not interfered in the colonization by S. aureus. In addition, we perceived that the use of polyurethane plate with silver is not indicated for infected wounds when patients had comorbidities such as hypertension and chronic venous insufficiency

Susceptibilidad antimicrobiana y genotipificación de Pseudomonas aeruginosa de pacientes con fibrosis quística y otras patologías en Cartagena (Colombia) / Antimicrobial susceptibility and genotypification of Pseudomonas aeruginosa from cystic fibrosis patients and other diseases in Cartagena (Colombia)

Objetivo: el objetivo de este estudio fue analizar el genotipo y susceptibilidad antimicrobiana de Pseudomonas aeruginosa de pacientes con fibrosis quística y otras patologías. Materiales y métodos: se analizaron 20 aislados de pacientes con fibrosis quística y 20 de pacientes con otras enfermedades por medio de la prueba de susceptibilidad antimicrobiana por microdilución en caldo y técnica del ADN polimorfo amplificado aleatorio. Resultados: se observó que los aislados de pacientes con fibrosis quística presentaron mayor resistencia (56 %) en comparación con aislados de pacientes sin fibrosis quística (25 %). Los antimicrobianos más efectivos en ambos grupos fueron cefepima, ceftriaxona y meropenem. Desde el punto de vista genotípico, se observa heterogeneidad entre las cepas de pacientes con fibrosis quística y dos grupos con cepas idénticas de origen hospitalario, lo que sugiere una posible transmisión cruzada. Conclusión: Concluimos que los porcentajes de resistencia de Pseudomonas aeruginosa en este estudio son altas, y este hallazgo se acentúa en el caso de pacientes con fibrosis quística, lo cual deja muy pocas opciones de tratamiento. La tipificación por técnica del ADN polimórfico amplificado aleatorio permitió conocer la variabilidad de genotipos para tener control sobre la transmisión de cepas, lo cual constituye un tópico de importancia en el sistema de salud y el mejoramiento de la calidad de vida de los pacientes.

Objective: Our aim was to analyze genotype and antimicrobial susceptibility of Pseudo-monas aeruginosa from cystic fibrosis patients and other diseases. Materials and methods: We analyzed 20 isolates from cystic fibrosis patients and 20 from patients with other diseases by dilution antimicrobial susceptibility test and random amplified polymorphic DNA technique. Results: We found that isolates from cystic fibrosis patients had higher resistance (56 %) than isolates from patients without cystic fibrosis (26 %). The most effective antimicrobi-als in both groups were cefepime, ceftriaxone and meropenem. With regard to the geno-type, we observed heterogeneity between strains from cystic fibrosis patients and two clus-ters with identical strains from hospital origin, suggesting a possible cross transmission. Conclusion: We concluded that the resistance rate of Pseudomonas aeruginosa in this study was high and this finding is accentuated in patients with cystic fibrosis, leaving few treatment options. Typification by random amplified polymorphic DNA technique allowed us to know the variability of genotypes to control strain transmission; this is an important topic to optimize health services and the quality of life of our patients.

Sensibilidade de isolados de candida spp. a antifúngicos por disco-difusão em ágar e microdiluição em caldo / Antifungal susceptibility of isolates of candida spp. by disk diffusion and broth microdilution

Os testes de sensibilidade aos antifúngicos realizados pelo método de disco-difusão em ágar são práticos e bem conhecidos pelos profissionais do laboratório de microbiologia, entretanto apresentam particularidades que os diferem dos testes realizados para bactérias. O objetivo deste trabalho foi comparar as técnicas de disco-difusão em ágar e microdiluição em caldo na determinação da sensibilidade in vitro de isolados de Candida spp. a antifúngicos. Foram analisados 63 isolados clínicos de leveduras, que incluíram as espécies Candida parapsilosis complex (n = 20), Candida albicans (n = 18), Candida tropicalis (n = 14), Candida glabrata (n = 4), Candida krusei (n = 4), Candida kefyr (n = 2) e Candida lusitaniae (n = 1). As técnicas de disco-difusão em ágar e de microdiluição em caldo foram utilizadas para testar a sensibilidade em relação aos antifúngicos fluconazol, itraconazol e anfotericina B. A sensibilidade ao voriconazol foi determinada somente pela técnica de disco-difusão. Os halos ao redor dos discos de fluconazol variaram de 14 mm a 50 mm, e a CIM de 0,125 µg/mL a 32 µg/mL; para itraconazol, os halos variaram de 9 mm a 27 mm e a CIM de 0,03 µg/mL a 0,25 µg/mL; para anfotericina B, 9 mm a 21mm e 0,5 µg/mL a 2 µg/mL, respectivamente; para voriconazol, o diâmetro dos halos variaram de 19 mm a 50 mm. Para as três espécies, C. albicans, C. parapsilosis e C. tropicalis, a técnica de disco-difusão apresentou boa concordância com a microdiluição, especialmente em relação ao fluconazol, representando, assim, um recurso importante para os laboratórios reportarem os resultados dos testes de sensibilidade dos isolados dessas espécies ao fluconazol.

Antimicrobial susceptibility tests performed by disk diffusion method are practical and well known by professionals that work in the microbiology laboratory. The disk diffusion methodology used to verify the susceptibility of fungi to antifungal agents, however, has characteristics that differ from the tests for bacteria. The objective of this study was to evaluate the disk diffusion method to determine the in vitro susceptibility to antifungal agents of Candida species. We analyzed 63 clinical isolates of yeasts, which included Candida parapsilosis complex species (n = 20), Candida albicans (n = 18), Candida tropicalis (n = 14), Candida glabrata (n = 4), Candida krusei (n = 4), Candida kefyr (n = 2) and Candida lusitaniae (n = 1). The susceptibility tests to antifungal drugs was performed by disk diffusion methods and broth microdilution for antifungal fluconazole, itraconazole and amphotericin B. Voriconazole was used to test the susceptibility only by the disk diffusion method. The inhibition halos of growth around disks of fluconazole ranged from 14 mm to 50 mm and the MIC from 0.125 µg/mL to 32 µg/mL, for itraconazole, halos ranged from 9 mm to 27 mm and the MIC from 0.03 µg/mL to 0.25 µg/mL, for amphotericin B, 9 mm to 21 mm and 0.5 µg/mL to 2 µg/mL, respectively. The diameter of voriconazole disks varied from 19 mm to 50 mm. For the three species, C. albicans, C. parapsilosis and C. tropicalis, the disk diffusion method showed good agreement with the microdilution, especially to fluconazole, thus representing an important resource for medical laboratories reporting results of susceptibility testing of isolates of these species to fluconazole.

Comparative evaluation of disk diffusion and E-test methods for clarithromycin susceptibility testing of Helicobacter pylori / 中华临床感染病杂志

Objective To compare disk diffusion with E-test methods for clarithromycin susceptibility testing of Helicobacter pylori (H.pylori).Methods A total of 44 strains of H.pylori were isolated from gastric mucosa biopsy from patients undergoing gastroscopic examination.Disk diffusion and E-test methods were used for clarithromycin susceptibility testing of H.pylori.The agreement of disk diffusion and E-test was assessed by linear regression analysis.Results The minimum inhibitory concentration (MIC) tested by E-test method ranged from 0.016 to 256 μg/mL,and drug resistance was observed in 12(27.3％) isolates.In range of 0-35 mm of inhibition diameter,the results of disk diffusion method were correlated well with the MICs obtained by E-test method (r2 =0.91,P ＜0.01).Regression analysis showed that with inhibition diameters≥ 18 mm as considered sensitive to clarithromycin and ≤ 15 mm as resistant,the agreement was 100％ between two methods.Conclusion The disk diffusion method is equivalent to the E-test method for clarithromycin susceptibility testing of H.pylori,which can be an alternative method for clinical application.

Evaluation of pH, calcium ion release and antimicrobial activity of a new calcium aluminate cement

This study evaluated the pH, calcium ion release and antimicrobial activity of EndoBinder (EB), containing different radiopacifiers: bismuth oxide (Bi2O3), zinc oxide (ZnO) or zirconium oxide (ZrO2), in comparison to MTA. For pH and calcium ion release tests, 5 specimens per group (n = 5) were immersed into 10 mL of distilled and deionized water at 37°C. After 2, 4, 12, 24, 48 h; 7, 14 and 28 days, the pH was measured and calcium ion release quantified in an atomic absorption spectrophotometer. For antimicrobial activity, the cements were tested against S. aureus, E. coli, E. faecalis and C. albicans, in triplicate. MTA presented higher values for pH and calcium ion release than the other groups, however, with no statistically significant difference after 28 days (p > 0.05); and the largest inhibition halos for all strains, with no significant difference (E. coli and E. faecalis) for pure EB and EB + Bi2O3 (p > 0.05). EB presented similar performance to that of MTA as regards pH and calcium ion release; however, when ZnO and ZrO2 were used, EB did not present antimicrobial activity against some strains.

Sucesso terapêutico da terbinafina em um caso de esporotricose / Successful treatment of terbinafine in a case of sporotrichosis

A esporotricose é uma infecção subaguda ou crônica, causada por fungos pertencentes ao Complexo Sporothrix. Relato do caso: esporotricose de localização nasal foi tratada com iodeto de potássio e como não se obteve sucesso, reiniciou-se o tratamento com associação de iodeto de potássio e itraconazol. Porém, ocorreu nova recidiva. As culturas dos exames micológicos foram submetidas a ensaios de atividade antifúngica in vitro para auxiliar no tratamento. A terbinafina foi o antifúngico que apresentou melhores resultados, por isso, o tratamento foi reiniciado com este antifúngico e, após dois anos do término do mesmo, não recidivou. Adicionalmente, ambas as culturas foram comparadas por RAPD, obtendo padrões de fragmentos distintos, indicando que os isolados são diferentes ou demonstrando um processo microevolutivo do microrganismo.

Sporotrichosis is a chronic subacute infection caused by fungi belonging to the Sporothrix Complex. In the present clinical case, nasal sporotrichosis was treated with potassium iodide. This was unsuccessful, and the treatment was restarted with a combination of potassium iodide and itraconazole. This however resulted in a further recurrence of the infection. The mycological cultures were tested in vitro for antifungal activity to assist in treatment. Terbinafine, an antifungal drug, produced the best results and was therefore used for the rest of the treatment course, with no recurrence after two years of its completion. In addition, both cultures were compared using RAPD and different fragment patterns were observed. This indicated that the isolates were either different or indicated a microevolutionary process of this microorganism.

Sensibilidad in vitro de hongos miceliales de aislamientos clínicos en pacientes con cáncer en el Instituto Nacional de Cancerología ESE / in vitro susceptibility of clinical isolates of filamentous fungi in patients with cancer in the Instituto Nacional de Cancerología

Introducción: la sensibilidad antifúngica in vitro en hongos filamentosos no ha tenido el mismo desarrollo que en levaduras. Se dispone de limitada información sobre la susceptibilidad en este tipo de aislamientos en Colombia. Materiales y métodos: se determinó la actividad in vitro de fluconazol, voriconazol, itraconazol, anfotericina B y caspofungina mediante el método de E-Test, de los géneros Aspergillus (36 A. fumigatus, 12 A. flavus, 9 A. niger, 6 A. terreus, 4 A. nidulans y 1 A. versicolor) e hifomicetes hialinos (9 Fusarium sp., 2 Geotrichum sp. y 2 Paecilomyces sp.), provenientes en su mayoría de lavados broncoalveolares (30%) y biopsias pulmonares (36%); 9% provenían de hemocultivos. Resultados: el perfil de resistencia general fue 28% para itraconazol, 15% para caspofungina, 14% para anfotericina B y 5% para voriconazol. En general, todos los aislamientos presentaron una sensibilidad disminuida para fluconazol e itraconazol. La mejor actividad farmacológica la presentaron voriconazol, caspofungina y anfotericina B. Fusarium sp. presentó una mayor actividad con el voriconazol. Se encontraron diferencias entre el tipo de micelio (Aspergillus vs no Aspergillus) y la susceptibilidad a voriconazol, anfotericina B y caspofungina. Conclusión: en general, los antimicóticos disponibles para el tratamiento de infecciones por miceliales muestran una sensibilidad disminuida in vitro en relación con el género y la especie identificada.

Introduction: fungal susceptibility against micelial fungi has not been developed at the same pace as susceptibility against yeasts. Scarce information is available about that kind of isolates in Colombia. Materials and methods: in vitro susceptibility against micelial isolates from patients with cancer was determined. The E-test method was used to find out susceptibility against fluconazole, voriconazole, itraconazole, amphotericin B, and caspofungin. Isolates of the genera Aspergillus (36 A. fumigatus, 12 A. flavus, 9 A. niger, 6 A. terreus, 4 A. nidulans and one A. versicolor isolate), Fusarium (n=9), Geotrichum and Paecilomyces (n=2 each one) obtained from patients with cancer were tested. These isolates were obtained from bronchoalveolar lavage (30%), pulmonary biopsies (36%) and bloodstream infections (9%). Results: The general pattern of resistance was 28% against intraconazole, 15% against caspofungin, 14% against amphotericin B, and 5% against voriconazole. In general, susceptibility against fluconazole and itraconazole showed a diminishing trend. Voriconazole, caspofungin, and amphotericin B showed the best pharmacologic potency. Fusarium sp. presented a higher activity level against voriconazole. There were differences in the susceptibility against voriconazole, anphotericin B, and caspofungin depending on the type of micelial isolate (Aspergillus vs. Non- Aspergillus). Conclusion: In general, the available antifungal treatments against mycelial fungi identified in the cancer center show diminished susceptibility.

Analysis of cefoxitin-sensitive, penicillin-resistant Staphylococcus tested by disk diffusion method / 中华检验医学杂志

Objective To evalue the ability of detecting the resistance of cefoxitin-sensitive,penicillin-resistant Staphylococcus by different methods and analyze the antibiotic susceptibility spectrum of coagulase-negative Staphylococcus which are non-mecA-mediated oxacillin resistance. Methods All the isolates were collected from Huashan hospital between 2007 and 2009. The isolates were recovered from various clinical sources, including respiratory tract, urine, secretion and sterile fluids samples. The oxacillin susceptibility of Staphylococcus aureus was determined by cefoxitin disk diffusion test, cefoxitin MIC test,oxacillin disk diffusion test and oxacillin MIC test Likewise, the oxacillin susceptibility of coagulasenegative Staphylococcus was determined by cefoxitin disk diffusion test and oxacillin MIC test. All the isolates with sensitive to cefoxitin were screened for the mec A gene by PCR Finally, the MIC of non-mecA-mediated oxacillin-resistant Staphylococcus were determined. Results Among 255 cefoxitin disk diffusion test sensitive and penicillin-resistant Staphylococcus aureus, 6 isolates were intermediated to oxacillin and 4 were resistant by oxacillin disk diffusion test, but all the isolates were sensitive by the cefoxitin disk diffusion test,cefoxitin MIC test and oxacillin MIC test. Among 75 cefoxitin disk diffusion test sensitive and penicillin-resistant coagulase-negative Staphylococcus, 16 isolates were resistant to oxacillin by oxacillin MIC method and 4 carried mecA gene. Among 12 non-mecA-mediated oxacillin-resistant Staphylococcus, the susceptible isolates of gentamicin is 10, clindamycin is 8, ciprofloxacin is 11, erythrornycin is 6, trimethoprim/sulfamethoxazo]e is 11 ,and cephalosporins, teicoplaninl, vancomycin, piperacillin/tazobactam, tetracycline are all 12. Conclusions The cefoxitin disk diffusion test can reliably predict mecA-mediated oxacillin resistant Staphylococcus aureus. It would be best to combine cefoxitin disk diffusion test and oxacillin MIC test to improve accuracy of detection of mecA-mediated oxacillin resistant coagulase-negative Staphylococcus.Furthermore, infections due to the non-mecA-mediated oxacillin resistant coagulase-negative Staphylococcus can be treated by penicillinase-stable penicillins, β-lactam/β-lactam inhibitor combinations, relevant cephems and carbapenems.

Influence of sulbactam content on susceptibility testing results of cefoperazone/sulbactam combination disks / 中华检验医学杂志

Objective To study the discrepancy influence of the sulbactam content on susceptibility testing results of cefoperazone/sulbactam combination disks.Methods Agar dilution method was used to determine MICs of cefoperazone,cefoperazone/sulbactam(2:1 and 1:1),and disk diffusion was used to detect the zone diameters of cefoperazone,cefoperazone/sulbactam(75/30 and 75/75μg/disk)disks against 534 clinical gram-negative isolates.The discrepancy within the results of MICs,zore diameters,the method of agar dilution and disk diffusion was analyzed.Results By standard agar dilution method,MIC_(50) of cefoperazone,cefoperazone/sulbactam(2:1 and 1:1)were 32,16,16μg/ml,and MIC_(90) of those were ≥256.128,64 μg/ml respectively.No statistic discrepancy was found for MICs between the ratios of 2:1 and 1:1 combination by Wilcoxon ranks test(Z=-0.248,P=0.804).Susceptibility rate,resistance rate,and intermediate rate with 75/30μg disk were 55.3%,24.5%and 20.2%respectively,which were similar to those determined by agar dilution method.Susceptibility rate,resistance rate,and intermediate rate(I%)with 75/75μg disk were 72.5%,12.4% and 15.1% respectively,compared with the susceptibility rate from 75/30μg disk was 17.2% higher.Statistic discrepancy were tested by paired t-test (t=21.613,P＜0.01)with two groups of whole strains' zone diameters from 75/30μg and 75/75μg disks,and resulting in the difference of susceptibility or resistance rates for ESBL-producing strains,Acinetobacter bauamnnii and Enterobacteriaceae without ESBL tested isolates.On the contrary,for Pseudomonas aeruginosa,Stenotrophomonas maltophilia and ESBL negative isolates,the zone diameters discrepancy was not statistically significant between the results from 75/30μg and 75/75μg disks.Conclusions There is no statistic discrepancy between the susceptibility results from cefoperazone/sulbactam(2:1 or 1:1 ratios)in dilution method and in diffusion method with 75/30μg disk.When the 75/75μg disk is used to be tested for ESBL-producing strains,Acinetobacter bauamnnii and other Enterobacteriaceae,the results should be shown with sulbactam content.

Estudio de susceptibilidad a tigeciclina: Influencia del agar Mueller-Hinton en el método de difusión en agar y validación del método E-test® para Acinetobacter baumannii / In vitro evaluation of tygecicline: Influence of Mueller-Hinton agar in agar diffussion methods and validation of E-test® for Acinetobacter baumannii

Tigecycline is a glicylcicline with broad antimicrobial spectrum. Susceptibility testing to this drug for Acinetobacter is difficult in hospitals due to the utilization of the disk diffusion method. FDA break points have shown an unacceptable rate of errors (23 percent) for disk diffusion versus broth microdilution in American studies and overcall of resistance depending on the brand of Mueller Hinton agar used. Modifications to these FDA break points have been proposed, but there is not enough evidence yet. Data from a multicenter study from Chile allowed the evaluation of the characteristics of the agar used for susceptibility testing and the utility of E-test as an alternative MIC method for Acinetobacter. The Mueller Hinton agar brand is an important factor that affects disk diffusion method results. There is very good correlation between broth microdilution and E-test for the susceptibility category as well as for MIC determination. The intermedíate and resistant results obtained with disk diffusion method should be checked by using E-test.

Tigeciclina es una glicilciclina de amplio espectro antimicrobiano. La determinación de la susceptibilidad a este fármaco presenta dificultades en el laboratorio asistencial al utilizar la técnica de difusión por disco para Acinetobacter spp. Los puntos de corte -según la (FDA- han mostrado una tasa inaceptable de errores (23 por ciento) en comparación con el método de micro-dilución en caldo en estudios americanos, diversas evaluaciones demuestran que existe una sobreestimación de resistencia in vitro dependiendo de las características del agar Mueller Hinton utilizado. Se han propuesto modificaciones a los puntos de corte pero no se han oficializado por insuficientes evidencias. Los datos de un estudio multicéntrico realizado en Chile permitieron evaluar la influencia de las distintas marcas de medios de cultivo en el tamaño de los halos y la utilidad de la epsilometría (E-test®) como método CIM para Acinetobacter sp. La marca de agar Mueller Hinton y otros factores propios del medio dificultan la determinación de la susceptibilidad a tigeciclina utilizando difusión por disco. Existe muy buena correlación entre la micro-dilución en caldo y el E- test®, tanto para la categoría de susceptibilidad como para la CIM. Por esto, se sugiere que los resultados intermedios o resistentes obtenidos por difusión en agar para A. baumannii sean comprobados mediante el uso de E-test®.

Antibacterial activity of Apis mellifera L. propolis collected in three regions of Kenya

The present study aimed at investigating the susceptibility of the microorganisms Pseudomonas aeruginosa, Salmonella typhi, Escherichia coli, Staphylococcus aureus, and Bacillus subtilis to ethanolic extracts of propolis (EEP) from three regions of Kenya (Taita, Tana and Samburu). Propolis was extracted using four different concentrations of ethanol: pure, 70 percent, 50 percent, and 30 percent. Ethanol (70 percent) and Streptomycin were used as controls. The agar diffusion method using filter paper disks was employed. Antibacterial activity was determined as an equivalent of the inhibition zones diameters (in millimeters) after incubation at 37°C for 24h. Significant differences in the antibacterial activities of propolis were observed among the three regions, depending on the test microorganisms and on the procedure used for the preparation of propolis extract. Bacillus subtilis and Staphylococcus aureus were the most susceptible bacteria and 70 percent EEP had the best antibacterial effect.(AU)