Crude extracts of an endophytic fungus attenuate the growth of pathogenic bacteria in aquaculture.

Background and Objectives: Endophytic fungi are believed to possess compounds as antibacterial agents. This study was designed to determine in vivo antibacterial activity of the crude extracts from Lasiodiplodia pseudotheobromae IBRL OS-64 against pathogenic bacteria. Materials and Methods: The qualitative and quantitative screenings were performed using agar plug and disk diffusion antimicrobial tests, respectively. Besides that, the MIC and MBC value of the extracts were determined using broth microdilution assay and morphological changes of the bacterial cells exposed to the extract were observed under Scanning Electron Microscope (SEM). Results: Agar plug diffusion assay revealed that V. parahaemolyticus ATCC 17802 and Exiguobacterium profundum IBRL MA6 were the most sensitive to the extract with the size of inhibition zones of 11 to ≤ 20 mm. The MIC and MBC values of the extract varied depending on the test bacteria. Observation through SEM revealed that the bacterial cells exposed to the extract experienced severe damage such as irregular shape with crumpled and shrunken cells which led to cell death. Conclusion: The data suggest that the crude extracts of L. pseudotheobromae IBRL OS-64 exert antibacterial activity against test bacteria and principally affect the cell wall in growing pathogenic bacterial cells.

Retentive efficacy, antimicrobial and cytotoxicity comparisons between different types of commercial and experimental denture adhesives with antifungal action.

The effect of the addition of nystatin and an alternative antifungal derived from pyrazoles in different commercial denture adhesives on their retentive efficacy, cytotoxicity and antimicrobial activity against Candida albicans was evaluated. Commercial denture adhesives were prepared with the inclusion of nystatin and 3,5-diaryl-4,5-dihydro-1H-pyrazole-1-carboximidamide (pyrazole) in three concentrations: 23.78 %w/w, 3.02 %w/w, and 0.31 %w/w (0.015 g, 0.0015 g, and 0.00015 g, respectively). The retentive efficacy was tested observing the influence of the medium, type of commercial denture type and the test condition (dipping). The antifungal action through disk diffusion and direct contact tests at 1, 4, 8 and 12 h and cytotoxic activity was evaluated in mouse fibroblasts (NIH/3T3) by the MTT reduction colorimetric assay. The addition of pyrazole and nystatin in commercial denture adhesives did not affect retentive efficacy rates and enhanced antifungal actions against Candida albicans. Results show a possibility of using denture adhesives as a delivery system for commercial antifungals (Nystatin) or pyrazole, with the second concentration (1,560 µg-3.02 %w/w) as the most efficient.

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.

Analysis of Eurasian Stone curlew (Burhinus oedicnemus) microbial flora reveals the presence of multi-drug resistant pathogens in agro-pastoral areas of Sicily (Italy).

Spread of multi-drug resistant (MDR) bacteria in natural environments pose a risk to human and animal health. Wild birds are considered to be reservoirs of human pathogens and vectors of antimicrobial resistance distribution in the environment. The aim of this study is to assess the occurrence of antibiotic resistant bacteria in isolates from bird specimens living in three agro-pastoral areas of the southeastern Sicily. We analyzed the microbiomes of the Eurasian Stone curlew Burhinus oedicnemus (Charadriiformes, Aves) and identified 91 Gram positive and 212 Gram negative strains, whose antimicrobial susceptibility to 11 and 9 antibiotic classes (respectively) was evaluated using agar disk diffusion test. Isolates showed significant levels of antimicrobial resistance, and a high percentage of MDR strains was found both between the Gram positive (49.4%) and the Gram negative (34.9%). Multi-drug resistance levels are higher among strains isolated in the beak and the eye than among enteric (faeces and cloaca) strains. Our results indicate high levels of MDR strains among wild bird populations, with a potential threat to wildlife and human populations.

Comparison of Escherichia coli antibiotic-resistance profiles and incidence of betalactamase phenotypes in three private health facilities in Perú / [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].

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.

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.

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.

Evaluation of Cumulative and Conditional Antibiotic Release from Vancomycin-Embedded Fibrin Sealant and Its Antibacterial Activity : An In Vitro Study.

OBJECTIVE: Fibrin sealants have been used for hemostasis, sealant for cerebrospinal fluid leakage, and adhesive barrier in neurosurgery. Further, as its clinical use and role of an effective drug delivery vehicle have been proposed. This study was performed to measure antibacterial activity and continuous local antibiotic release from different concentrations of vancomycin-impregnated fibrin sealant in vitro. METHODS: Antibacterial activity was investigated by disk diffusion test by measuring the diameter of the growth inhibition zone of bacteria (methicillin-resistant Staphylococcus aureus, ATCC29213) from vancomycin-embedded fibrin sealant disc diluted at five different concentrations (C1-C5; 8.33, 4.167, 0.83, 0.083, and 0.0083 mg/disc, respectively). Continuous and conditioned release of vancomycin concentration (for 2 weeks and for 5 days, respectively) were also measured using high-performance liquid chromatography (HPLC) method. To mimic the physiologic wound conditions with in vitro, conditioned vancomycin release in phosphate buffer solution (PBS) was measured and replaced PBS for five consecutive days, half a day or completely daily. RESULTS: In the disk diffusion test, the mean diameters of bacterial inhibition zone were 2.54±0.07 cm, 2.61±0.12 cm, and 2.13±0.15 cm (C1, C2, and C3 respectively) but 1.67±0.06 cm and 1.23±0.15 cm in C4 and C5, respectively. Continuous elution test elicited the peak release of vancomycin from the fibrin sealant at 48 hours, with continued release until 2 weeks. However, conditioned vancomycin release decreased to half or more on day 2, however, the sustainable release was measured over the therapeutic dose (10-20 µg/mL) for 5 days and 4 days in assays of half and total exchange of PBS. CONCLUSION: This study suggests that fibrin sealant can provide an efficient vehicle for antibiotic drug release in a wide range of neurosurgical procedures and the safe and effective therapeutic dose will be at the concentration embedded of 4.167 mg/disc or more of vancomycin.

Efficacy of Antimicrobials Against Multidrug-Resistant Acinetobacter baumannii from Patients in a Tertiary Care Hospital.

Introduction: In just two decades Acinetobacter baumannii has attained considerable importance, evolving from an insignificant organism to a leading pathogen especially in intensive care unit settings globally. Treatment options are already very limited and have almost run out due to the rapid emergence of antimicrobial resistance. Evaluation of antimicrobials that are currently in use to determine their effectiveness against multidrug-resistant (MDR) strains and developing newer options is of utmost importance. We thus set out to determine the efficacy of routinely used antibiotics against MDR A. baumannii. Materials and Methods: This was a cross-sectional study conducted at the Department of Microbiology, Army Medical College, National University of Medical Sciences (Rawalpindi, Pakistan) from December 2015 to June 2016. The organisms were identified on the basis of colony morphology, gram staining, catalase, oxidase, motility test, and API (analytical profile index) 20NE. The organisms were considered to be MDR when the isolate was found to be resistant to at least one agent in more than three antimicrobial groups. Antibiotic sensitivity was determined using the modified Kirby-Bauer disc diffusion method according to Clinical Laboratory Standards Institute (CLSI) guidelines. Results: The 77 isolates were found to have good sensitivity to tigecycline (94.8%) and minocycline (80.5%). Most of the isolates were resistant to other routinely used antibiotics. Conclusion: A few antibiotics, tigecycline and minocycline, are still effective against these MDR A. baumannii. We need to remain up to date regarding the efficacy of antibiotics to effectively treat patients with these MDR bacteria.

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.

Antimicrobial Effect of Different Sizes of Nano Zinc Oxide on Oral Microorganisms.

OBJECTIVES: The purpose of the present study was to evaluate the antimicrobial effect of various sizes and concentrations of zinc oxide (ZnO) nanoparticles on Streptococcus mutans (S. mutans), Enterococcus faecalis (E. faecalis), Lactobacillus fermentum (L. fermentum), and Candida albicans (C. albicans). MATERIALS AND METHODS: Solutions at the concentration of 10 µg/ml were prepared using 20-nm, 40-nm, and 140-nm nano ZnO (nZnO) powder. The antimicrobial effect of nZnO was determined using the disk diffusion method. The inhibition zone (mm) was measured using a ruler. Data were analyzed by analysis of variance (ANOVA) and the Bonferroni correction. The minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of nZnO were determined using the broth microdilution method in Mueller-Hinton Agar (MHA) for S. mutans and E. faecalis, De Man, Rogosa, and Sharpe (MRS) agar, and Sabouraud Dextrose Agar (SDA). RESULTS: The greatest inhibition zones were observed against S. mutans with 20-nm and 40-nm nZnO, while 140-nm nZnO formed the greatest inhibition zones against S. mutans and E. faecalis. The smallest inhibition zones were observed against C. albicans with the three nZnO particle sizes. The MICs for C. albicans with 40-nm and 140-nm particles and for L. fermentum with 140-nm particles were higher than 10 µg/ml. A significant correlation was found between the particle size and the antibacterial activity against S. mutans (P=0.00), L. fermentum, and E. faecalis (P<0.02). CONCLUSION: The antimicrobial activity of nZnO increases with decreasing the particle size. The greatest antimicrobial effect was observed against S. mutans and E. faecalis. S. mutans is more sensitive to the changes in the particle size compared to other bacteria.

COMPARISON OF TWO DIFFERENT METHODS FOR TIGECYCLINE SUSCEPTIBILITY TESTING IN ACINETOBACTER BAUMANNII.

- Tigecycline susceptibility testing (TST) presents a tremendous challenge for clinical microbiologists. Previous studies have shown that the Epsilometer test (E-test) and Vitek 2 automated system significantly overestimate the minimum inhibitory concentrations for tigecycline resistance compared to the broth microdilution method (BMM). This leads to very major errors or false susceptibility (i.e. the isolate is called susceptible when it is actually resistant). The aim of this study was to compare E-test against BMM for TST in carbapenem-resistant and carbapenem-susceptible Acinetobacter (A.) baumannii and to analyze changes in tigecycline susceptibility between two time periods (2009-2012 and 2013-2014), with BMM as the gold standard. Using the EUCAST criteria, the rate of resistance to tigecycline for the OXA-23 MBL-positive, OXA-23 MBL-negative and carbapenemase-negative strains for BMM was 54.5% (6/11), 29.4% (5/17) and 2.7% (1/37), respectively; the OXA-24/40 and OXA-58 producing organisms did not exhibit any resistance. With E-test, all OXA-23 MBL-positive organisms (11/11), 23.5% (4/17) of OXA-23 MBL-negative, and 4.1% of OXA-24/40 (3/74) strains displayed tigecycline resistance; there were no resistant strains among the OXA-58 and carbapenemase-negative isolates. Resistance emerged in the bacterial isolates from 2013 to 2014. Although tigecycline does not display cross-resistance, the highest rates of resistant A. baumannii isolates were observed among those producing VIM MBL, regardless of the testing method. These findings suggest that the commercial E-test does not provide reliable results for TST of A. baumannii. Further confirmation with the dilution method should be recommended, particularly in cases of serious infections.

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.

A cost-saving strategy for processing isolated uropathogens in community-acquired urinary tract infections.

Outpatient urine samples are among the most commonly processed in a microbiology laboratory, which involves a high economic burden. The aim of this study was compare cost and efficiency to process uropathogens between MicroScan system (2010-2011) versus a chromogenic medium and the disk diffusion method (2013-2014). In the first period, a total 9918 bacterial populations were isolated from urine samples. Annual estimated costs during 2010 and 2011 for processing were EUR 53,818 and EUR 57,306, respectively (EUR 111,124 total). In the second period, a total 11,728 bacterial isolates were processed, with annual estimated costs of EUR 21,078 and EUR 23,248, respectively (EUR 44,326 total). We included the cost for a laboratory technician (252h worked per year), estimated at EUR 2500 per year. The mean estimated savings were EUR 66,797 (60%).The identification by chromogenic media and antibiotic susceptibility patterns by disk diffusion method was similar to MicroScan in both study periods. Only some isolated Citrobacter spp., Enterobacter spp., Morganella morganii, and Providencia spp. were misidentified. The strategy reported here did not affect the quality of the results and yielded substantial cost savings.

Incidence and Characteristics of Scarlet Fever, South Korea, 2008-2015.

The incidence rate for scarlet fever in South Korea is rising. During 2008-2015, we collected group A Streptococcus isolates and performed emm and exotoxin genotyping and disk-diffusion antimicrobial tests. Scarlet fever in South Korea was most closely associated with emm types emm4, emm28, emm1, and emm3. In 2015, tetracycline resistance started increasing.

Antimicrobial Effects of Mastic Extract Against Oral and Periodontal Pathogens.

BACKGROUND: Various antimicrobial agents are widely used in the therapy of oral inflammatory diseases. However, their side effects and the appearance of drug resistance justify research on natural antimicrobial agents to target oral pathogens that are safe for the host. In the present study, antimicrobial properties of mastic extract on commensal and pathogenic oral bacteria, as well as its possible cytotoxic effect toward cells of epithelial and mesenchymal origin, were evaluated and compared with the common antimicrobial agents hydrogen peroxide (H2O2) and chlorhexidine digluconate (CHX). METHODS: Oral and periodontal pathogens (Porphyromonas gingivalis, Streptococcus mutans [Sm], Streptococcus oralis, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Prevotella intermedia, and Prevotella nigrescens) were treated with different concentrations of mastic extract, 3% H2O2, and 0.2% CHX, and evaluated with an agar diffusion test. The cytotoxic effect of mastic extract was tested on four cell lines of epithelial and mesenchymal origin (HaCaT, SaOS-2, MC3T3-E1, periodontal ligament [PDL] cells) by neutral red and 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide assay. RESULTS: Mastic extract led to significantly (P ≤0.016) increased inhibition of the tested periodontal pathogens compared with H2O2. No effect of mastic extract was observed on Sm. Mastic extract showed beneficial effects on cell viability because viability values of tested cells were significantly (P ≤0.016) lower for cells treated with CHX and H2O2 compared with mastic extract-treated cells after stimulation for 2, 4, and 6 hours. CONCLUSION: The present data demonstrate mastic extract's inhibition of periodontal pathogens, as well as beneficial effects on cell viability, compared with H2О2, suggesting that it could be considered an alternative antibacterial agent in the prevention of periodontal disease.

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.

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

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.