Antifungal Activity of Morinda citrifolia Methanolic Extract against Candida albicans: An In Vitro Study.

Aim: Natural medicine used as an alternative and/or complementary treatment to counteract diseases is of great importance in public health. Therefore, the purpose of the present study was to assess the in vitro antifungal activity of Morinda citrifolia methanolic extract of peel, pulp, and seed against Candida albicans. Materials and Methods: The present study was experimental in vitro and cross-sectional. Eight replicates were prepared in Sabouraud dextrose agar with five wells each, where 0.12% chlorhexidine, distilled water, and methanolic extract of seed, peel, and pulp of Morinda citrifolia fruit were placed at concentrations of 10,690, 8,270, and 6,430 mg/mL, respectively, to evaluate sensitivity according to Duraffourd's scale. In addition, the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were determined by dilution and agar seeding method. Statistical analysis was performed by analysis of variance (ANOVA) and Tukey's post hoc test, considering a significance level of P < 0.05. Results: The inhibition halos of Morinda citrifolia methanolic extract of seed, peel, and pulp against Candida albicans measured on average 15.94, 11.94, and 11.56 mm, respectively. The MIC of seed, peel, and pulp extract were 1366.25, 2067.5, and 1607.5 mg/mL respectively, whereas the MFC for seed, peel, and pulp extract were 2672.50, 2067.5, and 3215 mg/mL, respectively. Moreover, seed extract presented significantly higher antifungal activity than peel and pulp (P < 0.001). Conclusions: Morinda citrifolia methanolic extract of peel, pulp, and seed showed fungistatic and fungicidal effect against Candida albicans, being this very sensitive to seed extract with a MIC of 1366.25 mg/mL and a MFC of 2672.5 mg/mL, which allows recommending the development of effective pharmacological formulations for the control of candidiasis.

Homology of Escherichia coli isolated from urine and vagina and their antimicrobial susceptibility in postmenopausal women with recurrent cystitis.

Uropathogenic Escherichia coli (UPEC) is a typical cystitis-causing organism that can migrate from the vagina to the bladder and cause recurrent cystitis (RC). Few reports have compared the characteristics of urinary and vaginal UPEC in patients with RC. We carried out molecular biological analyses of Escherichia coli (E. coli) strains and their antimicrobial susceptibility to assess the association between urinary and vaginally UPEC. We included E. coli isolated from urinary and vaginal samples at the onset of cystitis in postmenopausal women with RC between 2014 and 2019 in our hospital. Pulsed-field gel electrophoresis (PFGE) was performed using a restriction enzyme (Xba I). These sequences were compared with 17 antimicrobial susceptibilities determined by a micro-liquid dilution method. Multilocus sequence typing (MLST) and classification of extended-spectrum ß-lactamase (ESBL) genotypes by multiplex polymerase chain reaction (PCR) were performed on ESBL-producing E. coli. We analyzed 14 specimens (each seven urine and vaginal) from seven patients in total. On PFGE, the similarity of urinary and vaginal E. coli per patient ranged from 89.5 to 100 %, including four patients with 100 % matches. MLST demonstrated that 29 % (4/14 specimens) were strain sequence type 131. Two specimens contained ESBL-producing strains and identified the CTX-M-27 genotype for each specimen. For each patient, antimicrobial susceptibilities between urinary and vaginal E. coli were mostly identical. Thus, urinary- and vaginally-derived E. coli were identical in postmenopausal women with RC. Management targeting both urinary and vaginal UPEC is essential for RC, indicating the importance of a vagina-targeted approach.

Testing and Diagnosis of Clostridioides difficile Infection in Special Scenarios: A Systematic Review.

INTRODUCTION: Clostridioides difficile infection (CDI) is a clinical and laboratory diagnosis. Populations at higher risk of developing disease require a high clinical index of suspicion for laboratory testing to avoid incorrect assumptions of colonization. Common risk factors include recent antibiotic use, elderly (>65 years old), and immunocompromised patients. C. difficile assays should be ordered in an algorithm approach to diagnose an infection rather than colonization. Screening tests are widely available in hospital systems, but novel molecular testing may aid in diagnosis in patients with inconclusive or discordant antigen and toxin test results.  Methods: Data was extracted from PubMed, Scopus, and Cumulative Index of Nursing and Allied Health Literature (CINAHL) databases based on the keywords "clostridioides difficile", "toxin assay", and "toxic megacolon". The data extracted is based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) 2020 guidelines. A total of 27 reports were included in this systematic review. RESULTS: Testing patients with a significant gastrointestinal surgical history, hypogammaglobulinemia, inflammatory bowel disease, intensive care unit, and immunocompromised patients for CDI is highly recommended. Diarrhea in these subsets of patients requires correlation of clinical context and an understanding of assay results to avoid over- and under-treating. CONCLUSION: CDI should be considered in all patients with traditional risk factors. Heightened clinical suspicion of CDI is required in patients with hypogammaglobulinemia, transplant recipients, patients with gastrointestinal surgical history, and inflammatory bowel disease. Testing should be limited to patients with clinical manifestations of CDI to ensure a high pretest probability for test interpretation. Healthcare workers should adhere to testing algorithms to optimize yield in the appropriate clinical context. Diagnostic assays should follow a sequential, stepwise approach to categorize the toxin expression status of the bacteria accurately.

[Establishment and Evaluation of a Resazurin-Based Microdilution Assay for Microbial Sensitivity Test of Neisseria gonorrhoeae]. / æ·‹ç— å¥ˆç‘ŸèŒè¯æ•è¯•éªŒåˆƒå¤©é’å¾®é‡ç¨€é‡Šæ³•çš„å»ºç«‹ä¸Žè¯„ä»·.

Objective: To establish and evaluate a microbial sensitivity test method for Neisseria gonorrhoeae based on resazurin coloration. Methods: Based on the broth microdilution method, resazurin was added as a live bacteria indicator. WHO G, a WHO gonococcal reference strain, was used to optimize the incubation time for resazurin-stained bacteria and the color change was visually observed to obtain the results. Agar dilution method (the gold standard) and resazurin-based microdilution assay were used to determine the minimum inhibitory concentration (MIC) of azithromycin, ceftriaxone, and spectinomycin for 3 reference strains and 32 isolates of Neisseria gonorrhoeae. The results were analyzed based on essential agreement (EA), which reflected the consistency of the MIC values, category agreement (CA), which reflected the consistency in the determination of drug resistance, intermediary, and sensitivity, very major error (VME), which reflected false sensitivity, and major error (ME), which reflected pseudo drug resistance, to evaluate the accuracy of resazurin-based microdilution assay as a microbial sensitivity test of of Neisseria gonorrhoeae. CA and EA rates≥90% and VME and ME rates≤3% were found to be the acceptable performance rates. Results: The results obtained 6 hours after resazurin was added were consistent with those of the agar dilution method and the resazurin-based microdilution assay was established accordingly based on this parameter. The EA of resazurin-based microdilution assay for measuring the MIC results of azithromycin, ceftriaxone, and spectinomycin was 97.1%, 91.5%, and 94.3%, respectively, and the CA was 88.6%, 94.3%, and 94.3%, respectively. The VME was 0% for all three antibiotics, while the ME was 11.4%, 5.7%, and 5.7%, respectively. Conclusion: The resazurin-based microdilution assay established in this study showed good agreement with agar dilution method for measuring the MIC of antibiotics against Neisseria gonorrhoeae. Moreover, the sensitivity results of this method were highly reliable and could be easily obtained through naked eye observation. Nonetheless, the results of drug resistance should be treated with caution and the optimization of parameters should be continued.

Osteomyelitis by Microsporum canis and Staphylococcus spp. in cat (Felis catus) - case report.

BACKGROUND: Staphylococcus spp and Microsporum canis are zoonotic microorganisms which can cause infections and systemic diseases. The bone infection is usually caused by invasion of pathogen through the hematologic route. Mixed osteomyelitis caused by bacteria and fungi is rare, and to date, there have been no reports of mixed osteomyelitis with Staphylococcus spp. and Microsporum canis. CASE PRESENTATION: This essay reports an atypical presentation of mixed osteomyelitis (Staphylococcus spp. and Microsporum canis) in a domestic cat. A 15-month-old female Persian cat was presented to a veterinary service; the main complaint was the appearance of a nodule in the mandibular ventral rostral region. A radiographic exam performed on the animal showed proliferative and osteolytic bone lesions. The patient was submitted to a biopsy for histopathological evaluation, along with bacterial and fungal cultures. Results showed mixed osteomyelitis by Staphylococcus spp. and Microsporum canis. Microbial Sensitivity Test was performed to choose a more suitable treatment. Two surgical procedures were executed to resect and curette the lesion, and treatments with anti-inflammatory, antibiotic, and antifungal drugs were established, showing a positive clinical evolution. After 8 months of treatment, the patient's owner moved to a different city, and the animal was seen by other veterinarians, who followed along with the same treatment. However, due to complications and a diminishing quality of life over 4 years of diagnosis, the patient was euthanized. CONCLUSION: Given the above, mixed osteomyelitis is difficult to treat and can cause losses of life quality resulting death, especially in infections where M. canis is the agent causing the disease. Bacterial osteomyelitis is more frequently reported. But the lack of investigation of microorganisms other than bacteria, such as fungal cases, may imply in underdiagnosed cases. Treatment of osteomyelitis can be difficult considering the difficulties in isolating the pathological agent, resistance to the drug used, prolonged treatment time, and cost.

Important Recommendations Regarding the Test-and-Treat Strategy for Helicobacter pylori Infection in Children.

Helicobacter pylori is a commonly encountered pathogen in medical practice. It causes chronic gastritis in patients of different ages. Many published papers have provided different opinions on whether the test-and-treat strategy for H. pylori infection should be implemented in children. It is critical that the opinion in favor of this strategy was published in Europe, where ESPGHAN/NASPGHAN guidelines have not recommended the use of test-and-treat strategy for H. pylori in children. Herein, I propose my opinion regarding this debate using 4 main points. First, this strategy should be implemented in areas where its benefits outweigh the associated risks. Thus, if it is not tailored to the locality, the results of the test-and-treat strategy for H. pylori in children may be erroneous. Second, the association between H. pylori infection and other diseases such as asthma and other allergic diseases should not be factored in when considering the test-and-treat strategy. This is because these diseases are associated with H. pylori noninfection and not with its posteradication status. Third, there is evidence that H. pylori infection can significantly alter children's intestinal microbiota. Finally, gastroscopy should not be performed in all H. pylori-positive children, particularly if the drug susceptibility test is not available. In the near future, it will be possible to easily conduct drug susceptibility tests for H. pylori using fecal samples. I believe that this report may expand the test-and-treat strategy for children infected with asymptomatic H. pylori outside of Japan and prevent not only gastric cancer but also minor diseases such as iron deficiency anemia, chronic idiopathic thrombocytopenic purpura, and failure to thrive. Pediatricians are responsible for keeping children healthy not only during childhood but also in adulthood. Thus, even if there are only a few H. pylori-related diseases and no severe cases in children, it is necessary to take early measures to prevent problems (eg, incidence of gastric cancer) in adulthood.

Características epidemiológicas de la Osteomielitis aguda en niños internados en un hospital infeccioso de referencia

Introducción: La osteomielitis aguda es una infección del hueso que afecta principalmente a los niños y tiene generalmente diseminación hematógena, a veces asociada a un trauma. En la etiología influyen factores, como la edad, el estado inmunológico y las enfermedades concomitantes. En la mayoría de los casos, el principal agente etiológico es Staphylococcus aureus. Es importante el diagnóstico oportuno para evitar secuelas a mediano o largo plazo. Objetivo: Describir las características epidemiológicas de un grupo de pacientes con osteomielitis aguda. Métodos: Se realizó la revisión retrospectiva de los expedientes clínicos de pacientes egresados del servicio de pediatría del Instituto de Medicina Tropical, entre enero de 2016 y diciembre de 2020, con diagnóstico de osteomielitis aguda. Resultados: Los varones con osteomielitis corresponden al 67,8% del total de 59 casos registrados, en cuanto a los signos y síntomas, el dolor, la tumefacción y la impotencia funcional fueron predominantes, la fiebre se documentó en 49 (83,1%) pacientes, se registró antecedentes de cirugía en 37 (62,7%) de los pacientes y complicaciones en 42 (71,2%) de los pacientes, la complicación más frecuente fue osteomielitis crónica El sitio anatómico más frecuente fueron los miembros inferiores. El tratamiento empírico fue realizado con cefalosporinas de 3G en 72,9% de los pacientes, ya sea solo o combinado con clindamicina o vancomicina, un paciente con aislamiento de M. tuberculosis recibió tratamiento HRZE. Se aisló algún germen 44 pacientes (74,5%), el microorganismo predominante fue Staphylococcus aureus en 81,8 %, la mitad (52,3%) correspondieron a SAMR Se encontró una alta resistencia a oxacilina del 55,8% y un solo paciente resistente a clindamicina (2,2%). Conclusión Los hallazgos fueron similares a los reportados en la literatura en cuanto a etiología, sitio anatómico afectado y cobertura antibiótica.

Introduction: Acute osteomyelitis is a bone infection that mainly affects children and generally has hematogenous spread, sometimes associated with trauma. The etiology is influenced by factors such as age, immune status, and comorbidities. In most cases, the main etiologic agent is Staphylococcus aureus. Timely diagnosis is important to avoid sequelae in the medium or long term. Objective: To describe the epidemiological characteristics of a group of patients with acute osteomyelitis. Methods: A retrospective review of the clinical records of patients discharged from the pediatric service of the Institute of Tropical Medicine was carried out between January 2016 and December 2020, with a diagnosis of acute osteomyelitis. Results: Men with osteomyelitis correspond to 67.8% of the total of 59 registered cases, in terms of signs and symptoms, pain, swelling and functional impotence were predominant, fever was documented in 49 (83.1%) patients, a history of surgery was recorded in 37 (62.7%) of the patients and complications in 42 (71.2%) of the patients, the most frequent complication was chronic osteomyelitis The most frequent anatomical site was the lower limbs. Empirical treatment was performed with 3G cephalosporins in 72.9% of the patients, either alone or in combination with clindamycin or vancomycin. One patient with M. tuberculosis isolation received HRZE treatment. Some germ was isolated in 44 patients (74.5%), the predominant microorganism was Staphylococcus aureus in 81.8%, half (52.3%) corresponded to MRSA. A high resistance to oxacillin of 55.8% and a only patient resistant to clindamycin (2.2%). Conclusion The findings were similar to those reported in the literature in terms of etiology, affected anatomical site, and antibiotic coverage.

A previously unreported case of suppurative thrombophlebitis due to Clavispora lusitaniae.

We report a case with Clavispora lusitaniae-induced purulent thrombophlebitis. The patient had multiple risk factors for the development of fungal thrombophlebitis including surgical procedure, mechanical ventilation, admission to intensive care unit, total parenteral nutrition and long-term antimicrobial therapy in addition to the insertion of central venous catheter. The symptoms finally improved by a combination therapy of micafungin and flucytosine, but the therapy did not rapidly resolve candidemia. The appropriate antifungal therapy for C. lusitaniae-induced purulent thrombophlebitis is uncertain. Further study is desired to seek the appropriate therapy for the disease.

European Committee on Antimicrobial Susceptibility Testing-Recommended Rapid Antimicrobial Susceptibility Testing of Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus From Positive Blood Culture Bottles.

Background: Early diagnosis and treatment are important for a good prognosis of bloodstream infections. The European Committee on Antimicrobial Susceptibility Testing (EUCAST) recommends rapid antimicrobial susceptibility testing (RAST) based on the disk diffusion methodology for 4, 6, and 8 hours of incubation. We evaluated EUCAST-RAST of Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus from positive blood culture bottles. Methods: Twenty strains of E. coli, K. pneumoniae, and S. aureus were tested using EUCAST-RAST. Ten antimicrobial agents against E. coli and K. pneumoniae and four agents against S. aureus were tested. The diameter of the inhibition zone (mm) was compared with the minimal inhibitory concentration (µg/mL) obtained using the Sensititre AST system (TREK Diagnostic Systems, East Grinstead, UK). Results: For E. coli, the percentage of total categorical agreement (CA) was 69.5% at 4 hours, and 87% at 8 hours. For K. pneumoniae, the total CA was 89% at 4 hours, and 95.5% at 6 hours. For S. aureus, the total CA was 100% after 4 hours. Discrepancies were observed mainly for E. coli with ß-lactam antimicrobial agents, and the numbers of errors decreased over time. Conclusions: EUCAST-RAST for K. pneumoniae and S. aureus met the United States Food and Drug Administration criteria at 6 and 4 hours, respectively, whereas that for E. coli did not meet the criteria for up to 8 hours. RAST can shorten the turn-around testing time by more than one day; therefore, if applied accurately according to laboratory conditions, antimicrobial agent results can be reported faster.

Characteristics and Outcomes of Patients with Delftia acidovorans Infections: a Retrospective Cohort Study.

Delftia acidovorans (D. acidovorans) is a Gram-negative bacteria and an uncommon cause of human infections. This retrospective cohort study investigated clinical and microbiological characteristics and outcomes of patients with D. acidovorans infections. We included patients with culture-confirmed D. acidovorans infections attending Rigshospitalet, during 2002-2020. Fifty-nine patients with a median interquartile ranges (IQR) age of 47 (15-67) years were included. Thirty-five (59%) were males, and 57 (97%) had at least one comorbidity, including 25 (42%) with solid or hematologic malignancies. Eight (14%) were admitted to ICU, and 15 (25%) died within 365 days after infection. Persistent infection was found in 4 (6.8%) patients, and 41 (70%) had polymicrobial cultures, mainly with Pseudomonas spp. and Stenotrophomonas maltophilia. More than 85% of the D. acidovorans isolates were susceptible to meropenem or ceftazidime. Although, 88% and 62% of the isolates were resistant to gentamicin and colistin, respectively. D. acidovorans infections mainly affect patients with preexisting comorbidities, including malignancies. In the first year, all-cause mortality is considerable, polymicrobial cultures are common, and meropenem or cephalosporins with antipseudomonal activity could be the antibiotics of choice. IMPORTANCE Delftia acidovorans (D. acidovorans) is a Gram-negative bacteria that can cause infection in immunocompetent and immunocompromised individuals. The current knowledge comes mainly from case reports and case series. In this retrospective cohort study, we found that D. acidovorans infections mainly affect male patients with preexisting comorbidities, including malignancies. Persistent infections were not common, and most of the patients had polymicrobial cultures, mainly with Pseudomonas spp. and Stenotrophomonas maltophilia. More than 85% of the D. acidovorans isolates were susceptible to meropenem or ceftazidime. In contrast, 88% and 62% of the isolates were resistant to gentamicin and colistin, respectively.

Caracterización molecular de genes de virulencia (lmb, bca y rib) y de resistencia a macrólidos (ermB, ermTR y mefA) en aislamientos clínicos de Streptococcus agalactiae / Molecular characterization of virulence (lmb, bca and rib) and macrolid resistance genes (ermB, ermTR and mefA) in clinical isolates of Streptococcus agalactiae

RESUMEN El objetivo del estudio fue identificar molecularmente los genes de virulencia y resistencia a macrólidos en aislamientos clínicos de Streptococcus agalactiae (EGB), recuperados en 2019 a partir de secreción vaginal (n=9) y orina (n=22), en dos establecimientos de salud de Lima. La identificación y susceptibilidad antimicrobiana se determinaron por el sistema automatizado Vitek® 2, se confirmó la identificación fenotípicamente; la resistencia a macrólidos por el método D-test; la identificación de genes de virulencia (lmb, bca y rib) y de resistencia a macrólidos (ermB, ermTR y mefA) por reacción en cadena de la polimerasa (PCR). El fenotipo y genotipo de resistencia a macrólidos predominante fue cMLSb (12/31) y ermB (11/31), y el gen de virulencia más frecuente fue lmb (23/31). Todos fueron sensibles a penicilina, ampicilina y vancomicina. Estos hallazgos muestran la necesidad de implementar estudios de epidemiología molecular que permitan un adecuado conocimiento y seguimiento de EGB en el Perú.

ABSTRACT The aim of the study was to molecularly identify virulence and macrolide resistance genes in clinical isolates of Streptococcus agalactiae (GBS), recovered in 2019 from vaginal discharge (n=9) and urine (n=22), from two health facilities in Lima. Identification and antimicrobial susceptibility were determined by the Vitek® 2 automated system, identification was confirmed phenotypically; macrolide resistance was determined by the D-test method. Identification of virulence genes (lmb, bca and rib) and macrolide resistance genes (ermB, ermTR and mefA) was carried out by polymerase chain reaction (PCR). The predominant macrolide resistance phenotype and genotype were cMLSb (12/31) and ermB (11/31); the most frequent virulence gene was lmb (23/31). All were sensitive to penicillin, ampicillin and vancomycin. These findings show the need to implement molecular epidemiology studies that allow adequate knowledge and follow-up of GBS in Peru.

Comparative evaluation of susceptibility testing methods for colistin and polymyxin B on isolates of Enterobacteriaceae / 中华检验医学杂志

Objective:To provide a promising and optimal laboratory susceptibility-testing method for the clinical usage of antibiotic (polymyxin), four susceptibility-testing methods were performed and the broth microdilution (BMD) was chosen as the gold standard.Methods:A total number of eighty-eight nonduplicate clinical Enterobacteriaceae specimes were collected from January to December of 2019 in the First Affiliated Hospital, Fujian Medical University. Among the clinical specimens, of which six strains were positive for mcr-1. The minimal inhibitory concentration (MIC) of polymyxin of the clinical specimens were examined by the following methods: (1) broth microdilution, (2) colistin broth disk elution, (3) Vitek-2?, (4)BD PhoenixTM,(5)commercial broth microdilution. With BMD as reference, essential agreement (EA), categorical agreement(CA), very major error(VME) and major error (ME) of polymyxins for different methods were analyzed. The Kappa-consistency testing, paired Chi-square testing and the Spearman-rank correlation testing were used to analyze the consistency between the four antimicrobial susceptibility testing methods and the gold standard.Results:Taking broth microdilution as reference, the EA of colistin broth disk elution, Vitek-2?, BD PhoenixTM, commercial broth microdilution were 94.32% (83/88), 92.05% (81/88), 90.90% (80/88), and 96.59%(85/88), respectively. The CA of all the four methods were 100% (88/88). No VME and ME were recorded for four methods. Moreover, the consistency between four susceptibility testing methods and the gold standard is acceptable (Kappa values=1, P<0.001, McNemar test P=1 and r>0.5, P<0.05). Conclusions:In the present work, four susceptibility testing methods all met the standards recommended jointly by the Clinical and Laboratory Standards Institute and European Committee on Antimicrobial Susceptibility Testing, of which the performance of the commercial broth microdilution and CBDE fared relatively well. Thus, these four methods could be routinely used in clinical microbiology laboratory of our hospital for colistin and polymyxin B susceptibility testing.

Prospective Study of Irrational Prescription of Teicoplanin in a Large Academic Hospital: A Dilemma of Antimicrobial Resistance.

OBJECTIVE: Teicoplanin is an antibiotic used to treat severe Gram-positive infections, especially those caused by methicillin-resistant Staphylococcus aureus (MRSA). In this study, we aimed to evaluate the pattern of teicoplanin rational prescribing to identify the factors which affected rational utilization. In addition, the teicoplanin minimum inhibitory concentration (MIC) was assessed in randomly selected isolates. METHODS: In this descriptive-analytical prospective study, a total of 256 patients were randomly selected to evaluate the pattern of teicoplanin use. The required data were gathered to assess the appropriateness of teicoplanin usage. Also, 100 teicoplanin Etests were used for measuring the MIC. FINDINGS: The results showed that the appropriateness rate of teicoplanin usage was 21.9%. The mean MIC was 2.24 ± 5.47 mg/L for the MRSA cultures (33 cultures), including 32 sensitive cultures (97%). In addition, the mean MIC was 28.71 ± 8.29 mg/L for the vancomycin-resistant enterococci (VRE) cultures (67 cultures), including five sensitive cultures (7.5%). Moreover, the analysis revealed that only the hospitalization ward was statistically significantly related to irrational usage (P = 0.014). CONCLUSION: The high prevalence of the inappropriate use of teicoplanin will lead to the development of antimicrobial resistance. Furthermore, the high rate of VRE cultures resistant to teicoplanin proves that teicoplanin has no advantage over vancomycin for treating VRE infections. Finally, we recommend guidelines' development for the appropriate administration of teicoplanin.

Sensibilidad in vitro de cepas chilenas de Cryptococcus neoformans de origen clínico / In vitro susceptibility of Chilean Cryptococcus neoformans strains of clinical source

Resumen Introducción: Las infecciones por levaduras del género Cryptococcus afectan principalmente a pacientes con déficit de la inmunidad mediada por células. Han sido escasos los estudios de sensibilidad realizados para este género en Chile. Objetivos: Determinar la sensibilidad in vitro de Cryptococcus sp a antifúngicos de uso habitual y evaluar la concordancia esencial entre sensibilidad determinada por microdilución en caldo y por difusión en agar con tiras comerciales. Materiales y Método: Estudio descriptivo de 21 cepas aisladas desde liquido céfalo-raquídeo y sangre. Las CIM50 y CIM90 para fluconazol, voriconazol y anfotericina B se determinaron por microdilución en caldo (Sensititre Yeast One®) y por difusión en agar con tiras comerciales (MIC Test Strips). Resultados: Todas las cepas correspondieron a C. neoformans. Los rangos de CIM50 y CIM90 para cada antifúngico estudiado fueron amplios por ambos métodos. La concordancia esencial entre microdilución y difusión en agar con tiras comerciales fue de 24, 62 y 29% para fluconazol, voriconazol y anfotericina B, respectivamente. Conclusiones: La prueba de Sensititre Yeast One® y la de difusión en agar con tiras comerciales, MIC Test Strips, tienen una pobre concordancia esencial para fluconazol y anfotericina B.

Abstract Background: Cryptococcus yeast infections primarily affect immunocompromised patients. There have been few susceptibility studies conducted for this genus in Chile. Aims: To determine the in vitro susceptibility to commonly used antifungals and evaluate the concordance between susceptibility determined by microdilution in broth and commercially available strips. Methods: Descriptive study of 21 Cryptococcus strains, isolated from cerebrospinal fluid and blood. The MIC50 and MIC90 for fluconazole, voriconazole and amphotericin B was determined by broth microdilution (Sensititre Yeast One®) and by commercial drug sensitivity strips (MIC Test Strips). Results: All strains corresponded to C. neoformans. The ranges of MIC50 and MIC90 for each antifungal studied were wide by both methods. The essential agreement between Sensititre Yeast One test and strips was 24, 62 and 29% for fluconazole, voriconazole and amphotericin B, respectively. Conclusions: The Sensititre Yeast One test and MIC Test Strips exhibited poor essential concordance, especially for fluconazole and amphotericin B.

Strong antimicrobial activity of collinin and isocollinin against periodontal and superinfectant pathogens in vitro.

Periodontitis pathogenesis involves activation of host immune responses triggered by microbial dysbiosis. Therefore, controlling periodontal pathogens in-vivo is a main goal of periodontal therapy. New antimicrobials might help to control periodontal infection and improve treatment outcomes at "the dark times" of increasing antibiotic resistance. Here, we determined the biological activity of collinin and isocollinin against 8 bacterial strains. Antimicrobial activity of collinin and isocollinin, chlorhexidine digluconate (CHX) and sodium hypochlorite (NaClO) was evaluated against clinically relevant periodontal bacteria, like Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Fusobacterium nucleatum, Prevotella intermedia, Dialister pneumosintes strains and superinfectants like Escherichia coli, Staphylococcusaureus, and Pseudomonasaeruginosa strains. A broth microdilution test was carried out to determine the minimum inhibitory concentration of collinin and isocollinin against those strains, and bacterial viability was determined by resazurin assay at diverse concentration and exposure times. P. gingivalis was the most susceptible strain to collinin and isocollinin (MIC 2.1 µg/mL and 4.2 µg/mL respectively). Other periodontal pathogens showed MICs <17 µg/mL for collinin and MICs between 20 and 42 µg/mL for isocollinin, whereas CHX and NaClO showed MICs of 62 and 326 µg/mL, respectively. Collinin and isocollinin also exhibited antimicrobial activity against superinfectant bacteria (MIC < 21 and < 42 µg/mL, respectively). Overall, collinin and isocollinin showed a remarkable antibacterial activity against relevant periodontal and superinfective bacteria, especially against P. gingivalis (MIC 2.1 µg/mL and 4.2 µg/mL respectively) and the highly virulent P. aeruginosa (MIC 5.2 and 20.8 µg/mL, respectively).

Recovery of aerobic and anaerobic bacteria from patients with acute appendicitis using blood culture bottles / Recuperación de bacterias aerobias y anaerobias de pacientes con apendicitis aguda mediante botellas de hemocultivo

Introduction: Acute appendicitis is the first cause of acute abdomen, however, there is a little information about the associated bacteria and its sensibility profile. Objetive: To identify and to determine the resistance pattern of aerobic and anaerobic bacteria isolated in periapendicular fluid cultures taken in patients with acute appendicitis and to establish the proportions of isolates according to the clinical phase. Materials and methods: A descriptive and prospective study was undertaken at the Hospital Universitario de San José (Bogotá, Colombia) of patients older than sixteen years of age, undergoing an open appendectomy. A sample of periappendiceal fluid was taken, which was deposited directly into aerobic and anaerobic blood culture bottles. Results: One hundred and fifty-four patients were included. The overall positivity of cultures was 87% (n=1344); 77% (n=118) for aerobes and 51% (n=79) for anaerobes. The proportion of positive cultures was lower in the uncomplicated appendicitis cases as compared to the complicated ones (80% (66/83) vs. 95%(67/71), p = 0.003). The microorganisms isolated most frequently were: Escherichia coli (53%) (n=84); Bacteroides spp. (25%) (n=25); Propionibacterium acnes (21%) (n=21); coagulase negative Staphylococci (17%) (n=27); Enterococcus spp. (11%) (n=15), and Fusobacterium spp. (11%) (n=11). The sensitivity of E. coli to ampicillin/sulbactam was 30%. The sensitivity of Bacteroides spp. to clindamycin and ampicillin/sulbactam was 91%. All anaerobe isolates were sensitive to piperacillin/tazobactam, ertapenem, meropenem and metronidazole. Conclusions: Intraoperative cultures in acute appendicits are relevant in order to determine the local epidemiological pattern and to establish prophylactic and therapeutic antibiotics for this pathology; direct inoculation in blood culture bottles allows a high recovery of microorganisms.

Introduccción. La apendicitis aguda es la primera causa de abdomen agudo; sin embargo, poco se conoce sobre las bacterias asociadas y su perfil de sensibilidad. Objetivo. Identificar y determinar el patrón de resistencia de las bacterias aerobias y anaerobias aisladas en cultivo de líquido periapendicular tomado de los pacientes con apendicitis aguda, y establecer la proporción de bacterias según la fase clínica. Materiales y métodos. Se llevó a cabo un estudio descriptivo y prospectivo en el Hospital Universitario de San José de Bogotá (Colombia), en pacientes mayores de 16 años sometidos a apendicectomía abierta. Se tomaron muestras de líquido periapendicular, las cuales se sembraron directamente en botellas de hemocultivos para aerobios y anaerobios. Resultados. Se incluyeron 154 pacientes. Del total de cultivos, el 87 % (n=134) fueron positivos: 77 % (n=118) para aerobios y 51 % (n=79) para anaerobios. La proporción de cultivos positivos fue inferior en los casos de apendicitis no complicada, en comparación con aquellos de apendicitis complicada (80 % (66/83) Vs. 95 % (67/71); p=0,003). Los microorganismos aislados con mayor frecuencia fueron: Escherichia coli (53 %) (n=84), Bacteroides sp. (25 %) (n=25), Propionibacterium acnes (21 %) (n=21), Staphylococci coagulasa negativo (17 %) (n=27), Enterococcus sp. (10 %) (n=15) y Fusobacterium sp. (11 %) (n=11). La sensibilidad de E. coli a la amplicilina sulbactam fue de 30 %. La sensibilidad de Bacteroides spp. a la clindamicina y la ampicilina sulbactam fue de 91 %. El 100 % de los anaerobios fueron sensibles a piperacilina tazobactam, ertapenem, meropenem y metronidazol. Conclusiones. Los cultivos intraoperatorios son pertinentes en la apendicitis para determinar el patrón epidemiológico local, y establecer los antibióticos profilácticos y terapéuticos para esta enfermedad. Su siembra directa en botellas de hemocultivo permite una gran recuperación de microorganismos.

Direct antimicrobial susceptibility testing from positive blood culture bottles in laboratories lacking automated antimicrobial susceptibility testing systems.

BACKGROUND: Timely initiation of appropriate antimicrobial can improve the outcome in terms of reduced morbidity and mortality in addition to reduced health-care costs. Availability of early preliminary Antimicrobial Susceptibility Test (AST) report will be useful in directing antimicrobial therapy. The aim of the study was to correlate AST by disc diffusion method, directly from positively flagged blood culture bottles, with the AST by automated method. METHODS: A total of 144 aerobic blood culture bottles flagged positive by the automated blood culture system were processed. The bacteria were pelleted by two-step centrifugation of the broth from the bottle and used to make a smear for Gram stain as well as an inoculum for antimicrobial sensitivity testing by Kirby Bauer disc diffusion method. Automated identification and AST were also carried out. RESULTS: On direct staining, 94 samples showed gram-negative bacilli, 39 showed gram-positive cocci, and 11 showed yeasts or polymicrobial growth. In the case of gram-negative bacteria, there was 99% categorical agreement between direct sensitivity testing and automated sensitivity testing with 1% disagreement. Among the gram-positive cocci, there was 96% categorical agreement with 4% disagreement between the two methods. CONCLUSION: High degree of agreement between the two methods is promising and applicable to situations where automated sensitivity testing is not available. Even if the systems are available, this method would prove useful as an adjunct to standard AST reporting. This sensitivity report can be generated earlier than the conventional AST, enabling choice of appropriate antimicrobial.

Low antimicrobial resistance rates of Mycobacterium tuberculosis complex between 2000 and 2015 in Gipuzkoa, northern Spain.

OBJECTIVE: Although the incidence of tuberculosis (TB) has declined, TB drug resistance remains a major problem. The TB rate in Gipuzkoa (northern Spain) is higher than the European average. The objective of this study was to determine the antimicrobial susceptibility of 1855 Mycobacterium tuberculosis complex isolates (94.5% of confirmed cases between 2000 and 2015). METHODS: Susceptibility testing was performed using the agar proportion method and a commercial broth system (MGIT 960). In isoniazid- or rifampicin-resistant strains, we studied genetic determinants of drug resistance and genotype (MIRU-VNTR). RESULTS: The annual mean incidence of TB was 24.5 cases per 100,000 population on average, and tended to decrease. The multidrug-resistant TB rate was 0.5% (9/1855), and no extensively drug-resistant TB strains were detected. Rates of resistance to isoniazid and rifampicin were 3.9% (range, 3.4-4.3%) and 0.6% (range, 0.4-1.4%), respectively. TB resistance was more common among foreign-born individuals and those who had received previous TB treatment. Genotyping of 102 resistant strains showed predominance of the Euro-American lineage, although 4/9 multidrug-resistant strains had Eastern lineages (2 East African-Indian, and 2 East Asian [Beijing]). CONCLUSIONS: In Gipuzkoa, with a moderate incidence of TB, resistance was very low, mostly being detected among individuals who were born abroad or who had a history of TB treatment.

[Analysis of etiology and in vitro drug susceptibility of fungal keratitis in northern China].

Objective: To analyze the etiology, drug susceptibility and etiological changes of fungal keratitis so as to to provide reference for clinical diagnosis and treatment. Methods: Retrospective case series study. A total of 5 654 copies of specimens for fungal culture were collected from Outpatients suspected of fungal keratitis in Beijing Tongren Ophthalmic Center during January 2007 and December 2016. Specimens were inoculated on Potato dextrose agar (PDA) slant medium, incubated at 30℃ and 40% humidity constantly for 3 to 10 days. Filamentous fungi isolated from positive cultures were identified according to morphological traits. Yeast-like fungi were identified by API 20 C Aux. Drug susceptibility tests were performed by using ROSCO disk diffusion method, which included natamycin, terbinafine, itraconazole, fluconazole, amphotericin B, voriconazole. The Chi-square test and Spearman correlation analysis were performed using SPSS 20.0 software. Results: The positive rate of culture was 26.1%. In positive patients, the ratio of male to female was 1.77∶1, and the age range was 11 days to 95 years. The mean age was 49.0±16.9 years. Among isolated fungi, Fusarium sp. was the most common genus, accounting for 53.5% (789 strains) , followed by 17.5% of Aspergillus sp. (259 stains) and 13.8% of Alternaria sp. (203 stains). The sensitive rate of natamycin was 92.3% (410 strains) , followed by that of terbinafine as 78.5% (1 093 strains), voriconazole as 41.0% (338 strains), amphotericin B as 40.7% (553 strains). Conclusion: The predominant patients of fungal keratitis are adult and male. Fusarium sp., Aspergillus sp., and Alternaria sp. are three common species in northern china with seasonal changes in their proportion. Natamycin should be the preferred drug for empirical treatment. The preferred empirical treatment for Aspergillus sp. is terbinafine. Fluconazole should not be used alone due to the high resistant rate. (Chin J Ophthalmol, 2018, 54: 432-436).

Clinical Evaluation of QMAC-dRAST for Direct and Rapid Antimicrobial Susceptibility Test with Gram-Positive Cocci from Positive Blood Culture Bottles / 대한임상미생물학회지

BACKGROUND: Timely intervention in the treatment of bloodstream infection is important for prescription of appropriate antimicrobials. With prompt determination of the antimicrobial susceptibility of a causative agent, rapid antimicrobial susceptibility test (AST) can help select the appropriate antimicrobial therapy. This clinical study is for evaluation of the clinical performance of the QMAC-dRAST for rapid AST directly from positive blood culture (PBC)s with Gram-positive cocci. METHODS: A total of 115 PBC samples with Gram-positive organisms (76 Staphylococcus spp. and 39 Enterococcus spp.) were evaluated by the QMAC-dRAST system, and their pure culture isolates were evaluated by the MicroScan WalkAway (Beckman Coulter, USA) as the comparative AST system. Thirteen antimicrobial agents were included, and the agreement and discrepancy rates of the QMAC-dRAST system (Quantamatrix Inc., Republic of Korea) compared to the MicroScan WalkAway were calculated. To resolve discrepancies, the broth microdilution method was performed. RESULTS: The QMAC-dRAST system exhibited a categorical agreement rate of 94.9% (1,126/1,187) and an essential agreement rate of 98.3% (1,167/1,187). The QMAC-dRAST system yielded very major (false-susceptible) errors at 1.0% (5/485), major (false-resistant) errors at 1.3% (9/693), and minor errors at 4.0% (47/1,187) compared to the MicroScan WalkAway. The QMAC-dRAST system significantly eliminated 30 hours of total turnaround time by combination of direct inoculation of PBC and an image-based approach. CONCLUSION: The results of the QMAC-dRAST system were highly accurate. Thereby, the QMAC-dRAST may provide essential information to accelerate therapeutic decisions for earlier and adequate antibiotic treatment and patient management in clinical settings.