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Sample records for antifungal susceptibility testing

  1. Antifungal susceptibility testing method for resource constrained laboratories

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    Khan S

    2006-01-01

    Full Text Available Purpose: In resource-constrained laboratories of developing countries determination of antifungal susceptibility testing by NCCLS/CLSI method is not always feasible. We describe herein a simple yet comparable method for antifungal susceptibility testing. Methods: Reference MICs of 72 fungal isolates including two quality control strains were determined by NCCLS/CLSI methods against fluconazole, itraconazole, voriconazole, amphotericin B and cancidas. Dermatophytes were also tested against terbinafine. Subsequently, on selection of optimum conditions, MIC was determined for all the fungal isolates by semisolid antifungal agar susceptibility method in Brain heart infusion broth supplemented with 0.5% agar (BHIA without oil overlay and results were compared with those obtained by reference NCCLS/CLSI methods. Results: Comparable results were obtained by NCCLS/CLSI and semisolid agar susceptibility (SAAS methods against quality control strains. MICs for 72 isolates did not differ by more than one dilution for all drugs by SAAS. Conclusions: SAAS using BHIA without oil overlay provides a simple and reproducible method for obtaining MICs against yeast, filamentous fungi and dermatophytes in resource-constrained laboratories.

  2. EVALUATION OF VITEK 2 SYSTEM FOR CLINICAL IDENTIFICATION OF CANDIDA SPECIES AND THEIR ANTIFUNGAL SUSCEPTIBILITY TEST

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    Mohan,, V.; Ram Murugan

    2016-01-01

    BJECTIVES 1. To evaluate the Vitek 2 system for clinical identification of Candida species and their antifungal susceptibility test; 2. To study the incidence of various types of Candida species in this part of Tamilnadu. METHODS Samples collected from different wards were subjected for culture, isolation and identification of Candida Species and Antifungal Susceptibility testing by Vitek System. Vitek 2 test was carried out in Apollo Specialty Hospital Lab Services, Madurai....

  3. Flow cytometry susceptibility testing for conventional antifungal drugs and Comparison with the NCCLS Broth Macrodilution Test

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    M.J. Najafzadeh

    2009-08-01

    Full Text Available Introduction: During the last decade, the incidence of fungal infection has been increased in many countries. Because of the advent of resistant to antifungal agents, determination of an efficient strategic plan for treatment of fungal disease is an important issue in clinical mycology. Many methods have been introduced and developed for determination of invitro susceptibility tests. During the recent years, flow cytometry has developed to solving the problem and many papers have documented the usefulness of this technique. Materials and methods: As the first step, the invitro susceptibility of standard PTCC (Persian Type of Culture Collection strain and some clinical isolates of Candida consisting of Candida albicans, C. dubliniensis, C. glabrata, C. kefyer and C. parapsilosis were evaluated by macrodilution broth method according to NCCLS (National Committee for Clinical Laboratory Standards guidelines and flow cytometry susceptibility test. Results:  The data indicated that macro dilution broth methods and flow cytometry have the same results in determination of MIC (Minimum Inhibitory Concentration for amphotericin B, clotrimazole, fluconazole, ketoconazole and miconazole in C. albicans PTCC 5027 as well as clinical Candida isolates, such as C.albicans, C.dubliniensis, C.glabrata C.kefyr, and C.parapsilosis. Discussion: Comparing the results obtained by macrodilution broth and flow cytometry methods revealed that flow cytometry was faster. It is suggested that flow cytometry susceptibility test can be used as a powerful tool for determination of MIC and administration of the best antifungal drug in treatment of patients with Candida infections.

  4. Detection and antifungal susceptibility testing of oral Candida dubliniensis from human immunodeficiency virus-infected patients

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    Chunchanur Sneha

    2009-10-01

    Full Text Available Context: Candida dubliniensis, an opportunistic yeast that has been implicated in oropharyngeal candidiasis (OPC in patients infected with Human Immunodeficiency Virus (HIV may be under-reported due to its similarity with Candida albicans. Resistance to Fluconazole is often seen in C. dubliniensis isolates from clinical specimens. Aims: To know the prevalence of C. dubliniensis in OPC in patients infected with HIV and their antifungal susceptibility pattern. Settings and Design: One hundred and thirty-two HIV seropositive individuals and 50 healthy controls were included in the study. Materials and Methods: Two oral swabs were collected from the site of the lesion from 132 HIV-infected patients. Oral rinse was obtained from 50 healthy controls. Samples were inoculated on Sabouraud′s dextrose agar (SDA medium and on HiCrome Candida Differential Agar (CHROM agar medium. Isolates were speciated by standard tests. Dark green-colored, germ tube positive isolates, which failed to grow at 420C and negative for xylose assimilation were identified as C. dubliniensis. Antifungal susceptibility test was performed by Macro broth dilution technique (National Committee for Clinical Laboratory Standards guidelines. Results and Conclusions: From 132 patients, 22 (16.3% C. dubliniensis were isolated; samples from healthy controls did not reveal their presence. Antifungal susceptibility test showed higher resistance among C. dubliniensis isolates to azoles compared to C. albicans. Five (22.7% isolates of C. dubliniensis were resistant to Fluconazole followed by four (18.2% to Ketoconazole. This study emphasizes the importance of identification and antifungal susceptibility testing of C. dubliniensis in HIV-infected patients.

  5. STUDY OF CHARACTERIZATION & ANTIFUNGAL SUSCEPTIBILITY TESTING OF CLINICALLY SIGNIFICANT CANDIDA SPECIES

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    Geeta

    2014-05-01

    Full Text Available BACKGROUND: Over the last two decades Candida has accounted for the most serious opportunistic infections especially in the immuno-compromised individuals. Candida species have emerged as important causes of invasive infections & the rates of resistance to standard antifungal therapies are on the rise. Awareness regarding fungal infections has compelled the clinicians and laboratories to lay more emphasis on the detection of fungi; as speciation and antifungal tests are not routinely done. Over the past decade significant progress has been made with standardization of the methods for antifungal susceptibility testing, correlation between in-vitro results & patient outcome. OBJECTIVES: The aim of this study was to isolate, identify & determine the susceptibility pattern of clinically significant Candida species and study the spectrum of Non-albicans Candida species, thus contributing to overall reduction in the cost of treatment and duration of hospital stay. METHODS: The study was carried out at department of microbiology MVJ Medical Hospital Bangalore for one year from Aug 2010 – July 2011. 50 Candida species which were isolated from various clinical specimens were included in the study. They were identified by using various media & identification methods. Antifungal susceptibility testing was done on Yeast nitrogen base agar by disk diffusion method & analyzed. RESULTS: Non-albicans Candida (NAC emerged as the commonest species with [39(22%] causing fungal infection followed by Candida albicans [11(22%]. Among the NAC isolates Candida tropicalis was predominant followed by Candida krusei, Candida glabrata & Candida guilliermondi. CONCLUSION: Studying the speciation & susceptibility patterns of Candida will help us understand the etio-pathology and might assist in better patient care

  6. Process analysis of variables for standardization of antifungal susceptibility testing of nonfermentative yeasts.

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    Zaragoza, Oscar; Mesa-Arango, Ana C; Gómez-López, Alicia; Bernal-Martínez, Leticia; Rodríguez-Tudela, Juan Luis; Cuenca-Estrella, Manuel

    2011-04-01

    Nonfermentative yeasts, such as Cryptococcus spp., have emerged as fungal pathogens during the last few years. However, standard methods to measure their antifungal susceptibility (antifungal susceptibility testing [AST]) are not completely reliable due to the impaired growth of these yeasts in standard media. In this work, we have compared the growth kinetics and the antifungal susceptibilities of representative species of nonfermentative yeasts such as Cryptococcus neoformans, Cryptococcus gattii, Cryptococcus albidus, Rhodotorula spp., Yarrowia lipolytica, Geotrichum spp., and Trichosporon spp. The effect of the growth medium (RPMI medium versus yeast nitrogen base [YNB]), glucose concentration (0.2% versus 2%), nitrogen source (ammonium sulfate), temperature (30°C versus 35°C), shaking, and inoculum size (10(3), 10(4), and 10(5) cells) were analyzed. The growth rate, lag phase, and maximum optical density were obtained from each growth experiment, and after multivariate analysis, YNB-based media demonstrated a significant improvement in the growth of yeasts. Shaking, an inoculum size of 10(5) CFU/ml, and incubation at 30°C also improved the growth kinetics of organisms. Supplementation with ammonium sulfate and with 2% glucose did not have any effect on growth. We also tested the antifungal susceptibilities of all the isolates by the reference methods of the CLSI and EUCAST, the EUCAST method with shaking, YNB under static conditions, and YNB with shaking. MIC values obtained under different conditions showed high percentages of agreement and significant correlation coefficient values between them. MIC value determinations according to CLSI and EUCAST standards were rather complicated, since more than half of isolates tested showed a limited growth index, hampering endpoint determinations. We conclude that AST conditions including YNB as an assay medium, agitation of the plates, reading after 48 h of incubation, an inoculum size of 10(5) CFU/ml, and

  7. Multilaboratory Evaluation of In Vitro Antifungal Susceptibility Testing of Dermatophytes for ME1111

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    Chaturvedi, V.; Diekema, D.; Ostrosky-Zeichner, L.; Rennie, R.; Walsh, T.; Wengenack, N.; Fothergill, A.; Wiederhold, N.

    2015-01-01

    ME1111 is a novel small molecule antifungal agent under development for the topical treatment of onychomycosis. Standardization of the susceptibility testing method for this candidate antifungal is needed. Toward this end, 8 independent laboratories determined the interlaboratory reproducibility of ME1111 susceptibility testing. In addition, we subsequently identified 2 strains as quality control (QC) isolates for the method. In the reproducibility study, 5 blinded clinical strains each of Trichophyton rubrum, Trichophyton mentagrophytes, and Epidermophyton floccosum were tested, while the QC study tested 6 blinded T. rubrum or T. mentagrophytes ATCC strains. Testing was performed in frozen microtiter panels according to the Clinical and Laboratory Standards Institute (CLSI) M38-A2 methodology. In the reproducibility study, 9 of 15 clinical strains showed interlaboratory agreement of >90% at the 80% inhibition endpoint, with a range of agreement of 76.2% to 100%. In the QC study, 4 of the 6 ATCC strains showed interlaboratory agreement of >90%. ME1111 demonstrated excellent interlaboratory agreement when tested against dermatophytes. Based on this data, the CLSI Subcommittee on Antifungal Susceptibility Tests approved the susceptibility testing of ME1111 against dermatophytes according to M38-A2 methodology, which stipulates RPMI 1640 as the test medium, an inoculum size of 1 to 3 × 103 CFU/ml, and an incubation time and temperature of 96 h at 35°C. The MIC endpoint should be 80% inhibition compared with the growth control. T. rubrum ATCC MYA-4438 and T. mentagrophytes ATCC 28185 were selected as QC isolates, with an acceptable range of 0.12 to 1 μg/ml for the two strains. PMID:26719434

  8. Spectrophotometric reading of EUCAST antifungal susceptibility testing of Aspergillus fumigatus

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    Meletiadis, J; Leth Mortensen, K; Verweij, P E

    2017-01-01

    . METHODS: Eighty-eight clinical isolates of A. fumigatus were tested against four medical azoles (posaconazole, voriconazole, itraconazole, isavuconazole) and one agricultural azole (tebuconazole) with EUCAST E.Def 9.3. The visually determined MICs (complete inhibition of growth) were compared...

  9. Antifungal susceptibility testing of vaginal candida isolates: the broth microdilution method

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    Mahmoudi Rad M

    2010-02-01

    Full Text Available "n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: Vulvovaginal candidiasis is a common mucosal infection among immunocompetent, healthy women, and is caused by opportunistic yeasts that belong to genus Candida. In this study, we isolated and identified the Candida species in the vagina of patients who admitted in Gynecology Department of Mahdieh Hospital in Tehran, Iran to evaluate the in vitro activities of fluconazole, miconazole, itraconazole and flucytosine against 191 clinical Candida isolates by the NCCLS microdilution method."n"nMethods: 191 Candida were isolated from vaginal secretions and identified with conventional mycological methods in the diagnosis of Candida species. The identity of all strains was confirmed genotypically by multiplex PCR. In vitro susceptibility testing of vaginal Candida isolates was performed by the NCCLS broth microdilution method. The results were read at 48 h."n"nResults: Most C. albicans isolates (>90% were sensitive in vitro to the antifungal agents tested. Most C. glabrata isolates showed sensitivity to miconazole and then flucytosine while they were more resistant to Itraconazole and fluconazole. Many isolates of C. tropicalis were susceptible to miconazole and then fluconazole. They showed a little resistance to

  10. EVALUATION OF VITEK 2 SYSTEM FOR CLINICAL IDENTIFICATION OF CANDIDA SPECIES AND THEIR ANTIFUNGAL SUSCEPTIBILITY TEST

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    Mohan

    2016-06-01

    Full Text Available BJECTIVES 1. To evaluate the Vitek 2 system for clinical identification of Candida species and their antifungal susceptibility test; 2. To study the incidence of various types of Candida species in this part of Tamilnadu. METHODS Samples collected from different wards were subjected for culture, isolation and identification of Candida Species and Antifungal Susceptibility testing by Vitek System. Vitek 2 test was carried out in Apollo Specialty Hospital Lab Services, Madurai. The cost per test is Rs. 200 (Subsidized rate. The expenses for the lab tests (Vitek were borne by the author himself. RESULTS 124 samples were collected from urine, sputum, blood, pus and wounds. Candida albicans formed 43% of the samples. Among the 57% of Non-Candida albicans, Candida tropicalis formed 42%, Candida krusei formed 6%, Candida guilliermondii formed 4%, Candida inconspicua, Candida parapsilosis, Candida glabrata, Candida rugosa and Candida lusitaniae formed 1% each. Candida albicans and C. tropicalis showed high sensitivity to Voriconazole, Flucytosine, Amphotericin B and Fluconazole. CONCLUSION Candida tropicalis was identified as the most common Candida non-albicans species. Candida albicans and C. tropicalis showed high sensitivity to Voriconazole, Flucytosine, Amphotericin B and Fluconazole. This study was helpful to treat Candida albicans and Non-Candida albicans species patients accurately and earlier by Vitek method.

  11. A case of Candida famata sepsis in a very low birth weight infant successfully treated with fluconazole following antifungal susceptibility testing

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    Shilpee Raturi

    2015-01-01

    This case report highlights the growing number of cases arising due to nonalbicans Candida infections in the neonatal intensive care units and the usefulness of antifungal susceptibility testing in deciding optimal antifungal therapy and preventing the emergence of drug resistance.

  12. Antifungal susceptibility testing of Exophiala spp.: a head-to-head comparison of amphotericin B, itraconazole, posaconazole and voriconazole.

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    Fothergill, Annette W; Rinaldi, Michael G; Sutton, Deanna A

    2009-02-01

    Frequently, diseases caused by black yeasts are chronic in nature with a high morbidity. In addition, these infections are often fatal and relapse is common, even after prolonged treatment. Although the CLSI Document M38-A outlines methods for antifungal susceptibility testing of moulds, Exophiala spp. are not directly discussed. In an effort to determine the antifungal susceptibility patterns of Exophiala spp. we tested 160 clinical isolates against amphotericin B, itraconazole, posaconazole, and voriconazole in a head-to-head comparison. Posaconazole and itraconazole were the most active in vitro with MICs falling well below the achievable serum levels typically observed with standard dosing regimens.

  13. Susceptibility of Clinical Candida Species Isolates to Antifungal Agents by E-Test, Southern Iran: A Five Year Study

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    A Alborzi

    2011-12-01

    Full Text Available Background and Objectives: The incidence of fungal infections in immunocompromised patients, especially by Candida species, has increased in recent years. This study was designed to identify Candida species and determine antifungal susceptibility patterns of 595 yeast strains isolated from various clinical specimens.Material and Methods: Identification of the isolates were determined by the API 20 C AUX kit and antifungal susceptibilities of the species to fluconazole, amphotericin B, ketoconazole, itraconazole, voriconazole, and caspofungin were determined by the agar-based E-test method.Results: Candida albicans (48% was the most frequently isolated species, followed by Candida kruzei (16.1%, Candida glabrata (13.5%, Candida kefyr (7.4%, Candida parapsilosis (4.8%, Candida tropicalis (1.7% and other species (8.5%. Resistance varies depending on the species and the respective antifungal agents. Comparing the MIC90 for all the strains, the lower MIC90 was observed for caspofungin (0.5 μg/ml. The MIC90 for all Candida species were 64 μg/ml for fluconazole, 0.75 μg/ml for amphotericin B, 4 μg/ml for ketoconazole, 4 μg/ml for itraconazole, and 2 μg/ml for voriconazole.Conclusions: Species definition and determination of antifungal susceptibility patterns are advised for the proper management and treatment of patients at risk for systemic candidiasis. Resistance to antifungal agents is an alarming sign for the emerging common nosocomial fungal infections.

  14. In vitro susceptibility testing of dermatophytes isolated in Goiania, Brazil, against five antifungal agents by broth microdilution method.

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    Araújo, Crystiane Rodrigues; Miranda, Karla Carvalho; Fernandes, Orionalda de Fatima Lisboa; Soares, Ailton José; Silva, Maria do Rosário Rodrigues

    2009-01-01

    The antifungal activities of fluconazole, itraconazole, ketoconazole, terbinafine and griseofulvin were tested by broth microdilution technique, against 60 dermatophytes isolated from nail or skin specimens from Goiania city patients, Brazil. In this study, the microtiter plates were incubated at 28 masculineC allowing a reading of the minimal inhibitory concentration (MIC) after four days of incubation for Trichophyton mentagrophytes and five days for T. rubrum and Microsporum canis. Most of the dermatophytes had uniform patterns of susceptibility to the antifungal agents tested. Low MIC values as 0.03 microg/mL were found for 33.3%, 31.6% and 15% of isolates for itraconazole, ketoconazole and terbinafine, respectively.

  15. Candidemia: Speciation and Antifungal susceptibility testing from a Tertiary Care Hospital in Maharashtra, India

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    Nageswari Rajesh Gandham

    2016-01-01

    Full Text Available Introduction: Invasive fungal infections are an important nosocomial health problem, candidal infections being the most common. Candidal bloodstream infections (BSIs account for one-fourth of the nosocomial BSIs. Candida albicans once the most common has now been replaced by nonalbicans Candida, accounting for majority of cases of candidal infections. The knowledge and understanding of specific risk factors, epidemiology, prophylactic measures, and outcomes with relation to speciation are changing rapidly. Emergence of antifungal resistance is a cause of concern. With this background, a prospective study was undertaken. Aims and Objectives: To isolate Candida spp. from blood cultures of patients with clinically diagnosed or suspected septicemia; to identify and speciate Candida isolates and carry out the antifungal susceptibility test; to determine incidence of candidemia; to find out the crude mortality rate in candidemia patients. Materials and Methods: Blood cultures received in the Department of Microbiology from various critical care units over a 1-year period were included in this study. Samples were cultured on Sabouraud dextrose agar and routine bacteriological cultures. Speciation of suspected Candida growth was done by conventional and automated system. Results: A total of 225 out of 839 blood cultures were culture positive. A total of 196 were bacterial pathogens. Candida spp. were isolated in 29 samples. These included 10 cases of Candida tropicalis, 8 Candida parapsilosis, and three cases each of C. albicans and Candida krusei. Fluconazole resistance was seen (minimum inhibitory concentration ≥64 μg/ml in 4 isolates, 2 of C. krusei and 2 of Candida haemulonii. Incidence of candidemia was 3.46%. Crude mortality rate in the present study was 24%.

  16. [Evaluation of a new method for antifungal drugs susceptibility testing to yeasts].

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    Ishigaki, S; Kawakami, S; Ono, Y; Miyazawa, Y

    2000-03-01

    We compared the Etest with a broth microdilution method (FP panel), performed according to the National Committee for modified Clinical Laboratory Standards (NCCLS) document M27-P guidelines, for determining the MICs of 81 clinical isolates of yeasts (7 Candida albicans, 8 Candida glabrata, 10 Candida parapsilosis, 6 Pichia anomala, 10 Candida tropicalis, 4 Candida guilliermondii, 4 Candida krusei, 6 Trichosporon cutaneum, 5 Candida ciferrii, 3 Candida famata, 4 Candida norvegensis, 2 Rhodotorula rubra, 3 Candida lusitaniae, 2 Candida curvata, 1 Candida inconspicua, 1 Candida intermedia, 1 Candida colliculosa, 1 Cryptococcus spp, 1 Tricosporon capitatum, 1 Pichia ohmeri, 1 Saccharomyces cerevisiae). The Etest results for 6 ATCC standard strains correlated well with reference MICs except those of flucytosine (5-FC) for C. krusei, which tended to be 1 to 2 log2 dilution higher than the MIC range determined by NCCLS guidelines. However, the best agreement between the results for clinical isolates was seen with 5-FC (100% agreement [Within +/- 2 log2 dilutions] between the results of the two methods). There was a 91.4% agreement between the results of the two methods with amphotericin B (Etest MICs tended to be 1 to 2 log2 dilution lower than those of the FP panel). The Etest results with litraconazole for clinical isolates except C. tropicalis were similar to MICs of the FP panel (Etest for C. tropicalis showed 1 to 2 log2 dilution lower than FP panel). Also, the Etest results with fluconazole for clinical isolates except C. tropicalis were similar of 1 log2 dilution higher than MICs of the FP panel (Etest for C. tropicalis showed more than 2 log2 dilution lower than FP panel). These results showed a good level of overall agreement between the Etest method and the broth microdilution test (FP panel). Since the Etest is a less laborintensive and much simpler method, it appears to be a useful procedure for testing the susceptibility of yeasts to antifungal agents.

  17. Antifungal susceptibility testing of Aspergillus species complex in the Clinical Laboratory: how to do it, when to do it, and how to interpret it

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    Esther Manso

    2014-12-01

    Full Text Available The emergence of drug resistance in fungal pathogens has a profound impact on human health given limited number of antifungal drugs. Antifungal resistance in Aspergillus spp. infection can be encountered in the antifungal drug-exposed patient due to selection of intrinsically resistant species or isolates with acquired resistance belonging to species that are normally susceptible. Resistance to triazoles is not common in Aspergillus spp., however, triazole resistance in A. fumigatus appears to be increasing in several European countries in recent years and can be clinically relevant. The Clinical and Laboratory Standards Institute and European Committee on Antimicrobial Susceptibility Testing have developed breakpoints and epidemiological cutoff values that are now established for Aspergillus spp. Clinical microbiology laboratories will be employed commercial susceptibility assays, rather than reference broth microdilution methods and comparative studies are particularly important.

  18. Isolation and antifungal susceptibility testing of Trichosporon asahii in Ceará, Brazil

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    Everardo Albuquerque Menezes

    2012-02-01

    Full Text Available Trichosporon spp. are yeasts capable of causing invasive disease, which mainly affect immunocompromised patients. A clinical strain of T. asahii was isolated from the blood cultures of patients admitted to the General Hospital of Fortaleza. Susceptibility tests were conducted by disk diffusion and broth microdilution. The isolated strain of T. asahii was resistant to fluconazole. The patient used amphotericin B and caspofungin in order to facilitate the microbiological cure. It was the first isolation and identification of T. asahii in blood culture in Ceará, Brazil.

  19. Comparative analysis of the Vitek 2 antifungal susceptibility system and E-test with the CLSI M27-A3 broth microdilution method for susceptibility testing of Indian clinical isolates of Cryptococcus neoformans.

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    Tewari, Aarti; Behera, Bijayani; Mathur, Purva; Xess, Immaculata

    2012-06-01

    The emergence of antifungal resistance among Cryptococcus neoformans isolates is a matter of great concern. The Clinical and Laboratory Standards Institute (CLSI) broth microdilution reference method (BMD) for antifungal susceptibility testing of C. neoformans is tedious and time-consuming. Consequently, there is a greater need for a reproducible in vitro susceptibility testing method for use in clinical microbiology laboratories. By random amplified polymorphic DNA analysis, the 62 Indian clinical isolates were characterized as Cryptococcus neoformans var. grubii. We evaluated the susceptibilities of these isolates for amphotericin B (AMB) and fluconazole (FLC) by two commercial techniques, i.e., Vitek 2 and E-test against the CLSI M27-A3 BMD. The essential agreement (EA) between the Vitek 2 and E-test with the reference procedure for FLC was similar (82.2%). For AMB, EA of 92 and 76% was obtained with E-test and Vitek 2. Excellent categorical agreement (CA) (98.3% and 100% by Vitek 2 and E-test, respectively) was obtained for AMB. The CA for FLC was 81 and 77.4% by Vitek 2 and E-test. We conclude that both E-test and Vitek 2 system have acceptable levels of accuracy for susceptibility testing of both the drugs. Both of them could identify fluconazole-resistant strains. Vitek 2 could be used for testing susceptibility of voriconazole and 5-flucytosine also at the same time.

  20. Antifungal susceptibility testing of Candida in the Clinical Laboratory: how to do it, when to do it, and how to interpret it

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    Esther Manso

    2014-06-01

    Full Text Available Significant changes in the management of fungaemia have occurred in the last decade with increased use of fluconazole prophylaxis, of empirical treatment and of echinocandins as first-line agents for documented disease. The emergence of drug resistance in fungal pathogens has a profound impact on human health given limited number of antifungal drugs. Antifungal resistance in Candida may be either intrinsic or acquired and may be encountered in the antifungal drug exposed but also the antifungal drug naïve patient The variation in resistance rates between centers emphasizes that it is essential to have knowledge of the local Candida species distribution and antifungal resistance rates to guide initial therapy for Candida BSI. Moreover, all Candida isolates from blood and normally sterile sites should be identified to the species level. The Clinical and Laboratory Standards Institute and European Committee on Antimicrobial Susceptibility Testing have developed breakpoints and epidemiological cutoff values that are now established for Candida spp. Clinical microbiology laboratories will be employed commercial susceptibility assays, rather than reference broth microdilution methods and comparative studies are particularly important. Vitek 2®, Etest® and Sensititre YeastOne® provided a high degree of essential agreement and comparable sensitivity and specificity to BMD-RPMI for identifying resistance to azole and echinocandins in Candida spp.

  1. Comparison of EUCAST and CLSI broth microdilution methods for the susceptibility testing of 10 systemically active antifungal agents when tested against Candida spp.

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    Pfaller, Michael A; Castanheira, Mariana; Messer, Shawn A; Rhomberg, Paul R; Jones, Ronald N

    2014-06-01

    The antifungal broth microdilution (BMD) method of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) was compared with Clinical and Laboratory Standards Institute (CLSI) BMD method M27-A3 for amphotericin B, flucytosine, anidulafungin, caspofungin, micafungin, fluconazole, isavuconazole, itraconazole, posaconazole, and voriconazole susceptibility testing of 357 isolates of Candida. The isolates were selected from global surveillance collections to represent both wild-type (WT) and non-WT MIC results for the azoles (12% of fluconazole and voriconazole results were non-WT) and the echinocandins (6% of anidulafungin and micafungin results were non-WT). The study collection included 114 isolates of Candida albicans, 73 of C. glabrata, 76 of C. parapsilosis, 60 of C. tropicalis, and 34 of C. krusei. The overall essential agreement (EA) between EUCAST and CLSI results ranged from 78.9% (posaconazole) to 99.6% (flucytosine). The categorical agreement (CA) between methods and species of Candida was assessed using previously determined CLSI epidemiological cutoff values. The overall CA between methods was 95.0% with 2.5% very major (VM) and major (M) discrepancies. The CA was >93% for all antifungal agents with the exception of caspofungin (84.6%), where 10% of the results were categorized as non-WT by the EUCAST method and WT by the CLSI method. Problem areas with low EA or CA include testing of amphotericin B, anidulafungin, and isavuconazole against C. glabrata, itraconazole, and posaconazole against most species, and caspofungin against C. parapsilosis, C. tropicalis, and C. krusei. We confirm high level EA and CA (>90%) between the 2 methods for testing fluconazole, voriconazole, and micafungin against all 5 species. The results indicate that the EUCAST and CLSI methods produce comparable results for testing the systemically active antifungal agents against the 5 most common species of Candida; however, there are several areas where additional

  2. Genetic diversity of Aspergillus species isolated from onychomycosis and Aspergillus hongkongensis sp. nov., with implications to antifungal susceptibility testing.

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    Tsang, Chi-Ching; Hui, Teresa W S; Lee, Kim-Chung; Chen, Jonathan H K; Ngan, Antonio H Y; Tam, Emily W T; Chan, Jasper F W; Wu, Andrea L; Cheung, Mei; Tse, Brian P H; Wu, Alan K L; Lai, Christopher K C; Tsang, Dominic N C; Que, Tak-Lun; Lam, Ching-Wan; Yuen, Kwok-Yung; Lau, Susanna K P; Woo, Patrick C Y

    2016-02-01

    Thirteen Aspergillus isolates recovered from nails of 13 patients (fingernails, n=2; toenails, n=11) with onychomycosis were characterized. Twelve strains were identified by multilocus sequencing as Aspergillus spp. (Aspergillus sydowii [n=4], Aspergillus welwitschiae [n=3], Aspergillus terreus [n=2], Aspergillus flavus [n=1], Aspergillus tubingensis [n=1], and Aspergillus unguis [n=1]). Isolates of A. terreus, A. flavus, and A. unguis were also identifiable by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The 13th isolate (HKU49(T)) possessed unique morphological characteristics different from other Aspergillus spp. Molecular characterization also unambiguously showed that HKU49(T) was distinct from other Aspergillus spp. We propose the novel species Aspergillus hongkongensis to describe this previously unknown fungus. Antifungal susceptibility testing showed most Aspergillus isolates had low MICs against itraconazole and voriconazole, but all Aspergillus isolates had high MICs against fluconazole. A diverse spectrum of Aspergillus species is associated with onychomycosis. Itraconazole and voriconazole are probably better drug options for Aspergillus onychomycosis.

  3. Antifungal susceptibility profile of cryptic species of Aspergillus.

    Science.gov (United States)

    Alastruey-Izquierdo, Ana; Alcazar-Fuoli, Laura; Cuenca-Estrella, Manuel

    2014-12-01

    The use of molecular tools has led to the description of new cryptic species among different Aspergillus species complexes. Their frequency in the clinical setting has been reported to be between 10 and 15%. The susceptibility to azoles and amphotericin B of many of these species is low, and some of them, such as Aspergillus calidoustus or Aspergillus lentulus, are considered multi-resistant. The changing epidemiology, the frequency of cryptic species, and the different susceptibility profiles make antifungal susceptibility testing an important tool to identify the optimal antifungal agent to treat the infections caused by these species.

  4. Comparative Evaluation of the Vitek 2 Yeast Susceptibility Test and CLSI Broth Microdilution Reference Method for Testing Antifungal Susceptibility of Invasive Fungal Isolates in Italy: the GISIA3 Study▿

    Science.gov (United States)

    Borghi, Elisa; Iatta, Roberta; Sciota, Rita; Biassoni, Caterina; Cuna, Teresa; Montagna, Maria Teresa; Morace, Giulia

    2010-01-01

    The newly available AST-YS01 Vitek 2 cards were evaluated, and the results were compared with those obtained by the CLSI M27-A2 microdilution reference method. Clinical fungal isolates, including 614 isolates of Candida spp., 10 Cryptococcus neoformans isolates, 1 Geotrichum capitatum isolate, and 2 quality control strains, were tested for their susceptibilities to amphotericin B, fluconazole, and voriconazole using both methods. The majority of fungal isolates were susceptible to all antifungal agents tested: the MIC90 values determined by the Vitek 2 and CLSI methods were 0.5 and 1 μg/ml, respectively, for amphotericin B; 8 and 16 μg/ml, respectively, for fluconazole; and <0.12 and 0.25 μg/ml, respectively, for voriconazole. Overall there was excellent categorical agreement (CA) between the methods (99.5% for amphotericin B, 92% for fluconazole, 98.2% for voriconazole), but discrepancies were observed within species. The CAs for fluconazole were low for Candida glabrata and Candida krusei when the results of the CLSI method at 48 h were considered. Moreover, the fully automated commercial system did not detect the susceptibility of Cryptococcus neoformans to voriconazole. The Vitek 2 system can be considered a valid support for antifungal susceptibility testing of fungi, but testing of susceptibility to agents not included in the system (e.g., echinocandins and posaconazole) should be performed with other methods. PMID:20631105

  5. Antifungal susceptibility testing of yeast isolated from corneal infections Teste de susceptibilidade a antifúngicos de leveduras isoladas de infecções corneais

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    Vera Lucia Degaspare Monte Mascaro

    2003-10-01

    Full Text Available PURPOSE: To report the antifungal susceptibility profile of yeast isolates obtained from cases of keratitis. METHODS: Susceptibility testing of 15 yeast strains isolated from corneal infections to amphotericin B, fluconazole, itraconazole and ketoconazole was performed using the NCCLS broth microdilution assay. RESULTS: Most episodes of eye infections were caused by Candida albicans. The antifungal drugs tested showed the following minimal inhibitory concentration values against yeast isolates: 0.125-0.5 µg/ml for amphotericin B; 0.125->64.0 µg/ml for fluconazole; 0.015-1.0 µg/ml for itraconazole and 0.015-0.125 µg/ml for ketoconazole. Despite the fact that all Candida isolates were judged to be susceptible to azoles, one isolate showed a minimal inhibitory concentration value significantly higher than a 90% minimal inhibitory concentration of all tested isolates. Rhodotorula rubra was resistant to fluconazole and itraconazole. CONCLUSIONS: Despite the fact that most yeast isolates from corneal infections are usually susceptible to amphotericin B and azoles, they exhibit a wide range of minimal inhibitory concentration values for antifungal drugs. The identification of strains at species level and their susceptibility pattern to antifungal drugs should be considered before determining the concentration to be used in topical antifungal formulations in order to optimize therapeutic response in eye infections.OBJETIVO: Relatar resultados e avaliar a aplicabilidade do teste de suscetibilidade a antifúngicos de leveduras isoladas de infecções corneais oculares. MÉTODOS: Realizou-se teste de suscetibilidade pelo método de microdiluição em caldo, padronizado pelo NCCLS-EUA, em 15 amostras de leveduras de infecções corneanas a anfotericina B, fluconazol, itraconazol e ketoconazol. RESULTADOS: A maioria dos episódios de infecção corneal foi causada por Candida albicans. As drogas antifúngicas testadas exibiram valores de concentra

  6. Taxonomy and antifungal susceptibility of clinically important Rasamsonia species

    DEFF Research Database (Denmark)

    Houbraken, J.; Giraud, S.; Meijer, M.

    2013-01-01

    In recent years, Geosmithia argillacea has been increasingly reported in humans and animals and can be considered an emerging pathogen. The taxonomy of Geosmithia was recently studied, and Geosmithia argillacea and related species were transferred to the new genus Rasamsonia. The diversity among...... a set of Rasamsonia argillacea strains, including 28 clinical strains, was studied, and antifungal susceptibility profiles were generated. Data obtained from morphological studies and from phylogenetic analyses of internal transcribed spacer (ITS) and partial _-tubulin and calmodulin sequences revealed...... was the least active of the antifungals tested. The phenotypically similar species R. brevistipitata and R. cylindrospora had different antifungal susceptibility profiles, and this indicates that correct species identification is important to help guide appropriate antifungal therapy....

  7. Antifungal susceptibilities of Candida species isolated from urine culture.

    Science.gov (United States)

    Toka Özer, Türkan; Durmaz, Süleyman; Yula, Erkan

    2016-09-01

    Candida spp. are the most common opportunistic mycosis worldwide. Although Candida albicans is the most common cause of urinary tract infections, the frequency of non-albicans Candida species is increasing with common use of antifungal in the prophylaxis and treatment. This may lead to difficulties in treatment. Antifungal tests should be applied with identification of species for effective treatment. In this study, identification of Candida species isolated from urine culture and investigation of susceptibility of these strains to amphotericin B, flucytosine, fluconazole, voriconazole was aimed. In this study, 58 Candida strains isolated from urine cultures at Osmaniye State Hospital between January 2012 and April 2013 were included. Urine culture and antifungal susceptibility tests were applied. Incidence rate of Candida spp. was determined as C. albicans (56.9%), Candida glabrata (20.6%), Candida tropicalis (10.3%), Candida parapsilosis (7%), Candida krusei (3.4%), Candida kefyr (1.8%). Most of the isolates were susceptible to amphotericin B, flucytosine, fluconazole, voriconazole. Twenty three (39.7%) Candida strains were isolated from internal medical branches and Intensive Care Unit and 12 (20.6%) from the Surgical Medical Branches. C. albicans and C. glabrata species were isolated most frequently as a candiduria factor in this hospital between January 2012 and April 2013. The analysis of antifungal susceptibility profile shows no significant resistance to antifungals.

  8. Multicenter Comparison of the Vitek 2 Antifungal Susceptibility Test with the CLSI Broth Microdilution Reference Method for Testing Caspofungin, Micafungin, and Posaconazole against Candida spp.▿

    Science.gov (United States)

    Peterson, Jess F.; Pfaller, Michael A.; Diekema, Daniel J.; Rinaldi, Michael G.; Riebe, Katherine M.; Ledeboer, Nathan A.

    2011-01-01

    The performance of the automated Vitek 2 (bioMérieux, Inc., Marcy l'Etoile, France) antifungal susceptibility system was compared to that of broth microdilution (BMD) for the determination of MICs of various antifungal drugs. A total of 112 challenge strains and 755 clinical isolates of Candida spp. were tested against caspofungin and micafungin. An additional 452 clinical isolates of Candida albicans were tested against posaconazole. Reference BMD MIC endpoints were established after 24 h of incubation for caspofungin and micafungin and after 48 h of incubation for posaconazole. Essential agreements (EAs) between the Vitek 2 and BMD methods for caspofungin and micafungin were 99.5% and 98.6%, respectively. EA between the Vitek 2 and BMD methods was 95.6% for posaconazole. The overall categorical agreements (CAs) between the Vitek 2 system and BMD were 99.8% for caspofungin, 98.2% for micafungin, and 98.1% for posaconazole. The Vitek 2 system reliably determined caspofungin and micafungin MICs among Candida spp. and posaconazole MICs among C. albicans isolates and demonstrated excellent quantitative and qualitative agreement with the reference BMD method. PMID:21430096

  9. Multicenter Comparison of the VITEK 2 Antifungal Susceptibility Test with the CLSI Broth Microdilution Reference Method for Testing Amphotericin B, Flucytosine, and Voriconazole against Candida spp.▿

    Science.gov (United States)

    Pfaller, M. A.; Diekema, D. J.; Procop, G. W.; Rinaldi, M. G.

    2007-01-01

    A fully automated commercial antifungal susceptibility test system (VITEK 2; bioMérieux, Inc., Hazelwood, MO) was compared in three different laboratories with the Clinical and Laboratory Standards Institute (formerly the NCCLS) reference broth microdilution method (BMD) by testing 2 quality control strains, 10 reproducibility strains, and 426 isolates of Candida spp. against amphotericin B, flucytosine, and voriconazole. Reference BMD MIC endpoints were established after 24 and 48 h of incubation. VITEK 2 system MIC endpoints were determined spectrophotometrically after 9.1 to 27.1 h of incubation (mean, 12 to 14 h). Excellent essential agreement (within 2 dilutions) between the VITEK 2 system and the 24- and 48-h BMD MICs was observed for all three antifungal agents: amphotericin B, 99.1% and 97%, respectively; flucytosine, 99.1% and 98.8%, respectively; and voriconazole, 96.7% and 96%, respectively. Both intra- and interlaboratory agreements were >98% for all three drugs. The overall categorical agreements between the VITEK 2 system and BMD for flucytosine and voriconazole were 98.1 to 98.6% at the 24-h BMD time point and 96.9 to 97.4% at the 48-h BMD time point. The VITEK 2 system reliably detected flucytosine and voriconazole resistance among Candida spp. and demonstrated excellent quantitative and qualitative agreement with the reference BMD method. PMID:17913927

  10. ANTIFUNGAL SUSCEPTIBILITY TESTING AND GENOTYPING CHARACTERIZATION OF Cryptococcus neoformans AND gattii ISOLATES FROM HIV-INFECTED PATIENTS OF RIBEIRÃO PRETO, SÃO PAULO, BRAZIL

    Science.gov (United States)

    FIGUEIREDO, Thais Pandini; de LUCAS, Rosymar Coutinho; CAZZANIGA, Rodrigo Anselmo; FRANÇA, Carolina Nunes; SEGATO, Fernando; TAGLIALEGNA, Rafael; MAFFEI, Claudia Maria Leite

    2016-01-01

    SUMMARY Cryptococcosis is a leading invasive fungal infection in immunocompromised patients. Considering the high prevalence and severity of these infections in immunocompromised patients attended at HC-FMRP-USP, the present research aimed to characterize the clinical isolates of Cryptococcus strains by biochemical and molecular methods and evaluate antifungal susceptibility of clinical isolates. Fifty isolates from 32 HIV-positive patients were obtained at HC-FMRP-USP. Most of the isolates (78.1%) were identified as C. neoformans, and 100% of C. neoformans and C. gattii strains were susceptible to amphotericin B, ketoconazole and fluconazole. All isolates were classified as serotype A (grubbii variety) by PCR and most of them were characterized in mating type MATa. PCR analysis of specific M13 microsatellite sequence revealed that VNI type was predominant among C. neoformans, while VGII was predominant among C. gattii. The strains did not show a significant resistance to the antifungals tested, and Canavanine-Glycine-Bromthymol Blue Agar (CGB) proved to be a reliable test presenting a good correlation with the molecular characterization. C. neoformans isolated from disseminated infections in the same patient showed molecular identity when different anatomical sites were compared; besides, the studied strains did not present a significant increase in resistance to antifungal agents. In addition, the homogeneity of the molecular types and detection of the mating types suggested a low possibility of crossing among the strains. PMID:27680174

  11. [Yeasts in domestic animals: species identification and susceptibility to antifungals].

    Science.gov (United States)

    Hamal, Petr; Koukalová, Dagmar

    2010-02-01

    Yeasts frequently colonize various kinds of domestic animals, but may also cause serious diseases. The aim of this study was to identify yeast isolates collected from dogs, cows and pigs, and to determine their in vitro antifungal susceptibility. Fifty-six yeast isolates from dogs (n = 24), cows (n = 20), and pigs (n = 12) were investigated. Appearance of colonies grown on Sabouraud agar, micromorphology on rice agar, as well as assimilation and fermentation of various carbon and nitrogen sources were evaluated. Susceptibility to six antifungals (flucytosine, amphotericin B, miconazole, ketoconazole, itraconazole and fluconazole) was determined semiquantitatively using the commercially available Fungitest kit (Bio-Rad Laboratories). Ten yeast species were identified in dogs with relatively even distribution. On the other hand, cow and pig were clearly dominated by Candida krusei (from 7 species) and Candida rugosa (from 5 species), respectively. Further, most of yeast isolates exhibited good susceptibility to the antifungals tested particularly to amphotericin B, ketoconazole and itraconazole. Based on results, it can be concluded that significant differences in the species spectrum and distribution were documented between groups of yeasts from dogs, cows and pigs. This is probably due to different environmental conditions and the endogenous origin of the yeast isolates. Mostly good susceptibility to systemic antifungals should positively influence the therapy of diseases caused by yeasts in veterinary medicine.

  12. Comparison of a glucose consumption based method with the CLSI M38-A method for testing antifungal susceptibility of Trichophyton rubrum and Trichophyton mentagrophytes

    Institute of Scientific and Technical Information of China (English)

    ZHANG Jing; CHEN Jian; HUANG Huai-qiu; XI Li-yan; LAI Wei; XUE Ru-zeng; ZHANG Xiao-hui; CHEN Rong-zhang

    2010-01-01

    Background The prevalence of dermatophytoses and the development of new antifungal agents has focused interest on susceptibility tests of dermatophytes. The method used universally for susceptibility tests of dermatophytes was published as document (M38-A) in 2002 by the Clinical and Laboratory Standards Institute (CLSI), dealing with the standardization of susceptibility tests in filamentous fungi, though not including dermatophytes especially. However, it is not a very practical method for the clinical laboratory in routine susceptibility testing. In this test, we developed a novel rapid susceptibility assay --glucose consumption method (GCM) for dermatophytes.Methods In this study, we investigated the antifungal susceptibilities of dermatophytes to itraconazole (ITC),voriconazole (VOC), econazole nitrate (ECN) and terbinafine (TBF) by glucose consumption method (GCM), in comparison to the Clinical and Laboratory Standards Institute (CLSI) M38-A method. Twenty-eight dermatophyte isolates,including Trichophyton rubrum ( T. Rubrum) (n=14) and Trichophyton mentagrophytes ( T. Mentagrophytes) (n=14), were tested. In the GCM, the minimum inhibitory concentrations (MICs) were determined spectrophotometrically at 490 nm after addition of enzyme substrate color mix. For the CLSI method, the MICs were determined visually.Results Comparison revealed best agreement for TBF against T. Mentagrophytes and T. Rubrum, since MIC range, MIC50, and MIC90 were identical from two methods. However, for ITC and VOC, GCM showed wider MIC ranges and higher MICs than CLSI methods in most isolates. For ECN against T. Rubrum, high MICs were tested by GCM (0.125-16 μg/ml) but not M38-A method (0.5-1 μg/ml). The overall agreements for all isolates between the two methods within one dilution and two dilutions for ITC, VOC, ECN and TBF was 53.6% and 75.0%, 57.1% and 75.0%, 82.1% and 89.3%, and 85.7 and 85.7%, respectively.Conclusion Measurement of glucose uptake can predict the susceptibility

  13. Sporothrix schenckii complex in Iran: Molecular identification and antifungal susceptibility.

    Science.gov (United States)

    Mahmoudi, Shahram; Zaini, Farideh; Kordbacheh, Parivash; Safara, Mahin; Heidari, Mansour

    2016-08-01

    Sporotrichosis is a global subcutaneous fungal infection caused by the Sporothrix schenckii complex. Sporotrichosis is an uncommon infection in Iran, and there have been no phenotypic, molecular typing or antifungal susceptibility studies of Sporothrix species. This study aimed to identify nine Iranian isolates of the S. schenckii complex to the species level using colony morphology, carbohydrate assimilation tests, and PCR-sequencing of the calmodulin gene. The antifungal susceptibilities of these Sporothrix isolates to five antifungal agents (amphotericin B (AMB), voriconazole (VRC), itraconazole (ITC), fluconazole (FLC), and terbinafine (TRB)) were also evaluated according to the M27-A3 and M38-A2 protocols of the Clinical and Laboratory Standards Institute for yeast and mycelial phases, respectively. Five of seven clinical isolates were identified as S. schenckii, and two clinical and two environmental isolates were identified as S. globosa. This is the first report of S. globosa in Iran. There was significant agreement (73%) between the results of the phenotypic and genotypic identification methods. TRB and ITC were the most effective antifungals against the Sporothrix isolates. The minimum inhibitory concentration (MIC) values of TRB for the yeast and mycelial phases of S. schenckii differed significantly. There was also a significant difference in the minimum fungicidal concentration (MFC) values of AMB and TRB for the two phases. Considering the low efficacy of VRC and FLC and the wide MIC ranges of AMB (1-16 μg/ml and 1-8 μg/ml for yeast and mycelial forms, respectively) observed in the present study, in vitro antifungal susceptibility testing should be performed to determine appropriate therapeutic regimens.

  14. [A study for testing the antifungal susceptibility of yeast by the Japanese Society for Medical Mycology (JSMM) method. The proposal of the modified JSMM method 2009].

    Science.gov (United States)

    Nishiyama, Yayoi; Abe, Michiko; Ikeda, Reiko; Uno, Jun; Oguri, Toyoko; Shibuya, Kazutoshi; Maesaki, Shigefumi; Mohri, Shinobu; Yamada, Tsuyoshi; Ishibashi, Hiroko; Hasumi, Yayoi; Abe, Shigeru

    2010-01-01

    The Japanese Society for Medical Mycology (JSMM) method used for testing the antifungal susceptibility of yeast, the MIC end point for azole antifungal agents, is currently set at IC(80). It was recently shown, however that there is an inconsistency in the MIC value between the JSMM method and the CLSI M27-A2 (CLSI) method, in which the end- point was to read as IC(50). To resolve this discrepancy and reassess the JSMM method, the MIC for three azoles, fluconazole, itraconazole and voriconazole were compared to 5 strains of each of the following Candida species: C. albicans, C. glabrata, C. tropicalis, C. parapsilosis and C. krusei, for a total of 25 comparisons, using the JSMM method, a modified JSMM method, and the CLSI method. The results showed that when the MIC end- point criterion of the JSMM method was changed from IC(80) to IC(50) (the modified JSMM method) , the MIC value was consistent and compatible with the CLSI method. Finally, it should be emphasized that the JSMM method, using a spectrophotometer for MIC measurement, was superior in both stability and reproducibility, as compared to the CLSI method in which growth was assessed by visual observation.

  15. SUSCEPTIBILITY OF CANDIDA SPECIES TO ANTIFUNGAL DRUGS IN WESTERN INDIA

    Directory of Open Access Journals (Sweden)

    Geeta M Vaghela

    2015-06-01

    Full Text Available Introduction: The increase in candidaemia is associated with high mortality. A shift has been observed in the relative frequency of each Candida spp. isolated from blood. Options of the antifungal drugs available for treatment of systemic and invasive candidiasis are restricted to polyenes, allylamines, azoles and recently developed echinocandin class of molecules. A rise in the incidence of antifungal resistance to Candida spp. has also been reported over the past decade. Studies on prevalence of infections and antifungal susceptibility testing are useful in deciding clinical strategies. Aims: To do species level identification and detect resistance, if any, among Indian clinical isolates of C. albicans. Methodology: From total 135 patients from a tertiary care hospital of Gujarat, Candida species were isolated from different clinical specimens. The growth of Candida on Sabouraud's dextrose agar was confirmed by Gram staining in which gram positive budding fungal cells were observed. Then its growth was examined for colony morphology on Sabouraud's dextrose agar and chlamydospore production on Corn meal tween 80 agar. Germ tube tests and other biochemical tests like sugar fermentation, sugar assimilation and urease test were performed to identify the species of Candida. Antifungal susceptibility testing was performed by NCCLS M44-A Disc diffusion method. Results: Out of total 135 samples, C. Albicans were isolated from 52 (38.5%. Among Non Albican Candid (NAC, Candida glabrata was 36 (26.7% followed by Candida tropicalis 25(18.5%. C. albicans was found resistant to Fluconazole, Itraconazole and Amphotericine B in 3.8%, 3.8% and 1.9% cases respectively. For NAC, resistance of Fluconazole, Itraconazole and Amphotericine B was found in 4.8%, 3.6% and 2.4% cases respectively. [Natl J Med Res 2015; 5(2.000: 122-126

  16. Evaluation of Etest and macrodilution broth method for antifungal susceptibility testing of Candida sp strains isolated from oral cavities of AIDS patients

    Directory of Open Access Journals (Sweden)

    SILVA Maria do Rosário R.

    2002-01-01

    Full Text Available A comparison of the Etest and the reference broth macrodilution susceptibility test for fluconazole, ketoconazole, itraconazole and amphotericin B was performed with 59 of Candida species isolated from the oral cavities of AIDS patients. The Etest method was performed according to the manufacturer's instructions, and the reference method was performed according to National Committee for Clinical Laboratory Standards document M27-A guidelines. Our data showed that there was a good correlation between the MICs obtained by the Etest and broth dilution methods. When only the MIC results at ± 2 dilutions for both methods were considered, the agreement rates were 90.4% for itraconazole, ketoconazole and amphotericin B and 84.6% for fluconazole of the C. albicans tested. In contrast, to the reference method, the Etest method classified as susceptible three fluconazole-resistant isolates and one itraconazole-resistant isolate, representing four very major errors. These results indicate that Etest could be considered useful for antifungal sensitivity evaluation of yeasts in clinical laboratories.

  17. Human Immunodefeciency Virus Associated Cryptococcal Meningitis at a Tertiary Care Centre: Diagnostic Tools and Antifungal Susceptibility Testing

    Science.gov (United States)

    Munivenkataswamy, Rashmi; Gopi, Anjana; Usman, Shaik Mohammed; Jagadeesh

    2013-01-01

    Context: Cryptococcal meningitis has emerged as a leading cause of the infectious morbidity and mortality in HIV sero-reactive subjects and it is the second most common cause of the opportunistic neuroinfections in it. As this is a indistinguishable from other causes of meningitis, its early diagnosis is the key to the therapeutic success. Objectives: This study was undertaken to know the incidence of Cryptococcal meningitis in HIV sero–reactive individuals and to assess the role of the microbiological parameters in its specific diagnosis, with a perspective of evaluating the anti–fungal resistance. Material and Methods: A total of 66 CSF samples from suspected cases of meningoencephalitis were subjected to standard microbiological procedures. The Cryptococcal isolates were identified by microscopy, the cultural characteristics, melanin production on Niger Seed agar, urea hydrolysis, the Nitrate assimilation test and by capsular antigen detection by latex agglutination. The Cryptoccal isolates were further biotyped by using Canavanine–Glycine–Bromothymol blue agar. The Minimum Inhibitory Concentrations (MIC) of Amphotericin B and Fluconazole for the isolates were detected. Results: The incidence of Cryptococcal meningitis in our study group was 18.2% (12/66). The Cryptococcal antigen was detected in all the 12 cases, whereas microscopy was positive only in 9 cases and Cryptococcus was isolated by culture in 10 cases. All the isolates were sensitive to Amphotericin B and 90% of the isolates were sensitive to Fluconazole. The CD4counts ranged between 22-138 cells /μl. Conclusion: A high incidence of Cryptococcal meningitis in HIV sero-reactive subjects necessitates the importance of a precise and an early microbiological diagnosis for better management of such subjects. Due to the growing concern of emerging drug resistance, the testing for the anti–fungal susceptibility has to be encouraged in all the cases. PMID:24086857

  18. Comparison of the Vitek 2 Antifungal Susceptibility System with the Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) Broth Microdilution Reference Methods and with the Sensititre YeastOne and Etest Techniques for In Vitro Detection of Antifungal Resistance in Yeast Isolates ▿ ‖

    Science.gov (United States)

    Cuenca-Estrella, Manuel; Gomez-Lopez, Alicia; Alastruey-Izquierdo, Ana; Bernal-Martinez, Leticia; Cuesta, Isabel; Buitrago, Maria J.; Rodriguez-Tudela, Juan L.

    2010-01-01

    The commercial technique Vitek 2 system for antifungal susceptibility testing of yeast species was evaluated. A collection of 154 clinical yeast isolates, including amphotericin B- and azole-resistant organisms, was tested. Results were compared with those obtained by the reference procedures of both the CLSI and the European Committee on Antimicrobial Susceptibility Testing (EUCAST). Two other commercial techniques approved for clinical use, the Etest and the Sensititre YeastOne, were included in the comparative exercise as well. The average essential agreement (EA) between the Vitek 2 system and the reference procedures was >95%, comparable with the average EAs observed between the reference procedures and the Sensititre YeastOne and Etest. The EA values were >97% for Candida spp. and stood at 92% for Cryptococcus neoformans. Intraclass correlation coefficients (ICC) between the commercial techniques and the reference procedures were statistically significant (P < 0.01). Percentages of very major errors were 2.6% between Vitek 2 and the EUCAST technique and 1.6% between Vitek 2 and the CLSI technique. The Vitek 2 MIC results were available after 14 to 18 h of incubation for all Candida spp. (average time to reading, 15.5 h). The Vitek 2 system was shown to be a reliable technique to determine antifungal susceptibility testing of yeast species and a more rapid and easier alternative for clinical laboratories than the procedures developed by either the CLSI or EUCAST. PMID:20220169

  19. Antifungal susceptibility profiles of 1698 yeast reference strains revealing potential emerging human pathogens.

    Directory of Open Access Journals (Sweden)

    Marie Desnos-Ollivier

    Full Text Available New molecular identification techniques and the increased number of patients with various immune defects or underlying conditions lead to the emergence and/or the description of novel species of human and animal fungal opportunistic pathogens. Antifungal susceptibility provides important information for ecological, epidemiological and therapeutic issues. The aim of this study was to assess the potential risk of the various species based on their antifungal drug resistance, keeping in mind the methodological limitations. Antifungal susceptibility profiles to the five classes of antifungal drugs (polyens, azoles, echinocandins, allylamines and antimetabolites were determined for 1698 yeast reference strains belonging to 992 species (634 Ascomycetes and 358 Basidiomycetes. Interestingly, geometric mean minimum inhibitory concentrations (MICs of all antifungal drugs tested were significantly higher for Basidiomycetes compared to Ascomycetes (p<0.001. Twenty four strains belonging to 23 species of which 19 were Basidiomycetes seem to be intrinsically "resistant" to all drugs. Comparison of the antifungal susceptibility profiles of the 4240 clinical isolates and the 315 reference strains belonging to 53 shared species showed similar results. Even in the absence of demonstrated in vitro/in vivo correlation, knowing the in vitro susceptibility to systemic antifungal agents and the putative intrinsic resistance of yeast species present in the environment is important because they could become opportunistic pathogens.

  20. 卢立康唑醇质体体外抗真菌活性研究%The antifungal susceptibility test in vitro of luliconazole ethosomes

    Institute of Scientific and Technical Information of China (English)

    樊建峰; 林碧雯; 李恒进

    2016-01-01

    Objective To study antifungal susceptibility of luliconazole ethosomes in vitro .Methods In vitro ,antifungal activity of luliconazole ethosome,liposome,hydroalcoholic solution:MICs were determined using the micro-broth dilution as-say.Results The MIC of the luliconazole ethosome was lower than that of liposome form or hydroalcoholic solution form.The MIC of luliconazole ethosome to Trichophyton spp.was lower than the sensitive degree to Candida spp.Conclusions In vitro ,the antifungal effect of luliconazole ethosome is better than luliconazole liposome or its hydroalcoholic solution.%目的:检测卢立康唑醇质体体外抗菌活性。方法采用微量液基稀释法检测卢立康唑醇质体、脂质体、45%水醇溶液对皮肤癣菌、念珠菌的体外抗菌活性。结果卢立康唑醇质体对皮肤癣菌及念珠菌的 MIC 值低于脂质体及水醇溶液,且对毛癣菌属较念珠菌属的 MIC 值更低。结论卢立康唑醇质体的体外抗菌性能优于脂质体及水醇溶液。

  1. Antimycotics susceptibility testing of dermatophytes

    Directory of Open Access Journals (Sweden)

    Arsić-Arsenijević Valentina

    2010-01-01

    Full Text Available Dermatophytes are moulds that produce infections of the skin, hair and nails of humans and animals. The most common forms among these infections are onychomycosis and tinea pedis affecting 20% of world population. These infections are usually chronic. The treatment of dermatophytoses tends to be prolonged partly because available treatments are not very effective. Antifungal drug consumption and public health expenditure are high worldwide, as well as in Serbia. For adequate therapy, it is necessary to prove infection by isolation of dermatophytes and to test the antifungal susceptibility of isolates. Susceptibility testing is important for the resistance monitoring, epidemiological research and to compare in vitro activities of new antifungal agents. The diffusion and dilution methods of susceptibility tests are used, and technical issues of importance for the proper performance and interpretation of test results are published in the document E.DEF 9.1 (EUCAST and M38-A2 (CLSI. The aim of our paper is to promptly inform the public about technical achievements in this area, as well as the new organization of laboratory for medical mycology in our country. The formation of laboratory networks coordinated by the National Reference Laboratory for the cause of mycosis need to enable interlaboratory studies and further standardization of methods for antifungal susceptibility testing of dermatophytes, reproducibility of tests and clinical correlation monitoring (MIK values and clinical outcome of dermatophytosis. The importance of the new organization is expected efficient improvement in the dermatophytosis therapy at home, better quality of patient's life and the reduction of the cost of treatment.

  2. Antifungal susceptibility profile of Trichosporon isolates: correlation between CLSI and etest methodologies

    OpenAIRE

    2010-01-01

    The aim of the present study was to evaluate the antifungal susceptibility profile of Trichosporon species isolated from different sources employing the Clinical and Laboratory Standards Institute (CLSI) method and E-test method. Thirty-four isolates of Trichosporon spp. and six CBS reference samples were tested for their susceptibility to Amphotericin B, 5-flucytosine, Fluconazole, Itraconazole, Voriconazole and Terbinafine. All species showed high Minimun Inhibitory Concentrations (MIC) for...

  3. Epidemiology and antifungal susceptibilities of yeasts causing vulvovaginitis in a teaching hospital.

    Science.gov (United States)

    Gamarra, Soledad; Morano, Susana; Dudiuk, Catiana; Mancilla, Estefanía; Nardin, María Elena; de Los Angeles Méndez, Emilce; Garcia-Effron, Guillermo

    2014-10-01

    Vulvovaginal candidiasis is one of the most common mycosis. However, the information about antifungal susceptibilities of the yeasts causing this infection is scant. We studied 121 yeasts isolated from 118 patients with vulvovaginal candidiasis. The isolates were identified by phenotypic and molecular methods, including four phenotypic methods described to differentiate Candida albicans from C. dubliniensis. Antifungal susceptibility testing was performed according to CLSI documents M27A3 and M27S4 using the drugs available as treatment option in the hospital. Diabetes, any antibacterial and amoxicillin treatment were statistically linked with vulvovaginal candidiasis, while oral contraceptives were not considered a risk factor. Previous azole-based over-the-counter antifungal treatment was statistically associated with non-C.albicans yeasts infections. The most common isolated yeast species was C. albicans (85.2 %) followed by C. glabrata (5 %), Saccharomyces cerevisiae (3.3 %), and C. dubliniensis (2.5 %). Fluconazole- and itraconazole-reduced susceptibility was observed in ten and in only one C. albicans strains, respectively. All the C. glabrata isolates showed low fluconazole MICs. Clotrimazole showed excellent potency against all but seven isolates (three C. glabrata, two S. cerevisiae, one C. albicans and one Picchia anomala). Any of the strains showed nystatin reduced susceptibility. On the other hand, terbinafine was the less potent drug. Antifungal resistance is still a rare phenomenon supporting the use of azole antifungals as empirical treatment of vulvovaginal candidiasis.

  4. Antifungal Susceptibility of Candida Biofilms: Unique Efficacy of Amphotericin B Lipid Formulations and Echinocandins

    OpenAIRE

    Kuhn, D M; T. George; CHANDRA, J; P. K. Mukherjee; Ghannoum, M A

    2002-01-01

    Biofilms, likely the predominant mode of device-related microbial infection, exhibit resistance to antimicrobial agents. Evidence suggests that Candida biofilms have dramatically reduced susceptibility to antifungal drugs. We examined antifungal susceptibilities of Candida albicans and Candida parapsilosis biofilms grown on a bioprosthetic model. In addition to conventional agents, we determined if new antifungal agents (triazoles, amphotericin B lipid formulations, and echinocandins) have ac...

  5. IDENTIFICATION OF CANDIDA SPECIES FROM CLINICAL SAMPLES AND THEIR ANTIFUNGAL SUSCEPTIBILITY PATTERNS

    Directory of Open Access Journals (Sweden)

    Bhaskar

    2015-09-01

    Full Text Available OBJECTIVE AND BACKGROUND : The incidence of Candida infections has increased dramatically over the past few decades due to increase in the number of population susceptible to fungal infections. With multiple antifungal ag ents that are available and recovery of clinical isolates that exhibit inherent or developed resistance to commonly used antifungal agents, it has become imperative to do susceptibility testing routinely. The study was done to determine the predisposing fa ctors, species incidence and susceptibility pattern of Candida isolates to commonly use d antifungal agents. METHODS: A total of 108 Candida species were recovered from symptomatic clinical cases. Candida isolates were speciated by germ tube test, chlamydospore formation on corn meal agar and color produced on chromogenic media. Antifungal susceptibility test was done by disk diffusion method for nystatin, fluconazole, itraconazole, voriconazole and amphotericin - B. RESULTS: Candida albicans is the m ost frequently isolated species. However, non - albicans Candida species, taken as a group has predominated in clinical samples. Chromogenic agar medium showed good correlation in species identification in comparison with conventional germ tube test and chla mydospore formation on corn meal agar. C. albicans (41, C. tropicalis (33, C. krusei (30 and C. glabrata (04 were isolated. Candida species showed 95.4% susceptibility to amphotericin - B, 77.8% to voriconazol e, 69.4% to nystatin, 64.1% to f luconazole an d 63.9% to itraconazole. CONCLUSION : Increasing incidence of non - albicans species infection. Chromogenic medium can be used for species identification. Increasing resistance of Candida species to commonly used antifungal agents.

  6. Antifungal susceptibility and virulence factors of clinically isolated dermatophytes in Tehran, Iran

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    Mohammad Ali Afshari

    2016-03-01

    Full Text Available Background and Objectives: Dermatophytes possess a wide array of virulence factors and various antifungal susceptibility patterns which influence their pathogenesis in humans and animals. The aim of this study was to evaluate antifungal suscep- tibility and keratinase and proteinase activity of 49 dermatophyte strains from the genera Microsporum, Trichophyton and Epidermophyton which were isolated from human cases of dermatophytosis.Materials and Methods: Forty-nine dermatophyte strains isolated from clinical samples were cultured on general and spe- cific culture media. Keratinase and proteinase activity was screened on solid mineral media and confirmed in liquid cultures. Drug susceptibility toward azoles (fluconazole, ketoconazole and itraconazole, griseofulvin and terbinafine was evaluated using disk diffusion method on Mueller-Hinton agar and minimum inhibitory concentrations (MICs were determined using microbroth dilution assay according to the Clinical and Laboratory Standards Institute (CLSI guidelines.Results: Our results indicated that clinically isolated dermatophytes from 7 major species produced keratinase and protein- ase at different extents. The mean keratinase and proteinase activity was reported as 6.69 ± 0.31 (U/ml and 2.10 ± 0.22 (U/ ml respectively. Disk diffusion and microbroth dilution (MIC results of antifungal susceptibility testing showed that ke- toconazole was the most effective drug against Epidermophyton floccosum and Trichophyton mentagrophytes, itraconazole against T. rubrum and E. floccosum, and griseofulvin and terbinafine against Trichophyton verrucosum. Our results showed that all dermatophyte isolates were resistant to fluconazole. Overall, ketoconazole and itraconazole were the most effective drugs for all dermatophyte species tested.Conclusion: Our results showed that antifungal susceptibility testing is an urgent need to select drugs of choice for treatment of different types of dermatophytosis and

  7. Species distribution & antifungal susceptibility pattern of oropharyngeal Candida isolates from human immunodeficiency virus infected individuals

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    Partha Pratim Das

    2016-01-01

    Results: From the 59 culture positive HIV seropositive cases, 61 Candida isolates were recovered; Candidaalbicans (n=47, 77.0%, C. dubliniensis (n=9, 14.7%, C. parapsilosis (n=2, 3.2%, C. glabrata (n=2, 3.2%, and C. famata (n=1, 1.6%. Candida colonization in HIV-seropositive individuals was significantly higher than that of HIV-seronegative (control group. Antifungal susceptibility testing revealed (n=6, 9.3% C. albicans isolates resistant to voriconazole and fluconazole by disk-diffusion method whereas no resistance was seen by Fungitest method. Interpretation & conclusions: C. albicans was the commonest Candida species infecting or colonizing HIV seropositive individuals. Oropharyngeal Candida isolates had high level susceptibility to all the major antifungals commonly in use. Increased level of immunosuppression in HIV-seropositives and drug resistance of non-albicans Candida species makes identification and susceptibility testing of Candida species necessary in different geographical areas of the country.

  8. Distribution and antifungal susceptibility of Candida species causing nosocomial candiduria.

    Science.gov (United States)

    Ozhak-Baysan, Betil; Ogunc, Dilara; Colak, Dilek; Ongut, Gozde; Donmez, Levent; Vural, Tumer; Gunseren, Filiz

    2012-07-01

    The aim of the study was to investigate the distribution of Candida species isolated from urine specimens of hospitalized patients in Akdeniz University Hospital, Antalya, Turkey, as well as their susceptibilities to antifungal agents. A total of 100 patients who had nosocomial candiduria between March 2003 and May 2004 at the facility were included in the study. Organisms were identified by conventional methods and the use of API ID 32C strips. Susceptibilities of the isolates to amphotericin B were determined by Etest, whereas the minimum inhibitory concentration (MIC) values of these same strains to fluconazole, voriconazole and caspofungin were assessed using the broth microdilution method. The most common species recovered was C. albicans 44% of all yeasts, followed by C. tropicalis (20%), C. glabrata (18%), C. krusei (6%), C. famata (5%), C. parapsilosis (4%), C. kefyr (2%) and C. guilliermondii (1%). A total of nine (9%) of the isolates, including five C. krusei and four C. glabrata isolates were susceptible dose-dependent (SDD) to fluconazole. In constrast, only two C. glabrata and one C. krusei isolates were resistant to this antifungal. The voriconazole MICs for all Candida isolates were ≤0.5 μg/ml, except for one C. glabrata isolate with a MIC value of 2 μg/ml. Among all isolates, 94% were susceptible to amphotericin B with MIC values of Candida urinary tract infections.

  9. Genotyping of Fusarium Isolates from Onychomycoses in Colombia: Detection of Two New Species Within the Fusarium solani Species Complex and In Vitro Antifungal Susceptibility Testing.

    Science.gov (United States)

    Guevara-Suarez, Marcela; Cano-Lira, José Francisco; de García, María Caridad Cepero; Sopo, Leticia; De Bedout, Catalina; Cano, Luz Elena; García, Ana María; Motta, Adriana; Amézquita, Adolfo; Cárdenas, Martha; Espinel-Ingroff, Ana; Guarro, Josep; Restrepo, Silvia; Celis, Adriana

    2016-04-01

    Fusariosis have been increasing in Colombia in recent years, but its epidemiology is poorly known. We have morphologically and molecularly characterized 89 isolates of Fusarium obtained between 2010 and 2012 in the cities of Bogotá and Medellín. Using a multi-locus sequence analysis of rDNA internal transcribed spacer, a fragment of the translation elongation factor 1-alpha (Tef-1α) and of the RNA-dependent polymerase subunit II (Rpb2) genes, we identified the phylogenetic species and circulating haplotypes. Since most of the isolates studied were from onychomycoses (nearly 90 %), we carried out an epidemiological study to determine the risk factors associated with such infections. Five phylogenetic species of the Fusarium solani species complex (FSSC), i.e., F. falciforme, F. keratoplasticum, F. lichenicola, F. petroliphilum, and FSSC 6 as well as two of the Fusarium oxysporum species complex (FOSC), i.e., FOSC 3 and FOSC 4, were identified. The most prevalent species were FOSC 3 (38.2%) followed by F. keratoplasticum (33.7%). In addition, our isolates were distributed into 23 haplotypes (14 into FOSC and nine into FSSC). Two of the FSSC phylogenetic species and two haplotypes of FSSC were not described before. Our results demonstrate that recipients of pedicure treatments have a lower probability of acquiring onychomycosis than those not receiving such treatments. The antifungal susceptibility of all the isolates to five clinically available agents showed that amphotericin B was the most active drug, while the azoles exhibited lower in vitro activity.

  10. Antifungal susceptibility of Candida biofilms: unique efficacy of amphotericin B lipid formulations and echinocandins.

    Science.gov (United States)

    Kuhn, D M; George, T; Chandra, J; Mukherjee, P K; Ghannoum, M A

    2002-06-01

    Biofilms, likely the predominant mode of device-related microbial infection, exhibit resistance to antimicrobial agents. Evidence suggests that Candida biofilms have dramatically reduced susceptibility to antifungal drugs. We examined antifungal susceptibilities of Candida albicans and Candida parapsilosis biofilms grown on a bioprosthetic model. In addition to conventional agents, we determined if new antifungal agents (triazoles, amphotericin B lipid formulations, and echinocandins) have activities against Candida biofilms. We also explored effects of preincubation of C. albicans cells with subinhibitory concentrations (sub-MICs) of drugs to see if they could modify subsequent biofilm formation. Finally, we used confocal scanning laser microscopy (CSLM) to image planktonic- and biofilm-exposed blastospores to examine drug effects on cell structure. Candida biofilms were formed on silicone elastomer and quantified by tetrazolium and dry weight (DW) assays. Susceptibility testing of fluconazole, nystatin, chlorhexidine, terbenafine, amphotericin B (AMB), and the triazoles voriconazole (VRC) and ravuconazole revealed resistance in all Candida isolates examined when grown as biofilms, compared to planktonic forms. In contrast, lipid formulations of AMB (liposomal AMB and AMB lipid complex [ABLC]) and echinocandins (caspofungin [Casp] and micafungin) showed activity against Candida biofilms. Preincubation of C. albicans cells with sub-MIC levels of antifungals decreased the ability of cells to subsequently form biofilm (measured by DW; P formulations.

  11. In Vitro Susceptibility of Aflatoxigenic and Non-aflatoxigenic Aspergillus flavus Strains to Conventional Antifungal Agents

    Directory of Open Access Journals (Sweden)

    Mahmoud Mahmoudi

    2012-12-01

    Full Text Available Presently appearance of resistance to antifungal agents among Aspergillus species is dramatically increasing. The objective of this study was to look at the in vitro activities of antifungal drugs against Iranian clinical (from nail, bronchoalveolar lavage, paranasal sinus isolated A. flavus strains. The susceptibility of 45 aflatoxigenic and non-aflatoxigenic Aspergillus flavus strains were evaluated to six antifungal agents (caspofungin, itraconazole, amphotericin B, ketoconazole, fluconazole, nystatin using CLSI M38-A2 broth microdilution method. The results indicated that 57.1%, 28.6% of aflatoxigenic and 25.8%, 6.5% of non-aflatoxigenic isolates were susceptible to caspofungin, amphotericin B respectively. All isolates but one aflatoxigenic strain were sensitive to ketoconazole. All 45 strains showed to be resistant to nystatin. Also 64.28%, 92.9% of aflatoxigenic and 64.51%, 100% of non-aflatoxigenic isolates were resistant to fluconazole and itraconazole in ranking order. There was no statistically significant difference between the susceptibilities of aflatoxigenic and non-aflatoxigenic strains of A. flavus to tested antifungal agents

  12. In vitro resistance of clinical Fusarium species to amphotericin B and voriconazole using the EUCAST antifungal susceptibility method

    NARCIS (Netherlands)

    Taj-Aldeen, Saad J; Salah, Husam; Al-Hatmi, Abdullah M S; Hamed, Manal; Theelen, Bart; van Diepeningen, Anne D; Boekhout, Teun; Lass-Flörl, Cornelia

    2016-01-01

    Susceptibility testing using the EUCAST-AFST method against 39 clinical Fusarium strains consecutively collected from local and invasive infections during the last 10years assessed the in vitro activities of amphotericin B (AmB) and triazole antifungal agents. In addition, the susceptibility pattern

  13. Assessment of the antifungal susceptibility of Malassezia pachydermatis in various media using a CLSI protocol.

    Science.gov (United States)

    Cafarchia, Claudia; Figueredo, Luciana A; Favuzzi, Vincenza; Surico, Mariannna R; Colao, Valeriana; Iatta, Roberta; Montagna, Maria Teresa; Otranto, Domenico

    2012-10-12

    The microdilution antifungal method (CLSI BMD, M27-A3) was used for testing the antifungal susceptibility of Malassezia species. However, optimal broth media that allow sufficient growth of M. pachydermatitis and produce reliable and reproducible MICs using the CLSI BMD protocol are yet to be established. In this study, the susceptibility of M. pachydermatis isolates to ketoconazole (KTZ), itraconazole (ITZ) and fluconazole (FLZ) was evaluated in vitro by the CLSI BMD test using Christensen's urea broth (CUB) and mRPMI 1640 containing lipid supplementation, Sabouraud dextrose broth with 1% tween 80 (SDB), and Dixon broth (DXB). A FLZ-resistant M. pachydermatis was generated in vitro and tested under the same conditions. A good growth of M. pachydermatis incubated for 48 and 72 h, respectively, was observed in CUB, SDB and DXB and not in mRPMI 1640 (p64 mg/L was observed only when the FLZ-resistant M. pachydermatis isolate was tested in SDB. Based on the results obtained herein, culture in SDB, stock inoculum suspensions of 1-5 × 10(6)CFU/ml, and an incubation time of 48 h are proposed as optimal conditions for the evaluation of the in vitro antifungal susceptibility of M. pachydermatis using a modified CLSI BMD protocol.

  14. Candida species isolated from urine specimens and antifungal susceptibility in hospitalized patients

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    Mustafa Altay Atalay

    2014-01-01

    [¤]CONCLUSION[|] In our hospital, C. albicans was the most frequently isolated Candida species from urine cultures, however, C. glabrata was found as the second most frequent species. As a result, in parallel to the increase of patient population who are at risk for Candida infections, the necessity of doing epidemiological studies for identification of species and susceptibility tests including new antifungal agents was concluded.[¤

  15. [In vitro susceptibility of isolates of Paracoccidioides spp complex to systemic antifungals using the microdilution method].

    Science.gov (United States)

    Cermehol, Julman R; Alvarado, Primavera; Mendoza, Mireya; Herndndez, Isabel; Cuestal, De

    2015-09-01

    Broth microdilution, the reference method recommended by the Clinical Laboratory Standards Institute (CLSI), is not available for use with dimorphic fungi, such as those of the Paracoccidioides genus. In this work, in vitro susceptibility of the Paracoccidioides complex (n=19) to systemic antifungals: amphotericin B, 5-flucytosine, ketoconazole, itraconazole, fluconazole, voriconazole and caspofungin, was evaluated using the microdilution method (Document M27-A3, M27-S3), with some modifications such as: culture time in Sabouraud dextrose agar (7-10 days), RPMI 1640 medium supplemented with 2% glucose and the incubation time (7, 8 and 18 days). The sensitivity in vitro was variable; the majority of Paracoccidioides isolates was susceptible to ketoconazol (73.7%), followed by voriconazole (68.4%), itraconazole (63.1%), amphotericin B (52.6%), fluconazole (47.4%), 5-flucytosine (42.1%) and caspofungin (5%). The overall resistance was mainly to caspofungin (94.7%), followed by 5-flucytosine (52.6%) and amphotericin B (47.4%). Fifty-three percent of the isolates were susceptible-dose dependent to fluconazole followed by itraconazole (15.7%) and 5-fluorocytosine (5.3%). Amphotericin B, itraconazole and voriconazole were the most potent antifungal drugs against Paracoccidioides spp (CMI: 0.03-1 microg/mL). Based on these results, we tentatively propose a microdilution assay protocol for susceptibility testing of Paracoccidioides spp to antifungal drugs. This method may be clinically useful to predict resistance, even though further studies are needed.

  16. Antifungal susceptibility of invasive yeast isolates in Italy: the GISIA3 study in critically ill patients

    Directory of Open Access Journals (Sweden)

    Mussap Michele

    2011-05-01

    Full Text Available Abstract Background Yeasts are a common cause of invasive fungal infections in critically ill patients. Antifungal susceptibility testing results of clinically significant fungal strains are of interest to physicians, enabling them to adopt appropriate strategies for empiric and prophylactic therapies. We investigated the antifungal susceptibility of yeasts isolated over a 2-year period from hospitalised patients with invasive yeast infections. Methods 638 yeasts were isolated from the blood, central venous catheters and sterile fluids of 578 patients on general and surgical intensive care units and surgical wards. Etest strips and Sensititre panels were used to test the susceptibility of the isolates to amphotericin B, anidulafungin, caspofungin, fluconazole, itraconazole, posaconazole and voriconazole in 13 laboratories centres (LC and two co-ordinating centres (CC. The Clinical and Laboratory Standards Institute (CLSI reference broth microdilution method was used at the CCs for comparison. Results Etest and Sensititre (LC/CC MIC90 values were, respectively: amphotericin B 0.5/0.38, 1/1 mg/L; anidulafungin 2/1.5 and 1/1 mg/L; caspofungin 1/0.75 and 0.5/0.5 mg/L; fluconazole 12/8 and 16/16 mg/L; itraconazole 1/1.5, 0.5/0.5 mg/L; posaconazole 0.5 mg/L and voriconazole 0.25 mg/L for all. The overall MIC90 values were influenced by the reduced susceptibility of Candida parapsilosis isolates to echinocandins and a reduced or lack of susceptibility of Candida glabrata and Candida krusei to azoles, in particular fluconazole and itraconazole. Comparison of the LC and CC results showed good Essential Agreement (90.3% for Etest and 92.9% for Sensititre, and even higher Categorical Agreement (93.9% for Etest and 96% for Sensititre; differences were observed according to the species, method, and antifungal drug. No cross-resistance between echinocandins and triazoles was detected. Conclusions Our data confirm the different antifungal susceptibility

  17. Determination of antifungal susceptibility patterns among the environmental isolates of Aspergillus fumigatus in Iran

    Science.gov (United States)

    Mohammadi, Faezeh; Dehghan, Parvin; Nekoeian, Shahram; Hashemi, Seyed Jamal

    2016-01-01

    Background: In recent years, triazole-resistant environmental isolates of Aspergillus fumigatus have emerged in Europe and Asia. Azole resistance has been reported in patients who are treated with long-term azole therapy or exposure of the fungus spores to the azole fungicides used in agriculture. To date, a wide range of mutations in A. fumigatus have been described conferring azole-resistance, which commonly involves modifications in the cyp51A gene. We investigated antifungal susceptibility pattern of environmental isolates of A. fumigatus. Materials and Methods: In this study, 170 environmental samples collected from indoors surfaces of three hospitals in Iran. It was used β-tubulin gene to confirm the all of A. fumigatus isolates, which was identified by conventional methods. Furthermore, the antifungal susceptibility of itraconazole, voriconazole, and posaconazole was investigated using broth microdilution test, according to European Committee on Antimicrobial Susceptibility testing reference method. Results: From a total of 158 environmental molds fungi obtained from the hospitals, 58 isolates were identified as A. fumigatus by amplification of expected size of β-tubulin gene (~500 bp). In this study, in vitro antifungal susceptibility testing has shown that there were not high minimum inhibitory concentration values of triazole antifungals in all of the 58 environmental isolates of A. fumigatus. Conclusion: Our findings demonstrated that there was not azole-resistant among environmental isolates of A. fumigatus. Medical triazoles compounds have structural similarity with triazole fungicide compounds in agriculture, therefore, resistance development through exposure to triazole fungicide compounds in the environment is important but it sounds there is not a serious health problem in drug resistance in environmental isolates in Iran. PMID:27656605

  18. Antifungal susceptibilities of non-Aspergillus filamentous fungi causing invasive infection in Australia: support for current antifungal guideline recommendations.

    Science.gov (United States)

    Halliday, Catriona L; Chen, Sharon C-A; Kidd, Sarah E; van Hal, Sebastian; Chapman, Belinda; Heath, Christopher H; Lee, Andie; Kennedy, Karina J; Daveson, Kathryn; Sorrell, Tania C; Morrissey, C Orla; Marriott, Deborah J; Slavin, Monica A

    2016-10-01

    Antifungal susceptibilities of non-Aspergillus filamentous fungal pathogens cannot always be inferred from their identification. Here we determined, using the Sensititre(®) YeastOne(®) YO10 panel, the in vitro activities of nine antifungal agents against 52 clinical isolates of emergent non-Aspergillus moulds representing 17 fungal groups in Australia. Isolates comprised Mucorales (n = 14), Scedosporium/Lomentospora spp. (n = 18) and a range of hyaline hyphomycetes (n = 9) and other dematiaceous fungi (n = 11). Excluding Verruconis gallopava, echinocandins demonstrated poor activity (MICs generally >8 mg/L) against these moulds. Lomentospora prolificans (n = 4) and Fusarium spp. (n = 6) demonstrated raised MICs to all antifungal drugs tested, with the lowest being to voriconazole and amphotericin B (AmB), respectively (geometric mean MICs of 3.4 mg/L and 2.2 mg/L, respectively). All Scedosporium apiospermum complex isolates (n = 14) were inhibited by voriconazole concentrations of ≤0.25 mg/L, followed by posaconazole and itraconazole at ≤1 mg/L. Posaconazole and AmB were the most active agents against the Mucorales, with MIC90 values of 1 mg/L and 2 mg/L, respectively, for Rhizopus spp. For dematiaceous fungi, all isolates were inhibited by itraconazole and posaconazole concentrations of ≤0.5 mg/L (MIC90, 0.12 mg/L and 0.25 mg/L, respectively), but voriconazole and AmB also had in vitro activity (MIC90, 0.5 mg/L and 1 mg/L, respectively). Differences in antifungal susceptibility within species and between species within genera support the need for testing individual patient isolates to guide therapy. The Sensititre(®) YeastOne(®) offers a practical alternative to the reference methodology for susceptibility testing of moulds.

  19. OTYPIC CHARACTERIZATION AND ANTIFUNGAL SUSCEPTIBILITY PATTERN OF CANDIDA SP ISOLATED FROM A TERTIARY CARE CENTER

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    Rudramurthy

    2014-02-01

    Full Text Available ACT: Candida , a yeast like ubiquitous fungus , is an endogenous species which produces commonest fungal infection; Candidiasis. Resistance to antifungal agents is an alarming sign for the emerging common nosocomial candidiasis. MATERIALS AND METHODS: Various types of specimens we re collected from the c linically suspected cases of candidiasis. Isolation and characterization of candida sp . was done by standard procedures. Antifungal susceptibility was done by disc diffusion method. RESULT: The candida was isolated from various clinical specimens , vaginal swab (24.66% , skin scraping (13.33% oral swabs (12.66% , ear swabs ( 11.33% , nail scraping (10% , and pus from diabetes foot ulcer and post - operative wound infection ( 8% , sputum ( 6% , urine (4.66% , stool ( 4% , blood ( 2.66% , and eye swabs ( 2.66%. Amon g different species of candida isolated C.albicans was the predominant species (79.33% followed by C tropicalis (19.33% and C.Guilliermondii (1.33%. Antifungal resistance of different species of candida was higher to fluconazole . The least resistance wa s seen with amphotericin - B (1.33%. CONCLUSION: The increased isolation of candida species and development of resistance to commonly used antifungal drugs requires careful interpretation and the in vitro susceptibility testing. This facilitates better pat ient care.

  20. Antifungal susceptibilities and identification of species of the Sporothrix schenckii complex isolated in Brazil.

    Science.gov (United States)

    Ottonelli Stopiglia, Cheila Denise; Magagnin, Cibele Massotti; Castrillón, Mauricio Ramírez; Mendes, Sandra Denise Camargo; Heidrich, Daiane; Valente, Patricia; Scroferneker, Maria Lúcia

    2014-01-01

    Sporotrichosis is a subacute or chronic mycosis caused worldwide by the dimorphic species complex, Sporothrix schenckii. We studied 85 isolates recovered in Brazil to verify their identification and evaluate their in vitro antifungal susceptibility patterns. Based on phenotypic tests (microscopic features, ability to grow at 30°C and 37°C, colony diameters, as well as assimilation of sucrose and raffinose) and molecular assays (amplification of a fragment of the calmodulin gene), the strains were identified as S. schenckii, S. brasiliensis and S. globosa, with a predominance of S. schenckii isolates. There was 37.7% disagreement between the phenotypic and genotypic identification methodologies. In general, terbinafine was the most active drug, followed by ketoconazole and itraconazole, and the less active fluconazole and voriconazole. Five isolates (one S. globosa and four S. schenckii) were found to be itraconazole-resistant strains but, in general, there were no differences in the in vitro antifungal susceptibility profiles among the Sporothrix species.

  1. [Species distribution and antifungal susceptibility of Candida spp. causing superficial mycosis. Coro, Falcon state, Venezuela].

    Science.gov (United States)

    Saúl-García, Yotsabeth; Humbría-García, Leyla; Hernández-Valles, Rosaura

    2015-09-01

    Candida species other than C. albicans are often described as causative agents of superficial mycosis and are more resistant to treatment with azoles. In order to determine the distribution of species and in vitro antifungal susceptibility of Candida spp., one ambispective study, which analyzed 18 yeast isolates obtained from samples from patients diagnosed with superficial mycosis, was performed. Taxonomic identification was performed by macroscopic visualization of the growth characteristics in chromogenic agar and by conventional methods. The susceptibility to fluconazole and voriconazole was evaluated by the disc diffusion method. Most of the isolates (88.8%), came from nail samples. C. parapsilosis was the most common species, followed by C. tropicalis, C. albicans and C. krusei, which confirmed the prevalence of non-albicans species as a cause of superficial mycoses. The pattern of susceptibility to fluconazole and voriconazole was similar: all isolates of C. parapsilosis and C. albicans were susceptible, while 83.3% of C. tropicalis showed sensitivity to both antifungals. C. krusei, fluconazole-resistant species showed intermediate susceptibility io voriconazole. The use of chromogenic agar allowed to detect mixed infections in nail samples, involving Candida spp. and C. tropicalis in one case, the latter with resistance to both fluconazole and voriconazole. The results demonstrate the importance of species identification and susceptibility testing to avoid therapeutic failures in superficial mycoses.

  2. Antifungal susceptibilities of bloodstream isolates of Candida species from nine hospitals in Korea: application of new antifungal breakpoints and relationship to antifungal usage.

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    Eun Jeong Won

    Full Text Available We applied the new clinical breakpoints (CBPs of the Clinical and Laboratory Standards Institute (CLSI to a multicenter study to determine the antifungal susceptibility of bloodstream infection (BSI isolates of Candida species in Korea, and determined the relationship between the frequency of antifungal-resistant Candida BSI isolates and antifungal use at hospitals. Four hundred and fifty BSI isolates of Candida species were collected over a 1-year period in 2011 from nine hospitals. The susceptibilities of the isolates to four antifungal agents were determined using the CLSI M27 broth microdilution method. By applying the species-specific CBPs, non-susceptibility to fluconazole was found in 16.4% (70/428 of isolates, comprising 2.6% resistant and 13.8% susceptible-dose dependent isolates. However, non-susceptibility to voriconazole, caspofungin, or micafungin was found in 0% (0/370, 0% (0/437, or 0.5% (2/437 of the Candida BSI isolates, respectively. Of the 450 isolates, 72 (16.0% showed decreased susceptibility to fluconazole [minimum inhibitory concentration (MIC ≥4 μg/ml]. The total usage of systemic antifungals varied considerably among the hospitals, ranging from 190.0 to 7.7 defined daily dose per 1,000 patient days, and fluconazole was the most commonly prescribed agent (46.3%. By Spearman's correlation analysis, fluconazole usage did not show a significant correlation with the percentage of fluconazole resistant isolates at hospitals. However, fluconazole usage was significantly correlated with the percentage of fluconazole non-susceptible isolates (r = 0.733; P = 0.025 or the percentage of isolates with decreased susceptibility to fluconazole (MIC ≥4 μg/ml (r = 0.700; P = 0.036 at hospitals. Our work represents the first South Korean multicenter study demonstrating an association between antifungal use and antifungal resistance among BSI isolates of Candida at hospitals using the new CBPs of the CLSI.

  3. Antifungal susceptibility profile of Trichosporon isolates: correlation between CLSI and etest methodologies

    Science.gov (United States)

    Lemes, Raquel M.L.; Lyon, Juliana P.; Moreira, Leonardo M.; de Resende, Maria Aparecida

    2010-01-01

    The aim of the present study was to evaluate the antifungal susceptibility profile of Trichosporon species isolated from different sources employing the Clinical and Laboratory Standards Institute (CLSI) method and E-test method. Thirty-four isolates of Trichosporon spp. and six CBS reference samples were tested for their susceptibility to Amphotericin B, 5-flucytosine, Fluconazole, Itraconazole, Voriconazole and Terbinafine. All species showed high Minimun Inhibitory Concentrations (MIC) for Itraconazole and susceptibility to Fluconazole, The comparison among the results obtained by the CLSI method and E-test revealed larger discrepancies among 5-flucytosine and Itraconazole. The present work provides epidemiological data that could influence therapeutic choices. Furthermore, the comparison between different methodologies could help to analyze results obtained by different laboratories. PMID:24031497

  4. Antifungal susceptibility profile of Trichosporon isolates: correlation between CLSI and etest methodologies

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    Raquel M.L. Lemes

    2010-06-01

    Full Text Available The aim of the present study was to evaluate the antifungal susceptibility profile of Trichosporon species isolated from different sources employing the Clinical and Laboratory Standards Institute (CLSI method and E-test method. Thirty-four isolates of Trichosporon spp. and six CBS reference samples were tested for their susceptibility to Amphotericin B, 5-flucytosine, Fluconazole, Itraconazole, Voriconazole and Terbinafine. All species showed high Minimun Inhibitory Concentrations (MIC for Itraconazole and susceptibility to Fluconazole, The comparison among the results obtained by the CLSI method and E-test revealed larger discrepancies among 5-flucytosine and Itraconazole. The present work provides epidemiological data that could influence therapeutic choices. Furthermore, the comparison between different methodologies could help to analyze results obtained by different laboratories.

  5. Statistical analyses of correlation between fluconazole MICs for Candida spp. assessed by standard methods set forth by the European Committee on Antimicrobial Susceptibility Testing (E.Dis. 7.1) and CLSI (M27-A2).

    NARCIS (Netherlands)

    Rodriguez-Tudela, J.L.; Donnelly, J.P.; Pfaller, M.A.; Chryssantou, E.; Warn, P.; Denning, D.W.; Espinel-Ingroff, A.; Barchiesi, F.; Cuenca-Estrella, M.

    2007-01-01

    The European Committee on Antimicrobial Susceptibility Testing (EUCAST) Subcommittee on Antifungal Susceptibility Testing recently published a standard for determining the susceptibility of fermentative yeasts to antifungals. From the beginning, the EUCAST and its North American counterpart, the CLS

  6. In Vitro Antifungal Susceptibility of Oral Candida Species from Iranian HIV Infected Patients

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    A A Khaksar

    2012-05-01

    Full Text Available Background: Oropharyngeal candidiasis and antifungal drug resistance are major problems in HIV positive patients. The increased reports of antifungal resistance and expanding therapeutic options prompted the determination of antifungal susceptibility profile of Candida species isolates in Iranian patients living with HIV/AIDS (PLWHA in the present study. Methods: One hundred fifty oral samples from Iranian HIV positive patients were obtained and cultured on CHROMagar and Sabourauds dextrose agar. All isolates were identified according to assimilation profile, germ tube, colony color and other conventional methods. Disk diffusion testing and Broth Microdilution of six antifungal agents were performed according to the methods described in CLSI. Results: Candida albicans (50.2% was the most frequent isolated yeast, followed by C. glabrata (22%. Non-Candida albicans species were isolated from 71 (61% positive cultures. 25.7% of Candida albicans isolates were resistant to fluconazole (MIC≥64 µg/ml as were 21.9% and 16.4% to ketoconazole and clotrimazole (MIC>0.125 µg/ml, respectively. Resistance to polyene antifungals including amphotericin B and nystatin, and caspofungin were scarce. 57.7% of candida glabrata isolates were resistant to fluconazole, 31% to ketoconazole and 35% to clotrimazole. Conclusion: Screening for antifungal resistant candida isolates by disk diffusion or broth dilution methods in clinical laboratories is an ideal surveillance measure in the management of oral thrush in patients with HIV/AIDS. Although nystatin is widely used in clinical practice for HIV positive patients, there was no evidence of enhanced resistance to it. Regarding no resistance to caspofungin, its administration is suggested.

  7. Comparison of the Sensititre YeastOne colorimetric antifungal panel with CLSI microdilution for antifungal susceptibility testing of the echinocandins against Candida spp., using new clinical breakpoints and epidemiological cutoff values.

    Science.gov (United States)

    Pfaller, M A; Chaturvedi, V; Diekema, D J; Ghannoum, M A; Holliday, N M; Killian, S B; Knapp, C C; Messer, S A; Miskou, A; Ramani, R

    2012-08-01

    A commercially prepared dried colorimetric microdilution panel (Sensititre Yeast One, TREK Diagnostic Systems, Cleveland, OH, USA) was compared in 3 different laboratories with the Clinical and Laboratory Standards Institute (CLSI) reference microdilution method by testing 2 quality control strains, 25 reproducibility strains, and 404 isolates of Candida spp. against anidulafungin, caspofungin, and micafungin. Reference CLSI BMD MIC end points and YeastOne colorimetric end points were read after 24 h of incubation. Excellent (100%) essential agreement (within 2 dilutions) between the reference and colorimetric MICs was observed. Categorical agreement (CA) between the 2 methods was assessed using the new species-specific clinical breakpoints (CBPs): susceptible (S), ≤0.25 μg/mL; intermediate (I), 0.5 μg/mL; and resistant (R), ≥1 μg/mL, for C. albicans, C. tropicalis, and C. krusei, and ≤2 μg/mL (S), 4 μg/mL (I), and ≥8 μg/mL (R) for C. parapsilosis and all 3 echinocandins. The new CBPs for anidulafungin and caspofungin and C. glabrata are ≤0.12 μg/mL (S), 0.25 μg/mL (I), and ≥0.5 μg/mL (R), whereas those for micafungin are ≤0.06 μg/mL (S), 0.12 μg/mL (I), and ≥0.25 μg/mL (R). Due to the lack of CBPs for any of the echinocandins and C. lusitaniae, the epidemiological cutoff values (ECVs) were used for this species to categorize the isolates as wild-type (WT; MIC ≤ECV) and non-WT (MIC >ECV), respectively, for anidulafungin (≤2 μg/mL/>2 μg/mL), caspofungin (≤1 μg/mL/>1 μg/mL), and micafungin (≤0.5 μg/mL/>0.5 μg/mL). CA ranged from 93.6% (caspofungin) to 99.6% (micafungin) with less than 1% very major or major errors. The YeastOne colorimetric method remains comparable to the CLSI BMD reference method for testing the susceptibility of Candida spp. to the echinocandins when using the new (lower) CBPs and ECVs. Further study using defined fks mutant strains of Candida is warranted.

  8. CHARACTERIZATION AND ANTIFUNGAL SUSCEPTIBILITY PATTERN OF CANDIDA SPP . ISOLATED FROM CLINICAL SPECIMENS

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    Sagarika

    2015-05-01

    Full Text Available BACKGROUND: With the changing health scenario fungal infections have increased significantly, contributing to morbidity, mortality and health care cost. Candida is major human fungal pathogens that cause both superficial and deep tissue infections. With emergence of non - albicans Candida species, availability of advanced identification methods and antifungal resistance, the spectrum of candidiasis has changed. OBJ ECTIVE: The aim of our study was to identify the distribution of Candida species among clinical isolates, risk factors associated with candidiasis and their sensitivity pattern for common antifungal drugs. MATERIALS AND METHODS: One hundred thirty nine dif ferent clinical isolates of Candida were collected from indoor patients of a tertiary care centre of Gujarat from May 2009 to June 2010. Identification of Candida species and antifungal susceptibility testing was performed with miniAPI (Analytical Prophylactic Index (Biomerieux, France which is an automatic identification and susceptibility testing instrument. RESULTS: We found that the non ‑ albicans Candida were more prevalent than Candida albicans . Candida tropicalis (48.9% was the most common Candida spp. and also more resistant than that of C.albicans . C.albicans showed resistance against fluconazole (3.5% and itraconazole (8.8% whereas C.tropicalis were res istant to amphotericin B (10.3%, fluconazole (20.7%, itraconazole (32.3%, and voriconazole (23.5% and flucytosine (5.8%. Overall resistance rates of Candida for amphotericin B, fluconazole, itraconazole, and voriconazole and flucytosine were 6.4%, 15. 2%, 22.3%, 12.9%, 5% respectively. CONCLUSION: To achieve better clinical results species ‑ level identification of Candida spp. and their antifungal sensitivity testing should be performed.

  9. Microscopic Evaluation, Molecular Identification, Antifungal Susceptibility, and Clinical Outcomes in Fusarium, Aspergillus and, Dematiaceous Keratitis

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    Devarshi U. Gajjar

    2013-01-01

    Full Text Available Purpose. Fusarium, Aspergillus, and Dematiaceous are the most common fungal species causing keratitis in tropical countries. Herein we report a prospective study on fungal keratitis caused by these three fungal species. Methodology. A prospective investigation was undertaken to evaluate eyes with presumed fungal keratitis. All the fungal isolates (n=73 obtained from keratitis infections were identified using morphological and microscopic characters. Molecular identification using sequencing of the ITS region and antifungal susceptibility tests using microdilution method were done. The final clinical outcome was evaluated in terms of the time taken for resolution of keratitis and the final visual outcome. The results were analyzed after segregating the cases into three groups, namely, Fusarium, Aspergillus, and Dematiaceous keratitis. Results. Diagnosis of fungal keratitis was established in 73 (35.9% cases out of 208 cases. The spectra of fungi isolated were Fusarium spp. (26.6%, Aspergillus spp. (21.6%, and Dematiaceous fungi (11.6%. The sequence of the ITS region could identify the Fusarium and Aspergillus species at the species complex level, and the Dematiaceous isolates were accurately identified. Using antifungal agents such as fluconazole, natamycin, amphotericin B, and itraconazole, the minimum inhibitory concentrations (MICs for Fusarium spp. were >32 μg/mL, 4–8 μg/mL, 0.5–1 μg/mL, and >32 μg/mL, respectively. Antifungal susceptibility data showed that Curvularia spp. was highly resistant to all the antifungal agents. Overall, natamycin and amphotericin B were found to be the most effective antifungal agents. The comparative clinical outcomes in all cases showed that the healing response in terms of visual acuity of the Dematiaceous group was significantly good when compared with the Fusarium and Aspergillus groups (P<0.05. The time required for healing in the Fusarium group was statistically significantly less when

  10. In vitro antifungal susceptibility and molecular identity of 99 clinical isolates of the opportunistic fungal genus Curvularia

    NARCIS (Netherlands)

    Cunha, da K.C.; Sutton, D.A.; Fothergill, A.W.; Gené, J.; Cano, J.; Madrid, H.; Hoog, de G.S.; Crous, P.W.; Guarro, J.

    2013-01-01

    The in vitro antifungal susceptibility of a set of 99 clinical isolates of Curvularia was tested against 9 drugs using a reference microdilution method. The isolates had been identified previously to species level by comparing their ITS rDNA and glyceraldehyde-3-phosphate dehydrogenase gene sequence

  11. Sentinel surveillance of invasive candidiasis in Spain: epidemiology and antifungal susceptibility.

    Science.gov (United States)

    Nieto, M C; Tellería, O; Cisterna, R

    2015-01-01

    In order to know the epidemiology and the changes of antifungal resistance in invasive candidiasis (IC) we carried out this prospective study of Candida strains belonging to patients admitted to 26 Spanish hospitals from June 2011 to June 2012 diagnosed with IC. Clinical information and the identity of the Candida species were collected and antifungal susceptibility was tested using broth microdilution in five agents: amphotericin B, fluconazole, voriconazole, caspofungin and anidulafungin. A total of 705 cases-isolates were documented. Most of the patients suffered from candidemia and several underlying diseases and more than half of them were neutropenic or under immunosuppressive therapy, factors associated with higher mortality. Thirty percent of global mortality was documented. C. albicans was the most frequently isolated species, although an increase of non-C. albicans species was observed. Resistance to fluconazole was of 3.4%, lower than in previous years (6.3%). C. parapsilosis presented a higher MIC90 of echinocandins compared to other species.

  12. 口腔念珠菌病的菌群分布特点及体外抗真菌药敏试验研究%Oral candidiasis fl ora distribution and antifungal susceptibility test in vitro studies

    Institute of Scientific and Technical Information of China (English)

    张华; 董培

    2014-01-01

    ObjectiveAnalyzes the bacterial fl ora distribution characteristics of oral candidiasis, study in vitro antifungal susceptibility test.MethodsA random sample of oral candida patients 85 cases, isolates, and USES the chromogenic medium, identifi cation of bacterial fl ora distribution characteristics of isolates; In vitro antifungal susceptibility test, detection of candida sensitivity to fl uconazole and itraconazole.ResultsOf candida albicans fl uconazole s ensitive rate was (66.7%), signifi cantly higher than that of itraconazole sensitive rate of candida albicans (39.4%), and compare the differences between groups was statistically significant (x2 = 8.724,P = 0.016,P<0.05).ConclusionOral candida albicans candida as most, and has certain resistance phenomenon, white candida to fl uconazole, sensitivity is good.%目的:探讨分析口腔念珠菌病的菌群分布特点,研究体外抗真菌药敏试验。方法随机抽取口腔念珠菌病患者85例,分离菌株,并采用显色培养基培养,鉴定分离株的菌群分布特点;进行体外抗真菌药敏试验,检测念珠菌对氟康唑和伊曲康唑的敏感性。结果氟康唑对白色念珠菌的敏感率(66.7%)显著高于伊曲康唑对白色念珠菌的敏感率(39.4%),组间比较差异有统计学意义(x2=8.724,P=0.016,P<0.05)。结论口腔念珠菌以白色念珠菌最多,并有一定的耐药现象,白色假丝酵母菌对氟康唑敏感性好。

  13. Molecular differentiation and antifungal susceptibilities of Candida parapsilosis isolated from patients with bloodstream infections.

    Science.gov (United States)

    Tay, Sun Tee; Na, Shiang Ling; Chong, Jennifer

    2009-02-01

    The genetic heterogeneity and antifungal susceptibility patterns of Candida parapsilosis isolated from blood cultures of patients were investigated in this study. Randomly amplified polymorphic DNA (RAPD) analysis generated 5 unique profiles from 42 isolates. Based on the major DNA fragments of the RAPD profiles, the isolates were identified as RAPD type P1 (29 isolates), P2 (6 isolates), P3 (4 isolates), P4 (2 isolates) and P5 (1 isolate). Sequence analysis of the internal transcribed spacer (ITS) gene of the isolates identified RAPD type P1 as C. parapsilosis, P2 and P3 as Candida orthopsilosis, P4 as Candida metapsilosis, and P5 as Lodderomyces elongisporus. Nucleotide variations in ITS gene sequences of C. orthopsilosis and C. metapsilosis were detected. Antifungal susceptibility testing using Etests showed that all isolates tested in this study were susceptible to amphotericin B, fluconazole, ketoconazole, itraconazole and voriconazole. C. parapsilosis isolates exhibited higher MIC(50) values than those of C. orthopsilosis for all of the drugs tested in this study; however, no significant difference in the MICs for these two Candida species was observed. The fact that C. orthopsilosis and C. metapsilosis were responsible for 23.8 and 4.8 % of the cases attributed to C. parapsilosis bloodstream infections, respectively, indicates the clinical relevance of these newly described yeasts. Further investigations of the ecological niche, mode of transmission and virulence of these species are thus essential.

  14. Species Distribution and In Vitro Antifungal Susceptibility of Vulvovaginal Candida Isolates in China

    Science.gov (United States)

    Wang, Feng-Juan; Zhang, Dai; Liu, Zhao-Hui; Wu, Wen-Xiang; Bai, Hui-Hui; Dong, Han-Yu

    2016-01-01

    Background: Vulvovaginal candidiasis (VVC) was a common infection associated with lifelong harassment of woman's social and sexual life. The purpose of this study was to describe the species distribution and in vitro antifungal susceptibility of Candida species (Candida spp.) isolated from patients with VVC over 8 years. Methods: Species which isolated from patients with VVC in Peking University First Hospital were identified using chromogenic culture media. Susceptibility to common antifungal agents was determined using agar diffusion method based on CLSI M44-A2 document. SPSS software (version 14.0, Inc., Chicago, IL, USA) was used for statistical analysis, involving statistical description and Chi-square test. Results: The most common strains were Candida (C.) albicans, 80.5% (n = 1775) followed by C. glabrata, 18.1% (n = 400). Nystatin exhibited excellent activity against all species (<4% resistant [R]). Resistance to azole drugs varied among different species. C. albicans: clotrimazole (3.1% R) < fluconazole (16.6% R) < itraconazole (51.5% R) < miconazole (54.0% R); C. glabrata: miconazole (25.6% R) < clotrimazole (50.5% R) < itraconazole (61.9% R) < fluconazole (73.3% R); Candida krusei: clotrimazole (0 R) < fluconazole (57.7% R) < miconazole (73.1% R) < itraconazole (83.3% R). The susceptibility of fluconazole was noticeably decreasing among all species in the study period. Conclusions: Nystatin was the optimal choice for the treatment of VVC at present. The species distribution and in vitro antifungal susceptibility of Candida spp. isolated from patients with VVC had changed over time. PMID:27174323

  15. Species Distribution and In Vitro Antifungal Susceptibility of Vulvovaginal Candida Isolates in China

    Institute of Scientific and Technical Information of China (English)

    Feng-Juan Wang; Dai Zhang; Zhao-Hui Liu; Wen-Xiang Wu; Hui-Hui Bai; Han-Yu Dong

    2016-01-01

    Background:Vulvovaginal candidiasis (VVC) was a common infection associated with lifelong harassment of woman's social and sexual life.The purpose of this study was to describe the species distribution and in vitro antifungal susceptibility of Candida species (Candida spp.) isolated from patients with WC over 8 years.Methods:Species which isolated from patients with WC in Peking University First Hospital were identified using chromogenic culture media.Susceptibility to common antifungal agents was determined using agar diffusion method based on CLSI M44-A2 document.SPSS software (version 14.0,Inc.,Chicago,IL,USA) was used for statistical analysis,involving statistical description and Chi-square test.Results:The most common strains were Candida (C.) albicans,80.5% (n =1775) followed by C.glabrata,18.1% (n =400).Nystatin exhibited excellent activity against all species (<4% resistant [R]).Resistance to azole drugs varied among different species.C albicans:clotrimazole (3.1% R) < fluconazole (16.6% R) < itraconazole (51.5% R) < miconazole (54.0% R);C.glabrata:miconazole (25.6% R) < clotrimazole (50.5% R) < itraconazole (61.9% R) < fluconazole (73.3% R);Candida krusei:clotrimazole (0 R) < fluconazole (57.7% R) < miconazole (73.1% R) < itraconazole (83.3% R).The susceptibility offluconazole was noticeably decreasing among all species in the study period.Conclusions:Nystatin was the optimal choice for the treatment of WC at present.The species distribution and in vitro antifungal susceptibility of Candida spp.isolated from patients with VVC had changed over time.

  16. Candiduria in adults and children: prevalence and antifungal susceptibility in outpatient of Jataí-GO

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    Izabela Alves de Sousa

    2014-08-01

    Full Text Available Introduction: The term candiduria refers to the presence of yeast in urine and Candida albicans is the most common agent. In general, routine laboratories do not perform identification and cultivation of yeast. Objectives: To determine the prevalence of Candida species and to evaluate the antifungal susceptibility of the species isolated in urine of outpatients Jataí-GO, between January-October 2013. Material and method: Urine samples containing fungal structures were plated out on Sabouraud agar with chloramphenicol. Differentiation was taken with the urease test, nitrogen and carbon sources assimilation, germ tube test, morphology on cornmeal agar and chromogenic agar cultivation. Susceptibility was evaluated at antifungal itraconazole, fluconazole, amphotericin B and ketoconazole. Results: 1,215 urine tests were performed, and 64 had fungal structures (5.3%. Two samples were lost, thus here we considered 62 isolates. From this total, 43 were identified as C. albicans (67.2 %, eight C. glabrata (12.5 %, five C. krusei (7.8%, three C. tropicalis (4.7%, and three could not determine the species (4.7%. Amphotericin B and ketoconazole inhibited 94.9% of the isolates. On the other hand, 55.9% and 54.2 % were resistant to itraconazole and fluconazole, respectively. The resistance rates of both fluconazole and itraconazole for C. glabrata and C. albicans, as fluconazole for C. albicans and C. krusei, showed significant differences (p < 0.05. Conclusion: These data demonstrate the importance of conducting a full identification and susceptibility to antifungal agents in samples with yeast infection.

  17. Spectrum and the In Vitro Antifungal Susceptibility Pattern of Yeast Isolates in Ethiopian HIV Patients with Oropharyngeal Candidiasis

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    Birhan Moges

    2016-01-01

    Full Text Available Background. In Ethiopia, little is known regarding the distribution and the in vitro antifungal susceptibility profile of yeasts. Objective. This study was undertaken to determine the spectrum and the in vitro antifungal susceptibility pattern of yeasts isolated from HIV infected patients with OPC. Method. Oral pharyngeal swabs taken from oral lesions of study subjects were inoculated onto Sabouraud Dextrose Agar. Yeasts were identified by employing conventional test procedures and the susceptibility of yeasts to antifungal agents was evaluated by disk diffusion assay method. Result. One hundred and fifty-five yeast isolates were recovered of which 91 isolates were from patients that were not under HAART and 64 were from patients that were under HAART. C. albicans was the most frequently isolated species followed by C. glabrata, C. tropicalis, C. krusei, C. kefyr, Cryptococcus laurentii, and Rhodotorula species. Irrespective of yeasts isolated and identified, 5.8%, 5.8%, 12.3%, 8.4%, 0.6%, and 1.3% of the isolates were resistant to amphotericin B, clotrimazole, fluconazole, ketoconazole, miconazole, and nystatin, respectively. Conclusion. Yeast colonization rate of 69.2% and 31% resistance to six antifungal agents was documented. These highlight the need for nationwide study on the epidemiology of OPC and resistance to antifungal drugs.

  18. Epidemiology and antifungal susceptibility of Candida species in a tertiary care hospital, Kolkata, India

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    Partha Bhattacharjee

    2016-06-01

    Conclusion: Species-level identification of Candida and their antifungal sensitivity testing should to be performed to achieve better clinical result and to select an appropriate and effective antifungal therapy. High resistance to antifungal agents is an alarming sign to the healthcare professionals.

  19. Candida vaginitis during contraceptive use: the influence of methods, antifungal susceptibility and virulence patterns.

    Science.gov (United States)

    Güzel, A B; Küçükgöz-Güleç, U; Aydin, M; Gümral, R; Kalkanci, A; Ilkit, M

    2013-11-01

    No consensus exists about whether contraceptives cause an increased risk of vaginitis, including vulvovaginal candidosis (VVC). We investigated 495 women (252 who used contraceptives; 243 who did not) for the presence of VVC. Antifungal susceptibility testing was performed for five antifungal agents and for boric acid, and three virulence factors were also examined. We recovered 129 (26.1%) monofungal populations from vaginal samples of women with acute VVC (AVVC, n = 18), symptomatic recurrent VVC (RVVC, n = 22) and asymptomatic RVVC (n = 28), as well as of other contraceptive users who carried Candida in their vaginas (n = 61). It is important to note that the women who had VVC used the same contraceptive methods (p > 0.05). Candida albicans was the most common species isolated (45%), followed by C. glabrata (40.3%). Most of the vaginal yeast isolates exhibited low minimum inhibitory concentration levels for the five antifungals tested. However, this was not the case for boric acid. In addition, the yeast fungi that was derived from the AVVC and RVVC patients showed higher amounts of haemolytic activity than the yeast fungi found among the controls (p contraception does not predispose women to VVC (p > 0.05). Also, both host- and organism-related factors were required to achieve optimal clinical treatment for VVC.

  20. In vitro susceptibility of environmental isolates of Exophiala dermatitidis to five antifungal drugs.

    Science.gov (United States)

    Duarte, Ana Paula Miranda; Pagnocca, Fernando Carlos; Baron, Noemi Carla; Melhem, Marcia de Souza Carvalho; Palmeira, Gislene Aparecida; de Angelis, Dejanira de Franceschi; Attili-Angelis, Derlene

    2013-06-01

    Several dematiaceous fungi frequently isolated from nature are involved in cases of superficial lesions to lethal cerebral infections. Antifungal susceptibility data on environmental and clinical isolates are still sparse despite the advances in testing methods. The objective of this study was to examine the activities of 5-flucytosine, amphotericin B, itraconazole, voriconazole and terbinafine against environmental isolates of Exophiala strains by minimum inhibition concentration (MIC) determination. The strains were obtained from hydrocarbon-contaminated soil, ant cuticle and fungal pellets from the infrabuccal pocket of attine gynes. Broth microdilution assay using M38-A2 reference methodology for the five antifungal drugs and DNA sequencing for fungal identification were applied. Terbinafine was the most active drug against the tested strains. It was observed that amphotericin B was less effective, notably against Exophiala spinifera, also studied. High MICs of 5-flucytosine against Exophiala dermatitidis occurred. This finding highlights the relevance of studies on the antifungal resistance of these potential opportunistic species. Our results also contribute to a future improvement of the standard methods to access the drug efficacy currently applied to black fungi.

  1. In vitro susceptibility of antifungal drugs against Sporothrix brasiliensis recovered from cats with sporotrichosis in Brazil.

    Science.gov (United States)

    Brilhante, Raimunda Sâmia Nogueira; Rodrigues, Anderson Messias; Sidrim, José Júlio Costa; Rocha, Marcos Fábio Gadelha; Pereira, Sandro Antonio; Gremião, Isabella Dib Ferreira; Schubach, Tânia Maria Pacheco; de Camargo, Zoilo Pires

    2016-03-01

    Sporotrichosis is an important subcutaneous mycosis of humans and animals. Classically, the disease is acquired upon traumatic inoculation of Sporothrix propagules from contaminated soil and plant debris. In addition, the direct horizontal transmission of Sporothrix among animals and the resulting zoonotic infection in humans highlight an alternative and efficient rout of transmission through biting and scratching. Sporothrix brasiliensis is the most virulent species of the Sporothrix schenckii complex and is responsible for the long-lasting outbreak of feline sporotrichosis in Brazil. However, antifungal susceptibility data of animal-borne isolates is scarce. Therefore, this study evaluated the in vitro activity of amphotericin B, caspofungin, itraconazole, voriconazole, fluconazole, and ketoconazole against animal-borne isolates of S. brasiliensis. The susceptibility tests were performed through broth microdilution (M38-A2). The results show the relevant activity of itraconazole, amphotericin B, and ketoconazole against S. brasiliensis, with the following MIC ranges: 0.125-2, 0.125-4 and 0.0312-2 μg/ml, respectively. Caspofungin was moderately effective, displaying higher variation in MIC values (0.25-64 μg/ml). Voriconazole (2-64 μg/ml) and fluconazole (62.5-500 μg/ml) showed low activity against S. brasiliensis strains. This study contributed to the characterization of the in vitro antifungal susceptibility of strains of S. brasiliensis recovered from cats with sporotrichosis, which have recently been considered the main source of human infections.

  2. Antifungal Susceptibility Patterns of Candida Species Recovered from Endotracheal Tube in an Intensive Care Unit.

    Science.gov (United States)

    Baghdadi, Elham; Khodavaisy, Sadegh; Rezaie, Sassan; Abolghasem, Sara; Kiasat, Neda; Salehi, Zahra; Sharifynia, Somayeh; Aala, Farzad

    2016-01-01

    Aims. Biofilms formed by Candida species which associated with drastically enhanced resistance against most antimicrobial agents. The aim of this study was to identify and determine the antifungal susceptibility pattern of Candida species isolated from endotracheal tubes from ICU patients. Methods. One hundred forty ICU patients with tracheal tubes who were intubated and mechanically ventilated were surveyed for endotracheal tube biofilms. Samples were processed for quantitative microbial culture. Yeast isolates were identified to the species level based on morphological characteristics and their identity was confirmed by PCR-RFLP. Antifungal susceptibility testing was determined according to CLSI document (M27-A3). Results. Ninety-five strains of Candida were obtained from endotracheal tubes of which C. albicans (n = 34; 35.7%) was the most frequently isolated species followed by other species which included C. glabrata (n = 24; 25.2%), C. parapsilosis (n = 16; 16.8%), C. tropicalis (n = 12; 12.6%), and C. krusei (n = 9; 9.4%). The resulting MIC90 for all Candida species were in increasing order as follows: caspofungin (0.5 μg/mL); amphotericin B (2 μg/mL); voriconazole (8.8 μg/mL); itraconazole (16 μg/mL); and fluconazole (64 μg/mL). Conclusion. Candida species recovered from endotracheal tube are the most susceptible to caspofungin.

  3. Antifungal Susceptibility Patterns of Candida Species Recovered from Endotracheal Tube in an Intensive Care Unit

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    Elham Baghdadi

    2016-01-01

    Full Text Available Aims. Biofilms formed by Candida species which associated with drastically enhanced resistance against most antimicrobial agents. The aim of this study was to identify and determine the antifungal susceptibility pattern of Candida species isolated from endotracheal tubes from ICU patients. Methods. One hundred forty ICU patients with tracheal tubes who were intubated and mechanically ventilated were surveyed for endotracheal tube biofilms. Samples were processed for quantitative microbial culture. Yeast isolates were identified to the species level based on morphological characteristics and their identity was confirmed by PCR-RFLP. Antifungal susceptibility testing was determined according to CLSI document (M27-A3. Results. Ninety-five strains of Candida were obtained from endotracheal tubes of which C. albicans (n=34; 35.7% was the most frequently isolated species followed by other species which included C. glabrata (n=24; 25.2%, C. parapsilosis (n=16; 16.8%, C. tropicalis (n=12; 12.6%, and C. krusei (n=9; 9.4%. The resulting MIC90 for all Candida species were in increasing order as follows: caspofungin (0.5 μg/mL; amphotericin B (2 μg/mL; voriconazole (8.8 μg/mL; itraconazole (16 μg/mL; and fluconazole (64 μg/mL. Conclusion. Candida species recovered from endotracheal tube are the most susceptible to caspofungin.

  4. In vitro antifungal susceptibility profiles of Cryptococcus species isolated from HIV-associated cryptococcal meningitis patients in Zimbabwe.

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    Nyazika, Tinashe K; Herkert, Patricia F; Hagen, Ferry; Mateveke, Kudzanai; Robertson, Valerie J; Meis, Jacques F

    2016-11-01

    Cryptococcus neoformans is the leading cause of cryptococcosis in HIV-infected subjects worldwide. Treatment of cryptococcosis is based on amphotericin B, flucytosine, and fluconazole. In Zimbabwe, little is known about antifungal susceptibility of Cryptococcus. Sixty-eight genotyped Cryptococcus isolates were tested for antifungal profiles. Amphotericin B, isavuconazole, and voriconazole showed higher activity than other triazoles. Fluconazole and flucytosine were less effective, with geometric mean MICs of 2.24 and 2.67mg/L for C. neoformans AFLP1/VNI, 1.38 and 1.53mg/L for C. neoformans AFLP1A/VNB/VNII and AFLP1B/VNII, and 1.85 and 0.68mg/L for Cryptococcus tetragattii, respectively. A significant difference between flucytosine geometric mean MICs of C. neoformans and C. tetragattii was observed (P=0.0002). The majority of isolates (n=66/68; 97.1%) had a wild-type MIC phenotype of all antifungal agents. This study demonstrates a favorable situation with respect to the tested antifungals agents. Continued surveillance of antifungal susceptibility profiles is important due to the high burden of cryptococcosis in Africa.

  5. Comparison of the Vitek 2 yeast susceptibility system with CLSI microdilution for antifungal susceptibility testing of fluconazole and voriconazole against Candida spp., using new clinical breakpoints and epidemiological cutoff values.

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    Pfaller, Michael A; Diekema, Daniel J; Procop, Gary W; Rinaldi, Michael G

    2013-09-01

    A commercially available, fully automated yeast susceptibility test system (Vitek 2; bioMérieux, Marcy d'Etoile, France) was compared in 3 different laboratories with the Clinical and Laboratory Standards Institute (CLSI) reference microdilution (BMD) method by testing 2 quality control strains, 10 reproducibility strains, and 425 isolates of Candida spp. against fluconazole and voriconazole. Reference CLSI BMD MIC endpoints and Vitek 2 MIC endpoints were read after 24 hours and 9.1-27.1 hours incubation, respectively. Excellent essential agreement (within 2 dilutions) between the reference and Vitek 2 MICs was observed for fluconazole (97.9%) and voriconazole (96.7%). Categorical agreement (CA) between the 2 methods was assessed using the new species-specific clinical breakpoints (CBPs): susceptible (S) ≤2 μg/mL, susceptible dose-dependent (SDD) 4 μg/mL, and resistant (R) ≥8 μg/mL for fluconazole and Candida albicans, Candida tropicalis, and Candida parapsilosis and ≤32 μg/mL (SDD), ≥64 μg/mL (R) for Candida glabrata; S ≤0.12 μg/mL, SDD 0.25-0.5 μg/mL, R ≥1 μg/mL for voriconazole and C. albicans, C. tropicalis, and C. parapsilosis, and ≤0.5 μg/mL (S), 1 μg/mL (SDD), ≥2 μg/mL (R) for Candida krusei. The epidemiological cutoff value (ECV) of 0.5 μg/mL for voriconazole and C. glabrata was used to differentiate wild-type (WT; MIC ≤ ECV) from non-WT (MIC > ECV) strains of this species. Due to the lack of CBPs for the less common species, the ECVs for fluconazole and voriconazole, respectively, were used for Candida lusitaniae (2 μg/mL and 0.03 μg/mL), Candida dubliniensis (0.5 μg/mL and 0.03 μg/mL), Candida guilliermondii (8 μg/mL and 0.25 μg/mL), and Candida pelliculosa (4 μg/mL and 0.25 μg/mL) to categorize isolates of these species as WT and non-WT. CA between the 2 methods was 96.8% for fluconazole and 96.5% for voriconazole with less than 1% very major errors and 1.3-3.0% major errors. The Vitek 2 yeast susceptibility system

  6. First description of Candida nivariensis in Brazil: antifungal susceptibility profile and potential virulence attributes

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    Figueiredo-Carvalho, Maria Helena Galdino; Ramos, Livia de Souza; Barbedo, Leonardo Silva; Chaves, Alessandra Leal da Silva; Muramoto, Ilda Akemi; dos Santos, André Luis Souza; Almeida-Paes, Rodrigo; Zancopé-Oliveira, Rosely Maria

    2016-01-01

    This study evaluated the antifungal susceptibility profile and the production of potential virulence attributes in a clinical strain of Candida nivariensis for the first time in Brazil, as identified by sequencing the internal transcribed spacer (ITS)1-5.8S-ITS2 region and D1/D2 domains of the 28S of the rDNA. For comparative purposes, tests were also performed with reference strains. All strains presented low planktonic minimal inhibitory concentrations (PMICs) to amphotericin B (AMB), caspofungin (CAS), and voriconazole. However, our strain showed elevated planktonic MICs to posaconazole (POS) and itraconazole, in addition to fluconazole resistance. Adherence to inert surfaces was conducted onto glass and polystyrene. The biofilm formation and antifungal susceptibility on biofilm-growing cells were evaluated by crystal violet staining and a XTT reduction assay. All fungal strains were able to bind both tested surfaces and form biofilm, with a binding preference to polystyrene (p < 0.001). AMB promoted significant reductions (≈50%) in biofilm production by our C. nivariensis strain using both methodologies. This reduction was also observed for CAS and POS, but only in the XTT assay. All strains were excellent protease producers and moderate phytase producers, but lipases were not detected. This study reinforces the pathogenic potential of C. nivariensis and its possible resistance profile to the azolic drugs generally used for candidiasis management. PMID:26814644

  7. First description of Candida nivariensis in Brazil: antifungal susceptibility profile and potential virulence attributes

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    Maria Helena Galdino Figueiredo-Carvalho

    2016-01-01

    Full Text Available This study evaluated the antifungal susceptibility profile and the production of potential virulence attributes in a clinical strain of Candida nivariensis for the first time in Brazil, as identified by sequencing the internal transcribed spacer (ITS1-5.8S-ITS2 region and D1/D2 domains of the 28S of the rDNA. For comparative purposes, tests were also performed with reference strains. All strains presented low planktonic minimal inhibitory concentrations (PMICs to amphotericin B (AMB, caspofungin (CAS, and voriconazole. However, our strain showed elevated planktonic MICs to posaconazole (POS and itraconazole, in addition to fluconazole resistance. Adherence to inert surfaces was conducted onto glass and polystyrene. The biofilm formation and antifungal susceptibility on biofilm-growing cells were evaluated by crystal violet staining and a XTT reduction assay. All fungal strains were able to bind both tested surfaces and form biofilm, with a binding preference to polystyrene (p < 0.001. AMB promoted significant reductions (≈50% in biofilm production by our C. nivariensis strain using both methodologies. This reduction was also observed for CAS and POS, but only in the XTT assay. All strains were excellent protease producers and moderate phytase producers, but lipases were not detected. This study reinforces the pathogenic potential of C. nivariensis and its possible resistance profile to the azolic drugs generally used for candidiasis management.

  8. Susceptibility pattern of Malassezia species to selected plant extracts and antifungal agents

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    G Sibi

    2014-01-01

    Full Text Available Objective: Malassezia is associated with dandruff, seborrhoeic dermatitis, pityriasis versicolor folliculitis and atopic eczema. This study determined the susceptibility pattern of Malassezia furfur, M. globosa, M. obtusa, M. restricta, M. slooffiae and M. sympodialis isolated from patients diagnosed with dandruff against plant extracts and antifungal agents. Materials and Methods: Twenty aqueous plant extracts and five azole drugs were tested against the isolates by well diffusion and broth dilution method. Results: Among the plant extracts, Phyllanthus emblica (fruits, Hibiscus rosa sinensis (flowers and Acacia concinna (pods have demonstrated significant antidandruff activity. Minimum inhibitory concentration values revealed that ketoconazole as the most effective drug followed by itraconazole. Conclusion: M. furfur and M. globosa were found as the most susceptible organisms against the aqueous extracts of Phyllanthus emblica (fruits, Hibiscus rosa sinensis (flowers, Acacia concinna (pods and azole drugs.

  9. The clinical distribution and antifungal susceptibility tests of Candida species in urinary infection%泌尿系统感染假丝酵母菌属分布与药敏分析

    Institute of Scientific and Technical Information of China (English)

    王宏伟; 许俊华; 孙美玲; 李慧

    2015-01-01

    OBJECTIVE To investigate the characteristics ,distribution of fungi such as Candida in urine samples from patients with urinary infections and antifungal susceptibility ,so as to improve the level of clinical treatment . METHODS A retrospective analysis was conducted on 242 strains of fungi isolated and identified from urine sam‐ples from inpatients during Dec .1st ,2009 to Nov .30th ,2012 .The constituent ratio and the department distribu‐tion of strain species were calculated and the drug susceptibility of main pathogens were summarized and analyzed . The software SPSS17 .0 was used for statistical analysis .RESULTS Totally 242 fungi were detected ,mainly inclu‐ding Candida albicans (85 strains ,35 .1% ) .The pathogens were mainly distributed in the integrated ICU (80 strains ,33 .1% ) .C .albicans ,Candida tropicalis and Candida glabrata were sensitive to amphotericin B with the sensitive rate of 100 .0% .CONCLUSION The incidence of Candida urinary infections increases along with the increased survival rate of immunocompromised patients due to population aging and a variety of medical interven‐tion ,which should be paid much attention to by clinicians .Early antifungal susceptibility test and selection of anti‐fungal drugs accordingly can be helpful to control urinary infection caused by these fungi .%目的:研究泌尿系统感染患者尿液标本分离的假丝酵母菌属等真菌的感染特点、分布及其对抗真菌药物敏感性,为临床治疗提供参考依据。方法回顾性分析2009年12月1日-2012年11月30日住院患者尿液标本中分离鉴定242株真菌,计算菌种构成比、科室分离率,统计分析主要致病真菌的药敏结果,采用SPSS17.0软件进行统计分析。结果共检测出242株真菌,以白色假丝酵母为主,共85株35.1%;其分布科室以综合IC U为主,共80株占33.1%;白色假丝酵母菌、热带假丝酵母菌、光滑假丝酵母菌对两性霉素 B

  10. Molecular identification and antifungal susceptibility profiles of Candida parapsilosis complex species isolated from culture collection of clinical samples

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    Fábio Silvestre Ataides

    2015-08-01

    Full Text Available AbstractINTRODUCTION:Candida parapsilosis is a common yeast species found in cases of onychomycosis and candidemia associated with infected intravascular devices. In this study, we differentiated Candida parapsilosis sensu stricto, Candida orthopsilosis , and Candida metapsilosis from a culture collection containing blood and subungual scraping samples. Furthermore, we assessed the in vitro antifungal susceptibility of these species to fluconazole, itraconazole, voriconazole, posaconazole, amphotericin B, and caspofungin.METHODS:Differentiation of C. parapsilosis complex species was performed by amplification of the secondary alcohol dehydrogenase (SADH gene and digestion by the restriction enzyme Ban I. All isolates were evaluated for the determination of minimal inhibitory concentrations using Etest, a method for antifungal susceptibility testing.RESULTS:Among the 87 isolates, 78 (89.7% were identified as C. parapsilosis sensu stricto , five (5.7% were identified as C. orthopsilosis , and four (4.6% were identified as C. metapsilosis . Analysis of antifungal susceptibility showed that C. parapsilosis sensu strictoisolates were less susceptible to amphotericin B and itraconazole. One C. parapsilosis sensu stricto isolate was resistant to amphotericin B and itraconazole. Moreover, 10.2% of C. parapsilosis sensu stricto isolates were resistant to caspofungin. Two C. parapsilosis sensu strictoisolates and one C. metapsilosis isolate were susceptible to fluconazole in a dose-dependent manner.CONCLUSIONS:We reported the first molecular identification of C. parapsilosiscomplex species in State of Goiás, Brazil. Additionally, we showed that although the three species exhibited differences in antifungal susceptibility profiles, the primary susceptibility of this species was to caspofungin.

  11. Prevalence and antifungal susceptibility of Candida parapsilosis complex isolates collected from oral cavities of HIV-infected individuals.

    Science.gov (United States)

    Moris, D V; Melhem, M S C; Martins, M A; Souza, L R; Kacew, S; Szeszs, M W; Carvalho, L R; Pimenta-Rodrigues, M V; Berghs, H A M; Mendes, R P

    2012-12-01

    At present, few data are available on the prevalence and antifungal susceptibility of Candida parapsilosis complex isolates from HIV-infected individuals. The C. parapsilosis complex comprises three species, C. parapsilosis sensu stricto, C. metapsilosis and C. orthopsilosis. Fifteen of 318 Candida isolates were identified as members of the C. parapsilosis complex by PCR and restriction fragment length polymorphism (RFLP). The prevalence of C. parapsilosis complex isolates was 4.7 %, 2.2 % being identified as C. parapsilosis sensu stricto and 2.5 % as C. metapsilosis, while no C. orthopsilosis was isolated. This is believed to be the first study that has identified isolates of C. metapsilosis obtained from the oral cavity of HIV-infected individuals. Antifungal susceptibility tests indicated that all the isolates were susceptible to amphotericin B (AMB), fluconazole (FLC), ketoconazole (KTC), itraconazole (ITC), voriconazole (VRC) and caspofungin (CASPO). Although isolates of C. parapsilosis sensu stricto and C. metapsilosis were susceptible to FLC, isolates of C. metapsilosis showed a tendency for higher MICs (≥1.0 µg ml(-1)). Based upon the frequency of candidiasis and the fact that certain isolates of the C. parapsilosis complex respond differently to FLC therapy, our data may be of therapeutic relevance with respect to susceptibility and potential resistance to specific antifungal agents. Our data suggest that C. metapsilosis can be a human commensal; its importance as a pathogen has yet to be confirmed.

  12. Species distribution and in vitro antifungal susceptibility of oral yeast isolates from Tanzanian HIV-infected patients with primary and recurrent oropharyngeal candidiasis

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    Rijs Antonius JMM

    2008-08-01

    Full Text Available Abstract Background In Tanzania, little is known on the species distribution and antifungal susceptibility profiles of yeast isolates from HIV-infected patients with primary and recurrent oropharyngeal candidiasis. Methods A total of 296 clinical oral yeasts were isolated from 292 HIV-infected patients with oropharyngeal candidiasis at the Muhimbili National Hospital, Dar es Salaam, Tanzania. Identification of the yeasts was performed using standard phenotypic methods. Antifungal susceptibility to fluconazole, itraconazole, miconazole, clotrimazole, amphotericin B and nystatin was assessed using a broth microdilution format according to the guidelines of the Clinical and Laboratory Standard Institute (CLSI; M27-A2. Results Candida albicans was the most frequently isolated species from 250 (84.5% patients followed by C. glabrata from 20 (6.8% patients, and C. krusei from 10 (3.4% patients. There was no observed significant difference in species distribution between patients with primary and recurrent oropharyngeal candidiasis, but isolates cultured from patients previously treated were significantly less susceptible to the azole compounds compared to those cultured from antifungal naïve patients. Conclusion C. albicans was the most frequently isolated species from patients with oropharyngeal candidiasis. Oral yeast isolates from Tanzania had high level susceptibility to the antifungal agents tested. Recurrent oropharyngeal candidiasis and previous antifungal therapy significantly correlated with reduced susceptibility to azoles antifungal agents.

  13. Genetic diversity and antifungal susceptibility testing of Trichosporon asahii isolated of Intensive Care Units patients Diversidade genética e suscetibilidade a antifúngicos de Trichosporon asahii isolado de pacientes de Unidades de Terapia Intensiva

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    Rosana Bellan de Oliveira Silva

    2008-09-01

    Full Text Available Trichosporon asahii is an opportunistic pathogen, associated with a high mortality rate in immunocompromised patients. In this study, ten isolates, recovered from oral cavity and urine of patients in Intensive Care Units (ICU over six months, were identified by classical and molecular methods, typed by RAPD and tested in vitro for susceptibility to fluconazole, itraconazole, 5-flucytosine and amphotericin B. A total agreement between the identification of Trichosporon sp by PCR based on sequences of the Internal Transcribed Spacer Regions (ITS and on the sequences of small-subunit (SSU ribosomal DNA (rDNA was found. Randomly amplified of polymorphic DNA (RAPD, with primers P6 and M13, was used to determine the genomic profiles. The dendogram analysis indicated that almost all strains showed similarity >0.9 among them and all strains were multidrug-resistant. This study brings new results on the identification and genotyping of T. asahii isolated from Brazilian ICU patients and information about their antifungal drugs susceptibility.Trichosporon asahii é um patógeno oportunista que apresenta altos índices de mortalidade em pacientes imunocomprometidos. No presente trabalho, dez cepas foram isoladas da cavidade bucal e urina de pacientes internados na Unidade de Terapia Intensiva (UTI por seis meses. Todos os isolados foram identificados por métodos clássicos e moleculares, tipados por RAPD e testados in vitro quanto à sensibilidade ao fluconazol, itraconazol, 5-fluorocitosina e anfotericina B. Houve concordância total entre a identificação de Trichosporon sp por PCR baseado na seqüência da região ITS (Internal Transcribed Spacer e na seqüência da subunidade menor do DNA ribossômico (rDNA. Os perfis genéticos foram determinados por RAPD utilizando dois iniciadores P6 e M13. A análise do dendrograma mostrou que a maioria das amostras apresentou alta similaridade entre elas (>0.9 e todas foram multidrogas resistentes. Este estudo

  14. Prevalence, distribution and antifungal susceptibility profiles of Candida parapsilosis, Candida orthopsilosis and Candida metapsilosis bloodstream isolates.

    Science.gov (United States)

    Bonfietti, Lucas Xavier; Martins, Marilena dos Anjos; Szeszs, Maria Walderez; Pukiskas, Sandra Brasil Stolf; Purisco, Sonia Ueda; Pimentel, Fabiana Cortez; Pereira, Graziella Hanna; Silva, Dayane Cristina; Oliveira, Lidiane; Melhem, Marcia de Souza Carvalho

    2012-07-01

    The Candida parapsilosis group encompasses three species: C. parapsilosis, Candida orthopsilosis and Candida metapsilosis. These species are phenotypically indistinguishable, and molecular methods are needed for their detection. We analysed 152 unique blood culture isolates of the C. parapsilosis group obtained during 1997-2011. The isolates were screened by PCR amplification of the gene encoding secondary alcohol dehydrogenase, followed by digestion with the restriction enzyme BanI. Isolates with RFLP patterns distinct from those of the C. parapsilosis group were characterized as C. parapsilosis sensu stricto (90.8 %), C. orthopsilosis (8.6 %) and C. metapsilosis (0.6 %). Antifungal susceptibility tests indicated that all isolates were susceptible to itraconazole, amphotericin B and caspofungin. Although C. orthopsilosis and C. metapsilosis isolates were susceptible to fluconazole, higher MICs (≥2 mg l(-1)) were observed for C. orthopsilosis. Three isolates (2.0 %) of C. parapsilosis sensu stricto were resistant to voriconazole. Five C. parapsilosis isolates (3.3 %) were intermediate, and a single isolate (0.7 %) was resistant (MIC 16 mg l(-1)) to fluconazole. These data were confirmed using reference strains. It was observed that C. parapsilosis isolates were less susceptible to all triazoles, and this finding deserves further attention to assess the appearance of cross-resistance phenomena. In conclusion, C. metapsilosis and C. orthopsilosis are involved in a small but significant number of invasive infections in Brazil.

  15. Speciation and antifungal susceptibility of esophageal candidiasis in cancer patients in a tertiary care hospital in South India

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    J. Abirami Lakshmy

    2016-01-01

    Full Text Available Esophageal candidiasis is the most common opportunistic infection in patients with altered immunity such as Human Immunodeficiency Virus (HIV infection, cancer patients on chemotherapy and radiotherapy. Neutropenia, irradiation and chemotherapy will facilitate deeper mucosal invasion leading to esophageal candidiasis. Empirical treatment of esophageal candidiasis without antifungal susceptibility testing will lead to the emergence of drug resistant species increasing the morbidity and mortality associated with cancer. The present study aimed to study the frequency of esophageal candida in individuals with cancer, species level identification and antifungal susceptibility pattern. Scrapings of whitish appearing lesions were obtained from a total of thirty five cases of endoscopically identified esophageal candidiasis were obtained from cancer patients. Identification of the Candida isolates were done by cultivation in Sabouraud dextrose agar (SDA, Gram staining, germ tube test, colony morphology in Chrom agar and corn meal agar, sugar assimilation and fermentation tests. Antifungal susceptibility was done by Microbroth dilution method for Fluconazole, Itraconazole and Amphotericin B. We found that Candida albicans was the predominant species isolated followed by Candida tropicalis and Candida glabrata. Sensitivity rates were 94%, 96% and 100% for Fluconazole, Itraconazole and Amphotericin B. Species level identification of Candida isolated from esophageal candidiasis and their antifungal sensitivity testing should be performed for early identification of resistant strains and for promptly treating the cases there by preventing the dissemination of infection in case of immune-compromised individuals. Further the susceptibility pattern will facilitate therapeutic guidance especially in individuals prone to relapse. [J Med Allied Sci 2016; 6(1: 29-34

  16. In vitro antifungal susceptibility of Malassezia pachydermatis strains isolated from dogs with chronic and acute otitis externa.

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    Chiavassa, E; Tizzani, P; Peano, A

    2014-10-01

    Malassezia pachydermatis is a yeast that is frequently involved as a secondary/perpetuating factor in canine otitis externa. Topical therapies with different antifungal agents, mainly azole compounds, are generally successful in controlling the yeast overgrowth, but treatment failure and rapid recurrences are common. This study compared the in vitro antifungal susceptibility of M. pachydermatis isolates obtained from chronic and acute cases of otitis externa. The aim was to assess the possible onset of resistance mechanisms in isolates involved in long-lasting episodes with poor response to treatment. We evaluated the in vitro susceptibility to miconazole (MCZ) and clotrimazole (CTZ) of 42 isolates of M. pachydermatis obtained from dogs with chronic (group A, n = 25) and acute otitis (group B, n = 17), using a modified CLSI M27-A3 microdilution method. All isolates were inhibited by the antifungal agents employed, but Malassezia isolates from group A were significantly associated with higher minimum inhibitory concentration (MIC) values for both agents (Median MIC values: MCZ group A 2 µg/ml, group B 1 µg/ml; CTZ group A 8 µg/ml, group B 4 µg/ml). These findings prove that these isolates had a reduced in vitro susceptibility to the antifungal agents tested. However, it is unlikely that this could have any influence on the outcome of a topical treatment. Indeed, marketed products include concentrations of the tested agents that largely exceed even the highest MICs found in this study (in most cases at least 1,000 × the MIC, or greater). In conclusion, this study suggests that isolates of M. pachydermatis involved in chronic cases of canine external otitis and exposed to repeated antifungal treatments are unlikely to develop mechanisms of resistance of clinical relevance.

  17. [Neonatal Candida infections and the antifungal susceptibilities of the related Candida species].

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    Altuncu, Emel; Bilgen, Hülya; Cerikçioğlu, Nilgün; Ilki, Arzu; Ulger, Nurver; Bakır, Mustafa; Akman, Ipek; Ozek, Eren

    2010-10-01

    Among nosocomial infections in the newborns, the incidence of fungal infections has been rising over the last decades. Fluconazole has been a new option for treatment however, expanded use of the drug brought up the development of resistance. In this study, species of the Candida isolates from neonates with candida infections, their antifungal susceptibilities and the effectiveness of the therapy were evaluated. All the species of Candida isolates from blood, urine and sterile body fluids of 54 neonates and their antifungal susceptibilities were evaluated retrospectively over the 13-year period. Demographic characteristics, risk factors, infection foci, Candida species causing infection and their in vitro susceptibilities for fluconazole (FCZ) and amphotericin B (AMB) and treatment responses were analyzed. The antifungal susceptibility testing of isolates was performed by microdilution technique. The median birth weight and gestational age of the study groups were 1735 (660-3990) g and 33 (24-40) weeks, respectively. Among the patients, 19 (35%) were term, while 35 (65%) were preterm [Candida spp. were isolated mostly from blood samples (63%), followed by urine (46%), cerebrospinal fluid (CSF; 5%), peritoneal fluid (3%) and endotracheal aspirate (2%). Multifocal growth was determined in 10 (18%) cases. The isolated species were C.albicans (n =36) as being the most common isolate followed by C.parapsilosis (n = 12), C.tropicalis (n = 1), C.kefyr (n = 1), C.lusitaniae (n = 1), C.pelluculosa (n = 1) and Candida spp. (n = 2). Prior antibiotic use, long term hospitalization, total parenteral nutrition and use of lipid solutions, prematurity and catheter use were determined as the most frequently associated factors causing candidal infections. A congenital abnormality, mainly myeloschisis and hydrocephaly, was detected in 18 (33%) of the cases. Overall FCZ resistance rate was 5.5% and the rate of resistance according to the species was 2.8% for C.albicans and 11% for non

  18. Environmental isolation, biochemical identification, and antifungal drug susceptibility of Cryptococcus species

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    Valter Luis Iost Teodoro

    2013-12-01

    Full Text Available Introduction The incidence of opportunistic fungal infections has increased in recent years and is considered an important public health problem. Among systemic and opportunistic mycoses, cryptococcosis is distinguished by its clinical importance due to the increased risk of infection in individuals infected by human immunodeficiency virus. Methods To determine the occurrence of pathogenic Cryptococcus in pigeon excrement in the City of Araraquara, samples were collected from nine environments, including state and municipal schools, abandoned buildings, parks, and a hospital. The isolates were identified using classical tests, and susceptibility testing for the antifungal drugs (fluconazole, itraconazole, voriconazole, and amphotericin B independently was also performed. After collection, the excrement samples were plated on Niger agar and incubated at room temperature. Results A total of 87 bird dropping samples were collected, and 66.6% were positive for the genus Cryptococcus. The following species were identified: Cryptococcus neoformans (17.2%, Cryptococcus gattii (5.2%, Cryptococcus ater (3.5%, Cryptococcus laurentti (1.7%, and Cryptococcus luteolus (1.7%. A total of 70.7% of the isolates were not identified to the species level and are referred to as Cryptococcus spp. throughout the manuscript. Conclusions Although none of the isolates demonstrated resistance to antifungal drugs, the identification of infested areas, the proper control of birds, and the disinfection of these environments are essential for the epidemiological control of cryptococcosis.

  19. Biofilm production and antifungal susceptibility of co-cultured Malassezia pachydermatis and Candida parapsilosis isolated from canine seborrheic dermatitis.

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    Bumroongthai, K; Chetanachan, P; Niyomtham, W; Yurayart, C; Prapasarakul, N

    2016-07-01

    The yeasts Malassezia (M.) pachydermatis and Candida (C.) parapsilosis are often co-isolated in case of canine seborrhea dermatitis (SD) and also are emerging as opportunistic pathogens of immunocompromised human beings. Increased information about how their relationship results in biofilm production and an antifungal response would be useful to inform treatment and control. This study was designed to investigate biofilm production derived from co-culture of M. pachydermatis and C. parapsilosis from dog skin and to determine their in vitro antifungal susceptibility. We demonstrated that regardless of yeast strain or origin all single and dual cultures produced biofilms within 24 hours, and the greatest amount was present after 72 hours. Biofilm production from mixed cultures was greater than for single strains (P < .05). All sessile forms of the single and dual cultures were resistant to the tested antifungals itraconazole and ketoconazole, whereas planktonic forms were susceptible. The study suggests that dual cultures produce stronger biofilms that are likely to enhance persistence in skin lesions in dogs and result in greater resistance to antifungal treatment.

  20. Antifungal susceptibility and distribution of Candida spp. isolates from the University Hospital in the municipality of Dourados, State of Mato Grosso do Sul, Brazil

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    Adriana Araujo de Almeida

    2013-06-01

    Full Text Available Introduction Hospital infections caused by Candida spp. are a leading cause of morbidity and mortality in hospitalized patients, particularly those that are critically ill or immunocompromised. In this study, the distribution of Candida species in isolates from the University Hospital of the Federal University at Grande Dourados and their in vitro susceptibility to antifungal drugs were analyzed. Methods Yeasts were phenotypically identified using classical methodologies. Antifungal susceptibility tests to amphotericin B and fluconazole were performed using the broth microdilution technique. Results A total of 50 Candida isolates were obtained from hospitalized patients during the study period. We analyzed yeast isolates from urine (n=31; 62%, blood (n=12; 24%, and tracheal secretions (n=7; 14%. The following Candida species were identified: C. tropicalis (n=21; 42%, C. albicans (n=18; 36%, C. glabrata (n=10; 20%, and C. krusei (n=1; 2%. Antifungal susceptibility tests demonstrated that C. albicans was susceptible to both antifungal agents. However, 31.2% of the non-C. albicans Candida isolates displayed dose-dependent susceptibility to fluconazole, and 3.1% were resistant to amphotericin B. Conclusions In contrast to previous reports, our results indicated that C. tropicalis was the most commonly isolated yeast species among the hospital patients. The predominance of non-C. albicans Candida infections confirms the importance of species-level identification for implementing appropriate antifungal therapies.

  1. Species Distribution, Virulence Factors, and Antifungal Susceptibility Among Candida parapsilosis Complex Isolates Recovered from Clinical Specimens.

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    da Silva, Beatriz Virgínia; Silva, Larissa Beatriz; de Oliveira, Diego Batista Carneiro; da Silva, Paulo Roberto; Ferreira-Paim, Kennio; Andrade-Silva, Leonardo Euripides; Silva-Vergara, Mario León; Andrade, Anderson Assunção

    2015-12-01

    The Candida parapsilosis complex has emerged as an important fungal pathogen. In spite of this, relatively little is known about its characteristics. Thus, the purposes of this study were (1) to determine by BanI-RFLP-assay the occurrence of C. parapsilosis complex species among 81 clinical isolates primarily identified as C. parapsilosis; (2) to evaluate their in vitro production of virulence factors; and (3) to compare their susceptibility profiles, grown as planktonic cells and biofilms, against amphotericin B, fluconazole, voriconazole, and caspofungin by following the Clinical and Laboratory Standards Institute (CLSI) guidelines. Seventy-seven isolates (95%) were identified as C. parapsilosis sensu stricto, 2 (2.5%) as C. orthopsilosis, and 2 (2.5%) as C. metapsilosis. Protease activity was detected in 29 (37.7%) isolates of C. parapsilosis sensu stricto, whereas only 7 (9.1%) exhibited phospholipase activity. None of the C. metapsilosis or C. orthopsilosis was able to produce protease or phospholipase. Biofilm production was detected in 35 (43.2%) isolates, among which 33 were C. parapsilosis sensu stricto and 2 were C. orthopsilosis. Antifungal resistance was uncommon; only one C. metapsilosis was fluconazole resistant. However, biofilm-producing isolates showed a marked resistance to all antifungal agents tested, particularly to voriconazole. This knowledge could be of clinical relevance for guiding therapeutic decisions.

  2. Antifungal Susceptibility Analysis of Clinical Isolates of Candida parapsilosis in Iran

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    Ensieh LOTFALI

    2016-03-01

    Full Text Available Background: Candida parapsilosis is an emergent agent of invasive fungal infections. This yeast is one of the five most widespread yeasts concerned in invasive candidiasis.C. parapsilosis stands out as the second most common yeast species isolated from patients with bloodstream infections especially in neonates with catheter.Recently several reports suggested that its reduced susceptibility to azoles and polyene might become a cause for clinical concern, although C. parapsilosis is not believed to be intensely prone to the development of antifungal resistance.Methods: In the present report, One hundred and twenty clinical isolates of C. parapsilosis complex were identified and differentiated by using PCR-RFLP analysis. The isolates were then analyzed to determine their susceptibility profile to fluconazole (FLU, itraconazole (ITC and amphotericin B. The minimum inhibitory concentration (MIC results were analyzed according to the standard CLSI guide.Results: All of isolates were identified as C. parapsilosis. No C. metapsilosis and C. orthopsilosis strains were found. Evaluation of the antifungal susceptibility profile showed that only three (2.5% C. parapsilosis were resistant to fluconazole, three (2.5% C. parapsilosis were resistant to itraconazole and two (1.7% C. parapsilosis were amphotericin B resistant.Conclusion: Profiles in clinical isolates of C. parapsilosis can provide important information for the control of antifungal resistance as well as distribution and susceptibility profiles in populations. Keywords: Candida parapsilosis, Antifungal susceptibility, Resistant, Iran

  3. In vitro susceptibility testing of dermatophytes isolated from pediatric cases in Nigeria against five antifungals Teste de susceptibilidade in vitro de dermatófitos isolados de casos pediátricos na Nigéria contra cinco antifúngicos

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    E.I. Nweze

    2007-10-01

    Full Text Available The antifungal activities of itraconazole, ketoconazole, fluconazole, terbinafine and griseofulvin were tested by broth microdilution methods against 71 isolates of dermatophytes isolated from Nigerian children. Most drugs were very active against all the dermatophytes and the MIC 90 ranged from 0.03 to 8.0 µg/mL. This appears to be the first documented data on the antifungal susceptibility testing of isolates of dermatophytes from Nigerian children.Atividades antifúngicas de itraconazole, ketoconazole, fluconazole, terbinafine e griseofulvina foram testadas por métodos de microdiluição em caldo contra 71 isolados de dermatófitos de crianças nigerianas. A maioria das drogas foi muito ativa contra todos os dermatófitos e o MIC 90 variou de 0,03 a 8,0 µg/mL. Estes parecem ser os primeiros dados documentados sobre os testes de susceptibilidade antifúngica de isolados de dermatófitos de crianças nigerianas.

  4. Subgingival distribution of yeast and their antifungal susceptibility in immunocompetent subjects with and without dental devices.

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    Jewtuchowicz, Virginia M; Brusca, Maria I; Mujica, Maria T; Gliosca, Laura A; Finquelievich, Jorge L; Lovannitti, Cristina A; Rosa, Alcira C

    2007-01-01

    Yeasts colonize the subgingival biofilm, which becomes a reservoir that favors their reproduction. The purpose of the present work was to determine the prevalence of yeasts of the Candida genus in the subgingival biofilm of gingivoperiodontal disease patients, including users and non-users of dental devices, and their susceptibility to fluconazole and voriconazole. Samples of subgingival pockets of immunocompetent nonsmokers showing gingivitis and periodontitis were inoculated in a differential chromogenic medium. Sixty three percent of subjects used dental devices. Yeasts were identified and susceptibility to fluconazole and voriconazole was tested following CLSI M44-A standards. The prevalence of yeasts in the subgingival biofilm was 40% CI 95% (30.5-50.3); 10% were patients who did not use dental appliances. The most frequently observed yeasts were C. albicans, and C. parapsilosis, C. dubliniensis, C. tropicalis and C. guilliermondii. Only C. dubliniensis and C. guilliermondii showed resistance to azoles. The use of dental devices significantly increased the prevalence of yeasts in periodontal pockets inpatients presenting gingivitis. It is noteworthy that non albicans Candida species, such as C. dubliniensis and C. guilliermondii, considered emerging species, which have a diminished susceptibility to antifungal agents were found in the crevicular fluid of immunocompetent patients.

  5. Distribution, virulence attributes and antifungal susceptibility patterns of Candida parapsilosis complex strains isolated from clinical samples.

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    Tosun, Ilknur; Akyuz, Zeynep; Guler, Nejla Cebeci; Gulmez, Dolunay; Bayramoglu, Gulcin; Kaklikkaya, Nese; Arikan-Akdagli, Sevtap; Aydin, Faruk

    2013-07-01

    It was recently proposed that Candida parapsilosis represents a complex composed of three closely related species, i.e., C. parapsilosis sensu stricto, C. orthopsilosis, and C. metapsilosis. The aim of this study was to describe the distribution of C. parapsilosis complex isolates among clinical samples. We also evaluated antifungal susceptibility profiles, in vitro presence of lipase and secreted aspartyl proteinase, as well as their ability to grow in total parenteral nutrition (TPN) solution, and biofilm production. A total of 413 non-C. albicans Candida isolates were obtained from various clinical samples between 2010 and 2011 in a Turkish Tertiary Care Hospital. Of them, 42 were identified as members of the C. parapsilosis complex. Among these, 38 (90.5%) were C. parapsilosis sensu stricto, 3 (7.1%) C. metapsilosis, and 1 (2.4%) C. orthopsilosis. All isolates recovered from blood were found to be C. parapsilosis sensu stricto and C. metapsilosis. In phenotypic tests, all 42 isolates grew in TPN solution and, although 26.2% of C. parapsilosis sensu stricto-isolates were capable of forming biofilms in vitro, neither C. orthopsilosis nor C. metapsilosis isolates were able to do so. Acid proteinase activity was detected in 31% of isolates and lipase activity in 33%. All isolates were sensitive to voriconazole, caspofungin, and anidulafungin, with only a single C. parapsilosis sensu stricto isolate showing dose-dependent susceptible to fluconazole. While the number of C. metapsilosis and C. orthopsilosis isolates remained low, there were no significant differences in antifungal MIC as compared to C. parapsilosis sensu stricto.

  6. Susceptibility variation of Malassezia pachydermatis to antifungal agents according to isolate source

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    Caroline Borges Weiler

    2013-01-01

    Full Text Available Malassezia pachydermatisis associated with dermatomycoses and otomycosis in dogs and cats. This study compared the susceptibility of M. pachydermatis isolates from sick (G1 and healthy (G2 animals to azole and polyene antifungals using the M27-A3 protocol. Isolates from G1 animals were less sensitive to amphotericin B, nystatin, fluconazole, clotrimazole and miconazole.

  7. Prevalence, distribution, and antifungal susceptibility profiles of Candida parapsilosis, C. orthopsilosis, and C. metapsilosis in a tertiary care hospital.

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    Silva, Ana P; Miranda, Isabel M; Lisboa, Carmen; Pina-Vaz, Cidália; Rodrigues, Acácio G

    2009-08-01

    Candida parapsilosis, an emergent agent of nosocomial infections, was previously made up of a complex of three genetically distinct groups (groups I, II, and III). Recently, the C. parapsilosis groups have been renamed as distinct species: C. parapsilosis sensu stricto, C. orthopsilosis, and C. metapsilosis. In Portugal, no data pertaining to the distribution and antifungal susceptibility of these Candida species are yet available. In the present report, we describe the incidence and distribution of C. parapsilosis sensu stricto, C. orthopsilosis, and C. metapsilosis among 175 clinical and environmental isolates previously identified by conventional methods as C. parapsilosis. We also evaluated the in vitro susceptibilities of the isolates to fluconazole, voriconazole, posaconazole, amphotericin B, and two echinocandins, caspofungin and anidulafungin. Of the 175 isolates tested, 160 (91.4%) were identified as C. parapsilosis sensu stricto, 4 (2.3%) were identified as C. orthopsilosis, and 5 (2.9%) were identified as C. metapsilosis. Six isolates corresponded to species other than the C. parapsilosis group. Interestingly, all isolates from blood cultures corresponded to C. parapsilosis sensu stricto. Evaluation of the antifungal susceptibility profile showed that only nine (5.6%) C. parapsilosis sensu stricto strains were susceptible-dose dependent or resistant to fluconazole, and a single strain displayed a multiazole-resistant phenotype; two (1.3%) C. parapsilosis sensu stricto strains were amphotericin B resistant. All C. orthopsilosis and C. metapsilosis isolates were susceptible to azoles and amphotericin B. A high number of strains were nonsusceptible to the echinocandins (caspofungin and anidulafungin).

  8. Veronaea botryosa: molecular identification with amplified fragment length polymorphism (AFLP) and in vitro antifungal susceptibility.

    Science.gov (United States)

    Badali, Hamid; Yazdanparast, Seyed Amir; Bonifaz, Alexandro; Mousavi, Bita; de Hoog, G Sybren; Klaassen, Corné H W; Meis, Jacques F

    2013-06-01

    Inter- and intraspecific genomic variability of 18 isolates of Veronaea botryosa originating from clinical and environmental sources was studied using amplified fragment length polymorphism (AFLP). The species was originally described from the environment, but several severe cases of disseminated infection in apparently healthy individuals have been reported worldwide. All tested strains of V. botryosa, identified on the basis of sequencing and phenotypic and physiological criteria prior to our study, were confirmed by AFLP analysis, yielding a clear separation of V. botryosa as a rather homogeneous group from related species. In vitro antifungal susceptibility testing resulted in MIC90s across all strains in increasing order posaconazole (0.25 μg/ml), itraconazole (1 μg/ml), voriconazole (4 μg/ml), terbinafine (4 μg/ml), caspofungin (8 μg/ml), anidulafungin (8 μg/ml), isavuconazole (16 μg/ml), amphotericin B (16 μg/ml), and fluconazole (32 μg/ml). Overall, the isolates showed a uniform pattern of low MICs of itraconazole and posaconazole, but high MICs for remaining agents. The echinocandins (caspofungin and anidulafungin) had no activity against V. botryosa. There was no statistically significant difference between susceptibilities of environmental (n = 11) and clinical (n = 7) isolates of V. botryosa (P > 0.05).

  9. Antifungal Susceptibilities of Candida Species Causing Vulvovaginitis and Epidemiology of Recurrent Cases

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    Richter, Sandra S.; Galask, Rudolph P.; Messer, Shawn A.; Hollis, Richard J.; Diekema, Daniel J.; Pfaller, Michael A.

    2005-01-01

    There are limited data regarding the antifungal susceptibility of yeast causing vulvovaginal candidiasis, since cultures are rarely performed. Susceptibility testing was performed on vaginal yeast isolates collected from January 1998 to March 2001 from 429 patients with suspected vulvovaginal candidiasis. The charts of 84 patients with multiple positive cultures were reviewed. The 593 yeast isolates were Candida albicans (n = 420), Candida glabrata (n = 112), Candida parapsilosis (n = 30), Candida krusei (n = 12), Saccharomyces cerevisiae ( n = 9), Candida tropicalis (n = 8), Candida lusitaniae (n = 1), and Trichosporon sp. (n = 1). Multiple species suggesting mixed infection were isolated from 27 cultures. Resistance to fluconazole and flucytosine was observed infrequently (3.7% and 3.0%); 16.2% of isolates were resistant to itraconazole (MIC ≥ 1 μg/ml). The four imidazoles (econazole, clotrimazole, miconazole, and ketoconazole) were active: 94.3 to 98.5% were susceptible at ≤1 μg/ml. Among different species, elevated fluconazole MICs (≥16 μg/ml) were only observed in C. glabrata (15.2% resistant [R], 51.8% susceptible-dose dependent [S-DD]), C. parapsilosis (3.3% S-DD), S. cerevisiae (11.1% S-DD), and C. krusei (50% S-DD, 41.7% R, considered intrinsically fluconazole resistant). Resistance to itraconazole was observed among C. glabrata (74.1%), C. krusei (58.3%), S. cerevisiae (55.6%), and C. parapsilosis (3.4%). Among 84 patients with recurrent episodes, non-albicans species were more common (42% versus 20%). A ≥4-fold rise in fluconazole MIC was observed in only one patient with C. parapsilosis. These results support the use of azoles for empirical therapy of uncomplicated candidal vulvovaginitis. Recurrent episodes are more often caused by non-albicans species, for which azole agents are less likely to be effective. PMID:15872235

  10. In vitro resistance of clinical Fusarium species to amphotericin B and voriconazole using the EUCAST antifungal susceptibility method.

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    Taj-Aldeen, Saad J; Salah, Husam; Al-Hatmi, Abdullah M S; Hamed, Manal; Theelen, Bart; van Diepeningen, Anne D; Boekhout, Teun; Lass-Flörl, Cornelia

    2016-08-01

    Susceptibility testing using the EUCAST-AFST method against 39 clinical Fusarium strains consecutively collected from local and invasive infections during the last 10years assessed the in vitro activities of amphotericin B (AmB) and triazole antifungal agents. In addition, the susceptibility pattern of 12 reference strains from the CBS-KNAW Fungal Biodiversity Centre (CBS) was evaluated. In particular Fusarium petroliphilum and F. solani sensu lato were involved in disseminated infections and known for treatment failure. AmB displayed the lowest MICs followed by voriconazole VRC, posaconazole (POC). Itraconazole (ITC) showed high MIC values, displaying in vitro resistance. Clinical isolates were significantly (P Fusarium and that susceptibility testing is important and may improve the prognosis of these infections.

  11. Antifungal susceptibility and virulence attributes of animal-derived isolates of Candida parapsilosis complex.

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    Brilhante, Raimunda Sâmia Nogueira; de Jesus Santos Rodrigues, Terezinha; de Souza Collares Maia Castelo-Branco, Débora; Teixeira, Carlos Eduardo Cordeiro; de Brito Macedo, Ramila; Bandeira, Silviane Praciano; Pereira de Alencar, Lucas; Monteiro, André Jalles; de Aguiar Cordeiro, Rossana; de Jesus Pinheiro Gomes Bandeira, Tereza; Moreira, José Luciano Bezerra; Sidrim, José Júlio Costa; Rocha, Marcos Fábio Gadelha

    2014-11-01

    This study aimed to identify strains of the Candida parapsilosis complex isolated from animals, as well as to assess their in vitro antifungal susceptibility profile and in vitro production of virulence attributes. We used 28 isolates of C. parapsilosis sensu lato recovered from clinically healthy animals. The strains were characterized phenotypically, followed by molecular identification of the species through PCR-restriction enzyme analysis. The susceptibility of the strains to amphotericin B, itraconazole, voriconazole, fluconazole and caspofungin was assessed through broth microdilution. Additionally, the ability of the strains to produce biofilm, phospholipases and proteases was analysed. Molecular analysis showed 13 C. parapsilosis sensu stricto, 10 Candida orthopsilosis and five Candida metapsilosis strains. In vitro resistance to fluconazole was observed in three strains of C. parapsilosis sensu stricto and two C. metapsilosis. All tested strains were able to form biofilms and 23/28 isolates presented protease production, whilst none was able to produce phospholipases. Our study showed that C. parapsilosis sensu stricto and C. orthopsilosis are the most common species of the C. parapsilosis species complex and that these cryptic species present no significant phenotypical differences.

  12. Comparison of microdilution broth and disk diffusion for antifungal susceptibility testing against Candida spp. Comparação entre microdiluição e disco difusão para o teste de susceptibilidade aos antifúngicos contra Candida spp.

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    Carlla Lorena Façanha Silva

    2012-06-01

    Full Text Available The evolution of fungal resistance can be evaluated by testing susceptibility to antifungal agents. The protocols for susceptibility testing most common are Broth Microdilution (BMD, Disk Diffusion (DD and E-test. The objective of this study was to compare the susceptibility tests conducted by the BMD and DD methods for amphotericin B and fluconazole against Candida spp. isolated in Fortaleza/CE. Were used 40 C. albicans, 50 C. tropicalis and 47 C. parapsilosis isolated from blood and urine samples of patients treated at Fortaleza General Hospital. The tests were conducted by the BMD and DD methods according to protocols M27-A3 and M44-A2 of the CLSI. These methods were compared and the percentages of agreement were calculated. For amphotericin B and fluconazole was a predominance of strains with reduced susceptibility. The percentage of agreement for the methods tested was above 97%. No serious errors were detected. The use of DD for performing antifungal susceptibility testing can be applied in routine laboratory tests, since one technique is easy, inexpensive, and reliable when compared to the BMD without compromising the results for the strains of C. albicans, C. tropicalis and C. parapsilosis.A evolução da resistência fúngica pode ser avaliada pelo teste de susceptibilidade aos antifúngicos. Os protocolos mais utilizados para testes de susceptibilidade são a micro diluição em caldo (MDC, disco difusão (DD e E-test. O objetivo principal deste estudo foi comparar os testes de sensibilidade realizados pelos métodos de MDC e DD para a anfotericina B e o fluconazol contra cepas de Candida spp. isoladas em Fortaleza/CE. Foram utilizadas 40 C. albicans, 50 C. tropicalis e 47 C. parapsilosis, isoladas de amostras de sangue e urina de pacientes atendidos no Hospital Geral de Fortaleza. Os testes foram realizados pelos métodos MDC e DD, de acordo com os protocolos M27-A3 e M44-A2 do CLSI. As metodologias foram comparadas e calculadas os

  13. Antifungal activity of essential oils from Iranian plants against fluconazole-resistant and fluconazole-susceptible Candida albicans

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    Sharifzadeh, Aghil; Shokri, Hojjatollah

    2016-01-01

    Objective: The purpose of this study was to assay the antifungal activity of selected essential oils obtained from plants against both fluconazole (FLU)-resistant and FLU-susceptible C. albicans strains isolated from HIV positive patients with oropharyngeal candidiasis (OPC). Materials and Methods: The essential oils were obtained by hydrodistillation method from Myrtus communis (My. communis), Zingiber officinale roscoe (Z. officinale roscoe), Matricaria chamomilla (Ma. chamomilla), Trachyspermum ammi (T. ammi) and Origanum vulgare (O. vulgare). The susceptibility test was based on the M27-A2 methodology. The chemical compositions of the essential oils were obtained by gas chromatography- mass spectroscopy (GC-MS). Results: In GC-MS analysis, thymol (63.40%), linalool (42%), α-pinene (27.87%), α-pinene (22.10%), and zingiberene (31.79%) were found to be the major components of T. ammi, O. vulgare, My. communis, Ma. chamomilla and Z. officinale roscoe, respectively. The results showed that essential oils have different levels of antifungal activity. O. vulgare and T. ammi essential oils were found to be the most efficient (P<0.05). The main finding was that the susceptibilities of FLU-resistant C. albicans to essential oils were higher than those of the FLU-susceptible yeasts. Conclusion: Results of this study indicated that the oils from medicinal plants could be used as potential anti FLU-resistant C. albicans agents. PMID:27222835

  14. Antifungal susceptibility survey of 2,000 bloodstream Candida isolates in the United States.

    Science.gov (United States)

    Ostrosky-Zeichner, Luis; Rex, John H; Pappas, Peter G; Hamill, Richard J; Larsen, Robert A; Horowitz, Harold W; Powderly, William G; Hyslop, Newton; Kauffman, Carol A; Cleary, John; Mangino, Julie E; Lee, Jeannette

    2003-10-01

    Candida bloodstream isolates (n = 2,000) from two multicenter clinical trials carried out by the National Institute of Allergy and Infectious Diseases Mycoses Study Group between 1995 and 1999 were tested against amphotericin B (AMB), flucytosine (5FC), fluconazole (FLU), itraconazole (ITR), voriconazole (VOR), posaconazole (POS), caspofungin (CFG), micafungin (MFG), and anidulafungin (AFG) using the NCCLS M27-A2 microdilution method. All drugs were tested in the NCCLS-specified RPMI 1640 medium except for AMB, which was tested in antibiotic medium 3. A sample of isolates was also tested in RPMI 1640 supplemented to 2% glucose and by using the diluent polyethylene glycol (PEG) in lieu of dimethyl sulfoxide for those drugs insoluble in water. Glucose supplementation tended to elevate the MIC, whereas using PEG tended to decrease the MIC. Trailing growth occurred frequently with azoles. Isolates were generally susceptible to AMB, 5FC, and FLU. Rates of resistance to ITR approached 20%. Although no established interpretative breakpoints are available for the candins (CFG, MFG, and AFG) and the new azoles (VOR and POS), they all exhibited excellent antifungal activity, even for those strains resistant to the other aforementioned agents.

  15. Nectar yeasts of the Metschnikowia clade are highly susceptible to azole antifungals widely used in medicine and agriculture.

    Science.gov (United States)

    Álvarez-Pérez, Sergio; de Vega, Clara; Pozo, María I; Lenaerts, Marijke; Van Assche, Ado; Herrera, Carlos M; Jacquemyn, Hans; Lievens, Bart

    2016-02-01

    The widespread use of azole antifungals in medicine and agriculture and the resulting long-persistent residues could potentially affect beneficial fungi. However, there is very little information on the tolerance of non-target environmental fungi to azoles. In this study, we assessed the susceptibility of diverse plant- and insect-associated yeasts from the Metschnikowia clade, including several ecologically important species, to widely used medical and agricultural azoles (epoxiconazole, imazalil, ketoconazole and voriconazole). A total of 120 strains from six species were tested. Minimum inhibitory concentrations (MICs) were determined by the EUCAST broth microdilution procedure after some necessary modifications were made. The majority of species tested were highly susceptible to epoxiconazole, ketoconazole and voriconazole (>95% of strains showed MICs ≤ 0.125 mg l(-1)). Most strains were also very susceptible to imazalil, although MIC values were generally higher than for the other azoles. Furthermore, certain Metschnikowia reukaufii strains displayed a 'trailing' phenotype (i.e. showed reduced but persistent growth at antifungal concentrations above the MIC), but this characteristic was dependent on test conditions. It was concluded that exposure to azoles may pose a risk for ecologically relevant yeasts from the Metschnikowia clade, and thus could potentially impinge on the tripartite interaction linking these fungi with plants and their insect pollinators.

  16. Antifungal Susceptibility Test of 506 Clinical Candida Albicans Isolates to Fluconazole and Itraconazole In Vitro%白念珠菌506株对氟康唑和伊曲康唑的体外药敏试验

    Institute of Scientific and Technical Information of China (English)

    景伟芳; 李春莉; 王惠平

    2011-01-01

    目的 了解白念珠菌临床株对氟康唑和伊曲康唑的体外药物敏感性.方法 收集鉴定白念珠菌临床株,采用美国国家临床实验室标准化研究所CLSI(即以前的NCCLS)推荐的M27-A2微量肉汤稀释法,进行氟康唑和伊曲康唑的MIC值测定.结果 共收集白念珠菌506株,检测出氟康唑耐药株3株,剂量依赖性敏感株1株,耐药率为0.59%;伊曲康唑耐药株18株,剂量依赖性敏感株269株,耐药率为3.56%;其中1株对氟康唑和伊曲康唑交叉耐药.结论 白念珠菌对唑类药物有耐药及交叉耐药现象的发生;伊曲康唑耐药率高于氟康唑.%Objective To investigate the susceptibility of C. albicans isolates to fluconazole and itraconazole in vitro.Methods C. albicans was collected and identified from clinical isolates. M27-A2 Broth microdilution method, recommended by the Clinical and Laboratory Standards Institute ( CLSI), was used to assess the susceptibility of clinical C. albicans to fluconazole and itraconazole. Results A total of 506 clinical C.albicans isolates were collected. Three of the isolates were resistant to fluconazole, and one was susceptible-dose dependent to fluconazole. The resistant rate was 0. 59%. Eighteen isolates were resistant to itraconazole, and 269 were susceptible-dose dependent to itraconazole. The resistant rate was 3.56%. There was one isolates presenting cross-resistant to both fluconazole and itraconazole. Conclusion Some of the clinical C. albicans isolates do resist or cross-resist to antifungal azoles. The resistant rate of ltraconazole is higher than fluconazole.

  17. Antifungal Susceptibility Analysis of Clinical Isolates of Candida parapsilosis in Iran

    Science.gov (United States)

    LOTFALI, Ensieh; KORDBACHEH, Parivash; MIRHENDI, Hossein; ZAINI, Farideh; GHAJARI, Ali; MOHAMMADI, Rasoul; NOORBAKHSH, Fatemeh; MOAZENI, Maryam; FALLAHI, Aliakbar; REZAIE, Sassan

    2016-01-01

    Background: Candida parapsilosis is an emergent agent of invasive fungal infections. This yeast is one of the five most widespread yeasts concerned in invasive candidiasis. C. parapsilosis stands out as the second most common yeast species isolated from patients with bloodstream infections especially in neonates with catheter. Recently several reports suggested that its reduced susceptibility to azoles and polyene might become a cause for clinical concern, although C. parapsilosis is not believed to be intensely prone to the development of antifungal resistance. Methods: In the present report, One hundred and twenty clinical isolates of C. parapsilosis complex were identified and differentiated by using PCR-RFLP analysis. The isolates were then analyzed to determine their susceptibility profile to fluconazole (FLU), itraconazole (ITC) and amphotericin B. The minimum inhibitory concentration (MIC) results were analyzed according to the standard CLSI guide. Results: All of isolates were identified as C. parapsilosis. No C. metapsilosis and C. orthopsilosis strains were found. Evaluation of the antifungal susceptibility profile showed that only three (2.5%) C. parapsilosis were resistant to fluconazole, three (2.5%) C. parapsilosis were resistant to itraconazole and two (1.7%) C. parapsilosis were amphotericin B resistant. Conclusion: Profiles in clinical isolates of C. parapsilosis can provide important information for the control of antifungal resistance as well as distribution and susceptibility profiles in populations. PMID:27141494

  18. Molecular typing and antifungal susceptibility of Exophiala isolates from patients with cystic fibrosis.

    Science.gov (United States)

    Packeu, Ann; Lebecque, Patrick; Rodriguez-Villalobos, Hector; Boeras, Anca; Hendrickx, Marijke; Bouchara, Jean-Philippe; Symoens, Françoise

    2012-09-01

    The black yeast Exophiala dermatitidis is a frequent agent of colonization of the lungs of patients with cystic fibrosis (CF). A total of 71 clinical isolates of Exophiala from 13 patients were identified at the species level by sequencing the internal transcribed spacer (ITS) regions 1 and 2 of the rDNA genes and typed by random amplification of polymorphic DNA (RAPD), using two different primers, BG-2 and ERIC-1. In vitro susceptibility of these isolates to some systemic antifungal drugs was investigated using the CLSI method. Almost all the isolates were identified as E. dermatitidis, but long-term colonization with the closely related species E. phaeomuriformis was observed in one patient. No clustering was found according to the geographical origin of the isolates, the isolation date or the antifungal susceptibility. Variations were seen in the susceptibility of studied isolates to antifungals but most of them exhibited low susceptibility to amphotericin B and although some patients were successively colonized by two distinct genotypes, most of the isolates were distributed in patient-specific clusters. This phenomenon may be due to genomic variations of E. dermatitidis in the lung environment of CF patients. These results are typical of colonization of the airways of patients by a poorly distributed environmental fungus, which occupies particular reservoirs that need to be defined.

  19. Fungal peritonitis in patients undergoing peritoneal dialysis (PD) in Brazil: molecular identification, biofilm production and antifungal susceptibility of the agents.

    Science.gov (United States)

    Giacobino, Juliana; Montelli, Augusto Cezar; Barretti, Pasqual; Bruder-Nascimento, Ariane; Caramori, Jacqueline Teixeira; Barbosa, Luciano; Bagagli, Eduardo

    2016-10-01

    This paper presents data on fungal peritonitis (FP) in patients undergoing peritoneal dialysis (PD) at the University Hospital of Botucatu Medical School, São Paulo, Brazil. In a total of 422 patients, 30 developed FP, from which the medical records and the fungal isolates of 23 patient cases were studied. All patients presented abdominal pain, cloudy peritoneal effluent, needed hospitalization, had the catheter removed and were treated with fluconazole or fluconazole plus 5-flucitosine; six of them died due to FP. Concerning the agents, it was observed that Candida parapsilosis was the leading species (9/23), followed by Candida albicans (5/23), Candida orthopsilosis (4/23), Candida tropicalis (3/23), Candida guilliermondii (1/23), and Kodamaea ohmeri (1/23). All the isolates were susceptible to amphotericin B, voriconazole and caspofungin whereas C. albicans isolates were susceptible to all antifungals tested. Resistance to fluconazole was observed in three isolates of C. orthopsilosis, and dose-dependent susceptibility to this antifungal was observed in two isolates of C. parapsilosis and in the K. ohmeri isolate. Biofilm production estimates were high or moderate in most isolates, especially in C. albicans species, and low in C. parapsilosis species, with a marked variation among the isolates. This Brazilian study reinforces that FP in PD is caused by a diverse group of yeasts, most prevalently C. parapsilosis sensu stricto species. In addition, they present significant variation in susceptibility to antifungals and biofilm production, thus contributing to the complexity and severity of the clinical features.

  20. Antifungal susceptibility test of Paecilomyces hepiali by CLSI M38-A2 broth dilution method%CLSI M38-A2肉汤稀释法对蝙蝠蛾拟青霉(Paecilomyces hepiali)的药敏检测

    Institute of Scientific and Technical Information of China (English)

    赵婷; 姚粟; 李辉; 薛强伟; 程池

    2012-01-01

    [目的]对保健食品生产菌株蝙蝠蛾拟青霉(Paecilomyces hepiali)进行6种常见抗真菌药物:伊曲康唑、酮康唑、伏立康唑、环吡酮胺、5-氟胞嘧啶、氟康唑的药敏检测.[方法]参考美国国家临床实验室标准化委员会CLSI M38-A2《肉汤稀释法抗真菌药敏试验参考方案》.[结果]质控菌株MIC值均在参考范围之内,蝙蝠蛾拟青霉(P.hepiali)对以上6种药物的MIC值分别为1 mg/L、0.5 mg/L、0.25 mg/L、≥128 mg/L、64 mg/L、2 mg/L.[结论]M38-A2方案适合用来测定蝙蝠蛾拟青霉(P.hepiali)对抗真菌药物的敏感性,但获得菌株耐药敏感性的判别标准,仍需进一步的试验数据.%[Objective] To test filamentous fungi antifungal susceptibility, this study determined the minimal inhibitory concentration (MIC) values of 6 antifungal agents: Itraconazole (ITC), Ketoconazole (KET), Voriconazole (VRC), Ciclopirox (CIC), 5-Flurocytosine (5FC), Fluconazole (FLU) to the strain Paecilomyces hepiali which usually used in health food industry.[Methods] Method of Clinical Laboratory Standards Institute (CLSI) M38-A2 was referred.[Results] The results showed that the quality control strain Candida parapsilosis ATCC 22019T MIC values were within the reference range, the strain P.hepiali MIC values for the 6 antifungal agents were 1 mg/L, 0.5 mg/L, 0.25 mg/L, ≥128 mg/L, 64 mg/L, 2 mg/L respectively.[Conclusion] M38-A2 method could be applied to determine the susceptibility of P.hepiali to antifungal agents.But, more tests needed to acquire the standard interpretation of the strains susceptibility.

  1. Multilocus phylogeny and antifungal susceptibility of Aspergillus section Circumdati from clinical samples and description of A. pseudosclerotiorum sp. nov.

    Science.gov (United States)

    A multilocus phylogenetic study was carried out to assess the species distribution in a set of 34 clinical isolates of Aspergillus section Circumdati from the USA and their in vitro antifungal susceptibility were determined against eight antifungal drugs. The genetic markers used were ITS, BenA, CaM...

  2. In vitro susceptibility testing of dermatophytes isolated in Goiania, Brazil, against five antifungal agents by broth microdilution method Teste de suscetibilidade in vitro de dermatófitos isolados em Goiânia, Brasil, contra cinco agentes antifúngicos pelo método de microdiluição em caldo

    Directory of Open Access Journals (Sweden)

    Crystiane Rodrigues Araújo

    2009-02-01

    Full Text Available The antifungal activities of fluconazole, itraconazole, ketoconazole, terbinafine and griseofulvin were tested by broth microdilution technique, against 60 dermatophytes isolated from nail or skin specimens from Goiania city patients, Brazil. In this study, the microtiter plates were incubated at 28 ºC allowing a reading of the minimal inhibitory concentration (MIC after four days of incubation for Trichophyton mentagrophytes and five days for T. rubrum and Microsporum canis. Most of the dermatophytes had uniform patterns of susceptibility to the antifungal agents tested. Low MIC values as 0.03 µg/mL were found for 33.3%, 31.6% and 15% of isolates for itraconazole, ketoconazole and terbinafine, respectively.Atividades antifúngicas de fluconazol, itraconazol, cetoconazol, terbinafina e griseofulvina foram testadas pelo método de microdiluição em caldo contra 60 isolados de dermatófitos. Os resultados mostraram que todos os isolados produziram crescimento claramente detectável a 28 ºC e a concentração inibitória mínima (CIM foi determinada após quatro dias de incubação para Trichophyton mentagrophytes e cinco dias para T. rubrum e Microsporum canis. A maioria dos isolados teve um padrão uniforme de suscetibilidade para os agentes antifúngicos testados. Baixos valores de CIM como 0,03 µg/mL foram encontrados para 33,3%, 31,6% e 15% dos isolados para itraconazol, cetoconazol e terbinafina, respectivamente.

  3. Lack of effect of cell-wall targeted antibacterials on biofilm formation and antifungal susceptibility of Candidaspecies

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    Gisela Myrian de Lima Leite

    2014-09-01

    Full Text Available The use of central venous catheters (CVC and broad-spectrum antibacterials are among the main risk factors for the development of candidemia in patients admitted to intensive care units (ICU. It is known that some antibacterials increase the resistance of these yeasts to azole antifungals. Thus, the aim of this research was to determine whether yeast present in CVC colonizations previously exposed to cell-wall targeted antibacterials benefit from a reduction in susceptibility to fluconazole and voriconazole, facilitating their ability to form biofilms. Candida albicans, C. tropicalis, C. glabrata, C. parapsilosis and C. guilhermondii were seeded into antibacterial (cefepime, meropenem, vancomycin, and piperacillin-tazobactam gradient plates produced in Mueller-Hinton Agar. The susceptibility to fluconazole and voriconazole and the biofilm formation of the yeasts were tested before and after exposure to the antibacterials. None of the antibacterials exerted a significant effect on the in vitro susceptibility of the yeasts to the antifungal agents or on their ability to form biofilms. These results suggest that increased candidemia in ICU patients is not attributable to possible alterations in the yeasts, but is more likely caused by a weakening of the patient's general condition after long exposure to infection.

  4. Oral Candidiasis among Cancer Patients Attending a Tertiary Care Hospital in Chennai, South India: An Evaluation of Clinicomycological Association and Antifungal Susceptibility Pattern.

    Science.gov (United States)

    Jayachandran, Abirami Lakshmy; Katragadda, Radhika; Thyagarajan, Ravinder; Vajravelu, Leela; Manikesi, Suganthi; Kaliappan, Shanmugam; Jayachandran, Balaji

    2016-01-01

    Oropharyngeal candidiasis is one of the common manifestations seen in cancer patients on cytotoxic therapy and invasion into deeper tissues can occur if not treated promptly. Emergence of antifungal drug resistance is of serious concern owing to the associated morbidity and mortality. The present study aims at evaluation of clinicomycological association and antifungal drug susceptibility among the 180 recruited patients with cancer on chemotherapy and/or radiotherapy with signs or symptoms suggestive of oral candidiasis. Speciation and antifungal susceptibility was done by Microbroth dilution method for fluconazole, Itraconazole, and Amphotericin B as per standard microbiological techniques. Chi-square test was used for statistical analysis (p < 0.05 was considered statistically significant). Candida albicans was the predominant species isolated (94) (58%) followed by Candida tropicalis (34) (20.9%). Fluconazole and Itraconazole showed an overall resistance rate of 14% and 14.8%, respectively. All the isolates were susceptible to Amphotericin B. There was a significant association between the presence of dry mouth and isolation of Candida (p < 0.001). Such clinicomicrobiological associations can help in associating certain symptoms with the isolation of Candida. Species level identification with in vitro antifungal susceptibility pattern is essential to choose the appropriate drug and to predict the outcome of therapy.

  5. 萎缩糜烂型口腔扁平苔藓患者不同基因型白色念珠菌的药敏研究%Antifungal susceptibility test of genotypes of Candida albicans from patients with atrophic or erosive oral lichen planus

    Institute of Scientific and Technical Information of China (English)

    李景媛; 孙红英; 章强强

    2011-01-01

    PURPOSE: To study the antifungal susceptibility of genotypes of Candida albicans from patients with atrophic or erosive oral lichen planus and provide evidence for the treatment of candidiasis. METHODS: Polymerase chain reaction(PCR) was adopted to analyze 101 Candida albicans which were isolated from atrophic or erosive oral lichen planus.Microdilution broth method was carried out for antifungal susceptibility test. SPSS16.0 software package was used for Chi-square test. RESULTS: A total of 101 strains of Candida albicans were divided into three types,39 were genotype A strains, 17 genotype B stains and 45 genotype C stains.Strains of genotype A were significantly more resistant to 5-fluorocytosine than strains of genotypes B and C (P0.05). CONCLUSIONS: There is a correlation between Candida albicans genotypes and antifungal susceptibility. The use of antifungal agent should be based on the genotypes and antifungal susceptibility test of Candida albicans. For the treatment of candidiasis in patients with atrophic or erosive oral lichen planus, the value of nystatin should be addressed.%目的:对萎缩糜烂型口腔扁平苔藓不同基因型白色念珠菌进行药物敏感性研究,为临床有效治疗萎缩糜烂型口腔扁平苔藓白色念珠菌感染提供依据.方法:采用聚合酶链反应(PCR)对101株萎缩糜烂型口腔扁平苔藓白色念珠菌进行基因分型,然后采用微量稀释法测定白色念珠菌不同基因型对4种临床常用抗真菌药物的敏感性.采用SPSS16.0软件包对数据进行x检验.结果:101株白色念珠菌中,A型39株,B型17株,C型45株.A型对5-氟胞嘧啶耐药率显著高于B、C型(P0.05).结论:萎缩糜烂型口腔扁平苔藓白色念珠菌不同基因型对抗真菌药物的耐药性有差异,治疗时应根据基因分型和药敏试验结果合理选用抗真菌药物.治疗萎缩糜烂型口腔扁平苔藓伴发白色念珠菌感染时,制霉菌素的临床价值应该引起充分重视.

  6. Virulence Attributes and Antifungal Susceptibility Profile of Opportunistic Fungi Isolated from Ophthalmic Infections.

    Science.gov (United States)

    Sav, Hafize; Ozdemir, Havva Gül; Altınbas, Rabiye; Kiraz, Nuri; Ilkit, Macit; Seyedmousavi, Seyedmojtaba

    2016-10-01

    Investigations of both virulence factors and antifungal susceptibility profiles are crucial for understanding the pathogenesis and prognosis of ophthalmic mycoses. In this study, we investigated the in vitro antifungal susceptibility of amphotericin B (AMB), voriconazole (VRC), and natamycin (NAT) against a set of 50 fungal isolates obtained from patients with ocular mycoses using the Clinical and Laboratory Standards Institute broth microdilution method. In addition, putative virulence factor, such as secretory phospholipases and proteinases, and biofilm formation activity were analyzed. The geometric means (GMs) of the minimum inhibitory concentrations (MICs) of the antifungals across all isolates were the following (in increasing order): VRC (0.70 μg/mL), AMB (0.81 μg/mL), and NAT (1.05 μg/mL). The highest activity against 14 Aspergillus strains was exhibited by VRC (GM MIC: 0.10 μg/mL), followed by AMB and NAT (GM MICs: 0.21 and 0.27 μg/mL), respectively. However, for 12 Fusarium spp., the GM MIC of VRC (2.66) was higher than those of NAT and AMB (GM MICs 1.3 and 0.8 μg/mL, respectively). Proteinase and phospholipase activity were observed in 30 % and 42 % of the isolates, respectively, whereas only 8 % of the isolates were able to produce biofilms. Phospholipase activity was observed in all Fusarium isolates, but not in any of the Aspergillus isolates. In contrast, biofilm-forming capability was detected in 25 % of the Fusarium isolates, but none of the Aspergillus isolates. The differences in the MICs of AMB, VRC, and NAT, biofilm-forming ability and proteinase and phospholipase activities among the isolates were not significant (p > 0.05). Overall, our study suggests no significant correlation between the antifungal susceptibility profiles and virulence attributes of ocular fungal isolates.

  7. Optimization of Antifungal Extracts from Ficus hirta Fruits Using Response Surface Methodology and Antifungal Activity Tests

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    Chuying Chen

    2015-10-01

    Full Text Available The fruits of Ficus hirta (FH display strong antifungal activity against Penicillium italicum and Penicillium digitatum. In order to optimize the extraction conditions of antifungal extracts from FH fruit, various extraction parameters, such as ethanol concentration, extraction time, solvent to solid ratio and temperature, were chosen to identify their effects on the diameters of inhibition zones (DIZs against these two Penicillium molds. Response surface methodology (RSM was applied to obtain the optimal combination of these parameters. Results showed that the optimal extraction parameters for maximum antifungal activity were: 90% (v/v ethanol concentration, 65 min extraction time, 31 mL/g solvent to solid ratio and 51 °C temperature. Under the abovementioned extraction conditions, the experimental DIZs values obtained experimentally were 57.17 ± 0.75 and 39.33 ± 0.82 mm, which were very close to the values of 57.26 and 39.29 mm predicted by the model. Further, nine kinds of phytopathogens were tested in vitro to explore the antifungal activity of the FH extracts. It was found for the first time that the FH extracts showed significant inhibition on the growth of P. italicum, A. citri, P. vexans, P. cytosporella and P. digitatum.

  8. Genetic relatedness and antifungal susceptibility profile of Candida albicans isolates from fungaemia patients.

    Science.gov (United States)

    Costa-de-Oliveira, Sofia; Sousa, Inês; Correia, Alexandra; Sampaio, Paula; Pais, Célia; Rodrigues, Acácio Gonçalves; Pina-Vaz, Cidália

    2011-04-01

    A prospective study to assess fungaemia was conducted for 12 months at a Portuguese University Hospital. A total of 35 Candida albicans isolates obtained from 12 patients with fungaemia were compared by a multiplex PCR system using four microsatellite loci. Blood isolates were evaluated against concomitant isolates from urine, lower respiratory secretions and central venous catheters, as well as with successive isolates recovered from recurrent episodes of fungaemia. The data analyzed included the department of admission, underlying diseases and antifungal therapy. The susceptibility phenotypes of all isolates to amphotericin B, fluconazole, itraconazole, voriconazole and caspofungin were determined according to the CLSI M27-A3 protocol. We observed a high degree of similarity between successive blood isolates and between blood and concomitant isolates from other sites of the same patient. This is suggestive of the recurrence of fungaemia and was due to the same strain, possibly as a result of the failure of antifungal therapy. The genetic similarity observed between some strains isolated from different patients suggested the likelihood that they were hospital acquired. Distinct patients were infected by the same strain at different time periods, and an increase in antifungal resistance was observed over time for some of these strains. Hospital-acquired exogenous nosocomial infections can be associated with higher risks of antifungal resistance and need to be closely monitored. Particular attention should also be given to endogenous non-blood Candida isolates which can be critical in high risk patients, as they often can become invasive in immunodeficient individuals.

  9. Antifungal Susceptibility of 205 Candida spp. Isolated Primarily during Invasive Candidiasis and Comparison of the Vitek 2 System with the CLSI Broth Microdilution and Etest Methods▿

    Science.gov (United States)

    Bourgeois, N.; Dehandschoewercker, L.; Bertout, S.; Bousquet, P.-J.; Rispail, P.; Lachaud, L.

    2010-01-01

    Infections due to Candida spp. are frequent, particularly in immunocompromised and intensive care unit patients. Antifungal susceptibility tests are now required to optimize antifungal treatment given the emergence of acquired antifungal resistance in some Candida species. An antifungal susceptibility automated method, the Vitek 2 system (VK2), was evaluated. VK2 was compared to the CLSI broth microdilution reference method and the Etest procedure. For this purpose, 205 clinical isolates of Candida spp., including 11 different species, were tested for fluconazole, voriconazole, and amphotericin B susceptibility. For azoles, essential agreement ranged from 25% to 100%, depending on the method used and the Candida species tested. Categorical agreements for all of the species averaged 92.2% and ranged from 14.3 to 100%, depending on the 24-h or 48-h MIC reading by the Etest and CLSI methods and on the Candida species. Results obtained for Candida albicans showed excellent categorical and essential agreements with the two comparative methods. For Candida glabrata, the essential agreement was high with the CLSI method but low with the Etest method, and several very major errors in interpretation were observed between VK2 and the Etest method for both azoles. Low MICs of fluconazole were obtained for all of the Candida krusei isolates, but the VK2 expert software corrected all of the results obtained to resistant. Amphotericin B results showed MICs of ≤1 mg/liter for 201 (VK2), 190 (CLSI), and 202 (Etest) isolates. The AST-YS01 Vitek 2 card system (bioMérieux) is a reliable and practical standardized automated antifungal susceptibility test. Nevertheless, more assays are needed to better evaluate C. glabrata fluconazole sensitivity. PMID:19889902

  10. Molecular epidemiology and antifungal susceptibility of Candida parapsilosis sensu stricto, Candida orthopsilosis, and Candida metapsilosis in Taiwan.

    Science.gov (United States)

    Chen, Yee-Chun; Lin, Yu-Hsin; Chen, Kuo-Wei; Lii, Joanne; Teng, Hwa-Jen; Li, Shu-Ying

    2010-11-01

    Candida parapsilosis was recently reclassified into 3 closely related species, C. parapsilosis sensu stricto, Candida orthopsilosis, and Candida metapsilosis. Variation in susceptibility characteristics and prevalence of the 3 genomic species could have therapeutic and epidemiologic implications. The aim of this study is to characterize the genetic and antifungal susceptibility profiles of 97 C. parapsilosis isolates from 71 patients. Among the 71 nonduplicate isolates, 85.9% (61/71) were identified as C. parapsilosis sensu stricto, 5.6% (4/71) as C. metapsilosis, and 8.5% (6/71) as C. orthopsilosis species based on sequences of the internal transcribed spacer (ITS) region. The delineation of these 3 species is concordant with that achieved by pulsed-field gel electrophoresis of BssHII restriction fragments at 75% similarity. Antifungal susceptibility tests showed that most isolates were susceptible to flucytosine, azoles, amphotericin B, and echinocandins, whereas 3 C. metapsilosis isolates from 1 patient showed resistance and susceptible-dose dependence to fluconazole. The C. metapsilosis isolates exhibited significantly higher MIC values to both fluconazole and voriconazole than those of C. parapsilosis sensu stricto and C. orthopsilosis. On the other hand, the C. metapsilosis isolates showed significantly lower MIC values on 24 h to caspofungin than those of C. parapsilosis sensu stricto and C. orthopsilosis. For micafungin, the isolates of C. parapsilosis sensu stricto had significantly higher MIC values on 24 h than those of C. orthopsilosis and C. metapsilosis. Compared to Candida albicans, mutations from proline to alanine were identified on the hot spot 1 of Fks1 in all these C. parapsilosis sensu lato isolates regardless of their MIC levels. Some of the C. orthopsilosis and C. metapsilosis isolates expressed the isoleucine to valine substitution on the hot spot 2 region. However, the amino acid variations in these isolates did not correlate to their MIC

  11. Epidemiology and Antifungal Susceptibility of Bloodstream Fungal Isolates in Pediatric Patients: a Spanish Multicenter Prospective Survey ▿

    Science.gov (United States)

    Pemán, Javier; Cantón, Emilia; Linares-Sicilia, María José; Roselló, Eva María; Borrell, Nuria; Ruiz-Pérez-de-Pipaon, María Teresa; Guinea, Jesús; García, Julio; Porras, Aurelio; García-Tapia, Ana María; Pérez-del-Molino, Luisa; Suárez, Anabel; Alcoba, Julia; García-García, Inmaculada

    2011-01-01

    Data on fungemia epidemiology and antifungal susceptibility of isolates from children are scarce, leading frequently to pediatric empirical treatment based on available adult data. The present study was designed to update the epidemiological, mycological, and in vitro susceptibility data on fungal isolates from children with fungemia in Spain. All fungemia episodes were identified prospectively by blood culture over 13 months at 30 hospitals. Tests of susceptibility to amphotericin B, flucytosine, fluconazole, itraconazole, posaconazole, voriconazole, anidulafungin, caspofungin, and micafungin were performed at participant institutions by a microdilution colorimetric method. New species-specific clinical breakpoints for fluconazole, voriconazole, and echinocandins were also applied. A total of 203 episodes of fungemia in 200 children were identified. A higher proportion of fungal isolates was from general wards than intensive care units (ICU). Candida parapsilosis (46.8%), Candida albicans (36.5%), Candida tropicalis (5.9%), Candida glabrata (3.9%), and Candida guilliermondii (2.5%) were the leading species. C. parapsilosis was the predominant species except in neonates. C. albicans was the most frequent in neonatal ICU settings (51.9%). Intravascular catheter (79.3%), surgery (35%), prematurity (30%), and neutropenia (11%) were the most frequent predisposing factors. Most Candida isolates (95.1%) were susceptible to all antifungals. When the new species-specific clinical breakpoints were applied, all C. parapsilosis isolates were susceptible to echinocandins except one, which was micafungin resistant. This is the largest published series of fungemia episodes in the pediatric setting. C. parapsilosis is the most prevalent species in Spain, followed by C. albicans and C. tropicalis. Resistance to azole and echinocandin agents is extremely rare among Candida species. The fluconazole resistance rate in Spain has decreased in the last 10 years. PMID:22012014

  12. Exogenous tyrosol inhibits planktonic cells and biofilms of Candida species and enhances their susceptibility to antifungals.

    Science.gov (United States)

    Cordeiro, Rossana de A; Teixeira, Carlos E C; Brilhante, Raimunda S N; Castelo-Branco, Débora S C M; Alencar, Lucas P; de Oliveira, Jonathas S; Monteiro, André J; Bandeira, Tereza J P G; Sidrim, José J C; Moreira, José Luciano Bezerra; Rocha, Marcos F G

    2015-06-01

    Tyrosol is a quorum-sensing molecule of Candida albicans able to induce hyphal development in the early and intermediate stages of biofilm growth. In the present study, we evaluated the effect of high concentrations of exogenous tyrosol on planktonic cells and biofilms of C. albicans (n = 10) and C. tropicalis (n = 10), and investigated whether tyrosol could be synergic to antifungals that target cellular ergosterol. Antifungal susceptibility and drug interaction against planktonic cells were investigated by the broth microdilution method. Tyrosol was able to inhibit planktonic cells, with MIC values ranging from 2.5 to 5.0 mM for both species. Synergism was observed between tyrosol/amphotericin B (11/20 strains), tyrosol/itraconazole (18/20 strains) and tyrosol/fluconazole (18/20 strains). Exogenous tyrosol alone or combined with antifungals at both 10 × MIC and 50 × MIC were able to reduce biofilm of both Candida species. Mature biofilms were susceptible to tyrosol alone at 50 × MIC or combined with amphotericin at both 10 × MIC and 50 × MIC. On the other hand, tyrosol plus azoles at both 10 × MIC and 50 × MIC enhanced biofilm growth.

  13. Antifungal susceptibility and molecular typing of 115 Candida albicans isolates obtained from vulvovaginal candidiasis patients in 3 Shanghai maternity hospitals.

    Science.gov (United States)

    Ying, Chunmei; Zhang, Hongju; Tang, Zhenhua; Chen, Huifen; Gao, Jing; Yue, Chaoyan

    2016-05-01

    In our multicenter study, we studied the distribution of Candida species in vulvovaginal candidiasis patients and investigated antifungal susceptibility profile and genotype of Candida albicans in vaginal swab. A total of 115 Candida albicans strains were detected in 135 clinical isolates. Minimum inhibitory concentration determinations showed that 83% and 81% of the 115 Candida albicans strains were susceptible to fluconazole and voriconazole. Randomly amplified polymorphic DNA analysis (RAPD) was applied to identify clonally related isolates from different patients at the local level. All tested strains were classified into genotype A (77.4%), genotype B (18.3%), and genotype C (4.3%). Genotype A was further classified into five subtypes and genotype B into two subtypes.Candida albicans was the dominant pathogen of vulvovaginal candidiasis, the majority belonging to genotype A in this study. Exposure to azoles is a risk factor for the emergence of azole resistance among Candida albicans isolated from VVC patients.

  14. Molecular Characterization and In Vitro Antifungal Susceptibility of 316 Clinical Isolates of Dermatophytes in Iran.

    Science.gov (United States)

    Ansari, Saham; Hedayati, Mohammad T; Zomorodian, Kamiar; Pakshir, Keyvan; Badali, Hamid; Rafiei, Abdollah; Ravandeh, Mostafa; Seyedmousavi, Seyedmojtaba

    2016-02-01

    Dermatophytosis is a common mycotic infection of the skin, nail, and hair, associated with major public health concern worldwide. Various species of dermatophytes show significant differences in susceptibility to antifungals. Here, we present the antifungal susceptibility of a large collection of molecularly identified dermatophyte isolates obtained from tropical region of south of Iran. A total of 9485 patients clinically suspected to have cutaneous fungal infections were examined. Dermatophytosis was confirmed in 1502 cases by direct microscopy and culture. Three hundred and sixteen isolates recovered in culture were identified to species level using PCR sequencing of ITS region and RFLP. Tinea corporis was the most prevalent type of clinical manifestation (35.2 %), followed by tinea cruris (17 %), tinea capitis (12.8 %), tinea pedis (11.3 %), tinea manuum (11 %), tinea unguium (6.9 %), and tinea barbae (5.8 %). Trichophyton interdigitale was the most common isolate (49.36 %), followed by Trichophyton rubrum (18.98 %), Epidermophyton floccosum (13.29 %), Microsporum canis (9.17 %), Arthroderma benhamiae (T. anamorph of A. benhamiae; 5.38 %), and Trichophyton tonsurans (3.79 %). Overall, irrespective of the geographical region, terbinafine was the most potent antifungal against all isolates, with an MIC range of 0.002-0.25 μg/mL, followed by itraconazole (0.004-0.5 μg/mL), griseofulvin (0.125-8 μg/mL), and fluconazole (4-128 μg/mL). Analysis of our data revealed a significant increase in the frequency of A.benhamiae, which definitely warrants further investigation to explore source of this infection in south of Iran. Moreover, terbinafine was the most effective antifungal against all isolates, in vitro.

  15. Antifungal activity of essential oils from Iranian plants against fluconazole-resistant and fluconazole-susceptible Candida albicans

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    Aghil Sharifzadeh

    2016-03-01

    Full Text Available Objectives: The purpose of this study was to assay the antifungal activity of selected essential oils obtained from plants against both fluconazole (FLU-resistant and FLU-susceptible C. albicans strains isolated from HIV positive patients with oropharyngeal candidiasis (OPC. Materials and Methods: The essential oils were obtained by hydrodistillation method from Myrtus communis (My. communis, Zingiber officinale roscoe (Z. officinale roscoe, Matricaria chamomilla (Ma. chamomilla, Trachyspermum ammi (T. ammi and Origanum vulgare (O. vulgare. The susceptibility test was based on the M27-A2 methodology. The chemical compositions of the essential oils were obtained by gas chromatography- mass spectroscopy (GC-MS. Results: In GC-MS analysis, thymol (63.40%, linalool (42%, α-pinene (27.87%, α-pinene (22.10%, and zingiberene (31.79% were found to be the major components of T. ammi, O. vulgare, My. communis, Ma. chamomilla and Z. officinale roscoe, respectively. The results showed that essential oils have different levels of antifungal activity. O. vulgare and T. ammi essential oils were found to be the most efficient (P

  16. Histoplasmosis in HIV-positive patients in Ceará, Brazil: clinical-laboratory aspects and in vitro antifungal susceptibility of Histoplasma capsulatum isolates.

    Science.gov (United States)

    Brilhante, Raimunda S N; Fechine, Maria A B; Mesquita, Jacó R L; Cordeiro, Rossana A; Rocha, Marcos F G; Monteiro, André J; Lima, Rita A C; Caetano, Érica P; Pereira, Juliana F; Castelo-Branco, Débora S C M; Camargo, Zoilo P; Sidrim, José J C

    2012-08-01

    This study contains a descriptive analysis of histoplasmosis in AIDS patients between 2006 and 2010 in the state of Ceará, Brazil. Additionally, the in vitro susceptibility of Histoplasma capsulatum isolates obtained during this period was assessed. We report 208 cases of patients with histoplasmosis and AIDS, describing the epidemiological, clinical, laboratory and therapeutic aspects. The in vitro antifungal susceptibility test was carried out by the microdilution method, according to Clinical and Laboratory Standards Institute, with H. capsulatum in the filamentous and yeast phases, against the antifungals amphotericin B, fluconazole, itraconazole, voriconazole and caspofungin. In 38.9% of the cases, histoplasmosis was the first indicator of AIDS and in 85.8% of the patients the CD4 cell count was lower than 100 cells/mm(3). The lactate dehydrogenase levels were high in all the patients evaluated, with impairment of hepatic and renal function and evolution to death in 42.3% of the cases. The in vitro susceptibility profile demonstrated there was no antifungal resistance among the isolates evaluated. There was a significant increase in the number of histoplasmosis cases in HIV-positive patients during the period surveyed in the state of Ceará, northeastern Brazil, but no antifungal resistance among the recovered isolates of H. capsulatum.

  17. Biofilm production and evaluation of antifungal susceptibility amongst clinical Candida spp. isolates, including strains of the Candida parapsilosis complex.

    Science.gov (United States)

    Melo, Analy S; Bizerra, Fernando C; Freymüller, Edna; Arthington-Skaggs, Beth A; Colombo, Arnaldo L

    2011-04-01

    Candida cells can form biofilms that frequently are sources of infections and are less susceptible to antifungal drugs. Some authors have reported that Candida orthopsilosis and Candida metapsilosis isolates are not able to produce biofilms in vitro and there are no studies available on biofilm susceptibility for these species to antifungals. The aims of this study were to (i) quantify Candida spp. biofilms in vitro, and (ii) test the in vitro susceptibilities of Candida spp. biofilms to fluconazole (FLC) and amphotericin B (AMB). Isolates studied included four Candida albicans, six C. tropicalis, seven C. parapsilosis, eight C. orthopsilosis, and five C. metapsilosis. We compared two different methods to evaluate biofilm production, i.e., crystal violet (CV) staining and XTT-reduction assays (XTT). Scanning electron microscopy (SEM) was used to observe high, medium and low biofilm producing isolates screened by these two methods. To determine the minimum biofilm eradication concentration (MBEC) for FLC and AMB, XTT-reduction assay was used to measure cell metabolic activity. Biofilm quantification by CV and XTT showed that C. tropicalis isolates were the highest biofilm producer, followed by C. albicans, C. parapsilosis, C. orthopsilosis and C. metapsilosis. Examination of SEM images revealed that the extent of biofilms formed by high, medium, and low producers was highly correlated to the results generated by CV assay. Biofilm of all the isolates evaluated were resistant to FLC (MBEC(80) ≥ 256 ug/ml) but, in general, susceptible to AMB, except for six C. parapsilosis strains (MBEC(80) ≥ 8 ug/ml).

  18. Species distribution and in vitro antifungal susceptibility profiles of yeast isolates from invasive infections during a Portuguese multicenter survey.

    Science.gov (United States)

    Faria-Ramos, I; Neves-Maia, J; Ricardo, E; Santos-Antunes, J; Silva, A T; Costa-de-Oliveira, S; Cantón, E; Rodrigues, A G; Pina-Vaz, C

    2014-12-01

    This is the first Portuguese multicenter observational and descriptive study that provides insights on the species distribution and susceptibility profiles of yeast isolates from fungemia episodes. Ten district hospitals across Portugal contributed by collecting yeast isolates from blood cultures and answering questionnaires concerning patients' data during a 12-month period. Molecular identification of cryptic species of Candida parapsilosis and C. glabrata complex was performed. The susceptibility profile of each isolate, considering eight of the most often used antifungals, was determined. Both Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) protocols were applied. The incidence of 240 episodes of fungemia was 0.88/1,000 admissions. Fifteen different species were found, with C. albicans (40 %) being the most prevalent, followed by C. parapsilosis (23 %) and C. glabrata (13 %). Most isolates were recovered from patients admitted to surgical wards or intensive care units, with 57 % being males and 32 % aged between 41 and 60 years. For both the CLSI and EUCAST protocols, the overall susceptibility rates ranged from 74 to 97 % for echinocandins and from 84 to 98 % for azoles. Important resistance rate discrepancies between protocols were observed in C. albicans and C. glabrata for echinocandins and in C. parapsilosis and C. tropicalis for azoles. Death associated with fungemia occurred in 25 % of the cases, with more than half of C. glabrata infections being fatal. The great number of Candida non-albicans is noteworthy despite a relatively low antifungal resistance rate. Studies like this are essential in order to improve empirical treatment guidelines.

  19. Species distribution and antifungal susceptibility profile of Candida spp. bloodstream isolates from Latin American hospitals

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    Godoy Patrício

    2003-01-01

    Full Text Available From March 1999 to March 2000, we conducted a prospective multicenter study of candidemia involving five tertiary care hospitals from four countries in Latin America. Yeast isolates were identified by classical methods and the antifungal susceptibility profile was determined according to the National Committee for Clinical Laboratory Standards microbroth assay method. During a 12 month-period we were able to collect a total of 103 bloodstream isolates of Candida spp. C. albicans was the most frequently isolated species accounting for 42% of all isolates. Non-albicans Candida species strains accounted for 58% of all episodes of candidemia and were mostly represented by C. tropicalis (24.2% and C. parapsilosis (21.3%. It is noteworthy that we were able to identify two cases of C. lusitaniae from different institutions. In our casuistic, non-albicans Candida species isolates related to candidemic episodes were susceptible to fluconazole. Continuously surveillance programs are needed in order to identify possible changes in the species distribution and antifungal susceptibility patterns of yeasts that may occurs after increasing the use of azoles in Latin American hospitals.

  20. CHANGING TRENDS OF CANDIDA ISOLATES AND THEIR ANTIFUNGAL SUSCEPTIBILITY PATTERN IN VULVOVAGINAL CANDIDIASIS CASES OF TRIPURA, NORTH EAST INDIA

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    Jhinuk Basu

    2015-11-01

    Full Text Available : BACKGROUND: Candida species are a part of the complex endogenous vaginal microflora, which under pathological condition cause vulvovaginal candidiasis. Worldwide it is the second most common cause of vaginitis after bacterial vaginosis and in India it accounts for 18-20% of clinical visits. AIM: To determine the spectrum of Candida species among the subjects of suspected vulvovaginal candidiasis with an objective to isolate and identify all the Candida species and determine their antifungal susceptibility pattern. SETTINGS AND DESIGN: The study was cross sectional observational, conducted on 275 symptomatic outdoor and admitted patients in the Department of Obstetrics and Gynaecology in a tertiary care hospital in Tripura between August 2012 and April 2015. MATERIALS AND METHODS: Discharge was collected from patients and identified by Grams staining and wet mount test. Culturing was done in Sabouraud dextrose agar followed by speciation through germ tube test, corn meal agar morphologies and chromogenic identification in HiCrome agar. Antifungal susceptibility was determined through E-test for Polyenes and Azoles. RESULT: Vulvovaginal candidiasis was found at a frequency of 25% with non-albicans Candida species (62% prevailing over C. albicans (38%. Highest frequency was observed in the 40–49 age group. Highest susceptibility was seen for fluconazole and least for ketoconazole. CONCLUSION: The frequency of vulvovaginal candidiasis was found to be higher than other prevalence studies from India. The study also showed increasing shift towards non-albicans Candida species from C. albicans. The reason for high frequency among the menopausal group and ectopic pregnancy cases could be topics for future studies. Amphotericin B and fluconazole were effective towards most species and hence could be used to treat the local population.

  1. Prevalence and antifungal susceptibility of Candida albicans and its related species Candida dubliniensis and Candida africana isolated from vulvovaginal samples in a hospital of Argentina.

    Science.gov (United States)

    Theill, Laura; Dudiuk, Catiana; Morano, Susana; Gamarra, Soledad; Nardin, María Elena; Méndez, Emilce; Garcia-Effron, Guillermo

    2016-01-01

    Candida africana taxonomical status is controversial. It was proposed as a separate species within the Candida albicans species complex; however, phylogenetic analyses suggested that it is an unusual variety of C. albicans. The prevalence of C. albicans-related species (Candida dubliniensis and C. africana) as vulvovaginal pathogens is not known in Argentina. Moreover, data on antifungal susceptibility of isolates causing vulvovaginal candidiasis is scarce. The aims of this study were to establish the prevalence of C. dubliniensis and C. africana in vaginal samples and to evaluate the antifungal susceptibilities of vaginal C. albicans species complex strains. We used a molecular-based method coupled with a new pooled DNA extraction methodology to differentiate C. dubliniensis and C. africana in a collection of 287 strains originally identified as C. albicans isolated from an Argentinian hospital during 2013. Antifungal susceptibilities to fluconazole, clotrimazole, itraconazole, voriconazole, nystatin, amphotericin B and terbinafine were evaluated by using the CLSI M27-A3 and M27-S4 documents. Of the 287 isolates, 4 C. dubliniensis and one C. africana strains (1.39% and 0.35% prevalence, respectively) were identified. This is the first description of C. africana in Argentina and its identification was confirmed by sequencing the ITS2 region and the hwp1 gene. C. dubliniensis and C. africana strains showed very low MIC values for all the tested antifungals. Fluconazole-reduced-susceptibility and azole cross-resistance were observed in 3.55% and 1.41% of the C. albicans isolates, respectively. These results demonstrate that antifungal resistance is still a rare phenomenon in this kind of isolates.

  2. Antifungal susceptibility analysis of berberine, baicalin, eugenol and curcumin on Candida albicans

    Institute of Scientific and Technical Information of China (English)

    Wu Jianhua; Wen Hai

    2009-01-01

    Objective: To analyze the antifungal effects of Chinese herb monomers, i.e. berberine, baicalin, eugenol and curcumin, on Candida albicans. Methods: After Candida albicans strain Y01-09 was incubated for 48 h in YEPD broth which contained different concentrations of Chinese herb components, the cell cycle, fluorescent intensity and the size of cell volume were detected by flow cytometry. Results: The 4 Chinese herb monomers could affect the cell cycle of Candida albicans in different ranges. The ratio of cells in S-G2-M period decreased as the agents concentration increased, indicating that the cell division was inhibited. The fluorescent intensity of Candida albicans cells became weaker after being incubated, which reflected the loss of DNA fragments. The higher the concentration was, the weaker the fluorescent intensity became. The cell size, cell diopter and particle size changed much as the agents concentration increased. Conclusion: Chinese herb monomers play the antifungal role in inhibiting cell division. FCM could be used to determine the susceptibility of antifungal agents.

  3. Biofilm formation by and antifungal susceptibility of Candida isolates from urine.

    Science.gov (United States)

    Jain, N; Kohli, R; Cook, E; Gialanella, P; Chang, T; Fries, B C

    2007-03-01

    Biofilm formation (BF) in the setting of candiduria has not been well studied. We determined BF and MIC to antifungals in Candida spp. isolates grown from urine samples of patients and performed a retrospective chart review to examine the correlation with risk factors. A total of 67 Candida spp. isolates were grown from urine samples from 55 patients. The species distribution was C. albicans (54%), C. glabrata (36%), and C. tropicalis (10%). BF varied greatly among individual Candida isolates but was stable in sequential isolates during chronic infection. BF also depended on the growth medium and especially in C. albicans was significantly enhanced in artificial urine (AU) compared to RPMI medium. In nine of the C. albicans strains BF was 4- to 10-fold higher in AU, whereas in three of the C. albicans strains and two of the C. glabrata strains higher BF was measured in RPMI medium than in AU. Determination of the MICs showed that planktonic cells of all strains were susceptible to amphotericin B (AMB) and caspofungin (CASPO) and that three of the C. glabrata strains and two of the C. albicans strains were resistant to fluconazole (FLU). In contrast, all biofilm-associated adherent cells were resistant to CASPO and FLU. The biofilms of 14 strains (28%) were sensitive to AMB (MIC(50) of Candida strains that varies greatly among clinical strains and is dependent on the growth medium. Resistance to AMB is associated with higher BF in AU, which may represent the more physiologic medium to test BF. Future studies should address whether in vitro BF can predict treatment failure in vivo.

  4. Geographic Distribution and Antifungal Susceptibility of the Newly Described Species Candida orthopsilosis and Candida metapsilosis in Comparison to the Closely Related Species Candida parapsilosis▿

    OpenAIRE

    Lockhart, Shawn R.; Messer, Shawn A.; Pfaller, Michael A.; Diekema, Daniel J.

    2008-01-01

    Candida orthopsilosis and Candida metapsilosis are recently described species, having previously been grouped with the more prevalent species Candida parapsilosis. Current literature contains very little data pertaining to the distributions and antifungal susceptibilities of these Candida species. We determined the species and antifungal susceptibilities of 1,929 invasive clinical isolates from the ARTEMIS antifungal surveillance program collected between 2001 and 2006 and identified as C. pa...

  5. Typing and antifungal susceptibility of the candida species isolated in geriatric patients at a tertiary care center

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    Sevgi Yılmaz Hancı

    2015-12-01

    Full Text Available Objective: In this study we aimed to detect, identification and distribution of the Candida species in blood and urine samples; and antifungal susceptibility of Candida bloodstream isolates in geriatric patients Methods: All Candida species isolated from blood and urine cultures samples of patients over than 65 years of during the period December 2011 -November 2014 in İzmir Tepecik Training and Research Hospital were enrolled in this study. For Candida species identification conventional methods, colony appearance in candida chromogenic agar and for some unidentified isolates API ID32C AUX (BioMérieux, France was used. Antifungal susceptibility testing of the isolates was performed with API ATB Fungus 3 (BioMérieux, France. Results: During the study period 681 Candida species were isolated in blood and urine samples of patients over than 65 years. When the sample species were evaluated, Candida species detected in 561(82.4% urine sample, and 120 (17.6% blood samples. Most isolated species were respectively detected C. albicans (47.0%, C.parapsilosis (16.0%, C.tropicalis (15.9%, C.glabrata (12.2%, C.kefy (3.7% and C.krusei (2.8%. C. albicans detected the most common in urine samples (51.7 %, on the other hand C.parapsilosis the most common species in blood samples (50.8%. In general surgery unit, C.tropicalis was found as the most common isolate. On the other hand C.albicans the most common species in the other units and services. C.parapsilosis rate in blood cultures increased with age. Antifungal susceptibility of the Candida bloodstream isolates were detected as 97.5% for flucytosine, 95.8% for amfotericin B, 82.5% for fluconazole, 91.7% for itraconazole and 77.5% for voriconazole. Conclusion: The most frequently isolated yeast species in blood and urinesamples of geriatric patients has been found C. albicans. C.parapsilosis especially frequently isolated from blood cultures and increasing the frequency with age. Voriconazole in Candida

  6. Candida parapsilosis (sensu lato) isolated from hospitals located in the Southeast of Brazil: Species distribution, antifungal susceptibility and virulence attributes.

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    Ziccardi, Mariangela; Souza, Lucieri O P; Gandra, Rafael M; Galdino, Anna Clara M; Baptista, Andréa R S; Nunes, Ana Paula F; Ribeiro, Mariceli A; Branquinha, Marta H; Santos, André L S

    2015-12-01

    Candida parapsilosis (sensu lato), which represents a fungal complex composed of three genetically related species - Candida parapsilosis sensu stricto, Candida orthopsilosis and Candida metapsilosis, has emerged as an important yeast causing fungemia worldwide. The goal of the present work was to assess the prevalence, antifungal susceptibility and production of virulence traits in 53 clinical isolates previously identified as C. parapsilosis (sensu lato) obtained from hospitals located in the Southeast of Brazil. Species forming this fungal complex are physiologically/morphologically indistinguishable; however, polymerase chain reaction followed by restriction fragment length polymorphism of FKS1 gene has solved the identification inaccuracy, revealing that 43 (81.1%) isolates were identified as C. parapsilosis sensu stricto and 10 (18.9%) as C. orthopsilosis. No C. metapsilosis was found. The geographic distribution of these Candida species was uniform among the studied Brazilian States (São Paulo, Rio de Janeiro and Espírito Santo). All C. orthopsilosis and almost all C. parapsilosis sensu stricto (95.3%) isolates were susceptible to amphotericin B, fluconazole, itraconazole, voriconazole and caspofungin. Nevertheless, one C. parapsilosis sensu stricto isolate was resistant to fluconazole and another one was resistant to caspofungin. C. parapsilosis sensu stricto isolates exhibited higher MIC mean values to amphotericin B, fluconazole and caspofungin than those of C. orthopsilosis, while C. orthopsilosis isolates displayed higher MIC mean to itraconazole compared to C. parapsilosis sensu stricto. Identical MIC mean values to voriconazole were measured for these Candida species. All the isolates of both species were able to form biofilm on polystyrene surface. Impressively, biofilm-growing cells of C. parapsilosis sensu stricto and C. orthopsilosis exhibited a considerable resistance to all antifungal agents tested. Pseudohyphae were observed in 67.4% and 80

  7. Ten-year study of species distribution and antifungal susceptibilities of Candida bloodstream isolates at a Brazilian tertiary hospital.

    Science.gov (United States)

    Bonfietti, L X; Szeszs, M W; Chang, M R; Martins, M A; Pukinskas, S R B S; Nunes, M O; Pereira, G H; Paniago, A M M; Purisco, S U; Melhem, M S C

    2012-12-01

    To describe the incidence and susceptibility profile of Candida bloodstream infections in a tertiary-care hospital, we performed a retrospective observational study from 1998 to 2007. Comorbidities and risk factors were compiled from all cases. In vitro susceptibility testing to fluconazole, itraconazole, voriconazole, and amphotericin B was performed for 100 isolates, and caspofungin was tested for C. parapsilosis complex. In a ten-year evaluation of candidemias, 44 % were caused by C. albicans, and species of the C. parapsilosis complex were the second most frequent agents (37 %). Other species presented lower incidences (C. tropicalis, 13 %, C. glabrata, 5 %, and C. krusei, 1 %). Neither C. dubliniensis nor C. metapsilosis were observed in this study. C. orthopsilosis (3 %) and C. parapsilosis stricto sensu (34 %) were also found. Species distribution was independent of catheterization, mechanical ventilation, or previous use of antifungals or corticoids. Parenteral nutrition administration was strongly related to C. glabrata infection, and the highest mortality (80 %) was observed in patients infected by this species. All C. albicans isolates showed high susceptibility to all tested drugs. However, two C. parapsilosis stricto sensu isolates presented high minimum inhibitory concentration (MIC) (4 mg/L each) to fluconazole, and one exhibited voriconazole MIC of 0.25 mg/L, highlighting the cross-resistance to these azoles. All isolates of C. tropicalis and C. glabrata showed no resistance to any drug tested. No difference was noted between C. parapsilosis and C. orthopsilosis susceptibilities to caspofungin. Our results suggest that resistance to amphotericin B, fluconazole, voriconazole, itraconazole, and caspofungin in Brazilian Candida bloodstream isolates is still uncommon.

  8. Correlation of anti-fungal susceptibility with clinical outcomes in patients with cryptococcal meningitis

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    Lee Chen-Hsiang

    2012-12-01

    Full Text Available Abstract Background This study aimed to investigate the correlation of minimum inhibiting concentrations (MICs, obtained by broth micro-dilution, and clinical response in patients with cryptococcal meningitis. Methods Using retrospective analyses covering the period 2001–2010, factors affecting clinical therapeutic cure in patients with cryptococcal meningitis 10 weeks after the start of anti-fungal therapy were identified. Specific emphasis was placed on the role of anti-fungal susceptibility. Results Of 46 patients with cryptococcal meningitis identified, 21 were cured after 10 weeks of treatment. Overall, 12 strains (26.1% were resistant to fluconazole (>8 μg/ml and 8 (17.4% had an MIC >1 μg/ml for amphotericin B. Twenty-three patients received combination amphotericin B and fluconazole as their initial antifungal therapy, 17 were given amphotericin B only, five received fluconazole only, and one received a combination of amphotericin B and flucytosine. After 2 weeks, all patients received fluconazole (400–600 mg daily for 8 weeks at least, then 200 mg daily thereafter. The presence of isolates resistant to fluconazole (MIC >8 μg/ml; 4.8% vs. 44%, p 8 μg/ml, was an independent predictor of therapeutic cure at 10-week evaluation (OR = 15.7; 95% CI: 1.8-135.9; p = 0.01, but higher MIC of amphotericin B (>1 μg/ml was not. Conclusions The MICs of fluconazole, determined by the CLSI method, may be a potential predictor of therapeutic cure in patients with cryptococcal meningitis.

  9. Geographic distribution and antifungal susceptibility of the newly described species Candida orthopsilosis and Candida metapsilosis in comparison to the closely related species Candida parapsilosis.

    Science.gov (United States)

    Lockhart, Shawn R; Messer, Shawn A; Pfaller, Michael A; Diekema, Daniel J

    2008-08-01

    Candida orthopsilosis and Candida metapsilosis are recently described species, having previously been grouped with the more prevalent species Candida parapsilosis. Current literature contains very little data pertaining to the distributions and antifungal susceptibilities of these Candida species. We determined the species and antifungal susceptibilities of 1,929 invasive clinical isolates from the ARTEMIS antifungal surveillance program collected between 2001 and 2006 and identified as C. parapsilosis using Vitek and conventional methods. Of the 1,929 isolates of presumed C. parapsilosis tested, 117 (6.1%) were identified as C. orthopsilosis and 34 (1.8%) as C. metapsilosis. The percentage of presumed C. parapsilosis isolates found to be C. orthopsilosis varied greatly by region, with the highest percentage (10.9%) from South America and the lowest (0.7%) from Africa. The MIC distributions of the C. orthopsilosis and C. metapsilosis isolates were statistically significantly lower than those of C. parapsilosis for all drugs except fluconazole, for which they were significantly higher (P orthopsilosis or C. metapsilosis isolates were fluconazole resistant, and all were susceptible to caspofungin, anidulafungin, and micafungin.

  10. Antifungal susceptibility of Candida albicans biofilms on titanium discs with different surface roughness.

    Science.gov (United States)

    Tsang, C S P; Ng, H; McMillan, A S

    2007-12-01

    Although it is well known that fungal biofilms have increased resistance to antimicrobial agents, limited information is available on the formation of candidal biofilms on implant surfaces with different surface roughness and their resistance to conventional antifungal therapy. In the current study, the effect of increasing the surface roughness of titanium discs on the susceptibility of Candida albicans biofilms to amphotericin B was determined. Grade I commercially pure titanium discs were sandblasted with 99.6% aluminium oxide of different grit sizes, producing surface roughness of 0.90, 1.88 and 3.82 microm (Groups A, B and C), respectively (P XTT assay. The 50% reduction in metabolic activity (50% RMA) of planktonic C. albicans (0.5 microg/mL) was much lower than those from Groups A, B and C (2, 16, 2 microg/mL, respectively), while the 50% RMA from Group B was three-fold higher than those from Groups A and C. In conclusion, difference in titanium surface roughness was associated with variations in the antifungal resistance of the candidal biofilm. Group C appeared to have an optimum surface roughness for biofilm resistance.

  11. Microdilution in vitro antifungal susceptibility of Exophiala dermatitidis, a systemic opportunist.

    Science.gov (United States)

    Badali, H; de Hoog, G S; Sudhadham, M; Meis, J F

    2011-11-01

    The in vitro activities of eight antifungal agents were determined against clinical (n = 63 genotype A, n = 3 genotype B) and environmental (n = 2 genotype A, n = 13 genotype B) strains of Exophiala dermatitidis. The resulting MIC(90)s for all strains (N = 81) were, in increasing order, as follows: posaconazole, 0.125 μg/ml; itraconazole, 0.25 μg/ml; voriconazole, 0.5 μg/ml; amphotericin B, 0.5 μg/ml; isavuconazole, 1 μ/ml; caspofungin, 8 μg/ml; anidulafungin, 8 μg/ml and fluconazole, 16 μg/ml. There were no significant differences in the patterns of susceptibility between genotypes A and B, environmental and clinical strains, isolates recovered from cutaneous and deep locations and strains from different geographical areas (P > 0.05). The difference in the MIC(90)s between each of these groups was not more than one dilution. The present study demonstrated that, based on in vitro activity, posaconazole and itraconazole have the highest activity against this fungus. In addition, voriconazole and the experimental broad-spectrum antifungal triazole, isavuconazole, both of which are available as intravenous preparations, have adequate activity against E. dermatitidis. However, in vivo efficacy remains to be determined.

  12. In vitro antifungal susceptibility profile and correlation of mycelial and yeast forms of molecularly characterized Histoplasma capsulatum strains from India

    NARCIS (Netherlands)

    Kathuria, S.; Singh, P.K.; Meis, J.F.G.M.; Chowdhary, A.

    2014-01-01

    The antifungal susceptibility profiles of the mycelial and yeast forms of 23 Histoplasma capsulatum strains from pulmonary and disseminated histoplasmosis patients in India are reported here. The MIC data of this dimorphic fungus had good agreement between both forms for azoles, amphotericin B, and

  13. Antifungal drug susceptibility and phylogenetic diversity among Cryptococcus isolates from dogs and cats in North America.

    Science.gov (United States)

    Singer, Lisa M; Meyer, Wieland; Firacative, Carolina; Thompson, George R; Samitz, Eileen; Sykes, Jane E

    2014-06-01

    Molecular types of the Cryptococcus neoformans/Cryptococcus gattii species complex that infect dogs and cats differ regionally and with host species. Antifungal drug susceptibility can vary with molecular type, but the susceptibility of Cryptococcus isolates from dogs and cats is largely unknown. Cryptococcus isolates from 15 dogs and 27 cats were typed using URA5 restriction fragment length polymorphism analysis (RFLP), PCR fingerprinting, and multilocus sequence typing (MLST). Susceptibility was determined using a microdilution assay (Sensititre YeastOne; Trek Diagnostic Systems). MICs were compared among groups. The 42 isolates studied comprised molecular types VGI (7%), VGIIa (7%), VGIIb (5%), VGIIc (5%), VGIII (38%), VGIV (2%), VNI (33%), and VNII (2%), as determined by URA5 RFLP. The VGIV isolate was more closely related to VGIII according to MLST. All VGIII isolates were from cats. All sequence types identified from veterinary isolates clustered with isolates from humans. VGIII isolates showed considerable genetic diversity compared with other Cryptococcus molecular types and could be divided into two major subgroups. Compared with C. neoformans MICs, C. gattii MICs were lower for flucytosine, and VGIII MICs were lower for flucytosine and itraconazole. For all drugs except itraconazole, C. gattii isolates exhibited a wider range of MICs than C. neoformans. MICs varied with Cryptococcus species and molecular type in dogs and cats, and MICs of VGIII isolates were most variable and may reflect phylogenetic diversity in this group. Because sequence types of dogs and cats reflect those infecting humans, these observations may also have implications for treatment of human cryptococcosis.

  14. Phylogeny of the clinically relevant species of the emerging fungus Trichoderma and their antifungal susceptibilities.

    Science.gov (United States)

    Sandoval-Denis, Marcelo; Sutton, Deanna A; Cano-Lira, José F; Gené, Josepa; Fothergill, Annette W; Wiederhold, Nathan P; Guarro, Josep

    2014-06-01

    A set of 73 isolates of the emerging fungus Trichoderma isolated from human and animal clinical specimens were characterized morphologically and molecularly using a multilocus sequence analysis that included the internal transcribed spacer (ITS) regions of the nuclear ribosomal DNA and fragments of the translation elongation factor 1 alpha (Tef1), endochitinase CHI18-5 (Chi18-5), and actin 1 (Act1) genes. The most frequent species was Trichoderma longibrachiatum (26%), followed by Trichoderma citrinoviride (18%), the Hypocrea lixii/Trichoderma harzianum species complex (15%), the newly described species Trichoderma bissettii (12%), and Trichoderma orientale (11%). The most common anatomical sites of isolation in human clinical specimens were the respiratory tract (40%), followed by deep tissue (30%) and superficial tissues (26%), while all the animal-associated isolates were obtained from superficial tissue samples. Susceptibilities of the isolates to eight antifungal drugs in vitro showed mostly high MICs, except for voriconazole and the echinocandins.

  15. Candida urinary tract infection and Candida species susceptibilities to antifungal agents.

    Science.gov (United States)

    Osawa, Kayo; Shigemura, Katsumi; Yoshida, Hiroyuki; Fujisawa, Masato; Arakawa, Soichi

    2013-11-01

    The purpose of this study is to review Candida isolation from urine of urinary tract infection (UTI) patients over the recent 3 years at the Kobe University Hospital. We recorded the type of strain, the department where the patient was treated such as the intensive care unit (ICU), and combined isolation of Candida with other microorganisms. We investigated Candida isolation and susceptibilities to antifungal agents and analyzed the risk factors for combined isolation with other microorganisms. The most frequently isolated Candida was Candida albicans, which showed good (100%) susceptibilities to 5-fluorocytosine (5-FC) and fluconazole (FLCZ) but not to voriconazole (VRCZ), followed by C. glabrata. ICU was the greatest source of Candida-positive samples, and the most relevant underlying diseases of ICU patients were pneumonia followed by renal failure and post liver transplantation status. Combined isolation with other bacteria was seen in 27 cases (42.9%) in 2009, 25 (33.3%) in 2010 and 31 (31.3%) in 2011 and comparatively often seen in non-ICU patients. Other candidas than C. albicans showed significantly decreased susceptibility to FLCZ over these 3 years (P=0.004). One hundred (97.1%) of 103 ICU cases were given antibiotics at the time of Candida isolation, and the most often used antibiotics were cefazolin or meropenem. In conclusion, C. albicans was representatively isolated in Candida UTI and showed good susceptibilities to 5-FC, FLCZ and VRCZ, but other candidas than C. albicans showed significantly decreased susceptibility to FLCZ in the change of these 3 years.

  16. Antifungal susceptibility of clinical and environmental isolates of Cryptococcus neoformans to four antifungal drugs determined by two techniques.

    Science.gov (United States)

    Moraes, E M P; Prímola, N S; Hamdan, Júnia Soares

    2003-06-01

    A total of 64 Cryptococcus neoformans strains, including clinical and environmental Brazilian isolates var. neoformans and var. gattii, were tested for susceptibility to amphotericin B, 5-flucytosine, fluconazole and itraconazole. The tests were performed according to the recommendations of National Committee of Clinical Laboratory Standards and the method of macrodilution in liquid medium of Shadomy et al. [Manual de Microbiologia Clínica, 4th ed. Buenos Aires: Editorial Medica Panamericana, 1987: 1229-38]. For most drugs there was a significant difference between the readings taken at 24 and 48 h with both methods. When the minimum inhibitory concentrations obtained by the two techniques were compared, significant differences were observed for amphotericin B and fluconazole. Overall, differences in drug susceptibility with respect to the origin of the isolates or the variety of the fungus were not observed. As an exception, the gattii variety exhibited a high resistance rate to amphotericin B when the technique of Shadomy et al. was applied, a fact possibly related to the greater difficulty for treatment of the disease caused by this fungal variety.

  17. Studies in Phylogeny, Development of Rapid IdentificationMethods, Antifungal Susceptibility, and Growth Rates of Clinical Strains of Sporothrix schenckii Complex in Japan.

    Science.gov (United States)

    Suzuki, Rumi; Yikelamu, Alimu; Tanaka, Reiko; Igawa, Ken; Yokozeki, Hiroo; Yaguchi, Takashi

    2016-01-01

    Sporotrichosis is a fungal infection caused by the Sporothrix species, which have distinct virulence profiles and geographic distributions. We performed a phylogenetic study in strains morphologically identified as Sporothrix schenckii from clinical specimens in Japan, which were preserved at the Medical Mycology Research Center, Chiba University. In addition, we examined the in vitro antifungal susceptibility and growth rate to evaluate their physiological features. Three hundred strains were examined using sequence analysis of the partial calmodulin gene, or polymerase chain reaction(PCR)method using newly designed species-specific primers; 291 strains were Sporothrix globosa and 9 strains were S. schenckii sensu stricto (in narrow sense, s. s.). S. globosa strains were further clustered into two subclades, and S. schenckii s. s. strains were divided into three subclades. In 38 strains of S. globosa for which antifungal profiles were determined, 4 strains (11%) showed high minimal inhibitory concentration (MIC) value for itraconazole. All tested strains of S. schenckii s. s. and S. globosa showed low sensitivity for amphotericin B. These antifungals are used for treatment of sporotrichosis when infection is severe. S. schenckii s. s. grew better than S. globosa; wherein S. globosa showed restricted growth at 35℃ and did not grow at 37℃. Our molecular data showed that S. globosa is the main causal agent of sporotrichosis in Japan. It is important to determine the antifungal profiles of each case, in addition to accurate species-level identification, to strategize the therapy for sporotrichosis.

  18. Comparison between E-test and CLSI broth microdilution method for antifungal susceptibility testing of Candida albicans oral isolates Comparação entre E-test e o método da microdiluição do CLSI para teste de susceptibilidade a antifúngicos de isolados orais de Candida albicans

    Directory of Open Access Journals (Sweden)

    Cristiane Yumi Koga-Ito

    2008-02-01

    Full Text Available Thirty Candida albicans isolated from oral candidosis patients and 30 C. albicans isolated from control individuals were studied. In vitro susceptibility tests were performed for amphotericin B, fluconazole, 5-flucytosine and itraconazole through the Clinical and Laboratorial Standards Institute (CLSI reference method and E test system. The results obtained were analyzed and compared. MIC values were similar for the strains isolated from oral candidosis patients and control individuals. The agreement rate for the two methods was 66.67% for amphotericin B, 53.33% for fluconazole, 65% for flucytosine and 45% for itraconazole. According to our data, E test method could be an alternative to trial routine susceptibility testing due to its simplicity. However, it can not be considered a substitute for the CLSI reference method.Trinta Candida albicans isoladas de pacientes portadores de candidose oral e 30 Candida albicans isoladas de indivíduos controle foram estudadas. Testes de susceptibilidade in vitro foram realizados com anfotericina B, fluconazol, 5-flucitosina e itraconazol pelo método do Clinical and Laboratorial Standars Institute (CLSI e por E-test. Os resultados obtidos foram analisados e comparados. Os valores de CIM foram semelhantes para amostras isoladas de pacientes portadores de candidose oral e indivíduos controle. A concordância entre os dois métodos foi de 66,7% para a anfotericina B, 53,33% para o fluconazol, 65% para a flucitosina e 45% para o itraconazol. De acordo com estes resultados, o método do E-test poderia ser uma alternativa para a triagem de casos de rotina pela sua simplicidade. Entretanto, este método não pode ser considerado como um substituto para o método de referência do CLSI.

  19. Utility of in-house fluconazole disk diffusion susceptibility testing in the treatment of candidemia

    Science.gov (United States)

    Kubiak, David W.; Farmakiotis, Dimitrios; Arons, Viktoria; Hollins, Randy M.; Rostas, Sara E.; Weiser, Linda M.; Baden, Lindsey R.; Marty, Francisco M.; Koo, Sophia

    2016-01-01

    Among 302 first candidemia episodes, 210 (69.6%) were initially treated with an echinocandin or polyene (E/P) antifungal drug. In 137 (72.5%) patients with fluconazole-susceptible isolates, treatment was changed to fluconazole based on disk diffusion susceptibility testing. Clinical outcomes were not compromised in patients receiving E/P who were de-escalated to fluconazole for treatment of candidemia based on disk diffusion results. PMID:26763713

  20. Utility of in-house fluconazole disk diffusion susceptibility testing in the treatment of candidemia.

    Science.gov (United States)

    Kubiak, David W; Farmakiotis, Dimitrios; Arons, Viktoria; Hollins, Randy M; Rostas, Sara E; Weiser, Linda M; Baden, Lindsey R; Marty, Francisco M; Koo, Sophia

    2016-03-01

    Among 302 first candidemia episodes, 210 (69.6%) were initially treated with an echinocandin or polyene (E/P) antifungal drug. In 137 (72.5%) patients with fluconazole-susceptible isolates, treatment was changed to fluconazole based on disk diffusion susceptibility testing. Clinical outcomes were not compromised in patients receiving E/P who were de-escalated to fluconazole for treatment of candidemia based on disk diffusion results.

  1. SUSCEPTIBILITY TEST FOR FUNGI: CLINICAL AND LABORATORIAL CORRELATIONS IN MEDICAL MYCOLOGY

    Directory of Open Access Journals (Sweden)

    Ana ALASTRUEY-IZQUIERDO

    2015-09-01

    Full Text Available SUMMARYDuring recent decades, antifungal susceptibility testing has become standardized and nowadays has the same role of the antibacterial susceptibility testing in microbiology laboratories. American and European standards have been developed, as well as equivalent commercial systems which are more appropriate for clinical laboratories. The detection of resistant strains by means of these systems has allowed the study and understanding of the molecular basis and the mechanisms of resistance of fungal species to antifungal agents. In addition, many studies on the correlation of in vitro results with the outcome of patients have been performed, reaching the conclusion that infections caused by resistant strains have worse outcome than those caused by susceptible fungal isolates. These studies have allowed the development of interpretative breakpoints for Candida spp. and Aspergillus spp., the most frequent agents of fungal infections in the world. In summary, antifungal susceptibility tests have become essential tools to guide the treatment of fungal diseases, to know the local and global disease epidemiology, and to identify resistance to antifungals.

  2. Antifungal Drug Susceptibility of Candida Species Isolated from HIV-Positive Patients Recruited at a Public Hospital in São Luís, Maranhão, Brazil

    Science.gov (United States)

    Terças, Ana L. G.; Marques, Sirlei G.; Moffa, Eduardo B.; Alves, Márcia B.; de Azevedo, Conceição M. P. S.; Siqueira, Walter L.; Monteiro, Cristina A.

    2017-01-01

    Oropharyngeal candidiasis is the most common fungal infection in hospitalized patients with acquired immune deficiency syndrome (AIDS). Its progression results in invasive infections, which are a significant cause of morbidity and mortality. This study aimed to quickly and accurately identify Candida spp. from oral mucosa of AIDS patients recruited at Presidente Vargas Hospital, in São Luís city, Brazil and to evaluate the sensitivity profile of these fungi to antifungals by using an automated system. Isolates were collected from oropharyngeal mucosa of 52 hospitalized AIDS patients, under anti-viral and antifungal therapies. Patients were included in research if they were HIV-positive, above 18 years of age and after obtaining their written consent. CHROMagar®Candida and the automated ViteK-2®system were used to isolate and identify Candida spp., respectively. Antifungal susceptibility testing was performed using the ViteK-2®system, complemented with the Etest®, using the drugs amphotericin B, fluconazole, flucytosine, and voriconazole. Oropharyngeal candidiasis had a high prevalence in these hospitalized AIDS patients (83%), and the most prevalent species was Candida albicans (56%). Antifungal susceptibility test showed that 64.7% of the Candida spp. were susceptible, 11.8% were dose-dependent sensitive, and 23.5% were resistant. All the Candida krusei and Candida famata isolates and two of Candida glabrata were resistant to fluconazole. Most of AIDS patients presented oropharyngeal candidiasis and C. albicans was the most frequently isolated species. The results showed high variability in resistance among isolated species and indicates the need to identify the Candida spp. involved in the infection and the need to test antifungal susceptibility as a guide in drug therapy in patients hospitalized with AIDS. This is the first relate about AIDS patients monitoring in a public hospital in São Luís concerning the precise identification and establishing of

  3. Isavuconazole and nine comparator antifungal susceptibility profiles for common and uncommon Candida species collected in 2012: application of new CLSI clinical breakpoints and epidemiological cutoff values.

    Science.gov (United States)

    Castanheira, Mariana; Messer, Shawn A; Rhomberg, Paul R; Dietrich, Rachel R; Jones, Ronald N; Pfaller, Michael A

    2014-08-01

    The in vitro activity of isavuconazole and nine antifungal comparator agents was assessed using reference broth microdilution methods against 1,421 common and uncommon species of Candida from a 2012 global survey. Isolates were identified using CHROMagar, biochemical methods and sequencing of ITS and/or 28S regions. Candida spp. were classified as either susceptible or resistant and as wild type (WT) or non-WT using CLSI clinical breakpoints or epidemiological cutoff values, respectively, for the antifungal agents. Isolates included 1,421 organisms from 21 different species of Candida. Among Candida spp., resistance to all 10 tested antifungal agents was low (0.0-7.9 %). The vast majority of each species of Candida, with the exception of Candida glabrata, Candida krusei, and Candida guilliermondii (modal MICs of 0.5 µg/ml), were inhibited by ≤0.12 µg/ml of isavuconazole (99.0 %; range 94.3 % [Candida tropicalis] to 100.0 % [Candida lusitaniae and Candida dubliniensis]). C. glabrata, C. krusei, and C. guilliermondii were largely inhibited by ≤1 µg/ml of isavuconazole (89.7, 96.9 and 92.8 %, respectively). Decreased susceptibility to isavuconazole was most prominent with C. glabrata where the modal MIC for isavuconazole was 0.5 µg/ml for those strains that were SDD to fluconazole or WT to voriconazole, and was 4 µg/ml for those that were either resistant or non-WT to fluconazole or voriconazole, respectively. In conclusion, these data document the activity of isavuconazole and generally the low resistance levels to the available antifungal agents in a large, contemporary (2012), global collection of molecularly characterized species of Candida.

  4. Molecular characterization and antifungal susceptibility of Cryptococcus neoformans strains collected from a single institution in Lima, Peru : Revista Iberoamericana De Micologia

    NARCIS (Netherlands)

    Bejar, V.; Tello, M.; Garcia, R.; Guevara, J. M.; Gonzales, S.; Vergaray, G.; Valencia, E.; Abanto, E.; Ortega-Loayza, A. G.; Hagen, F.; Gutierrez, E. L.

    2015-01-01

    Background: Cryptococcosis is a fungal infection with a worldwide distribution, mainly caused by Cryptococcus neoformans and Cryptococcus gattii. Aims: To molecularly characterize the mating-types, serotypes, genotypes and antifungal susceptibility profiles of a set of retrospectively isolated C. ne

  5. [Antifungal susceptibility profiles of Candida species to triazole: application of new CLSI species-specific clinical breakpoints and epidemiological cutoff values for characterization of antifungal resistance].

    Science.gov (United States)

    Karabıçak, Nilgün; Alem, Nihal

    2016-01-01

    The Clinical and Laboratory Standards Institute (CLSI) Subcommittee on Antifungal Susceptibility Testing has newly introduced species-specific clinical breakpoints (CBPs) for fluconazole and voriconazole. When CBPs can not be determined, wild-type minimal inhibitory concentration (MIC) distributions are detected and epidemiological cutoff values (ECVs) provide valuable means for the detection of emerging resistance. The aim of this study is to determine triazole resistance patterns in Candida species by the recently revised CLSI CBPs. A total of 140 Candida strains isolated from blood cultures of patients with invasive candidiasis hospitalized in various intensive care units in Turkey and sent to our reference laboratory between 2011-2012, were included in the study. The isolates were identified by conventional methods, and susceptibility testing was performed against fluconazole, itraconazole and voriconazole, by the 24-h CLSI broth microdilution (BMD) method. Azole resistance rates for all Candida species were determined using the new species-specific CLSI CBPs and ECVs criteria, when appropriate. The species distribution of the isolates were as follows; C.parapsilosis (n= 31 ), C.tropicalis (n= 26 ), C.glabrata (n= 21), C.albicans (n= 18), C.lusitaniae (n= 16), C.krusei (n= 16), C.kefyr (n= 9), C.guilliermondii (n= 2), and C.dubliniensis (n= 1). According to the newly determined CLSI CBPs for fluconazole and C.albicans, C.parapsilosis, C.tropicalis [susceptible (S), ≤ 2 µg/ml; dose-dependent susceptible (SDD), 4 µg/ml; resistant (R), ≥ 8 µg/ml], and C.glabrata (SDD, ≤ 32 µg/ml; R≥ 64 µg/ml) and for voriconazole and C.albicans, C.parapsilosis, C.tropicalis (S, ≤ 0.12 µg/ml; SDD, 0.25-0.5 µg/ml; R, ≥ 1 µg/ml), and C.krusei (S, ≤ 0.5 µg/ml; SDD, 1 µg/ml; R, ≥ 2 µg/ml), it was found that three of C.albicans, one of C.parapsilosis and one of C.glabrata isolates were resistant to fluconazole, while two of C.albicans and two of C

  6. Species Diversity, Antifungal Susceptibility, and Virulence Attributes of Candida Colonising the Oral Cavities of Adult Diabetic Patients

    OpenAIRE

    Deepa Anil Kumar; Sumathi Muralidhar; Krishna Biswas; Uma Banerjee; Seemi Farhat Basir; Luqman Ahmad Khan

    2014-01-01

    Oral candidiasis is a common occurrence in diabetic patients. Species of Candida isolated from these infections and their virulence pattern undergo changes over time and require periodic assessments. Objective of this study was to determine changes in the spectrum of Candida species colonizing oral cavity, their antifungal susceptibility patterns, and virulence attributes, in adult diabetic patients. Oral swabs were collected from 100 patients with diabetes mellitus (DM) and an equal number o...

  7. Dermatophyte susceptibilities to antifungal azole agents tested in vitro by broth macro and microdilution methods Suscetibilidade in vitro de dermatófitos a azóis pelos métodos macro e microdiluição em caldo

    Directory of Open Access Journals (Sweden)

    Emerson Roberto Siqueira

    2008-02-01

    Full Text Available The in vitro susceptibility of dermatophytes to the azole antifungals itraconazole, fluconazole and ketoconazole was evaluated by broth macro and microdilution methods, according to recommendations of the CLSI, with some adaptations. Twenty nail and skin clinical isolates, four of Trichophyton mentagrophytes and 16 of T. rubrum were selected for the tests. Itraconazole minimal inhibitory concentrations (MIC varied from Foi avaliada a suscetibilidade in vitro de dermatófitos aos antifúngicos itraconazol, fluconazol e cetoconazol, pelos métodos macro e microdiluição em caldo, de acordo com as recomendações do CLSI, com algumas modificações. Foram estudados 20 isolados clínicos de lesões de unha e pele, sendo quatro Trichophyton mentagrophytes e 16 T. rubrum. A concentração inibitória mínima (CIM para itraconazol variou de < 0,03 a 0,25 µg/mL pelo método da macrodiluição, e de < 0,03 a 0,5 µg/mL pela microdiluição em caldo; de 0,5 a 64 µg/mL e de 0,125 a 16 µg/mL para fluconazol, respectivamente, pela macro e microdiluição; e de < 0,03 a 0,5 µg/mL por ambos os métodos para cetoconazol. A concordância entre os dois métodos (considerando ± uma diluição foi de 70% para itraconazol, 45% para fluconazol e 85% para cetoconazol. Conclui-se que os isolados estudados foram inibidos por concentrações relativamente baixas dos antifúngicos testados, e os dois métodos apresentam boa concordância, especialmente para itraconazol e cetoconazol.

  8. Isothermal microcalorimetry: a novel method for real-time determination of antifungal susceptibility of Aspergillus species.

    NARCIS (Netherlands)

    Furustrand Tafin, U.; Clauss, M.; Hauser, P.M.; Bille, J.; Meis, J.F.G.M.; Trampuz, A.

    2012-01-01

    We evaluated microcalorimetry for real-time susceptibility testing of Aspergillus spp. based on growth-related heat production. The minimal heat inhibitory concentration (MHIC) for A. fumigatus ATCC 204305 was 1 mg/L for amphotericin B, 0.25 mg/L for voriconazole, 0.06 mg/L for posaconazole, 0.125 m

  9. Antimicrobial susceptibility and susceptibility testing of Mycoplasma hominis: a review.

    Science.gov (United States)

    Bygdeman, S M; Mårdh, P A

    1983-01-01

    The determination of the minimal growth-inhibiting concentration (MIC), the minimal metabolism-inhibiting concentration (MMC), and the minimal mycoplasmacidal concentration (MCC) of various antimicrobial compounds for Mycoplasma hominis is influenced by the pH of the test media, the inoculum size, and the incubation time, although each of these factors generally do not affect the minimal concentration more than fourfold. M. hominis is resistant to beta-lactam antibiotics, vancomycin, sulfonamides, trimethoprim, and polymyxin B. There are great differences in the susceptibility of M. hominis to various macrolide antibiotics. Thus the organism is resistant to erythromycin and oleandomycin, moderately resistant to tylosin and spiramycin, susceptible to josamycin as well as to another macrolide drug, labelled M-4365G. M. hominis is also highly susceptible to the macrolide-like compound rosaramicin and to the tetracyclines (although resistant strains occur). It is susceptible to lincomycin and clindamycin, and moderately susceptible to chloramphenicol and rifampicin. The aminoglycosides have limited activity against M. hominis.

  10. In vitro antifungal susceptibility patterns of Candida albicans from HIV and AIDS patients attending the Nylon Health District Hospital in Douala, Cameroon

    Directory of Open Access Journals (Sweden)

    Anna L. Njunda

    2012-05-01

    Full Text Available HIV and AIDS are major public health problems in Cameroon where the HIV prevalence is 5.5%. Candidiasis is the leading opportunistic mycosis in HIV and AIDS patients. The objective of this study was to determine the in vitro antifungal susceptibility pattern of Candida albicans in HIV and AIDS patients to eight antifungal agents in the Nylon Health District of Douala in Cameroon. Three hundred and four HIV and AIDS patients were recruited between March and August 2007 to participate in a cross-sectional study. All subjects who fulfilled the inclusion criteria were enrolled. Informed consent was obtained from all subjects before samples were collected. Three samples comprising oral swabs, vagina/urethra swabs and a mid-stream urine were collected from each subject. Specimens were cultured on sabouraud dextrose agar and C. albicans isolates were identified using the germ tube technique. The disk diffusion method was used for antifungal susceptibility testing using eight antifungal agents. The prevalence of candidiasis in the study population was 67.8% (95% CI: 62.5-73.1% and that of C. albicans was 42.8% (95% CI: 37.2-48.4%. Oral swabs had the highest prevalence of C. albicans followed by vaginal/urethral samples (52.6% vs. 29.7% respectively. Forty (30.8% subjects had C. albicans infection at more than one collection site. There was a statistically significant difference in the infectivity of C. albicans with age, sex and site of infection (P<0.05. C. albicans isolates were most sensitive to ketoconazole (80% followed by econazole (64.6% while fluconazole and 5-flurocytosin recorded the poorest sensitivities (22.9% vs 24.6%, respectively. There was a statistically significant difference in the sensitivity pattern of antifungal agents with respect to the site of isolation of the organism (P<0.05. Ketoconazole is the drug of choice for the treatment of C. albicans infection in HIV and AIDS patients in the Nylon Health District of Douala, Cameroon.

  11. Echinocandin and triazole antifungal susceptibility profiles for clinical opportunistic yeast and mold isolates collected from 2010 to 2011: application of new CLSI clinical breakpoints and epidemiological cutoff values for characterization of geographic and temporal trends of antifungal resistance.

    Science.gov (United States)

    Pfaller, Michael A; Messer, Shawn A; Woosley, Leah N; Jones, Ronald N; Castanheira, Mariana

    2013-08-01

    The SENTRY Antimicrobial Surveillance Program monitors global susceptibility and resistance rates of newer and established antifungal agents. We report the echinocandin and triazole antifungal susceptibility patterns for 3,418 contemporary clinical isolates of yeasts and molds. The isolates were obtained from 98 laboratories in 34 countries during 2010 and 2011. Yeasts not presumptively identified by CHROMagar, the trehalose test, or growth at 42°C and all molds were sequence identified using internal transcribed spacer (ITS) and 28S (yeasts) or ITS, translation elongation factor (TEF), and 28S (molds) genes. Susceptibility testing was performed against 7 antifungals (anidulafungin, caspofungin, micafungin, fluconazole, itraconazole, posaconazole, and voriconazole) using CLSI methods. Rates of resistance to all agents were determined using the new CLSI clinical breakpoints and epidemiological cutoff value criteria, as appropriate. Sequencing of fks hot spots was performed for echinocandin non-wild-type (WT) strains. Isolates included 3,107 from 21 Candida spp., 146 from 9 Aspergillus spp., 84 from Cryptococcus neoformans, 40 from 23 other mold species, and 41 from 9 other yeast species. Among Candida spp., resistance to the echinocandins was low (0.0 to 1.7%). Candida albicans and Candida glabrata that were resistant to anidulafungin, caspofungin, or micafungin were shown to have fks mutations. Resistance to fluconazole was low among the isolates of C. albicans (0.4%), Candida tropicalis (1.3%), and Candida parapsilosis (2.1%); however, 8.8% of C. glabrata isolates were resistant to fluconazole. Among echinocandin-resistant C. glabrata isolates from 2011, 38% were fluconazole resistant. Voriconazole was active against all Candida spp. except C. glabrata (10.5% non-WT), whereas posaconazole showed decreased activity against C. albicans (4.4%) and Candida krusei (15.2% non-WT). All agents except for the echinocandins were active against C. neoformans, and the

  12. Real-Time Optical Antimicrobial Susceptibility Testing

    DEFF Research Database (Denmark)

    Fredborg, Marlene; Andersen, Klaus R; Jørgensen, Erik;

    2013-01-01

    Rapid antibiotic susceptibility testing is in highly demand in health-care fields as antimicrobial resistant bacterial strains emerge and spread. Here we describe an optical screening system (oCelloScope), which based on time-lapse imaging of 96 bacteria-antibiotic combinations at a time, introdu......Rapid antibiotic susceptibility testing is in highly demand in health-care fields as antimicrobial resistant bacterial strains emerge and spread. Here we describe an optical screening system (oCelloScope), which based on time-lapse imaging of 96 bacteria-antibiotic combinations at a time......, introduces real-time detection of bacterial growth and antimicrobial susceptibility, with imaging material to support the automatically generated graphs. Automated antibiotic susceptibility tests of a monoculture showed statistically significant antibiotic effect within 6 minutes and within 30 minutes...

  13. INCIDENCE OF NON-CANDIDA ALBICANS IN PATIENTS WITH URINARY TRACT INFECTION WITH SPECIAL REFERENCE TO SPECIATIO N AND ANTIFUNGAL SUSCEPTIBILITY

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    Ragini Ananth

    2012-10-01

    Full Text Available ABSTRACT: BACKGROUND AND OBJECTIVES: Fungal urinary tract infections have become frequent, as a result of increased use of broad spec trum antibiotics, corticosteroids, immunosuppressive drugs and bladder catheters in acut e care settings. The associated risk factors which are seen in cases of candiduria are: antibiotic therapy, female gender, urinary catheterization, surgical procedure and extended hos pitalization. Candiduria has become a potential source of morbidity and mortality if untre ated. We undertook a prospective study to note the incidence of non-Candida albicans in patien ts with urinary tract infection with special reference to speciation, antifungal susceptibility an d the associated risk factors. METHODS: Candida species isolated from urine samples of patient s with urinary tract infection were subjected to speciation using standard yeast identif ication protocol and CHROM agar. Antifungal Susceptibility testing was done by the disc diffusio n method to amphotericin B and fluconazole. Clinical details and risk factors of the patients we re noted down. RESULTS: Among the 60 culture positive cases, six Candida species which wer e isolated are : C.tropicalis (66.66%, C.albicans (13.33%, C.parapsilosis (8.33%, C.glabr ata (6.66%, C.kefyr (3.33% and C.guilliermondii (1.66% The susceptibility pattern s howed, that of the 60 isolates, 40% were resistant to fluconazole. No resistance was seen to amphotericin B. CONCLUSION: Isolation of non-Candida albicans species was more than Candida a lbicans. Candida tropicalis was the predominant isolate. The following risk factors were noted: 43.33 % of the patients had diabetes mellitus, 30%had history of prolonged antib iotics (cephalosporin and aminoglycosides, 16.66% had underlying renal pathol ogy, 3.33% had post –renal transplant status, 1.66% were on steroids, 1.66%had pregnancy a nd 3.33% had no identifiable risk factors.20% patients had an indwelling catheter in them. The antifungal

  14. The clinical spectrum of Exophiala jeanselmei, with a case report and in vitro antifungal susceptibility of the species.

    Science.gov (United States)

    Badali, H; Najafzadeh, M J; van Esbroeck, M; van den Enden, E; Tarazooie, B; Meis, J F G M; de Hoog, G S

    2010-03-01

    Exophiala jeanselmei is clinically redefined as a rare agent of subcutaneous lesions of traumatic origin, eventually causing eumycetoma. Mycetoma is a localized, chronic, suppurative subcutaneous infection of tissue and contiguous bone after a traumatic inoculation of the causative organism. In advanced stages of the infection, one finds tumefaction, abscess formation and draining sinuses. The species has been described as being common in the environment, but molecular methods have only confirmed its occurrence in clinical samples. Current diagnostics of E. jeanselmei is based on sequence data of the Internal Transcribed Spacer (ITS) region of ribosomal DNA (rDNA), which sufficiently reflects the taxonomy of this group. The first purpose of this study was the re-identification of all clinical (n=11) and environmental strains (n=6) maintained under the name E. jeanselmei, and to establish clinical preference of the species in its restricted sense. Given the high incidence of eumycetoma in endemic areas, the second goal of this investigation was the evaluation of in vitro susceptibility of E.jeanselmei to eight conventional and new generations of antifungal drugs to improve antifungal therapy in patients. As an example, we describe a case of black grain mycetoma in a 43-year-old Thai male with several draining sinuses involving the left foot. The disease required extensive surgical excision coupled with intense antifungal chemotherapy to achieve cure. In vitro studies demonstrated that posaconazole and itraconazole had the highest antifungal activity against E. jeanselmei and E. oligosperma for which high MICs were found for caspofungin. However, their clinical effectiveness in the treatment of Exophiala infections remains to be determined.

  15. Antifungal susceptibility of Malassezia furfur in vitro tested by using modified ATB Fungus 3 method%改良ATB Fungus 3法检测糠秕马拉色菌对抗真菌药物的敏感性

    Institute of Scientific and Technical Information of China (English)

    陈东科

    2012-01-01

    目的 用改良ATB Fungus 3 法测定糠秕马拉色菌的药物敏感性,以了解临床患者呼吸道分离的糠秕马拉色菌对常用抗真菌药物的最低抑菌浓度(MIC).方法 在ATB F2半固体培养基中添加0.5%吐温40和0.5%吐温60,用ATB Fungus 3药敏板条对47株临床分离的糠秕马拉色菌进行MIC检测.结果 氟康唑MIC50为>128 μg/mL,MICRange为≤1.0~>128μg/mL;5-氟胞嘧啶MIC50为>16 μg/mL,MICRange为>16 μg/mL;沃尔康唑MIC50为8.0 μg/mL,MICRange为≤0.06~ >8.0μg/mL;两性霉素B MIC50为4.0 μg/mL,MICRange为≤0.5 ~ 16 μg/mL;伊曲康唑MIC50为1.0 μg/mL,MICRange为≤0.125 ~4.0μg/mL.呼吸道分离的糠秕马拉色菌对5种抗真菌药物的MIC值高于健康人皮肤分离的糠秕马拉色菌及标准菌株ATCC14521.结论 改良ATB Fungus 3法操作简便,结果重复性好且易观察.糠秕马拉色菌MIC值的升高可能与患者使用抗真菌药物有关.%Objective To determine the drug-susceptibility of Malassezia furfur by using modified ATB Fungus 3 method and evaluate the minimum inhibitory concentration (MIC) of clinical isolates of Malassezia furfur to antifungal agents. Methods Tween-40 and Tween-60 were added into ATB F2 semisolid culture media with final concentration of 0.5%. The MICs of 5 antifungal agents against 47 strains of Malassezia furfur were determined by ATB Fungus 3 method. Results The MIC50 of fluconazole,5-flucytosine,voricon-azole,amphotericin B and itraconazole against 47 Malassezia furfur strains were more than 128,more than 16,8. 0,4. 0 and 1. 0 μg/mL respectively,and the ranges of MIC were less than or equal to1. 0 to more than 128 μg/mL,more than 16μg/mL,less than or equal to 0. 06 to more than 8. 0 μg/mL,less than or equal to 0. 5 to 16 μg/mL,less than or equal to 0. 125 to 4.0 μg/mL. The results showed that the MIC values of 5 antifungal agents against clinical isolates of Malassezia furfur from respiratory tract were higher than those of

  16. Synergistic effects of tacrolimus and azole antifungal compounds in fluconazole-susceptible and fluconazole-resistant Candida glabrata isolates

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    Laura Bedin Denardi

    2015-03-01

    Full Text Available In vitro interaction between tacrolimus (FK506 and four azoles (fluconazole, ketoconazole, itraconazole and voriconazole against thirty clinical isolates of both fluconazole susceptible and -resistant Candida glabrata were evaluated by the checkerboard microdilution method. Synergistic, indifferent or antagonism interactions were found for combinations of the antifungal agents and FK506. A larger synergistic effect was observed for the combinations of FK506 with itraconazole and voriconazole (43%, followed by that of the combination with ketoconazole (37%, against fluconazole-susceptible isolates. For fluconazole-resistant C. glabrata, a higher synergistic effect was obtained from FK506 combined with ketoconazole (77%, itraconazole (73%, voriconazole (63% and fluconazole (60%. The synergisms that we observed in vitro, notably against fluconazole-resistant C. glabrata isolates, are promising and warrant further analysis of their applications in experimental in vivo studies.

  17. Susceptibility testing of Entamoeba histolytica

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    Cedeno, J.R.; Krogstad, D.J.

    1983-12-01

    The growth of Entamoeba histolytica in microtiter plates in vitro in a variety of environments with reduced oxygen tensions is reported. With 3% O/sub 2/, 3% CO/sub 2/, and 94% N/sub 2/, the parasite growth in microtiter plates was identical to that in screw-capped culture tubes, as measured by (/sup 3/H)thymidine incorporation and by quantitative parasite counts. There were no significant differences between the drug concentrations necessary to inhibit parasite growth by 50% based on (/sup 3/H)thymidine incorporation vs those defined by quantitative parasite counts for the 15 antimicrobial agents tested (including seven drugs used for the treatment of amebiasis). This technique provides a reproducible method to quantitate the activity of potential antiamebic agents in vitro. The isotopic method should be of particular value in defining the metabolism of the parasite and effects of antimicrobial agents on it, whereas the morphologic method may be more valuable for workers with limited resources available to them.

  18. Results from the ARTEMIS DISK Global Antifungal Surveillance Study, 1997 to 2005: an 8.5-Year Analysis of Susceptibilities of Candida Species and Other Yeast Species to Fluconazole and Voriconazole Determined by CLSI Standardized Disk Diffusion Testing▿

    Science.gov (United States)

    Pfaller, M. A.; Diekema, D. J.; Gibbs, D. L.; Newell, V. A.; Meis, J. F.; Gould, I. M.; Fu, W.; Colombo, A. L.; Rodriguez-Noriega, E.

    2007-01-01

    Fluconazole in vitro susceptibility test results for 205,329 yeasts were collected from 134 study sites in 40 countries from June 1997 through December 2005. Data were collected for 147,776 yeast isolates tested with voriconazole from 2001 through 2005. All investigators tested clinical yeast isolates by the CLSI M44-A disk diffusion method. Test plates were automatically read and results recorded with a BIOMIC image analysis system. Species, drug, zone diameter, susceptibility category, and quality control results were collected quarterly. Duplicate (same patient, same species, and same susceptible-resistant biotype profile during any 7-day period) and uncontrolled test results were not analyzed. Overall, 90.1% of all Candida isolates tested were susceptible (S) to fluconazole; however, 10 of the 22 species identified exhibited decreased susceptibility (<75% S) on the order of that seen with the resistant (R) species C. glabrata and C. krusei. Among 137,487 isolates of Candida spp. tested against voriconazole, 94.8% were S and 3.1% were R. Less than 30% of fluconazole-resistant isolates of C. albicans, C. glabrata, C. tropicalis, and C. rugosa remained S to voriconazole. The non-Candida yeasts (8,821 isolates) were generally less susceptible to fluconazole than Candida spp. but, aside from Rhodotorula spp., remained susceptible to voriconazole. This survey demonstrates the broad spectrum of these azoles against the most common opportunistic yeast pathogens but identifies several less common yeast species with decreased susceptibility to antifungal agents. These organisms may pose a future threat to optimal antifungal therapy and emphasize the importance of prompt and accurate species identification. PMID:17442797

  19. Identification and Antifungal Susceptibility Profiles of Candida nivariensis and Candida bracarensis in a Multi-Center Chinese Collection of Yeasts

    Science.gov (United States)

    Hou, Xin; Xiao, Meng; Chen, Sharon C.-A.; Wang, He; Yu, Shu-Ying; Fan, Xin; Kong, Fanrong; Xu, Ying-Chun

    2017-01-01

    Candida nivariensis and C. bracarensis are two emerging cryptic species within the C. glabrata complex. Thirteen of these isolates from 10 hospitals in China were studied for their species identification and antifungal susceptibilities. Phenotypic and molecular [rDNA ITS sequencing, D1/D2 sequencing and ITS sequencer-based capillary gel electrophoresis (SCGE)] and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) MS identification methods were compared for their performance in species identification. Twelve of 13 (92.3%) isolates were identified as C. nivariensis and one as C. bracarensis using ITS sequencing as the reference method. Results obtained by D1/D2 sequencing and ITS SCGE were concordant with ITS sequencing results for all (100%) isolates. SCGE was able to subtype 12 C. nivariensis into four ITS SCGE length types. All isolates failed to be identified by the Vitek MALDI-TOF MS system (bioMérieux), whilst the Bruker MS system (Bruker Daltoniks) correctly identified all C. nivariensis isolates but using a lowered (≥1.700) cut-off score for species assignment; the C. bracarensis isolate was identified but with score MALDI-TOF may be improved by adding mass spectral profiles (MSPs) into the current databases. The antifungal susceptibility profile of isolates should be monitored. PMID:28154553

  20. Apophysomyces elegans: epidemiology, amplified fragment length polymorphism typing, and in vitro antifungal susceptibility pattern.

    NARCIS (Netherlands)

    Chakrabarti, A.; Shivaprakash, M.R.; Curfs-Breuker, I.; Baghela, A.; Klaassen, C.H.; Meis, J.F.G.M.

    2010-01-01

    Apophysomyces elegans is an emerging pathogen in India. We planned the present study to analyze the clinical pattern of the disease, to perform molecular strain typing, and to determine the in vitro activities of eight antifungal drugs against A. elegans. A total of 16 clinical and two environmental

  1. 皮炎外瓶霉临床和环境分离株对6种常用抗真菌药物的敏感性测定%In vitro susceptibility testing of 16 strains of Exophiala dermatitidis to 6 commonly used antifungal agents

    Institute of Scientific and Technical Information of China (English)

    孙毅; 刘伟; 万喆; 陈伟; 李若瑜

    2010-01-01

    Objective To investigate the in vitro susceptibility of 16 strains of Exophiala dermatitidis to 6 commonly used antifungal agents. Methods The Glinical and Laboratory Standards Institute (CLSI)M27-A2 protocol was carried out to determine the MIGs of terbinafine (TRB), itraconazole (ITC), amphotericin B (AMB), fluconazole (FLC), voriconazole (VRC), and caspofungin (GAS) to 16 strains of E. dermatitidis, and E-test was performed to determine those of VRG, ITC and AMB. Besides, the minimal fungicidal concentrations (MFGs) of the above antifungal agents to the 16 strains of E. dermatitidis were further assessed.The activity of TRB in combination with ITC and VRG against E. dermatitidis was also estimated. Results The MIC ranges of TRB, VRC, ITC, AMB, FLC, and CAS were 0.125 - 0.25 mg/L, 0.25 - 0.5 mg/L, 2.0 mg/L,2.0 mg/L, 16 - 32 mg/L and 16 - 64 mg/L respectively as shown by M27-A2 microdilution assay, while the MIC ranges of VRG, ITG and AMB, as determined by E-test, were 0.032 - 0.094 mg/L, 0.047 - 0.5 mg/L and 0.125 - 3.0 mg/L, respectively. The MFC ranges of TRB, VRC, ITG, AMB and FLG were 0.125 - 0.5 mg/L,0.25 - 0.5 mg/L, 2.0 - 4.0 mg/L, 2.0 - 4.0 mg/L and 16 - 64 mg/L, respectively. No synergism in the acitivity against E. dermatitidis was observed for the combination of TBR with ITC or VRC. Conclusion E. dermatitidis is susceptible to TRB, ITC, AMB, and VRC, but less sensitive to both FLC and GAS.%目的 探讨临床分离的皮炎外瓶霉对6种常用抗真菌药物的敏感性.方法 参考美国临床实验室标准化研究所M27-A2方案测定特比萘芬(TRB)、伊曲康唑(ITC)、两性霉素B(AMB)、氟康唑(FLC)、伏立康唑(VRC)及卡泊芬净(CAS)对16株皮炎外瓶霉的最低抑菌浓度(MIC),同时以E-test法测定VRC、ITC和AMB对16株皮炎外瓶霉的MIC,并进一步测定每种药物的最低杀菌浓度(MFC).在此基础上,观察TRB与ITC及vRC联合应用时对皮炎外瓶霉的抗菌作用.结果 微量液基稀释法测得TRB

  2. In vitro susceptibility of Paracoccidioides brasiliensis yeast form to antifungal agents

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    Angela Restrepo

    1984-12-01

    Full Text Available A study was conducted to determine the susceptibility of P. brasiliensis yeast form to amphotericin B (A, ketoconazole (K, 5-fluorocytosine (5-FC and rifampin (R. The three isolates tested produced minimal inhibitory concentrations (MICs (mcg/ml in the following range: A: 0.09-0.18; K: 0.001-0.007; 5-FC: 62.5-250 and R: 40-80. The minimal fungicidal concentrations (MFC were several times higher than the corresponding MICs. Precise MFC for 5-FC were not obtained (> 500 mcg/ml. Combination of K plus A proved synergic, with the fractional inhibitory concentration (FIC indices revealing synergy when the drugs were combined at the 1 to 1 and 1 to 5 MIC ratios. R (40 mcg/ml appeared to antagonize K. These results indicate promise for the combined use of K plus A as a therapeutical regimen.

  3. Species distribution and in vitro antifungal susceptibility of oral yeast isolates from Tanzanian HIV-infected patients with primary and recurrent oropharyngeal candidiasis.

    NARCIS (Netherlands)

    Hamza, O.J.; Matee, M.I.N.; Moshi, M.J.; Simon, E.N.; Mugusi, F.; Mikx, F.H.M.; Palenstein Helderman, W.H. van; Rijs, A.J.M.M.; Ven, A.J.A.M. van der; Verweij, P.E.

    2008-01-01

    BACKGROUND: In Tanzania, little is known on the species distribution and antifungal susceptibility profiles of yeast isolates from HIV-infected patients with primary and recurrent oropharyngeal candidiasis. METHODS: A total of 296 clinical oral yeasts were isolated from 292 HIV-infected patients wit

  4. Epidemiology, species distribution, antifungal susceptibility and outcome of candidemia among Internal Medicine Wards of community hospitals of Udine province, Italy

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    Federico Silvestri

    2014-09-01

    Full Text Available Candidemia is an emerging problem among patients hospitalized in Internal Medicine Wards (IMW. We performed a retrospective study to assess the epidemiology, species distribution, antifungal susceptibility and outcome of candidaemia recorded over a 3-year period (2010-2012 among IMW of community hospitals of Udine province in Italy: forty-eight patients were identified, with an overall incidence of 1.44 cases/1000 hospital admissions/year. Candida albicans was the most frequent species, followed by Candida parapsilosis that accounted for 42.9% of Tolmezzo cases. All isolates were susceptible to amphotericin and caspofungin, while 11.4% of strains were not-susceptible to voriconazole and 14.3% to fluconazole. Crude mortality was 41.7%. In conclusion, in community hospitals overall incidence of candidemia is similar to tertiary care hospitals, but 80% of cases are detected in IMW. Candida species distribution is overlapping, but differences in local epidemiology were found and should be taken into consideration. No resistance to amphotericin and caspofungin was found while resistance to azoles was observed. Knowledge of this data might be useful when planning the best therapeutic strategy.

  5. Molecular characterisation and antifungal susceptibility of clinical Trichosporon isolates in India.

    Science.gov (United States)

    Rastogi, Vijaylatha; Honnavar, Prasanna; Rudramurthy, Shivaprakash M; Pamidi, Umabala; Ghosh, Anup; Chakrabarti, Arunaloke

    2016-08-01

    In Asian countries, Trichosporon infection is a well-known disease in Japan. In India, the infection is increasingly recognised. The study was conducted to characterise the clinical Trichosporon isolates from India by phenotypic and molecular techniques. A total of 31 Trichosporon clinical isolates, recovered from patients of 14 hospitals across India were sequenced (ITS and IGS1 regions of rDNA). In vitro drug susceptibility testing of the isolates was performed against amphotericin-B, fluconazole, itraconazole, voriconazole and posaconazole. IGS1, rather than ITS sequences, correctly identified the isolates: Trichosporon asahii, 20; Trichosporon ovoides, 3; Trichosporon inkin, 2; Trichosporon asteroides, 1; Trichosporon mucoides, 1; Trichosporon loubieri, 1; Trichosporon debeurmannianum, 1; and Trichosporon dermatis, 1. Trichosporon asahii genotype III was the most common type, followed by genotype I and VII. Both these targets did not help to identify one Trichosporon to the species level. Trichosporon debeurmannianum, T. dermatis and T. asteroides were isolated for the first time from a human disease in India. The minimum inhibitory concentrations for voriconazole and posaconazole were within effective range. The study highlights the presence of wide range of Trichosporon species causing infection in India. Voriconazole or posaconazole may be the better drugs to treat such patients.

  6. Candidemia by species of the Candida parapsilosis complex in children's hospital: prevalence, biofilm production and antifungal susceptibility.

    Science.gov (United States)

    Ruiz, Luciana da Silva; Khouri, Sonia; Hahn, Rosane Christine; da Silva, Eriques Gonçalves; de Oliveira, Vanessa Krummer Perinazzo; Gandra, Rinaldo Ferreira; Paula, Claudete Rodrigues

    2013-04-01

    Opportunistic infections are an increasingly common problem in hospitals, and the yeast Candida parapsilosis has emerged as an important nosocomial pathogen. The aims of this study were to determine and compare (i) the prevalence rate among C. parapsilosis complex organisms isolated from blood in a public children's hospital in São Paulo state, (ii) the ability of the complex C. parapsilosis species identified to produce biofilm and (iii) the antifungal susceptibility profiles. Forty-nine (49) specimens of isolated blood yeast were analyzed, previously identified as C. parapsilosis by conventional methods. After the molecular analysis, the isolates were characterized as C. parapsilosis sensu stricto (83.7 %), C. orthopsilosis (10.2 %) and C. metapsilosis (6.1 %). All species were able to form biofilm. The species with the highest biofilm production was C. parapsilosis sensu stricto, followed by C. orthopsilosis and further by C. metapsilosis. All of the strains have demonstrated similar susceptibility to fluconazole, caspofungin, voriconazole, cetoconazole and 5-flucytosine. Only one strain of C. parapsilosis was resistant to amphotericin B. Regarding itraconazole, 66.6 and 43.9 % isolates of C. metapsilosis and C. parapsilosis, respectively, have demonstrated to be susceptible dose-dependent, with one isolate of the latter species resistant to the drug. Candida parapsilosis sensu stricto has demonstrated to be the less susceptible, mainly to amphotericin B, caspofungin and "azoles" such as fluconazole. Therefore, C. metapsilosis and C. orthopsilosis are still involved in a restricted number of infections, but these data have become essential for there are very few studies of these species in Latin America.

  7. Distribution and antifungal susceptibility of Candida species causing candidemia from 1996 to 1999.

    Science.gov (United States)

    Cheng, Ming-Fang; Yu, Kwok-Woon; Tang, Ran-Bin; Fan, Yu-Hua; Yang, Yun-Liang; Hsieh, Kai-Sheng; Ho, Monto; Lo, Hsiu-Jung

    2004-01-01

    Susceptibilities to amphotericin B and fluconazole of 383 Candida species isolated from blood were determined. Candida albicans was the most common species (55.6%), followed by Candida parapsilosis (17.5%), Candida tropicalis (16.5%), Candida glabrata (5.2%), Candida guilliermondii (2.3%), and others (2.9%). All but three isolates, Candida ciferrii, C. tropicalis, and C. glabrata, one each, were susceptible to amphotericin B. A total of 367 (95.8%) and 15 (4.2%) isolates were susceptible and susceptible-dose dependent to fluconazole, respectively. Only one isolate, a C. glabrata, was resistant to fluconazole. Few patients (13%) having prior fluconazole treatments may explain the low rate of resistance to fluconazole in this study.

  8. Microfluidics for Antibiotic Susceptibility and Toxicity Testing

    Directory of Open Access Journals (Sweden)

    Jing Dai

    2016-10-01

    Full Text Available The recent emergence of antimicrobial resistance has become a major concern for worldwide policy makers as very few new antibiotics have been developed in the last twenty-five years. To prevent the death of millions of people worldwide, there is an urgent need for a cheap, fast and accurate set of tools and techniques that can help to discover and develop new antimicrobial drugs. In the past decade, microfluidic platforms have emerged as potential systems for conducting pharmacological studies. Recent studies have demonstrated that microfluidic platforms can perform rapid antibiotic susceptibility tests to evaluate antimicrobial drugs’ efficacy. In addition, the development of cell-on-a-chip and organ-on-a-chip platforms have enabled the early drug testing, providing more accurate insights into conventional cell cultures on the drug pharmacokinetics and toxicity, at the early and cheaper stage of drug development, i.e., prior to animal and human testing. In this review, we focus on the recent developments of microfluidic platforms for rapid antibiotics susceptibility testing, investigating bacterial persistence and non-growing but metabolically active (NGMA bacteria, evaluating antibiotic effectiveness on biofilms and combinatorial effect of antibiotics, as well as microfluidic platforms that can be used for in vitro antibiotic toxicity testing.

  9. Comparison of commercial methods and the CLSI broth microdilution to determine the antifungal susceptibility of Candida parapsilosis complex bloodstream isolates from three health institutions in Rio de Janeiro, Brazil.

    Science.gov (United States)

    Figueiredo-Carvalho, Maria Helena G; Barbedo, Leonardo S; Oliveira, Manoel M E; Brito-Santos, Fábio; Almeida-Paes, Rodrigo; Zancopé-Oliveira, Rosely M

    2014-08-01

    Two commercial methods, the Etest and Vitek 2, were compared with the Clinical and Laboratory Standards Institute broth microdilution method to determine the susceptibility of Candida parapsilosis complex to amphotericin B, caspofungin, fluconazole, voriconazole, and itraconazole. One-hundred bloodstream isolates of C. parapsilosis complex from three hospitals in Rio de Janeiro city, Brazil, between 1998 and 2006 were analyzed. C. parapsilosis sensu stricto (61 %) was the predominant species, followed by C. orthopsilosis (37 %) and C. metapsilosis (2 %). Most isolates were susceptible to the tested drugs. However, one C. parapsilosis sensu stricto isolate was considered resistant for amphotericin B. The essential agreement was 100 % between the methods, except for itraconazole (96.3 %). The categorical agreement varied for fluconazole and itraconazole by Etest and for amphotericin B and fluconazole by Vitek 2. This study reinforces the suitability of the commercial methods in routine clinical microbiology laboratories for antifungal susceptibility testing.

  10. Antifungal testing and high-throughput screening of compound library against Geomyces destructans, the etiologic agent of geomycosis (WNS in bats.

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    Sudha Chaturvedi

    Full Text Available Bats in the northeastern U.S. are affected by geomycosis caused by the fungus Geomyces destructans (Gd. This infection is commonly referred to as White Nose Syndrome (WNS. Over a million hibernating bats have died since the fungus was first discovered in 2006 in a cave near Albany, New York. A population viability analysis conducted on little brown bats (Myotis lucifugus, one of six bat species infected with Gd, suggests regional extinction of this species within 20 years. The fungus Gd is a psychrophile ("cold loving", but nothing is known about how it thrives at low temperatures and what pathogenic attributes allow it to infect bats. This study aimed to determine if currently available antifungal drugs and biocides are effective against Gd. We tested five Gd strains for their susceptibility to antifungal drugs and high-throughput screened (HTS one representative strain with SpectrumPlus compound library containing 1,920 compounds. The results indicated that Gd is susceptible to a number of antifungal drugs at concentrations similar to the susceptibility range of human pathogenic fungi. Strains of Gd were susceptible to amphotericin B, fluconazole, itraconazole, ketoconazole and voriconazole. In contrast, very high MICs (minimum inhibitory concentrations of flucytosine and echinocandins were needed for growth inhibition, which were suggestive of fungal resistance to these drugs. Of the 1,920 compounds in the library, a few caused 50%--to greater than 90% inhibition of Gd growth. A number of azole antifungals, a fungicide, and some biocides caused prominent growth inhibition. Our results could provide a theoretical basis for future strategies aimed at the rehabilitation of most affected bat species and for decontamination of Gd in the cave environment.

  11. Prospective multicenter study of the epidemiology, molecular identification, and antifungal susceptibility of Candida parapsilosis, Candida orthopsilosis, and Candida metapsilosis isolated from patients with candidemia.

    Science.gov (United States)

    Cantón, Emilia; Pemán, Javier; Quindós, Guillermo; Eraso, Elena; Miranda-Zapico, Ilargi; Álvarez, María; Merino, Paloma; Campos-Herrero, Isolina; Marco, Francesc; de la Pedrosa, Elia Gomez G; Yagüe, Genoveva; Guna, Remedios; Rubio, Carmen; Miranda, Consuelo; Pazos, Carmen; Velasco, David

    2011-12-01

    A 13-month prospective multicenter study including 44 hospitals was carried out to evaluate the epidemiology of Candida parapsilosis complex candidemia in Spain. Susceptibility to amphotericin B, flucytosine, fluconazole, itraconazole, voriconazole, posaconazole, anidulafungin, caspofungin, and micafungin was tested by the microdilution colorimetric method. A total of 364 C. parapsilosis complex isolates were identified by molecular methods: C. parapsilosis (90.7%), Candida orthopsilosis (8.2%), and Candida metapsilosis (1.1%). Most candidemias (C. parapsilosis, 76.4%; C. orthopsilosis, 70.0%; C. metapsilosis, 100%) were observed in adults. No C. orthopsilosis or C. metapsilosis candidemias occurred in neonates. C. parapsilosis was most frequent in adult intensive care unit (28.8%), surgery (20.9%), and internal medicine (19.7%) departments; and C. orthopsilosis was most frequent in hematology (28.6%), pediatrics (12.0%), and neonatology (11.5%) departments. The geographic distribution of C. orthopsilosis and C. metapsilosis was not uniform. According to CLSI clinical breakpoints, all C. orthopsilosis and C. metapsilosis isolates were susceptible to the nine agents tested. Resistance (MICs > 1 mg/liter) was observed only in C. parapsilosis: amphotericin B, posaconazole, itraconazole, and caspofungin (0.3% each), anidulafungin (1.9%), and micafungin (2.5%). Applying the new species-specific fluconazole and echinocandin breakpoints, the rates of resistance to fluconazole for C. parapsilosis and C. orthopsilosis increased to 4.8% and 0.3%, respectively; conversely, for C. parapsilosis they shifted from 1.9 to 0.6% (anidulafungin) and from 2.5 to 0.6% (micafungin). Our study confirms the different prevalence of C. parapsilosis complex candidemia among age groups: neither C. orthopsilosis nor C. metapsilosis was isolated from neonates; interestingly, C. metapsilosis was isolated only from adults and the elderly. The disparity in antifungal susceptibility among species

  12. Results from the ARTEMIS DISK Global Antifungal Surveillance Study, 1997 to 2007: a 10.5-year analysis of susceptibilities of Candida Species to fluconazole and voriconazole as determined by CLSI standardized disk diffusion.

    Science.gov (United States)

    Pfaller, M A; Diekema, D J; Gibbs, D L; Newell, V A; Ellis, D; Tullio, V; Rodloff, A; Fu, W; Ling, T A

    2010-04-01

    Fluconazole in vitro susceptibility test results for 256,882 isolates of Candida spp. were collected from 142 sites in 41 countries from June 1997 to December 2007. Data were collected for 197,619 isolates tested with voriconazole from 2001 to 2007. A total of 31 different species of Candida were isolated. Increased rates of isolation of the common non-albicans species C. glabrata (10.2% to 11.7%), C. tropicalis (5.4% to 8.0%), and C. parapsilosis (4.8% to 5.6%) were noted when the time periods 1997 to 2000 and 2005 to 2007 were compared. Investigators tested clinical isolates of Candida spp. by the CLSI M44-A disk diffusion method. Overall, 90.2% of Candida isolates tested were susceptible (S) to fluconazole; however, 13 of 31 species identified exhibited decreased susceptibility (ciferrii remained S to voriconazole. An increase in fluconazole resistance over time was seen with C. parapsilosis, C. guilliermondii, C. lusitaniae, C. sake, and C. pelliculosa. Among the emerging fluconazole-R species were C. guilliermondii (11.4% R), C. inconspicua (53.2% R), C. rugosa (41.8% R), and C. norvegensis (40.7% R). The rates of isolation of C. rugosa, C. inconspicua, and C. norvegensis increased by 5- to 10-fold over the 10.5-year study period. C. guilliermondii and C. rugosa were most prominent in Latin America, whereas C. inconspicua and C. norvegensis were most common in Eastern European countries. This survey identifies several less-common species of Candida with decreased susceptibility to azoles. These organisms may pose a future threat to optimal antifungal therapy and underscore the importance of prompt and accurate species identification and antifungal susceptibility testing.

  13. Results from the ARTEMIS DISK Global Antifungal Surveillance Study, 1997 to 2007: a 10.5-Year Analysis of Susceptibilities of Candida Species to Fluconazole and Voriconazole as Determined by CLSI Standardized Disk Diffusion ▿

    Science.gov (United States)

    Pfaller, M. A.; Diekema, D. J.; Gibbs, D. L.; Newell, V. A.; Ellis, D.; Tullio, V.; Rodloff, A.; Fu, W.; Ling, T. A.

    2010-01-01

    Fluconazole in vitro susceptibility test results for 256,882 isolates of Candida spp. were collected from 142 sites in 41 countries from June 1997 to December 2007. Data were collected for 197,619 isolates tested with voriconazole from 2001 to 2007. A total of 31 different species of Candida were isolated. Increased rates of isolation of the common non-albicans species C. glabrata (10.2% to 11.7%), C. tropicalis (5.4% to 8.0%), and C. parapsilosis (4.8% to 5.6%) were noted when the time periods 1997 to 2000 and 2005 to 2007 were compared. Investigators tested clinical isolates of Candida spp. by the CLSI M44-A disk diffusion method. Overall, 90.2% of Candida isolates tested were susceptible (S) to fluconazole; however, 13 of 31 species identified exhibited decreased susceptibility (ciferrii remained S to voriconazole. An increase in fluconazole resistance over time was seen with C. parapsilosis, C. guilliermondii, C. lusitaniae, C. sake, and C. pelliculosa. Among the emerging fluconazole-R species were C. guilliermondii (11.4% R), C. inconspicua (53.2% R), C. rugosa (41.8% R), and C. norvegensis (40.7% R). The rates of isolation of C. rugosa, C. inconspicua, and C. norvegensis increased by 5- to 10-fold over the 10.5-year study period. C. guilliermondii and C. rugosa were most prominent in Latin America, whereas C. inconspicua and C. norvegensis were most common in Eastern European countries. This survey identifies several less-common species of Candida with decreased susceptibility to azoles. These organisms may pose a future threat to optimal antifungal therapy and underscore the importance of prompt and accurate species identification and antifungal susceptibility testing. PMID:20164282

  14. Epidemiology and antifungal resistance in invasive candidiasis

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    Rodloff AC

    2011-04-01

    Full Text Available Abstract The epidemiology of Candida infections has changed over the last two decades: The number of patients suffering from such infections has increased dramatically and the Candida species involved have become more numerous as Candida albicans is replaced as an infecting agent by various non-C. albicans species (NAC. At the same time, additional antifungal agents have become available. The different Candida species may vary in their susceptibility for these various antifungals. This draws more attention to in vitro susceptibility testing. Unfortunately, several different test methods exist that may deliver different results. Moreover, clinical breakpoints (CBP that classify test results into susceptible, intermediate and resistant are controver- sial between CLSI and EUCAST. Therefore, clinicians should be aware that interpretations may vary with the test system being followed by the microbiological laboratory. Thus, knowledge of actual MIC values and pharmacokinetic properties of individual antifungal agents is important in delivering appropriate therapy to patients

  15. Species Diversity, Antifungal Susceptibility, and Virulence Attributes of Candida Colonising the Oral Cavities of Adult Diabetic Patients

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    Deepa Anil Kumar

    2014-01-01

    Full Text Available Oral candidiasis is a common occurrence in diabetic patients. Species of Candida isolated from these infections and their virulence pattern undergo changes over time and require periodic assessments. Objective of this study was to determine changes in the spectrum of Candida species colonizing oral cavity, their antifungal susceptibility patterns, and virulence attributes, in adult diabetic patients. Oral swabs were collected from 100 patients with diabetes mellitus (DM and an equal number of healthy controls. Specimens were cultured for Candida and species were identified, according to standard protocols. Of 100 diabetic patients, 42 were colonized by yeasts with C. albicans as the predominant species (51%. Non-Candida albicans Candida (NCAC species accounted for 47% of the specimens, with C. tropicalis being the commonest. Among healthy controls, 23 were colonized by Candida species, of which C. albicans was predominant. Results obtained indicate that C. albicans continues to be the predominant species in oral cavities of diabetic patients. Candidal carriage was significantly associated with duration of diabetes and fasting blood sugar levels. Virulence attributes, proteinase and phospholipase secretion, and biofilm formation were significantly higher in DM group.

  16. Conidial germination in Scedosporium apiospermum, S. aurantiacum, S. minutisporum and Lomentospora prolificans: influence of growth conditions and antifungal susceptibility profiles

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    Thaís Pereira de Mello

    2016-01-01

    Full Text Available In the present study, we have investigated some growth conditions capable of inducing the conidial germination in Scedosporium apiospermum, S. aurantiacum, S. minutisporum and Lomentospora prolificans. Germination in Sabouraud medium (pH 7.0, 37ºC, 5% CO2 showed to be a typically time-dependent event, reaching ~75% in S. minutisporum and > 90% in S. apiospermum, S. aurantiacum and L. prolificans after 4 h. Similar germination rate was observed when conidia were incubated under different media and pHs. Contrarily, temperature and CO2 tension modulated the germination. The isotropic conidial growth (swelling and germ tube-like projection were evidenced by microscopy and cytometry. Morphometric parameters augmented in a time-dependent fashion, evidencing changes in size and granularity of fungal cells compared with dormant 0 h conidia. In parallel, a clear increase in the mitochondrial activity was measured during the transformation of conidia-into-germinated conidia. Susceptibility profiles to itraconazole, fluconazole, voriconazole, amphotericin B and caspofungin varied regarding each morphotype and each fungal species. Overall, the minimal inhibitory concentrations for hyphae were higher than conidia and germinated conidia, except for caspofungin. Collectively, our study add new data about the conidia-into-hyphae transformation in Scedosporium and Lomentospora species, which is a relevant biological process of these molds directly connected to their antifungal resistance and pathogenicity mechanisms.

  17. Secondary metabolite profiles and antifungal drug susceptibility of Aspergillus fumigatus and closely related species, Aspergillus lentulus, Aspergillus udagawae, and Aspergillus viridinutans.

    Science.gov (United States)

    Tamiya, Hiroyuki; Ochiai, Eri; Kikuchi, Kazuyo; Yahiro, Maki; Toyotome, Takahito; Watanabe, Akira; Yaguchi, Takashi; Kamei, Katsuhiko

    2015-05-01

    The incidence of Aspergillus infection has been increasing in the past few years. Also, new Aspergillus fumigatus-related species, namely Aspergillus lentulus, Aspergillus udagawae, and Aspergillus viridinutans, were shown to infect humans. These fungi exhibit marked morphological similarities to A. fumigatus, albeit with different clinical courses and antifungal drug susceptibilities. The present study used liquid chromatography/time-of-flight mass spectrometry to identify the secondary metabolites secreted as virulence factors by these Aspergillus species and compared their antifungal susceptibility. The metabolite profiles varied widely among A. fumigatus, A. lentulus, A. udagawae, and A. viridinutans, producing 27, 13, 8, and 11 substances, respectively. Among the mycotoxins, fumifungin, fumiquinazoline A/B and D, fumitremorgin B, gliotoxin, sphingofungins, pseurotins, and verruculogen were only found in A. fumigatus, whereas auranthine was only found in A. lentulus. The amount of gliotoxin, one of the most abundant mycotoxins in A. fumigatus, was negligible in these related species. In addition, they had decreased susceptibility to antifungal agents such as itraconazole and voriconazole, even though metabolites that were shared in the isolates showing higher minimum inhibitory concentrations than epidemiological cutoff values were not detected. These strikingly different secondary metabolite profiles may lead to the development of more discriminative identification protocols for such closely related Aspergillus species as well as improved treatment outcomes.

  18. In vitro susceptibility testing of Aspergillus spp. against voriconazole, itraconazole, posaconazole, amphotericin B and caspofungin

    Institute of Scientific and Technical Information of China (English)

    SHI Jun-yan; WANG He; GUO Li-na; XU Ying-chun; SHI Yi; L(U) Huo-xiang; LIU Yong; ZHAO Wang-sheng; CHEN Dong-mei; XI Li-yan; ZHOU Xin

    2010-01-01

    Background During recent years, the incidence of serious infections caused by opportunistic fungi has increased dramatically due to alterations of the immune status of patients with hematological diseases, malignant tumors,transplantations and so forth. Unfortunately, the wide use of triazole antifungal agents to treat these infections has lead to the emergence of Aspergillus spp. resistant to triazoles. The present study was to assess the in vitro activities of five antifungal agents (voriconazole, itraconazole, posaconazole, amphotericin B and caspofungin) against different kinds of Aspergillus spp. that are commonly encountered in the clinical setting.Methods The agar-based Etest MIC method was employed. One hundred and seven strains of Aspergillus spp. (5 species) were collected and prepared according to Etest Technique Manuel. Etest MICs were determined with RPMI agar containing 2% glucose and were read after incubation for 48 hours at 35℃. MIC50, MIC90 and MIC range were acquired by Whonet 5.4 software.Results The MIC90 of caspofungin against A. fumigatus, A. flavus and A. nidulans was 0.094 μg/ml whereas the MIC90 against A. niger was 0.19 μg/ml. For these four species, the MlC90 of caspofungin was the lowest among the five antifungal agents. For A. terrus, the MIC90 of posaconazole was the lowest. For A. fumigatus and A. flavus, the MlC90in order of increasing was caspofungin, posaconazole, voriconazole, itraconazole, and amphotericin B. The MIC of amphotericin B against A. terrus was higher than 32 μg/ml in all 7 strains tested.Conclusions The in vitro antifungal susceptibility test shows the new drug caspofungin, which is a kind of echinocandins, has good activity against the five species of Aspergillus spp. and all the triazoles tested have better in vitro activity than traditional amphotericin B.

  19. Characterization of Candida spp. isolated from vaginal fluid: identification, antifungal susceptibility, and virulence profile - doi: 10.4025/actascihealthsci.v35i1.13557

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    Terezinha Inez Estivalet Svidzinski

    2013-03-01

    Full Text Available A total of 101 (20.0% yeast samples were isolated from vaginal fluids of 504 non-hospitalized patients in Maringá, Paraná State, Brazil and Candida albicans was more frequent specie (93.1% identified by seminested PCR method. All the isolates were susceptible to amphotericin B and nystatin, and 93.1% of them were susceptible to fluconazole. The acid proteinase, hemolytic and phospholipase activities were observed in 99.0, 90.0, and 88.0% of Candida spp., respectively. Around 67.0% of the strains had adherence indexes of 0.5 to 1.5 yeasts by Vero cell, and most of them showed a hydrophilic profile. Correlation studies indicated hydrophilic yeasts presented higher adherence index, proteinase, and phospholipase activities; and a positive correlation between all enzymes was also observed. In addition, the isolates with high hemolytic activity were less susceptible to fluconazole and amphotericin B. These results of Candida prevalence and antifungal susceptibility corroborate with literature’s datas and correlation between virulence factors and MIC values suggest Candida isolates from vaginal fluid less susceptible to antifungal and with higher extracellular enzymes production can be more virulent to cause tissue damage.  

  20. Antimicrobial resistance and susceptibility testing of anaerobic bacteria.

    Science.gov (United States)

    Schuetz, Audrey N

    2014-09-01

    Infections due to anaerobic bacteria can be severe and life-threatening. Susceptibility testing of anaerobes is not frequently performed in laboratories, but such testing is important to direct appropriate therapy. Anaerobic resistance is increasing globally, and resistance trends vary by geographic region. An overview of a variety of susceptibility testing methods for anaerobes is provided, and the advantages and disadvantages of each method are reviewed. Specific clinical situations warranting anaerobic susceptibility testing are discussed.

  1. [Molecular epidemiology and antifungal susceptibility of Candida species isolated from urine samples of patients in intensive care unit].

    Science.gov (United States)

    Yüksekkaya, Serife; Fındık, Duygu; Arslan, Uğur

    2011-01-01

    The aims of this study were to analyse the amphotericin B and fluconazole susceptibility and molecular epidemiology of Candida strains (Candida albicans, Candida tropicalis and Candida glabrata) isolated from the urine samples of patients hospitalized in the intensive care unit. Identification of the isolates was done according to microscopic morphology (chlamydospor, blastospor, pseudohyphae and true hyphae) on cornmeal agar, germ tube formation and carbohydrate assimilation patterns (API ID 32C bioMérieux, France). Antifungal susceptibilities of the isolates were determined by in vitro broth microdilution method recommended by Clinical and Laboratory Standards Institute (CLSI). To investigate the clonal relationship of the isolates, randomly amplified polymorphic DNA (RAPD) analysis was performed by using Cnd3 primer. Of the 56 Candida isolates minimum inhibitory concentration (MIC) ranges, MIC50 and MIC90 values for amphotericin B were 0.125-1 µg/ml, 0.125 and 0.5 µg/ml for C.albicans, 0.125-1 µg/ml, 0.25 and 1 µg/ml for C.tropicalis and 0.125-1 µg/ml, 0.25 and 1 µg/ml for C.glabrata, respectively. Fluconazole MIC ranges, MIC50 and MIC90 values were 0.25-4 µg/ml, 0.25 and 0.5 µg/ml for C.albicans, 0.25-16 µg/ml, 0.5 and 1 µg/ml for C.tropicalis and 0.5-64 µg/ml, 8 and 16 µg/ml for C.glabrata, respectively. For amphotericin B, none of the isolates had high MIC values (MIC > 1 µg/ml). While one of the C.glabrata isolates was resistant to fluconazole (MIC ≥ 64 µg/ml), one C.tropicalis and two C.glabrata isolates were dose-dependent susceptible (MIC: 16-32 µg/ml). The results of RAPD analysis indicated an exogenous spread from two clones for C.albicans, one clone for C.glabrata and one clone for C.tropicalis. This study underlines the importance of molecular epidemiological analysis of clinical samples together with hospital environmental samples in terms of Candida spp. To determine the exogenous origin for the related strains and to prevent

  2. CLSI broth microdilution method for testing susceptibility of Malassezia pachydermatis to thiabendazole

    Science.gov (United States)

    Nascente, Patrícia da Silva; Meinerz, Ana Raquel Mano; de Faria, Renata Osório; Schuch, Luiz Filipe Damé; Meireles, Mário Carlos Araújo; de Mello, João Roberto Braga

    2009-01-01

    Thiabendazole, classified as antiparasitic and also used as an antifungal drug, can be found as otological solution indicated for treatment of parasitic and fungal external otitis in small animals. Malassezia pachydermatis is a yeast recognized as a normal inhabitant on the skin and mucous membranes of dogs and cats. However, it is considered an opportunistic agent that causes external otitis and dermatitis in these animals. The aim of this study was to evaluate the in vitro effect of thiabendazole against 51 isolates of M. pachydermatis using the CLSI Broth Microdilution method that has been adapted for this yeast species (NCCLS, 2002). Based on this test, the Minimum Inhibitory Concentrations (MIC) of thiabendazol was calculated. Subsequently, the susceptibility of each isolate against this antifungal was determined. It was observed that the MIC of thiabendazole against M. pachydermatis ranged from 0.03 to > 4 µg/mL. A total of 13.7% of the isolates were found to be resistant, 47.1% were intermediate and 39.2% were sensitive to the drug. The rate of resistance of the yeasts against thiabendazole was similar to the results previously obtained with other antifungals, while the adapted broth microdilution technique used in this study proved to be efficient. PMID:24031347

  3. Antifungal susceptibility of oral Candida species among drug abusers%吸毒人群口腔假丝酵母药物敏感性分析

    Institute of Scientific and Technical Information of China (English)

    袁有华; 白丽; 武有聪; 钱金栿; 刘奇; 郭利军

    2011-01-01

    目的 了解吸毒人群口腔假丝酵母对常用抗真菌药物的敏感性,为临床治疗假丝酵母病提供参考资料.方法 采用美国临床标准化实验委员会推荐的微量稀释法测定75株假丝酵母对4种常用抗真菌药物两性霉素B(Amb)、5-氟胞嘧啶(5-FC)、氟康唑(FCZ)和酮康唑(KETO)的药物敏感性.结果 75株吸毒人群口腔假丝酵母对Amb、5-FC、FCZ和KETO的耐药率分别为0、4%、8%和13.3%;对FCZ和KETO的交叉耐药率为8%;非白色假丝酵母对FCZ的耐药率为23.5%,对KETO的耐药率为43.2%,高于白色假丝酵母的3.45%和5.2%,差异有统计学意义(P<0.05),2种假丝酵母对5-FC和Amb的耐药率差异无统计学意义.结论 吸毒人群口腔假丝酵母对Amb和5-FC的敏感性高于对FCZ和KETO;口腔假丝酵母对FCZ、KETO和5-FC有天然耐药株和对唑类药物的天然交叉耐药株,且非白色假丝酵母对FCZ和KETO的耐药率及交叉耐药率高于白色假丝酵母.%Objective To examine the antifungal susceptibility of oral Candida species among drug abusers and to provide reference for treatment of candidasis in clinic. Methods Totally 75 oral Candida species were tested for susceptibility of amphotericine B( Amb) ,5-flurocytosine(5-FC) ,fluconazole( FCZ) ,and ketonazole( KETO) with microdilution antifungal susceptibility test recommended by National Committee for Clinical Laboratory Standards ( NCCLS). Results The resistant rate of 75 oral Candida species from drug abusers for Amb,5-FC,FCZ,and KETO was 0,4% ,8% ,and 13.3% , respectively. The crosswise resistant rate of the strains for FCZ and KETO was 8%. The resistant rates of nan-Candida albicans to FCZ(23.5% ) and KETO(43. 2% ) were higher than those of Candida albicans with a significant difference. But the resistant rates to 5-FC and Amb were not significantly different for the two Candidas groups. Conclusion The susceptibility of oral Candida species from drug abusers for Amb and 5-FC is higher

  4. Sporothrix schenckii COMPLEX:SUSCEPTIBILITIES TO COMBINED ANTIFUNGAL AGENTS AND CHARACTERIZATION OF ENZYMATIC PROFILES.

    Science.gov (United States)

    Oliveira, Daniele Carvalho; de Loreto, Érico Silva; Mario, Débora Alves Nunes; Lopes, Paulo G Markus; Neves, Louise Vignolles; da Rocha, Marta Pires; Santurio, Janio Morais; Alves, Sydney Hartz

    2015-01-01

    Sporothrix schenckii was reclassified as a complex encompassing six cryptic species, which calls for the reassessment of clinical and epidemiological data of these new species. We evaluated the susceptibility of Sporothrix albicans(n = 1) , S. brasiliensis(n = 6) , S. globosa(n = 1), S. mexicana(n = 1) and S. schenckii(n = 36) to terbinafine (TRB) alone and in combination with itraconazole (ITZ), ketoconazole (KTZ), and voriconazole (VRZ) by a checkerboard microdilution method and determined the enzymatic profile of these species with the API-ZYM kit. Most interactions were additive (27.5%, 32.5% and 5%) or indifferent (70%, 50% and 52.5%) for TRB+KTZ, TRB+ITZ and TRB+VRZ, respectively. Antagonisms were observed in 42.5% of isolates for the TRB+VRZ combination. Based on enzymatic profiling, the Sporothrix schenckii strains were categorized into 14 biotypes. Leucine arylamidase (LA) activity was observed only for S. albicans and S. mexicana. The species S. globosa and S. Mexicana were the only species without β-glucosidase (GS) activity. Our results may contribute to a better understanding of virulence and resistance among species of the genus Sporothrix in further studies.

  5. Sporothrix schenckii COMPLEX: SUSCEPTIBILITIES TO COMBINED ANTIFUNGAL AGENTS AND CHARACTERIZATION OF ENZYMATIC PROFILES

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    Daniele Carvalho OLIVEIRA

    2015-08-01

    Full Text Available SUMMARY Sporothrix schenckiiwas reclassified as a complex encompassing six cryptic species, which calls for the reassessment of clinical and epidemiological data of these new species. We evaluated the susceptibility of Sporothrix albicans (n = 1 , S. brasiliensis (n = 6 , S. globosa (n = 1, S. mexicana(n = 1 and S. schenckii(n = 36 to terbinafine (TRB alone and in combination with itraconazole (ITZ, ketoconazole (KTZ, and voriconazole (VRZ by a checkerboard microdilution method and determined the enzymatic profile of these species with the API-ZYM kit. Most interactions were additive (27.5%, 32.5% and 5% or indifferent (70%, 50% and 52.5% for TRB+KTZ, TRB+ITZ and TRB+VRZ, respectively. Antagonisms were observed in 42.5% of isolates for the TRB+VRZ combination. Based on enzymatic profiling, the Sporothrix schenckii strains were categorized into 14 biotypes. Leucine arylamidase (LA activity was observed only for S. albicans and S. mexicana. The species S. globosaand S. mexicanawere the only species without β-glucosidase (GS activity. Our results may contribute to a better understanding of virulence and resistance among species of the genus Sporothrixin further studies.

  6. Sporothrix schenckii COMPLEX: SUSCEPTIBILITIES TO COMBINED ANTIFUNGAL AGENTS AND CHARACTERIZATION OF ENZYMATIC PROFILES

    Science.gov (United States)

    OLIVEIRA, Daniele Carvalho; de LORETO, Érico Silva; MARIO, Débora Alves Nunes; LOPES, Paulo G. Markus; NEVES, Louise Vignolles; da ROCHA, Marta Pires; SANTURIO, Janio Morais; ALVES, Sydney Hartz

    2015-01-01

    SUMMARY Sporothrix schenckiiwas reclassified as a complex encompassing six cryptic species, which calls for the reassessment of clinical and epidemiological data of these new species. We evaluated the susceptibility of Sporothrix albicans (n = 1) , S. brasiliensis (n = 6) , S. globosa (n = 1), S. mexicana(n = 1) and S. schenckii(n = 36) to terbinafine (TRB) alone and in combination with itraconazole (ITZ), ketoconazole (KTZ), and voriconazole (VRZ) by a checkerboard microdilution method and determined the enzymatic profile of these species with the API-ZYM kit. Most interactions were additive (27.5%, 32.5% and 5%) or indifferent (70%, 50% and 52.5%) for TRB+KTZ, TRB+ITZ and TRB+VRZ, respectively. Antagonisms were observed in 42.5% of isolates for the TRB+VRZ combination. Based on enzymatic profiling, the Sporothrix schenckii strains were categorized into 14 biotypes. Leucine arylamidase (LA) activity was observed only for S. albicans and S. mexicana. The species S. globosaand S. mexicanawere the only species without β-glucosidase (GS) activity. Our results may contribute to a better understanding of virulence and resistance among species of the genus Sporothrixin further studies. PMID:26422151

  7. Identificação molecular e suscetibilidade antifúngica de Candida parapsilosis isoladas no Ceará, Brasil Molecular identification and antifungal susceptibility of Candida parapsilosis isolates in Ceará, Brazil

    Directory of Open Access Journals (Sweden)

    Everardo Albuquerque Menezes

    2012-12-01

    Full Text Available INTRODUÇÃO: Candida parapsilosis é a segunda ou terceira levedura mais isolada de hemoculturas em várias partes do mundo. É um patógeno comumente isolado no Brasil e no Ceará. Apresenta capacidade de formar biofilmes em cateteres e outros dispositivos médicos e esses fatores contribuem para a disseminação dessa levedura. OBJETIVOS: Identificar e avaliar a suscetibilidade aos antifúngicos de C. parapsilosis isoladas de amostras de sangue e urina de pacientes atendidos em hospitais no Ceará. MÉTODOS: Foram isoladas e identificadas 57 cepas de C. parapsilosis. As cepas foram identificadas por testes fenotípicos e moleculares. A suscetibilidade foi avaliada pelo método de microdiluição em caldo (protocolo do Clinical and Laboratory Standards Institute [CLSI] M27-A3. Foram avaliados cinco antifúngicos (anfotericina B, caspofungina, fluconazol, itraconazol e voriconazol. RESULTADOS E CONCLUSÃO: As cepas foram identificadas como C. parapsilosis por testes fenotípicos e confirmadas pelos testes moleculares. Quanto ao perfil de sensibilidade, elas se mostraram sensíveis aos antifúngicos testados, sendo a resistência ainda um fenômeno raro entre cepas de C. parapsilosis isoladas no Ceará.INTRODUCTION: C. parapsilosis is the second or third most isolated yeast from blood cultures in various parts of the world. It is a commonly isolated pathogen in Brazil and Ceará. C. parapsilosis is liable to form biofilms on catheters and other medical devices, which contributes to the spread of this yeast. OBJECTIVE: The objective of this study was to identify and assess the antifungal susceptibility of C. parapsilosis isolates from blood and urine samples collected from patients in hospitals in Ceará. METHODS: We isolated and identified 57 strains of C. parapsilosis. The strains were identified by phenotypic and molecular tests. The susceptibility to antifungals was assessed by broth microdilution (Clinical and Laboratory Standards Institute

  8. Activity of a Long-Acting Echinocandin (CD101) and Seven Comparator Antifungal Agents Tested against a Global Collection of Contemporary Invasive Fungal Isolates in the SENTRY 2014 Antifungal Surveillance Program

    Science.gov (United States)

    Pfaller, Michael A.; Messer, Shawn A.; Rhomberg, Paul R.

    2017-01-01

    ABSTRACT The activity of CD101 and comparator antifungal agents against 606 invasive fungal isolates collected worldwide during 2014 was evaluated using the Clinical and Laboratory Standards Institute (CLSI) method. All Candida albicans (n = 251), Candida tropicalis (n = 51), Candida krusei (n = 16), and Candida dubliniensis (n = 11) isolates were inhibited by ≤0.12 μg/ml of CD101 and were susceptible or showed wild-type susceptibility to the other echinocandins tested. Five C. glabrata isolates (n = 100) displayed CD101 MIC values of 1 to 4 μg/ml, had elevated MICs of caspofungin (2 to >8 μg/ml), anidulafungin (2 to 4 μg/ml), and micafungin (2 to 4 μg/ml), and carried mutations on fks1 and fks2. Candida parapsilosis (n = 92) and Candida orthopsilosis (n = 10) displayed higher CD101 MIC values (ranges, 0.5 to 4 μg/ml and 0.12 to 2 μg/ml, respectively), and similar results were observed for the other echinocandins tested. Fluconazole resistance was noted among 11.0% of Candida glabrata isolates, 4.3% of C. parapsilosis isolates, and 2.0% of C. albicans and C. tropicalis isolates. The activity of CD101 against Aspergillus fumigatus (n = 56) was similar to that of micafungin and 2-fold greater than that of caspofungin but less than that of anidulafungin. These isolates had wild-type susceptibility to itraconazole, voriconazole, and posaconazole. The echinocandins had limited activity against Cryptococcus neoformans (n = 19). CD101 was as active as the other echinocandins against common fungal organisms recovered from patients with invasive fungal infections. The long half-life profile is very desirable for the prevention and treatment of serious fungal infections, especially in patients who can then be discharged from the hospital to complete antifungal therapy on an outpatient basis. PMID:28052853

  9. Alternative testing strategies for predicting developmental toxicity of antifungal compound

    NARCIS (Netherlands)

    Li, H.

    2016-01-01

    Determination of safe human exposure levels of chemicals in toxicological risk assessments largely relies on animal toxicity data. In these toxicity studies, the highest number of animals are used for reproductive and developmental toxicity testing. Because of economic and ethical reasons, there is

  10. BSAC standardized disc susceptibility testing method (version 11).

    Science.gov (United States)

    Howe, R A; Andrews, J M

    2012-12-01

    This article highlights key amendments incorporated into version 11 of the BSAC standardized disc susceptibility testing method, available as Supplementary data at JAC Online (http://jac.oxfordjournals.org/) and on the BSAC web site (http://bsac.org.uk/susceptibility/guidelines-standardized-disc-susceptibility-testing-method/). The basic disc susceptibility testing method remains unchanged, but there have been a number of alterations to the interpretive criteria for certain organism/drug combinations due to continuing harmonization with the EUCAST MIC breakpoints and constant efforts to improve the reliability and clinical applicability of the guidance.

  11. In vitro evaluation of Malassezia pachydermatis susceptibility to azole compounds using E-test and CLSI microdilution methods.

    Science.gov (United States)

    Cafarchia, Claudia; Figueredo, Luciana A; Iatta, Roberta; Colao, Valeriana; Montagna, Maria T; Otranto, Domenico

    2012-11-01

    Dermatitis caused by Malassezia spp., one of most common skin disease in dogs, requires prolonged therapy and/or high doses of antifungal agents. In the present study, the antifungal susceptibility of M. pachydermatis to ketoconazole (KTZ), fluconazole (FLZ), itraconazole (ITZ), posaconazole (POS) and voriconazole (VOR) was evaluated in vitro using both CLSI reference broth microdilution (CLSI BMD) and E-test. A total of 62 M. pachydermatis strains from dogs with and without skin lesions were tested. M. pachydermatis strains were susceptible to ITZ, KTZ and POS using both test methods, with the highest MIC found in tests of FLZ. Essential agreement between the two methods ranged from 87.1% (VOR) to 91.9% (ITZ), and categorical agreement from 74.2% (FLZ) to 96.8% (ITZ). Minor error discrepancies were observed between the two methods, with major discrepancies observed for KTZ. A higher MIC(50) value for FLZ was noted with M. pachydermatis genotype B. The MICs(50) of M. pachydermatis genotype B for KTZ, VOR and POS were higher in isolates from dogs with skin lesions than those in isolates from animals without skin lesions. The results suggest a link between genotypes of M. pachydermatis and in vitro drug susceptibility. The categorical agreement for both E-test and CLSI BMD methods found in this investigation confirms the E-test as a reliable diagnostic method for routine use in clinical mycology laboratories.

  12. Evaluation of Malassezia pachydermatis antifungal susceptibility using two different methods Avaliação da sensibilidade da Malassezia pachydermatis frente a antifúngicos através de duas técnicas

    Directory of Open Access Journals (Sweden)

    Patrícia da Silva Nascente

    2003-12-01

    Full Text Available Malassezia pachydermatis is recognized as a normal inhabitant and an opportunistic pathogen of the external ear canal and skin of dogs and cats. In especial clinical conditions, and mainly in the cases of therapeutic failure related to external otitis and dermatitis complicated by this yeast, it is recommended test susceptibility to antifungal drugs. The purpose of this work was to evaluate the susceptibility of 44 isolates of M. pachydermatis from the external ear canal and skin of dogs and cats using two different in vitro antifungal susceptibility methods: the Etest® and the broth microdilution method. Thirty-five samples were tested using the Etest®, twenty-four samples were tested using the broth microdilution method and fifteen samples were tested using both tests. The antifungal agents used were ketoconazole (KTZ, fluconazole (FLZ and itraconazole (ITZ. In the broth microdilution method the yeast was less susceptible to ITZ while KTZ had the strongest activity. On the other hand, the Etest® showed that M. pachydermatis was more susceptible to ITZ while the less active drug was FLZ. The simultaneous evaluation using both methods identified FLZ as the antifungal drug with the highest activity (64.3%, followed by KTZ (57.1% and ITZ (28.6%. These results showed that there is an urgent need to standardize of the values considered as parameters for growth inhibition of this yeast so a simple and efficient method can be used routinely in the laboratory practice.Malassezia pachydermatis é considerada um habitante normal e patógeno oportunista do meato acústico externo e tegumento cutâneo de cães e gatos. Em condições clínicas especiais e nos casos de fracasso terapêutico, comum em casos de otite externa ou dermatite complicadas por esta levedura, é recomendado testar a sensibilidade aos antifúngicos. O objetivo do trabalho foi avaliar a sensibilidade de 44 isolados de M. pachydermatis do meato acústico externo e do tegumento cut

  13. Broth dilution testing of Candida albicans susceptibility to ketoconazole.

    OpenAIRE

    Hughes, C E; Bennett, R L; Beggs, W H

    1987-01-01

    We performed a detailed investigation of the kinetics of ketoconazole activity in the setting of broth dilution testing of Candida albicans susceptibility. Turbidimetric readings reflected parallel quantitative colony counts. The method of endpoint determination markedly affected the results. Determinations of 50% inhibitory concentrations clearly separated the ketoconazole-resistant strains from the susceptible strains.

  14. Caspofungin Etest susceptibility testing of Candida species

    DEFF Research Database (Denmark)

    Arendrup, Maiken Cavling; Pfaller, Michael A; Schønheyder, Henrik Carl

    2012-01-01

    The purpose of this study was to evaluate the performance of caspofungin Etest and the recently revised CLSI breakpoints. A total of 497 blood isolates, of which 496 were wild-type isolates, were included. A total of 65/496 susceptible isolates (13.1%) were misclassified as intermediate (I) or re......) or resistant (R). Such misclassifications were most commonly observed for Candida krusei (73.1%) and Candida glabrata (33.1%). The revised breakpoints cannot be safely adopted for these two species....

  15. Species distribution and antifungal susceptibility of 143 Candida strains%143株念珠菌菌种分布及对5种抗真菌药物的敏感性分析

    Institute of Scientific and Technical Information of China (English)

    黄云昆; 朱雯梅; 高辉; 徐敏; 许丽虹; 王杨

    2011-01-01

    Objective To analyze the species distribution and antifungal susceptibility of 143 Candida strains isolated from clinical specimens. Methods A comprehensive study was carried out to analyze the species distribution of Candida spp. and their susceptibility to five commonly used antifungal agents. A total of 143 isolates were collected from Yan' an Hospital between 2005 and 2009. Antifungal susceptibility testing was conducted with ATB-Fungus-3 strip. Results Candida albicans accounted for 39. 2% (56/143), and non-albicans Candida spp. 60.8% (87/143), including Candida glabrata 24.5% (35/143), Candida tropicalis 7. 7% (11/143) and Candida parapsilosis 5.6% (8/143). Of the 143 Candida isolates, the overall percentage of strains susceptible to flucytosine, amphotericin B, fluconazole, itraconazole, and voriconazole was 86. 0%, 100%, 90.9%, 69.9%, and 93.5 %, respectively. About 91. l%, 100%, 96.4%, 82. 1 %, and 96.3% of the 56 Candida a lbicans strains were susceptible to these five antifungal agents. Of the 87 non-albicans Candida spp. isolates, the susceptibility rates were 82.8%, 100%, 87. 4%, 62. 1 %, and 92. 0%, respectively. Conclusions Candida albicans remains a common species in Candida infections. Non-albicans Candida spp. is also increasing. Candida albicans isolates are still highly susceptible to the common antifungal agents. Non-albicans Candida species are more resistant than Candida albicans. Clinical antifungal therapy should be based on the results of pathogen identification and antifungal susceptibility testing.%目的 分析临床标本中念珠菌属的菌种分布及对常用抗真菌药物的敏感性,为临床合理用药提供依据.方法 总结分析昆明市延安医院2005-2009年念珠菌属的菌种分布及其对5种常用抗真菌药物的敏感性.药敏试验采用ATB-Fungus-3微量稀释法.结果 在2005-2009年分离的共143株念珠菌中,白念珠菌占39.2%(56/143),非白念珠菌占60

  16. Comparison of European Committee on Antimicrobial Susceptibility Testing (EUCAST) and Etest Methods with the CLSI Broth Microdilution Method for Echinocandin Susceptibility Testing of Candida Species▿

    Science.gov (United States)

    Pfaller, M. A.; Castanheira, M.; Diekema, D. J.; Messer, S. A.; Moet, G. J.; Jones, R. N.

    2010-01-01

    The antifungal broth microdilution (BMD) method of the European Committee on Antibiotic Susceptibility Testing (EUCAST) and the Etest agar diffusion method were compared with the Clinical and Laboratory Standards Institute (CLSI) BMD method M27-A3 for anidulafungin, caspofungin, and micafungin susceptibility testing of 133 clinical isolates of Candida species. The isolates were characterized for the presence or absence of fks1 and/or fks2 gene mutations and included 34 isolates of C. glabrata (4 mutant strains), 32 of C. albicans (1 mutant strain), 25 of C. parapsilosis, 19 of C. guilliermondii, 12 of C. tropicalis (2 mutant strains), and 11 of C. krusei. Excellent essential agreement (EA; within 2 dilutions) between the CLSI and EUCAST and CLSI and Etest MIC results was observed. The overall EA between the EUCAST and CLSI results ranged from 89.5% (caspofungin) to 99.2% (micafungin), whereas the EA between the Etest and CLSI results ranged from 90.2% (caspofungin) to 93.2% (anidulafungin). The categorical agreement (CA) between methods for each antifungal agent was assessed using previously determined epidemiological cutoff values (ECVs). Excellent CA (>90%) was observed for all comparisons between the EUCAST and CLSI results with the exceptions of C. glabrata and caspofungin (85.3%) and C. krusei and caspofungin (54.5%). The CA between the Etest and CLSI results was also excellent for all comparisons, with the exception of C. krusei and caspofungin (81.8%). All three methods were able to differentiate wild-type (WT) strains from those with fks mutations. With anidulafungin as the test reagent, the CLSI method identified 5 of 7 mutant strains, whereas the EUCAST method and the Etest identified 6 of 7 mutant strains. With either caspofungin or micafungin as the test reagent, the CLSI method identified all 7 mutant strains and the EUCAST method identified 6 of 7 mutant strains. The Etest identified all 7 mutant strains using caspofungin as the reagent. All three

  17. Chromogenic medium for direct susceptibility testing of Candida spp. isolated from urine.

    Science.gov (United States)

    de Vasconcelos, Antônio Alexandre; Menezes, Everardo Albuquerque; Cunha, Francisco Afrânio

    2011-08-01

    Currently, there has been an increased frequency of fungal infections. Candida albicans and other Candida spp. have been proven to be major causes for urinary tract infection. Increased resistance to antifungals indicates the need to develop strategies in order to prevent the spread of resistance. Chromogenic medium have been proven to be useful in the detection of yeasts in clinical specimens containing mixed cultures of Candida. The aim of this study was to compare the results of antifungal susceptibility testing with fluconazole and amphotericin B on strains of Candida spp. isolated from urine, conducted on a Mueller-Hinton Agar with Glucose and Methylene Blue (MHAGMB) medium and on a Hicrome Candida® Agar with 2% Glucose (HCAG) medium. We used 40 samples of Candida spp. isolated from urine samples from inpatients and outpatients. The results showed that both media presented high rates of agreement, above 94%. The use of the HCAG medium decreases the release time of the results by 24-48 h, which may be decisive for initiating the correct drug treatment.

  18. ATB FUNGUS 3与CLSI M27-A2微量液基稀释法测定酵母菌药物敏感性的一致性比较%Comparative evaluation of ATB FUNGUS 3 procedure and CLSI M27-A2 broth microdilution method for antifungal susceptibility testing of pathogenic yeasts

    Institute of Scientific and Technical Information of China (English)

    刘伟; 乔建军; 马彦; 万喆; 李若瑜

    2009-01-01

    为92.4%、93.0%和93.6%.对于有判读折点的念珠菌,2种方法测定FLC、ITC、VRC、5-FC的一致性分别为81.7%、59.2%、90.9%和96.5%.结论 ATB FUNGUS 3方案操作简便,读取结果更加快速,除了FLC和ITC对于有判读折点的念珠菌的抑菌活性外,ATB FUNGUS 3与CLSI M27-A2方案具有良好的一致性.%to common antifungal drugs.

  19. Results from the ARTEMIS DISK Global Antifungal Surveillance Study, 1997 to 2007: 10.5-Year Analysis of Susceptibilities of Noncandidal Yeast Species to Fluconazole and Voriconazole Determined by CLSI Standardized Disk Diffusion Testing▿

    Science.gov (United States)

    Pfaller, M. A.; Diekema, D. J.; Gibbs, D. L.; Newell, V. A.; Bijie, H.; Dzierzanowska, D.; Klimko, N. N.; Letscher-Bru, V.; Lisalova, M.; Muehlethaler, K.; Rennison, C.; Zaidi, M.

    2009-01-01

    Fluconazole in vitro susceptibility test results determined by the CLSI M44-A disk diffusion method for 11,240 isolates of noncandidal yeasts were collected from 134 study sites in 40 countries from June 1997 through December 2007. Data were collected for 8,717 yeast isolates tested with voriconazole from 2001 through 2007. A total of 22 different species/organism groups were isolated, of which Cryptococcus neoformans was the most common (31.2% of all isolates). Overall, Cryptococcus (32.9%), Saccharomyces (11.7%), Trichosporon (10.6%), and Rhodotorula (4.1%) were the most commonly identified genera. The overall percentages of isolates in each category (susceptible, susceptible dose dependent, and resistant) were 78.0%, 9.5%, and 12.5% and 92.7%, 2.3%, and 5.0% for fluconazole and voriconazole, respectively. Less than 30% of fluconazole-resistant isolates of Cryptococcus spp., Cryptococcus albidus, Cryptococcus laurentii, Trichosporon beigelii/Trichosporon cutaneum, Rhodotorula spp., Rhodotorula rubra/Rhodotorula mucilaginosa, and Rhodotorula glutinis remained susceptible to voriconazole. Emerging resistance to fluconazole was documented among isolates of C. neoformans from the Asia-Pacific, Africa/Middle East, and Latin American regions but not among isolates from Europe or North America. This survey documents the continuing broad spectrum of activity of voriconazole against opportunistic yeast pathogens but identifies several of the less common species with decreased azole susceptibility. These organisms may pose a future threat to optimal antifungal therapy and emphasize the importance of prompt and accurate species identification. PMID:19005141

  20. In vitro Study on Antifungal Susceptibility of Vaginal Candida from Healthy Population in Dali%大理地区健康人阴道念珠菌体外药物敏感性研究

    Institute of Scientific and Technical Information of China (English)

    何伟; 陈财发; 姚毅; 朱文; 杨泽丽; 白丽

    2012-01-01

    目的:通过对分离自大理地区健康女性阴道念珠菌的药物敏感性分析,为临床抗真菌药物的选择提供依据。方法:采用微量液基稀释法,测定念珠菌对氟康唑(FCZ)、酮康唑(KCZ)、5-氟胞嘧啶(5-FC)和两性霉素B(AMB)4种常用抗真菌药物的敏感性(MIC值)。结果:45株健康人群阴道念珠菌对抗真菌药物FCZ、KCZ、5-FC、AMB的耐药率分别为:8.89%、6.67%、6.67%、2.22%,敏感率分别为:80%、84.44%、91.11%、97.78%,1株克柔念珠菌对FCZ、KCZ和5-FC同时耐药,另1株克柔念珠菌对KCZ和5-FC同时耐药。45株念珠菌对四种抗真菌药物的耐药率差异无统计学意义,敏感率除AMB的药物敏感性比FCZ好(P=0.008),AMB的药物敏感性比KCZ的好(P=0.027),其余药物的敏感率差异无统计学意义(P〉0.05)。结论:上述4种抗真菌药物均可选作阴道念珠菌病的治疗,相比而言,更推荐使用AMB,但应注意某些菌株的剂量依赖性敏感、耐药及交叉耐药现象。%Objective: To provide a basis for clinical selection of antifungal drugs through vaginal candida antifungal susceptibility test.Methods: We measured the vaginal candida antifungal susceptibility(minimal inhibitory concentration,MIC) to fluconazole(FCZ),ketoconazole(KCZ),5-fluorocytosine(5-FC) and amphotericin B(AMB) by using Broth microdilution method.Results: The resistance rates of 45 strains vaginal candida from healthy subjects to antifungal drug FCZ,KCZ,5-FC,AMB were 8.89%,6.67%, 6.67% and 2.22% respectively.And the sensitive rates were 80%,84.44%,91.11%,97.78% respectively.One strain of C.krusei is resistant to FCZ,KCZ and 5-FC while another strain was simultaneous resistant to KCZ and 5-FC.There were no significance differences in all 45 strains of candida in antifungal drugs resistant rate.The sensitivity of Candida to AMB was higher than that of FCZ(P=0.008).The sensitivity of Candida to AMB was higher than that of KCZ

  1. Experimental evaluation of antifungal and antiseptic agents against Rhodotorula spp.

    Science.gov (United States)

    Preney, L; Théraud, M; Guiguen, C; Gangneux, J P

    2003-12-01

    We studied the susceptibility of 21 strains of Rhodotorula rubra and nine strains of R. glutinis to eight antifungals and tested eight antiseptic agents on one strain of R. rubra. The tested strains were susceptible to ketoconazole, 5-fluorocytosine, amphotericin B, and nystatin, intermediate to econazole and resistant to fluconazole, itraconazole and miconazole. After 5-min contact, six of the eight antiseptic agents tested showed a fungicidal activity on the tested R. rubra strain.

  2. [Investigation of the correlation between biofilm forming ability of urinary Candida isolates with the use of urinary catheters and change of antifungal susceptibility in the presence of biofilm].

    Science.gov (United States)

    Aslan, Hacer; Gülmez, Dolunay

    2016-04-01

    Frequency of Candida species causing urinary tract infections is increasing, and this increase is outstanding in nosocomial urinary tract infections especially in intensive care units. The ability of biofilm formation that is contributed to the virulence of the yeast, plays a role in the pathogenesis of biomaterial-related infections and also constitutes a risk for treatment failure. The aims of this study were to compare biofilm forming abilities of Candida strains isolated from urine cultures of patients with and without urinary catheters, and to investigate the change of antifungal susceptibility in the presence of biofilm. A total of 50 Candida strains isolated from urine cultures of 25 patients with urinary catheters (10 C.tropicalis, 6 C.glabrata, 4 C.albicans, 4 C.parapsilosis, 1 C.krusei) and 25 without urinary catheters (8 C.tropicalis, 6 C.albicans, 4 C.krusei, 3 C.parapsilosis, 2 C.kefyr, 1 C.glabrata, 1 C.lusitaniae) were included in the study. Biofilm forming ability was tested by Congo red agar (CRA) and microplate XTT [2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] reduction methods. Fluconazole (FLU) and amphotericin B (AMP-B) susceptibilities of the isolates were determined by reference microdilution method recommended by Clinical and Laboratory Standards Institute for planktonic cells and by XTT reduction assay in case of biofilm presence. Biofilm formation was detected in 12 (24%) by CRA and 50 (100%) of the isolates by XTT reduction method. None of the C.albicans (n= 10) and C.tropicalis (n= 18) strains were detected as biofilm positive by CRA, however, these strains were strongly positive by XTT reduction method. No statistically significant correlation was detected between the presence of urinary catheter and biofilm forming ability of the isolate (p> 0.05). This might be caused by the advantage of biofilm forming strains in adhesion to bladder mucosa at the initial stages of infection. For all of the isolates in

  3. Rapid susceptibility testing and microcolony analysis of Candida spp. cultured and imaged on porous aluminum oxide.

    Directory of Open Access Journals (Sweden)

    Colin J Ingham

    Full Text Available BACKGROUND: Acquired resistance to antifungal agents now supports the introduction of susceptibility testing for species-drug combinations for which this was previously thought unnecessary. For pathogenic yeasts, conventional phenotypic testing needs at least 24 h. Culture on a porous aluminum oxide (PAO support combined with microscopy offers a route to more rapid results. METHODS: Microcolonies of Candida species grown on PAO were stained with the fluorogenic dyes Fun-1 and Calcofluor White and then imaged by fluorescence microscopy. Images were captured by a charge-coupled device camera and processed by publicly available software. By this method, the growth of yeasts could be detected and quantified within 2 h. Microcolony imaging was then used to assess the susceptibility of the yeasts to amphotericin B, anidulafungin and caspofungin (3.5 h culture, and voriconazole and itraconazole (7 h culture. SIGNIFICANCE: Overall, the results showed good agreement with EUCAST (86.5% agreement; n = 170 and E-test (85.9% agreement; n = 170. The closest agreement to standard tests was found when testing susceptibility to amphotericin B and echinocandins (88.2 to 91.2% and the least good for the triazoles (79.4 to 82.4%. Furthermore, large datasets on population variation could be rapidly obtained. An analysis of microcolonies revealed subtle effects of antimycotics on resistant strains and below the MIC of sensitive strains, particularly an increase in population heterogeneity and cell density-dependent effects of triazoles. Additionally, the method could be adapted to strain identification via germ tube extension. We suggest PAO culture is a rapid and versatile method that may be usefully adapted to clinical mycology and has research applications.

  4. [Antimicrobial susceptibility testing of anaerobic bacteria].

    Science.gov (United States)

    García-Sánchez, José E; García-Sánchez, Enrique; García-García, María Inmaculada

    2014-02-01

    The anaerobic bacteria resistance to antibiotics is increasing, and even has appeared against the most active of those, like metronidazol and carbapenems. This fact forces to make and periodical sensibility tests -at least in the most aggressive and virulent species, in cases that they are isolated from life locations and in the absence of therapeutic response- to check the local sensibility and to establish suitable empiric therapies, all based on multicentric studies carried out in order to this or well to check the activity of new antibiotics. For the laboratory routine, the easiest sensibility method is the E-test/MIC evaluator. Another alternative is microdilution, that's only normalized for Bacteroides. There are preliminary facts that allow the use of disc diffusion method in some species of Bacteroides and Clostridium. For the temporal and multicentric studies, the procedure is dilution in agar plate, the reference method.

  5. Susceptibility testing of fish cell lines for virus isolation

    DEFF Research Database (Denmark)

    Ariel, Ellen; Skall, Helle Frank; Olesen, Niels Jørgen

    2009-01-01

    compare susceptibility between cell lines and between lineages within a laboratory and between laboratories (Inter-laboratory Proficiency Test). The objective being that the most sensitive cell line and lineages are routinely selected for diagnostic purposes.In comparing cell lines, we simulated "non......-cell-culture-adapted" virus by propagating the virus in heterologous cell lines to the one tested. A stock of test virus was produced and stored at - 80 °C and tests were conducted biannually. This procedure becomes complicated when several cell lines are in use and does not account for variation among lineages. In comparing...... cell lineages, we increased the number of isolates of each virus, propagated stocks in a given cell line and tested all lineages of that line in use in the laboratory. Testing of relative cell line susceptibility between laboratories is carried out annually via the Inter-laboratory Proficiency Test...

  6. Disk diffusion susceptibility testing of ticarcillin plus clavulanic acid.

    Science.gov (United States)

    Fuchs, P C; Jones, R N; Barry, A L; Thornsberry, C

    1984-01-01

    Disk diffusion susceptibility testing of ticarcillin plus the beta-lactamase inhibitor clavulanic acid was performed on 489 clinical isolates, and the results were compared with reference broth microdilution susceptibilities. Four different disks containing 75 micrograms of ticarcillin plus 2.5, 5, 10, and 15 micrograms of clavulanic acid were evaluated. Based on test performance and clavulanic acid stability, the 75-10-micrograms disk is recommended. Interpretive criteria for ticarcillin as published by the National Committee for Clinical Laboratory Standards appear to be satisfactory for the combination drug, but because the number of ticarcillin-clavulanic acid-resistant isolates was small, this proposal must be considered only tentative. PMID:6715524

  7. Candida albicans bloodstream isolates in a German university hospital are genetically heterogenous and susceptible to commonly used antifungals.

    Science.gov (United States)

    Huyke, Johanna; Martin, Ronny; Walther, Grit; Weber, Michael; Kaerger, Kerstin; Bougnoux, Marie-Elisabeth; Elias, Johannes; Kurzai, Oliver

    2015-10-01

    From an eight-year-span, 99 Candida bloodstream isolates were collected at the University Hospital Wuerzburg, Germany. In this study, all strains were analyzed using molecular and phenotypic typing methods. Confirmatory species identification revealed three isolates that were initially diagnosed as C. albicans to be actually C. dubliniensis. Two isolates contained a mixed culture of C. albicans and C. glabrata, in one of the specimens both species could be separated while it was not possible to recover C. albicans in the other sample. The remaining 95 C. albicans isolates were profiled by multilocus sequence typing (MLST). Phylogenetic analyses showed a highly heterogenous collection of strains, associated with many different clades and constituting a set of new diploid sequence types (DST). For all strains with identical DST, patient data were reviewed for potential nosocomial transmission. In addition, all isolates were tested for their susceptibility to amphotericin B, caspofungin, fluconazole, itraconazole, posaconazole and voriconazole. No clinically relevant resistance could be detected. Furthermore, these data underline that correlation between minimal inhibitory concentrations for caspofungin and anidulafungin is low.

  8. Antimycobacterial susceptibility testing methods for natural products research

    Directory of Open Access Journals (Sweden)

    Juan Gabriel Bueno Sánchez

    2010-06-01

    Full Text Available The emergence of multidrug-resistant strains of Mycobacterium tuberculosis underscores the need of continuous developments on new and efficient methods to determine the susceptibility of isolates of M. tuberculosis in the search for novel antimicrobial agents. Natural products constitute an important source of new drugs, but design and implementation of antimycobacterial susceptibility testing methods are necessary for evaluate the different extracts and compounds. A number of biological assay methodologies are in current use, ranging from the classical disk diffusion and broth dilution assay format, to radiorespirometric (BACTEC, dye-based, and fluorescent/luminescence reporter assays. This review presents an analysis on the in vitro susceptibility testing methods developed for determinate antitubercular activity in natural products and related compounds (semi-synthetic natural products and natural products-derived compounds and the criteria to select the adequate method for determination of biological activity of new natural products.

  9. In Vitro Activity of a Novel Broad-Spectrum Antifungal, E1210, Tested against Aspergillus spp. Determined by CLSI and EUCAST Broth Microdilution Methods ▿

    Science.gov (United States)

    Pfaller, Michael A.; Duncanson, Frederick; Messer, Shawn A.; Moet, Gary J.; Jones, Ronald N.; Castanheira, Mariana

    2011-01-01

    E1210 is a first-in-class broad-spectrum antifungal that suppresses hyphal growth by inhibiting fungal glycophosphatidylinositol (GPI) biosynthesis. In the present study, we extend these findings by examining the activity of E1210 and comparator antifungal agents against Aspergillus spp. by using the methods of the Clinical and Laboratory Standards Institute (CLSI) and the European Committee for Antimicrobial Susceptibility Testing (EUCAST) to test wild-type (WT) as well as amphotericin B (AMB)-resistant (-R) and azole-R strains (as determined by CLSI methods). Seventy-eight clinical isolates of Aspergillus were tested including 20 isolates of Aspergillus flavus species complex (SC), 22 of A. fumigatus SC, 13 of A. niger SC, and 23 of A. terreus SC. The collection included 15 AMB-R (MIC, ≥2 μg/ml) isolates of A. terreus SC and 10 itraconazole-R (MIC, ≥4 μg/ml) isolates of A. fumigatus SC (7 isolates), A. niger SC (2 isolates), and A. terreus SC (1 isolate). Comparator antifungal agents included anidulafungin, caspofungin, amphotericin B, itraconazole, posaconzole, and voriconazole. Both CLSI and EUCAST methods were highly concordant for E1210 and all comparators. The essential agreement (EA; ±2 log2 dilution steps) was 100% for all comparisons with the exception of posaconazole versus A. terreus SC (EA = 91.3%). The minimum effective concentration (MEC)/MIC90 values (μg/ml) for E1210, anidulafungin, caspofungin, itraconazole, posaconazole, and voriconazole, respectively, were as follows for each species: for A. flavus SC, 0.03, ≤0.008, 0.12, 1, 1, and 1; for A. fumigatus SC, 0.06, 0.015, 0.12, >8, 1, and 4; for A. niger SC, 0.015, 0.03, 0.12, 4, 1, and 2; and for A. terreus SC, 0.06, 0.015, 0.12, 1, 0.5, and 1. E1210 was very active against AMB-R strains of A. terreus SC (MEC range, 0.015 to 0.06 μg/ml) and itraconazole-R strains of A. fumigatus SC (MEC range, 0.03 to 0.12 μg/ml), A. niger SC (MEC, 0.008 μg/ml), and A. terreus SC (MEC, 0.015

  10. Antifungal Production of a Strain of Actinomycetes spp Isolated from the Rhizosphere of Cajuput Plant: Selection and Detection of Exhibiting Activity Against Tested Fungi

    Directory of Open Access Journals (Sweden)

    Alimuddin A

    2015-11-01

    Full Text Available Actinomycetes are bacteria known to constitute a large part of the rhizosphere microbiota. Their isolation is an important step for screening of new bioactive compounds. Culturable actinomycetes populations from cajuput plant rhizosphere soils in Wanagama I Forest UGM Yogyakarta were collected to study about their antifungal activity. Among 17 of a total 43 isolates that showed activity were screened for producing antifungi substances. Screening for antifungal activity of isolates were performed with dual culture bioassay in vitro. One isolate that was designated as Streptomyces sp.GMR-22 was the strongest against all tested fungi and appeared promising for a sources of antifungal. Culture’s supernatant and mycelia were extracted with chloroform, ethyl acetate and methanol, respectively. Antifungal activity of crude extracts was tested by diffusion method against tested fungi. The result indicates that isolates of actinomycetes from cajuput plant rhizosphere could be an interesting sources of antifungal bioactive substances.

  11. Antifungal activity of traditional medicinal plants from Tamil Nadu, India

    Institute of Scientific and Technical Information of China (English)

    Duraipandiyan V; Ignacimuthu S

    2011-01-01

    Objective:To assess the antifungal activity of hexane, ethyl acetate and methanol extracts of 45 medicinal plants and to determine the minimum inhibitory concentration for each extract against human pathogenic fungi. Methods:A total of 45 medicinal plants were collected from different places of Tamil Nadu and identified. Hexane, ethyl acetate and methanol extracts of 45 medicinal plants were assessed for antifungal susceptibility using broth microdilution method. Two known antifungal agents were used as positive controls. Results: Most of the extracts inhibited more than four fungal strains. From the evaluation we found that ethyl acetate extracts inhibited large number of fungal growth. Hexane extracts also nearly showed the same level of inhibition against fungal growth. Methanol extracts showed the minimum antifungal activity. Among the 45 plants tested, broad spectrum antifungal activity was detected in Albizzia procera (A. procera), Atalantia monophylla, Asclepias curassavica, Azima tetracantha, Cassia fistula (C. fistula), Cinnomomum verum, Costus speciosus (C. speciosus), Nymphaea stellata, Osbeckia chinensis, Piper argyrophyllum, Punica granatum, Tinospora cordifolia and Toddalia asiatica (T. asiatica). Promising antifungal activity was seen in A. procera, C. speciosus, C. fistula and T. asiatica. Conclusions:It can be concluded that the plant species assayed possess antifungal properties. Further phytochemical research is needed to identify the active principles responsible for the antifungal effects of some of these medicinal plants.

  12. Agar dilution method for susceptibility testing of Neisseria gonorrhoeae

    Directory of Open Access Journals (Sweden)

    Marta C de Castillo

    1996-12-01

    Full Text Available The antibiotic susceptibilities of Neisseria gonorrhoeae isolates obtained from patients attending a clinic for sexually transmitted diseases in Tucumán, Argentina, were determined by the agar dilution method (MIC. 3.5% of the isolates produced ²-lactamase. A total of 96.5% of ²-lactamase negative isolates tested were susceptible to penicillin (MIC < 2 µgml-1; 14.03% of the tested isolates were resistant to tetracycline (MIC < 2 µgml-1, and 98% of the tested isolates were susceptible to spectinomycin (MIC < 64 µgml-1. The MICs for 95% of the isolates, tested for other drugs were: < 2 µgml-1 for cefoxitin, < 0.06 µgml-1 for cefotaxime, < 0.25 µgml-1 for norfloxacin, < 10 µgml-1 for cephaloridine, < 10 µgml-1 for cephalexin, and < 50 µgml-1 for kanamycin. Antibiotic resistance among N. gonorrhoeae isolates from Tucumán, Argentina, appeared to be primarily limited to penicillin and tetracycline, which has been a general use against gonorrhoeae in Tucumán since 1960. Periodic monitoring of the underlying susceptibility profiles of the N. gonorrhoeae strains prevalent in areas of frequent transmission may provide clues regarding treatment options and emerging of drug resistance.

  13. Radiated Susceptibility Tests in Thermal Vacuum Chambers for Space Systems

    Science.gov (United States)

    Anon Cancela, Manuel; Hernandez-Gomez, Daniel; Vazquez-Pascual, Mercedes; Lopez-Sanz, Daniel

    2016-05-01

    INTA EMC Area has a wide experience in performing Radiated Susceptibility (RS) tests according to civilian, military and aeronautical standards in Mode Tuned Chambers (MTC) for national and international projects; besides, INTA has two Thermal Vacuum Chamber (TVC) facilities in service for Space Systems tests. In order to perform RS tests to Space Systems in a more realistic environment, INTA EMC Area has stablished an internal research program to develop a procedure to perform this kind of tests inside a TVC as a Mode Tuned Chamber (MTC). In this paper the results of the TVC-04 validation measurements as a MTC are presented.

  14. Statistical Analyses of Correlation between Fluconazole MICs for Candida spp. Assessed by Standard Methods Set Forth by the European Committee on Antimicrobial Susceptibility Testing (E.Dis. 7.1) and CLSI (M27-A2)▿

    Science.gov (United States)

    Rodriguez-Tudela, Juan Luis; Donnelly, J. Peter; Pfaller, Michael A.; Chryssantou, Erja; Warn, Peter; Denning, David W.; Espinel-Ingroff, Ana; Barchiesi, Francesco; Cuenca-Estrella, Manuel

    2007-01-01

    The European Committee on Antimicrobial Susceptibility Testing (EUCAST) Subcommittee on Antifungal Susceptibility Testing recently published a standard for determining the susceptibility of fermentative yeasts to antifungals. From the beginning, the EUCAST and its North American counterpart, the CLSI, decided to work together in order to establish common standards. As part of this exercise, the susceptibility of a set of 475 yeast isolates was tested by both standards. The intraclass correlation coefficient and the equations defining the linear regression between both methods were estimated. Both methods produced very similar results, with an intraclass correlation coefficient of 0.954 (0.945 to 0.962), although linear regression analysis shows that the EUCAST standard resulted in slightly lower MICs. There were only eight isolates showing at least four twofold dilution MIC differences between both standards. After 24 h of incubation, the MICs obtained by the CLSI method were equivalent to those obtained by the EUCAST standard. In summary, both methods produce very similar MICs, indicating that methodology does not pose any obstacle to obtaining uniform standards for antifungal susceptibility testing of yeasts. PMID:17093015

  15. Antifungal susceptibility testing of clinical isolates of fluconazole-resistant Candida spp. and itraconazole-resistant Aspergillus funmigates to posaconazole%耐氟康唑念珠菌和耐伊曲康唑烟曲霉对泊沙康唑的敏感性测定

    Institute of Scientific and Technical Information of China (English)

    王雪洁; 万哲; 李若瑜; 刘伟

    2014-01-01

    目的:测定耐氟康唑念珠菌和耐伊曲康唑烟曲霉临床分离株对泊沙康唑的敏感性。方法参照美国临床实验室标准化研究所制定的 M27-A3和 M38-A2方案,测定从临床获得的11株耐氟康唑的念珠菌和3株耐伊曲康唑烟曲霉对泊沙康唑的 MIC 值。结果对于氟康唑耐药的念珠菌,泊沙康唑的 MIC 范围是0.125~1μg/ mL。对于伊曲康唑耐药烟曲霉,泊沙康唑的 MIC 范围是0.06~0.5μg/ mL。结论11株耐氟康唑的念珠菌和3株耐伊曲康唑烟曲霉均对泊沙康唑有效。%Objective To determine the in vitro susceptibility of fluconazole( FLU)-resistant Candida spp. and itraconazole (ITC)-resistant Aspergillus spp. to posaconazole(POS). Methods The MICs of FLU,ITC,voriconazole(VRC)and POS were determined according to CLSI M27-A3 and M38-A2 methods in eleven isolates of Candida spp. and three isolates of Aspergillus spp. Results MICs of FLU,ITC,VRC and POS against FLU-resistant Candida spp. were 64 μg/ mL,0. 5 ~ 16 μg/ mL,0. 25 ~ 2 μg/mL and 0. 125 ~ 1 μg/ mL. And MICs of ITC,VRC and POS against ITC-resistant Aspergillus spp. were ﹥ 16 μg/ mL,1 ~ 2 μg/ mL and 0. 25 ~ 0. 5 μg/ mL. Conclusion POS is effective to FLU-resistant Candida spp. and ITC-resistant Aspergillus spp. in vitro .

  16. Antifungal pharmacokinetics and pharmacodynamics.

    Science.gov (United States)

    Lepak, Alexander J; Andes, David R

    2014-11-10

    Successful treatment of infectious diseases requires choice of the most suitable antimicrobial agent, comprising consideration of drug pharmacokinetics (PK), including penetration into infection site, pathogen susceptibility, optimal route of drug administration, drug dose, frequency of administration, duration of therapy, and drug toxicity. Antimicrobial pharmacokinetic/pharmacodynamic (PK/PD) studies consider these variables and have been useful in drug development, optimizing dosing regimens, determining susceptibility breakpoints, and limiting toxicity of antifungal therapy. Here the concepts of antifungal PK/PD studies are reviewed, with emphasis on methodology and application. The initial sections of this review focus on principles and methodology. Then the pharmacodynamics of each major antifungal drug class (polyenes, flucytosine, azoles, and echinocandins) is discussed. Finally, the review discusses novel areas of pharmacodynamic investigation in the study and application of combination therapy.

  17. Multidrug-Resistant Candida auris Misidentified as Candida haemulonii: Characterization by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry and DNA Sequencing and Its Antifungal Susceptibility Profile Variability by Vitek 2, CLSI Broth Microdilution, and Etest Method.

    Science.gov (United States)

    Kathuria, Shallu; Singh, Pradeep K; Sharma, Cheshta; Prakash, Anupam; Masih, Aradhana; Kumar, Anil; Meis, Jacques F; Chowdhary, Anuradha

    2015-06-01

    Candida auris is a multidrug-resistant yeast that causes a wide spectrum of infections, especially in intensive care settings. We investigated C. auris prevalence among 102 clinical isolates previously identified as Candida haemulonii or Candida famata by the Vitek 2 system. Internal transcribed spacer region (ITS) sequencing confirmed 88.2% of the isolates as C. auris, and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) easily separated all related species, viz., C. auris (n = 90), C. haemulonii (n = 6), C. haemulonii var. vulnera (n = 1), and Candida duobushaemulonii (n = 5). The in vitro antifungal susceptibility was determined using CLSI broth microdilution (CLSI-BMD), the Vitek 2 antifungal susceptibility test, and the Etest method. C. auris isolates revealed uniformly elevated fluconazole MICs (MIC50, 64 μg/ml), and an alarming percentage of isolates (37%) exhibited elevated caspofungin MICs by CLSI-BMD. Notably, 34% of C. auris isolates had coexisting elevated MICs (≥2 μg/ml) for both fluconazole and voriconazole, and 10% of the isolates had elevated coexisting MICs (≥2 μg/ml) to two additional azoles, i.e., posaconazole and isavuconazole. In contrast to reduced amphotericin B MICs by CLSI-BMD (MIC50, 1 μg/ml) for C. auris, elevated MICs were noted by Vitek 2 (MIC50, 8 μg/ml), which were statistically significant. Candida auris remains an unnoticed pathogen in routine microbiology laboratories, as 90% of the isolates characterized by commercial identification systems are misidentified as C. haemulonii. MALDI-TOF MS proved to be a more robust diagnostic technique for rapid identification of C. auris. Considering that misleading elevated MICs of amphotericin B by the Vitek AST-YS07 card may lead to the selection of inappropriate therapy, a cautionary approach is recommended for laboratories relying on commercial systems for identification and antifungal susceptibility testing of rare yeasts.

  18. Multidrug-Resistant Candida auris Misidentified as Candida haemulonii: Characterization by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry and DNA Sequencing and Its Antifungal Susceptibility Profile Variability by Vitek 2, CLSI Broth Microdilution, and Etest Method

    Science.gov (United States)

    Kathuria, Shallu; Singh, Pradeep K.; Sharma, Cheshta; Prakash, Anupam; Masih, Aradhana; Kumar, Anil

    2015-01-01

    Candida auris is a multidrug-resistant yeast that causes a wide spectrum of infections, especially in intensive care settings. We investigated C. auris prevalence among 102 clinical isolates previously identified as Candida haemulonii or Candida famata by the Vitek 2 system. Internal transcribed spacer region (ITS) sequencing confirmed 88.2% of the isolates as C. auris, and matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) easily separated all related species, viz., C. auris (n = 90), C. haemulonii (n = 6), C. haemulonii var. vulnera (n = 1), and Candida duobushaemulonii (n = 5). The in vitro antifungal susceptibility was determined using CLSI broth microdilution (CLSI-BMD), the Vitek 2 antifungal susceptibility test, and the Etest method. C. auris isolates revealed uniformly elevated fluconazole MICs (MIC50, 64 μg/ml), and an alarming percentage of isolates (37%) exhibited elevated caspofungin MICs by CLSI-BMD. Notably, 34% of C. auris isolates had coexisting elevated MICs (≥2 μg/ml) for both fluconazole and voriconazole, and 10% of the isolates had elevated coexisting MICs (≥2 μg/ml) to two additional azoles, i.e., posaconazole and isavuconazole. In contrast to reduced amphotericin B MICs by CLSI-BMD (MIC50, 1 μg/ml) for C. auris, elevated MICs were noted by Vitek 2 (MIC50, 8 μg/ml), which were statistically significant. Candida auris remains an unnoticed pathogen in routine microbiology laboratories, as 90% of the isolates characterized by commercial identification systems are misidentified as C. haemulonii. MALDI-TOF MS proved to be a more robust diagnostic technique for rapid identification of C. auris. Considering that misleading elevated MICs of amphotericin B by the Vitek AST-YS07 card may lead to the selection of inappropriate therapy, a cautionary approach is recommended for laboratories relying on commercial systems for identification and antifungal susceptibility testing of rare yeasts. PMID:25809970

  19. Antimicrobial susceptibility testing in biofilm-growing bacteria.

    Science.gov (United States)

    Macià, M D; Rojo-Molinero, E; Oliver, A

    2014-10-01

    Biofilms are organized bacterial communities embedded in an extracellular polymeric matrix attached to living or abiotic surfaces. The development of biofilms is currently recognized as one of the most relevant drivers of persistent infections. Among them, chronic respiratory infection by Pseudomonas aeruginosa in cystic fibrosis patients is probably the most intensively studied. The lack of correlation between conventional susceptibility test results and therapeutic success in chronic infections is probably a consequence of the use of planktonically growing instead of biofilm-growing bacteria. Therefore, several in vitro models to evaluate antimicrobial activity on biofilms have been implemented over the last decade. Microtitre plate-based assays, the Calgary device, substratum suspending reactors and the flow cell system are some of the most used in vitro biofilm models for susceptibility studies. Likewise, new pharmacodynamic parameters, including minimal biofilm inhibitory concentration, minimal biofilm-eradication concentration, biofilm bactericidal concentration, and biofilm-prevention concentration, have been defined in recent years to quantify antibiotic activity in biofilms. Using these parameters, several studies have shown very significant quantitative and qualitative differences for the effects of most antibiotics when acting on planktonic or biofilm bacteria. Nevertheless, standardization of the procedures, parameters and breakpoints, by official agencies, is needed before they are implemented in clinical microbiology laboratories for routine susceptibility testing. Research efforts should also be directed to obtaining a deeper understanding of biofilm resistance mechanisms, the evaluation of optimal pharmacokinetic/pharmacodynamic models for biofilm growth, and correlation with clinical outcome.

  20. Molecular typing and antifungal susceptibility of clinical and environmental Cryptococcus neoformans species complex isolates in Goiania, Brazil.

    Science.gov (United States)

    Souza, L K H; Souza Junior, A H; Costa, C R; Faganello, J; Vainstein, M H; Chagas, A L B; Souza, A C M; Silva, M R R

    2010-01-01

    A total of 124 Cryptococcus isolates, including 84 clinical strains obtained from cerebrospinal fluid from AIDS patients and 40 environmental isolates from pigeon excreta and from Eucalyptus trees, were studied. The varieties, serotypes, phospholipase activity and molecular profile of these isolates were determined. Cryptococcus neoformans var. grubii serotype A was identified in 120 isolates and Cryptococcus gattii serotype B in four isolates. The clinical isolates showed higher phospholipase activity than environmental isolates. Similar patterns of in vitro susceptibility to amphotericin B, fluconazole, itraconazole and voriconazole and no resistance were found for all isolates. Molecular type VNI (C. neoformans var. grubii) was recovered in 80 clinical and 40 environmental isolates while the type VGIII (C. gattii) was found in four clinical isolates. This study demonstrated for the first time the molecular types of clinical and environmental Cryptococcus isolates in the midwest Brazil region.

  1. 含有额外拷贝黄曲霉cyp51同源基因的烟曲霉对抗真菌药物的敏感性测定%Antifungal susceptibility of the A.fumigatus transformants containing extra copies of A.flavus cyp51 gene homologues to the common antifungal drugs

    Institute of Scientific and Technical Information of China (English)

    刘伟霞; 孙毅; 万喆; 李若瑜; 刘伟

    2011-01-01

    Objective To investigate the effect of Aspergillus flavus cyp51 genes on antifungal susceptibility by cloning and constucting the extra copies of Aspergillus flavus cyp51 genes. Methods A.flavus cyp5l gene homologues were identified by tblastn searches inA. flavus genome database. PCR fragments composed of the 5'flanking sequence (approximately 1 000 bp) ofcyp51 ,cyp51 ORF, and its 3'flanking sequence (approximately 1 000 bp), were subcloned into shuttle plasmid pRG3-AMAl-NotI to produce recombinant plasmids. These plasmids and empty plasmid pRG3-AMA1-Notl were transformed into A.fumigatus strain AF293.1 (pyrG-) respectively to produce transformants. The Clinical Laboratory Standard Institute broth microdilution method M38-A2 and E-test method were used to assay the minimal inhibitory concentrations (MICs) of itraconazole ( ITC), voriconazole ( VRC), amphotericin B (AMB), and posaconazole (POS), or minimal effect concentration (MEC) of caspofungin (CAS), against these transformants. Results A. flavus genome contains three cyp51 gene homologues, cyp51A ,cyp51B and cyp51 C, of which the ORF size are 1 400-2 000 bp. When these genes were subcloned into shuttle plasmid pRG3-AMA1-NotI, we get plasmids pRG3-AMA1-CYP51 A, pRG3-AMA1-CYP51B and pRG3-AMA1-CYP51C. These plasmids and empty plasmid were transformed into A.fumigatus strain AF293.1 (pyrG-) to produce transformants rCYP51A, rCYP51B, rCYP51C and rpRG. The antifungal susceptibility of these A.fumigatus transformants to the antifungal drugs by broth microdilution assaying and E-test method showed that, rCYP51A and rCYP51B were cross-resistant to VRC and ITC, susceptible to both AM B and CAS; rCYP51C and rpRG were intermediate to ITC and VRC, susceptible to both A MB and CAS. Conclusion In A. fumigatus , extra copies of A.flavus ' cyp51A gene or cyp51B gene have effect on antifungal susceptibility to azoles, have no effect on AMB and CAS. Extra copy ofcyp51C has no obvious effect on all the tested drugs.%目的

  2. The calcineurin inhibitor cyclosporin A exhibits synergism with antifungals against Candida parapsilosis species complex.

    Science.gov (United States)

    Cordeiro, Rossana de Aguiar; Macedo, Ramila de Brito; Teixeira, Carlos Eduardo Cordeiro; Marques, Francisca Jakelyne de Farias; Bandeira, Tereza de Jesus Pinheiro Gomes; Moreira, José Luciano Bezerra; Brilhante, Raimunda Sâmia Nogueira; Rocha, Marcos Fábio Gadelha; Sidrim, José Júlio Costa

    2014-07-01

    Candida parapsilosis complex comprises three closely related species, C. parapsilosis sensu stricto, Candida metapsilosis and Candida orthopsilosis. In the last decade, antifungal resistance to azoles and caspofungin among C. parapsilosis sensu lato strains has been considered a matter of concern worldwide. In the present study, we evaluated the synergistic potential of antifungals and the calcineurin inhibitor cyclosporin A (Cys) against planktonic and biofilms of C. parapsilosis complex from clinical sources. Susceptibility assays with amphotericin, fluconazole, voriconazole, caspofungin and Cys were performed by microdilution in accordance with Clinical and Laboratory Standards Institute guidelines. Synergy testing against planktonic cells of C. parapsilosis sensu lato strains was assessed by the chequerboard method. Combinations formed by antifungals with Cys were evaluated against mature biofilms in microtitre plates. No differences in the antifungal susceptibility pattern among species were observed, but C. parapsilosis sensu stricto strains were more susceptible to Cys than C. orthopsilosis and C. metapsilosis. Synergism between antifungals and Cys was observed in C. parapsilosis sensu lato strains. Combinations formed by antifungals and Cys were able to prevent biofilm formation and showed an inhibitory effect against mature biofilms of C. parapsilosis sensu stricto, C. metapsilosis and C. orthopsilosis. These results strengthen the potential of calcineurin inhibition as a promising approach to enhance the efficiency of antifungal drugs.

  3. Evaluation of the disc diffusion method with a comparison study for fluconazole susceptibility of Candida strains

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Objective To performance susceptibility testing of antifungal agents. Due to the increasing number of resistant strains, susceptibility testing of antifungal agents is gaining importance. Methods We compared the results of standard macrotube dilution reference method with two different microdilution methods, as well as the disc diffusion method in order to test the susceptibility of 150 Candida strains to fluconazole. Results Overall correlation between microdilution and macrodilution methods was 86%. It was 91% between the Minimal Inhibitory Concentrations obtained from macrodilution and disc diffusion zone diameters. Conclusion The disc diffusion test was evaluated as a low-cost, reproducible, and efficient way of assessing the in vitro susceptibility of Candida strains to fluconazole.

  4. Multicenter evaluation of the new Vitek 2 yeast susceptibility test using new CLSI clinical breakpoints for fluconazole.

    Science.gov (United States)

    Pfaller, M A; Diekema, D J; Procop, G W; Wiederhold, N P

    2014-06-01

    A fully automated antifungal susceptibility test system recently updated to reflect the new species-specific clinical breakpoints (CBPs) of fluconazole for Candida (Vitek 2 AF03 yeast susceptibility test; bioMérieux, Inc., Durham, NC) was compared in three different laboratories with the Clinical and Laboratory Standards Institute (CLSI) reference broth microdilution (BMD) method by testing 2 quality control strains, 10 reproducibility strains (4 Candida species and 6 Cryptococcus neoformans strains), and 746 isolates of Candida species (702 isolates, 13 species) and 44 isolates of C. neoformans against fluconazole. Excellent essential agreement (EA) (within 2 dilutions) between the reference and Vitek 2 MICs was observed for fluconazole and Candida species (94.0%). The EA was lower for fluconazole and C. neoformans at 86.4%. The mean times to a result with the Vitek 2 test were 9.1 h for Candida species and 12.1 h for C. neoformans. Categorical agreement (CA) between the two methods was assessed by using the new species-specific CBPs. For less common species without fluconazole CBPs, the epidemiological cutoff values (ECVs) were used to differentiate wild-type (WT; MIC, ≤ ECV) from non-WT (MIC, >ECV) strains. The CAs between the two methods were 92.0% for Candida species (0.3% very major errors [VME] and 2.6% major errors [ME]) and 84.1% for C. neoformans (4.5% VME and 11.4% ME). The updated Vitek 2 AF03 IUO yeast susceptibility system is comparable to the CLSI BMD reference method for testing the susceptibility of clinically important yeasts to fluconazole when using the new (lower) CBPs and ECVs.

  5. Vancomycin-resistant enterococci: validation of susceptibility testing and in vitro activity of novel antibiotics

    DEFF Research Database (Denmark)

    Rathe, Mathias; Lise, Kristensen,; Ellermann-Eriksen, Svend;

    Vancomycin-resistant enterococci: validation of susceptibility testing and in vitro activity of novel antibiotics......Vancomycin-resistant enterococci: validation of susceptibility testing and in vitro activity of novel antibiotics...

  6. Susceptibilidad "in vitro" de cepas de Cryptococcus a 5 drogas antifungicas "In vitro" susceptibility of Cryptococcus strains to 5 antifungal drugs

    Directory of Open Access Journals (Sweden)

    A. J. Bava

    1989-10-01

    Full Text Available Se estudió la susceptibilidad "in vitro" de 24 cepas de 3 especies del género Cryptococcus a 5 drogas antifúngicas (anfotericina B, 5 fluorocitosina, ketoconazol, itraconazol y miconazol. Las mismas se agruparon según su especie, variedad y origen de aislamiento. Para determinar la concentración inhibitoria mínima (C.I.M. de cada droga se empleó el método de dilución en agar con el medio básico nitrogenado para levaduras, adicionado de glucosa. Se obtuvo además la media geométrica de estos valores para cada grupo y se comparó cada uno de ellos. Los resultados obtenidos fueron homogéneos con la sola excepción de las cepas de Cryptococcus sp (no neoformans, en las cuales se detectaron elevados valores de C.I.M. para la 5 fluorocitosina.A comparative study of the "in vitro" susceptibility of 24 Cryptococcus strains to 5 antifungal drugs (amphotericin B, 5 fluorocytosine, miconazole, itraconazole and ketoconazole, was carried out. These strains were grouped according to species, varieties and isolation's origins. The minimum inhibitory concentration (M.I.C. was determinated by the agar dilution technique in yeast nitrogen base agar with dextrose. The mean geometrical of the M.I.C. values of each group was compared with the others. The results obtained were homogeneous with the only exception of the "non neoformans" strains, in which, higher M.I.C. to 5 fluorocytosine values were detected.

  7. Antifungal activity of TiO2 photocatalysis against Penicillium expansum in vitro and in fruit tests.

    Science.gov (United States)

    Maneerat, Chamorn; Hayata, Yasuyoshi

    2006-03-15

    The antifungal activity of TiO2 photocatalytic reaction in the form of TiO2 powder and TiO2 coated on a plastic film against Penicillium expansum was investigated in vitro and in fruit tests. The mixture of P. expansum conidial suspension and TiO2 powder was added to potato dextrose agar (PDA) plates for vitro test. The TiO2 photocatalytic reaction reduced conidial germination of the fungal pathogen. It was found that the ability of the TiO2 photocatalytic reaction to suppress P. expansum growth correlated to the amount of TiO2 added. Lower numbers of viable colonies of P. expansum were observed with increasing amount of TiO2. Regardless of the kind of selected fruit inoculated with P. expansum, both TiO2 powder and TiO2-coated film exhibited antifungal activity to control fruit rot. Development of Penicillium rot in apple was significantly (P = 0.05) retarded by the TiO2 photocatalytic reaction. Similarly the TiO2 photocatalytic reaction was the only treatment where no tomato fruit rot was noticeable after 1 week of storage. TiO2-coated film also decreased brown lesions and Penicillium rot infection in lemons. The mean severity fruit rot scores (browning and softening flesh) were 3.2 and 1.9 for uncoated and TiO2-coated film, respectively. Our findings suggest that "TiO2 photocatalytic reaction" shows antifungal activity against P. expansum which may have potential for postharvest disease control.

  8. Prospective Multicenter Study of the Epidemiology, Molecular Identification, and Antifungal Susceptibility of Candida parapsilosis, Candida orthopsilosis, and Candida metapsilosis Isolated from Patients with Candidemia ▿

    OpenAIRE

    Cantón, Emilia; Pemán, Javier; Quindós, Guillermo; Eraso, Elena; Miranda-Zapico, Ilargi; Álvarez, María; Merino, Paloma; Campos-Herrero, Isolina; Marco, Francesc; de la Pedrosa, Elia Gomez G.; Yagüe, Genoveva; Guna, Remedios; Rubio, Carmen; Miranda, Consuelo; Pazos, Carmen

    2011-01-01

    A 13-month prospective multicenter study including 44 hospitals was carried out to evaluate the epidemiology of Candida parapsilosis complex candidemia in Spain. Susceptibility to amphotericin B, flucytosine, fluconazole, itraconazole, voriconazole, posaconazole, anidulafungin, caspofungin, and micafungin was tested by the microdilution colorimetric method. A total of 364 C. parapsilosis complex isolates were identified by molecular methods: C. parapsilosis (90.7%), Candida orthopsilosis (8.2...

  9. Prevalence rates and antifungal susceptibility profiles of the Candida parapsilosis species complex: results from a nationwide surveillance of candidaemia in Brazil.

    Science.gov (United States)

    Gonçalves, S S; Amorim, C S; Nucci, M; Padovan, A C B; Briones, M R S; Melo, A S A; Colombo, A L

    2010-07-01

    The genetically heterogeneous taxon Candida parapsilosis was recently reclassified into three species: Candida parapsilosis, Candida orthopsilosis and Candida metapsilosis. The prevalences of these species among 141 bloodstream isolates tested in Brazil were 88% for C. parapsilosis, 9% for C. orthopsilosis, and 3% for C. metapsilosis. Except for three C. orthopsilosis isolates that were considered resistant to 5-flucytosine, all isolates representing the different species of this complex were susceptible to polyenes, triazoles and caspofungin.

  10. Antifungal Susceptibility Patterns, In Vitro Production of Virulence Factors, and Evaluation of Diagnostic Modalities for the Speciation of Pathogenic Candida from Blood Stream Infections and Vulvovaginal Candidiasis

    Directory of Open Access Journals (Sweden)

    Chaitanya Tellapragada

    2014-01-01

    Full Text Available Candida spp. have emerged as successful pathogens in both invasive and mucosal infections. Varied virulence factors and growing resistance to antifungal agents have contributed to their pathogenicity. We studied diagnostic accuracy of HiCrome Candida Differential Agar and Vitek 2 Compact system for identification of Candida spp. in comparison with species-specific PCR on 110 clinical isolates of Candida from blood stream infections (54, 49% and vulvovaginal candidiasis (56, 51%. C. albicans (61% was the leading pathogen in VVC, while C. tropicalis (46% was prominent among BSIs. HiCrome Agar and Vitek 2 Compact had good measures of agreement (κ 0.826 and 0.895, respectively, in comparison with PCR. We also tested these isolates for in vitro production of proteinase, esterase, phospholipases, and biofilms. Proteinase production was more among invasive isolates (P=0.017, while phospholipase production was more among noninvasive isolates (P=0.001. There was an overall increase in the production of virulence factors among non-albicans Candida. Identification of clinical isolates of Candida up to species level either by chromogenic agar or by Vitek 2 Compact system should be routinely done to choose appropriate therapy.

  11. Multicenter Comparison of the VITEK 2 Yeast Susceptibility Test with the CLSI Broth Microdilution Reference Method for Testing Fluconazole against Candida spp.▿

    Science.gov (United States)

    Pfaller, M. A.; Diekema, D. J.; Procop, G. W.; Rinaldi, M. G.

    2007-01-01

    A fully automated commercial antifungal susceptibility test system (VITEK 2 yeast susceptibility test; bioMerieux, Inc., Hazelwood, Mo.) was compared in three different laboratories with Clinical and Laboratory Standards Institute (CLSI) reference broth microdilution (BMD) method by testing two quality control strains and a total of 426 isolates of Candida spp. (103 to 135 clinical isolates in each laboratory plus 80 challenge isolates in one laboratory) against fluconazole. Reference BMD MIC endpoints were established after 24 and 48 h of incubation. VITEK 2 endpoints were determined spectrophotometrically after 10 to 26 h of incubation (mean, 13 h). Excellent essential agreement (within two dilutions) between the VITEK 2 and the 24- and 48-h BMD MICs was observed. The overall agreement values were 97.9 and 93.7%, respectively. Both intra- and interlaboratory agreement was 100%. The overall categorical agreement between VITEK 2 and BMD was 97.2% at the 24-h BMD time point and 88.3% at the 48-h BMD time point. Decreased categorical agreement at 48 h was attributed to trailing growth observed with Candida glabrata. The VITEK 2 system reliably detected fluconazole resistance among Candida spp. and demonstrated excellent quantitative and qualitative agreement with the reference BMD method. PMID:17215344

  12. Candidemia in a Brazilian tertiary care hospital: species distribution and antifungal susceptibility patterns Candidemia em hospital terciário brasileiro: distribuição das espécies e padrões de susceptibilidade aos antifúngicos

    Directory of Open Access Journals (Sweden)

    Ana Graciela Ventura Antunes

    2004-10-01

    Full Text Available Recent studies have shown differences in the epidemiology of invasive infections caused by Candida species worldwide. In the period comprising August 2002 to August 2003, we performed a study in Santa Casa Complexo Hospitalar, Brazil, to determine Candida species distribution associated with candidemia and their antifungal susceptibility profiles to amphotericin B, fluconazole and itraconazole. Antifungal susceptibility was tested according to the broth microdilution method described in the NCCLS (M27A-2 method. Only one sample from each patient was analyzed (the first isolate. Most of the episodes had been caused by species other than C. albicans (51.6%, including C. parapsilosis (25.8%, C. tropicalis (13.3%, C. glabrata (3.3%, C. krusei (1.7%, and others (7.5%. Dose-dependent susceptibility to itraconazole was observed in 14.2% of strains, and dose-dependent susceptibility to fluconazole was found in 1.6%. Antifungal resistance was not found, probably related to low use of fluconazole. Further epidemiological surveillance is needed.Estudos realizados em diferentes países têm mostrado diferença na epidemiologia das infecções invasivas por Candida spp. No período de agosto de 2002 a agosto de 2003, foi conduzido estudo na Santa Casa Complexo Hospitalar, Porto Alegre, Brasil, para determinar a distribuição das espécies de Candida associadas a candidemia e o perfil de susceptibilidade das mesmas aos antifúngicos anfotericina B, fluconazol e itraconazol. Os testes de susceptibilidade foram realizados de acordo com a metodologia M27-A2 padronizada pelo NCCLS. Foi incluído no estudo o primeiro isolado de hemocultivo de cada paciente. A maioria dos episódios (51,6% ocorreu por espécies outras que C. albicans, incluindo C. parapsilosis (25,8%, C. tropicalis (13,3%, C. glabrata (3,3%, C. krusei (1,7% e outras espécies (7,5%. Não foi encontrada resistência aos antifúngicos testados, possivelmente devido ao baixo consumo de fluconazol na

  13. Antifungal compounds from cyanobacteria.

    Science.gov (United States)

    Shishido, Tânia K; Humisto, Anu; Jokela, Jouni; Liu, Liwei; Wahlsten, Matti; Tamrakar, Anisha; Fewer, David P; Permi, Perttu; Andreote, Ana P D; Fiore, Marli F; Sivonen, Kaarina

    2015-04-13

    Cyanobacteria are photosynthetic prokaryotes found in a range of environments. They are infamous for the production of toxins, as well as bioactive compounds, which exhibit anticancer, antimicrobial and protease inhibition activities. Cyanobacteria produce a broad range of antifungals belonging to structural classes, such as peptides, polyketides and alkaloids. Here, we tested cyanobacteria from a wide variety of environments for antifungal activity. The potent antifungal macrolide scytophycin was detected in Anabaena sp. HAN21/1, Anabaena cf. cylindrica PH133, Nostoc sp. HAN11/1 and Scytonema sp. HAN3/2. To our knowledge, this is the first description of Anabaena strains that produce scytophycins. We detected antifungal glycolipopeptide hassallidin production in Anabaena spp. BIR JV1 and HAN7/1 and in Nostoc spp. 6sf Calc and CENA 219. These strains were isolated from brackish and freshwater samples collected in Brazil, the Czech Republic and Finland. In addition, three cyanobacterial strains, Fischerella sp. CENA 298, Scytonema hofmanni PCC 7110 and Nostoc sp. N107.3, produced unidentified antifungal compounds that warrant further characterization. Interestingly, all of the strains shown to produce antifungal compounds in this study belong to Nostocales or Stigonematales cyanobacterial orders.

  14. Advances in the susceptibility testing of Sporothrix schenckii and treatment of sporotrichosis%孢子丝菌病药敏和治疗进展

    Institute of Scientific and Technical Information of China (English)

    蔡晴; 李珊山; 刘维达

    2010-01-01

    Spomtrichosis is a subcutaneous infection caused by Sporothrix schenckii,a dimorphic fungus.No unified standard is available at present for in vitro susceptibility testing of dimorphie fungi.Ahhoush saturated solution of potassium iodide.itraconazole and terbinafine have been commonly used in the treatment of sporotrichosis for many years,it is still essential to choose appropriate antifungals.In vitro susceptibility test is not only an important means to the estimate of antifungal activity but also a basis for drug selection.%孢子丝菌病是一种深部真菌病,其致病菌申克孢子丝菌为一种双相真菌,双相真菌的体外药敏试验至今仍无统一标准.饱和碘化钾溶液、伊曲康唑、特比萘芬为治疗孢子丝菌病的常用药物,临床中选择合适的抗真菌药物对治疗此病具有重要意义.体外药敏试验是评价抗真菌药物活性的重要方法,也是选择药物的依据之一.

  15. 外阴阴道念珠菌病的念珠菌种类及抗真菌药物敏感性分析%Species and Antifungal Susceptibility of Candida in Patients with Vulvovaginal Candidiasis

    Institute of Scientific and Technical Information of China (English)

    刘小平; 樊尚荣; 彭燕婷; 张慧萍

    2015-01-01

    Objective To determine species and antifungal drug susceptibility of Candida from patients with vulvovaginal candidiasis(VVC). Methods We enrolled 3 141 patients with VVC who received treatment in gynecological clinic of Peking University Shenzhen Hospital from April,2003 to September,2012. Specimens of Candida were collected from enrolled patients,API Candida system was used to identify the species,and Rosco Disk Diffusion method was employed to test drug susceptibility. Results A total of 3 182 strains of two species' candida were extracted from 41 patients. C. albicans was the predominant species(2 705 strains,85. 0%)in VVC,followed by C. glabrata(338 strains),C. parapsilosis(49 strains), C. tropicalis(31 strains),Saccharomyces cerevisiae(23 strains),C. krusei(15 strains),Candida famata(11 strains), Rhodotorula sp. ( 6 strains ), C. lusitaniae ( 2 strains ) and Candida humicola ( 2 strains ) . The drug resistance rate of C. albicans to fluconazole,itraconazole,miconazole,clotrimazole,and nystatin were 1. 1%(18/1 612),2. 2%(36/1 612), 4. 2%(68/1 612),0. 9%(14/1 612) and 0. Conclusion Candida albicans is the predominant Candida species in patients with vulvovaginal candidiasis. Resistance of vaginal C. albicans to antifungal agents is infrequent.%目的:明确外阴阴道念珠菌病( VVC)患者分离的念珠菌种类及对抗真菌药物的敏感性。方法选取2003年4月—2012年9月就诊于北京大学深圳医院妇科门诊的3141例VVC患者为研究对象。采集念珠菌标本进行培养,采用API Candida鉴定分离菌株,采用Rosco纸片扩散法进行抗真菌药敏试验。结果共分离出3182株念珠菌(从41例患者中分离出两种念珠菌)。其中白念珠菌分离最多,占85.0%(2705株),其次依次为光滑念珠菌(338株)、近平滑念珠菌(49株)、热带念珠菌(31株)、酿酒酵母菌(23株)、克柔念珠菌(15株)、无名念珠菌(11株)、红酵母菌(6

  16. Separation, determination and antifungal activity test of the products from a new Bacillus amyloliquefaciens.

    Science.gov (United States)

    Wang, Tao; Wu, Mian-Bin; Chen, Zheng-Jie; Lin, Jian-Pin; Yang, Li-Rong

    2016-01-01

    A new Bacillus amyloliquefaciens named ZJU-2011 was discovered, and the culture supernatant showed a strong inhibitory effect against Candida albicans. In this study, a novel method was developed to purify the antifungal compounds in high purity. The obtained products were analysed by high performance liquid chromatography and proven to be of high purity. Mass spectrometry showed that the molecular weights of the two bioactive components were 270 and 288, respectively, and their structures were determined to be bacilysin and chlorotetaine by using (1)H and (13)C nuclear magnetic resonance spectroscopy. To the best of our knowledge, this is the first time that B. amyloliquefaciens has been reported to produce bacilysin and chlorotetaine simultaneously. The minimum inhibitory concentration of chlorotetaine against six common fungal pathogens were determined to be in the range of 1.8-7.8 μg/mL.

  17. Molecular epidemiology and in-vitro antifungal susceptibility of Aspergillus terreus species complex isolates in Delhi, India: evidence of genetic diversity by amplified fragment length polymorphism and microsatellite typing.

    Directory of Open Access Journals (Sweden)

    Shallu Kathuria

    Full Text Available Aspergillus terreus is emerging as an etiologic agent of invasive aspergillosis in immunocompromised individuals in several medical centers in the world. Infections due to A. terreus are of concern due to its resistance to amphotericin B, in vivo and in vitro, resulting in poor response to antifungal therapy and high mortality. Herein we examined a large collection of molecularly characterized, geographically diverse A. terreus isolates (n = 140 from clinical and environmental sources in India for the occurrence of cryptic A. terreus species. The population structure of the Indian A. terreus isolates and their association with those outside India was determined using microsatellite based typing (STR technique and Amplified Fragment Length Polymorphism analysis (AFLP. Additionally, in vitro antifungal susceptibility of A. terreus isolates was determined against 7 antifungals. Sequence analyses of the calmodulin locus identified the recently described cryptic species A. hortai, comprising 1.4% of Aspergillus section Terrei isolates cultured from cases of aspergilloma and probable invasive aspergillosis not reported previously. All the nine markers used for STR typing of A. terreus species complex proved to be highly polymorphic. The presence of high genetic diversity revealing 75 distinct genotypes among 101 Indian A. terreus isolates was similar to the marked heterogeneity noticed in the 47 global A. terreus population exhibiting 38 unique genotypes mainly among isolates from North America and Europe. Also, AFLP analysis showed distinct banding patterns for genotypically diverse A. terreus isolates. Furthermore, no correlation between a particular genotype and amphotericin B susceptibility was observed. Overall, 8% of the A. terreus isolates exhibited low MICs of amphotericin B. All the echinocandins and azoles (voriconazole, posaconazole and isavuconazole demonstrated high potency against all the isolates. The study emphasizes the need of

  18. Frost Susceptibility of Soil, Review of Index Tests,

    Science.gov (United States)

    1981-12-01

    Schaible (1957) 36 36. Frost susceptibility criteria according to Floss 37 37 Ruhr University at Bo(hum frost susceptibility criteria according to I...freezing front causes ening can take place. alternating bands of soil and ice to form. The ex- It is important, then, that both kinds of frost ternal...particles and the uniformity early research concentrated on a narrow band of of the gradation of soil particle sizes less than heat extraction rates

  19. Comparison of the Broth Microdilution (BMD) Method of the European Committee on Antimicrobial Susceptibility Testing with the 24-Hour CLSI BMD Method for Testing Susceptibility of Candida Species to Fluconazole, Posaconazole, and Voriconazole by Use of Epidemiological Cutoff Values▿

    Science.gov (United States)

    Pfaller, M. A.; Espinel-Ingroff, A.; Boyken, L.; Hollis, R. J.; Kroeger, J.; Messer, S. A.; Tendolkar, S.; Diekema, D. J.

    2011-01-01

    The antifungal broth microdilution (BMD) method of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) was compared with CLSI BMD method M27-A3 for fluconazole, posaconazole, and voriconazole susceptibility testing of 1,056 isolates of Candida. The isolates were obtained in 2009 from more than 60 centers worldwide and included 560 isolates of C. albicans, 175 of C. glabrata, 162 of C. parapsilosis, 124 of C. tropicalis, and 35 of C. krusei. The overall essential agreement (EA) between EUCAST and CLSI results ranged from 96.9% (voriconazole) to 98.6% (fluconazole). The categorical agreement (CA) between methods and species of Candida was assessed using previously determined epidemiological cutoff values (ECVs). The ECVs (expressed as μg/ml) for fluconazole, posaconazole, and voriconazole, respectively, were as follows: 0.12, 0.06, and 0.03 for C. albicans; 32, 2, and 0.5 for C. glabrata; 2, 0.25, and 0.12 for C. parapsilosis; 2, 0.12, and 0.06 for C. tropicalis; 64, 0.5, and 0.5 for C. krusei. Excellent CA was observed for all comparisons between the EUCAST and CLSI results for fluconazole, posaconazole, and voriconazole, respectively, for each species: 98.9%, 93.6%, and 98.6% for C. albicans; 96.0%, 98.9%, and 93.7% for C. glabrata; 90.8%, 98.1%, and 98.1% for C. parapsilosis; 99.2%, 99.2%, and 96.8% for C. tropicalis; 97.1%, 97.1%, and 97.1% for C. krusei. We demonstrate high levels of EA and CA between the CLSI and EUCAST BMD methods for testing of triazoles against Candida when the MICs were determined after 24 h and ECVs were used to differentiate wild-type (WT) from non-WT strains. These results provide additional data in favor of the harmonization of these two methods. PMID:21227994

  20. Candida antifungal drug resistance in sub-Saharan African populations: A systematic review

    Science.gov (United States)

    Africa, Charlene Wilma Joyce; Abrantes, Pedro Miguel dos Santos

    2017-01-01

    Background: Candida infections are responsible for increased morbidity and mortality rates in at-risk patients, especially in developing countries where there is limited access to antifungal drugs and a high burden of HIV co-infection.  Objectives: This study aimed to identify antifungal drug resistance patterns within the subcontinent of Africa.  Methods: A literature search was conducted on published studies that employed antifungal susceptibility testing on clinical Candida isolates from sub-Saharan African countries using Pubmed and Google Scholar.  Results: A total of 21 studies from 8 countries constituted this review. Only studies conducted in sub-Saharan Africa and employing antifungal drug susceptibility testing were included. Regional differences in Candida species prevalence and resistance patterns were identified.  Discussion: The outcomes of this review highlight the need for a revision of antifungal therapy guidelines in regions most affected by Candida drug resistance.  Better controls in antimicrobial drug distribution and the implementation of regional antimicrobial susceptibility surveillance programmes are required in order to reduce the high Candida drug resistance levels seen to be emerging in sub-Saharan Africa. PMID:28154753

  1. Testing anti-fungal activity of a soil-like substrate for growing plants in bioregenerative life support systems

    Science.gov (United States)

    Nesterenko, E. V.; Kozlov, V. A.; Khizhnyak, S. V.; Manukovsky, N. S.; Kovalev, V. S.; Gurevich, Yu. L.; Liu, Hong; Xing, Yidong; Hu, Enzhu

    2009-10-01

    The object of this research is to study a soil-like substrate (SLS) to grow plants in a Bioregenerative Life Support System (BLSS). Wheat and rice straw were used as raw materials to prepare SLS. Anti-fungal activity of SLS using test cultures of Bipolaris sorokiniana, a plant-pathogenic fungus which causes wheat root rot was studied. Experiments were conducted with SLS samples, using natural soil and sand as controls. Infecting the substrates, was performed at two levels: the first level was done with wheat seeds carrying B. sorokiniana and the second level with seeds and additional conidia of B. sorokiniana from an outside source. We measured wheat disease incidence and severity in two crop plantings. Lowest disease incidence values were obtained from the second planting, SLS: 26% and 41% at the first and the second infection levels, respectively. For soil the values were 60% and 82%, respectively, and for sand they were 67% and 74%, respectively. Wheat root rot in the second crop planting on SLS, at both infection levels was considerably less severe (9% and 13%, respectively) than on natural soil (20% and 33%) and sand (22% and 32%). SLS significantly suppressed the germination of B. sorokiniana conidia. Conidia germination was 5% in aqueous SLS suspension, and 18% in clean water. No significant differences were found regarding the impact on conidia germination between the SLS samples obtained from wheat and rice straw. The anti-fungal activity in SLS increased because of the presence of worms. SLS also contained bacteria stimulating and inhibiting B. sorokiniana growth.

  2. Selection of media for antimicrobial susceptibility testing of fish pathogenic bacteria

    DEFF Research Database (Denmark)

    Dalsgaard, Inger

    2001-01-01

    The available data concerning antimicrobial susceptibility testing of fish pathogens showed that there is no consensus to the basal medium currently being employed. Different media recommended for susceptibility testing of human pathogens (Mueller-Hinton Agar, Tryptone Soya Agar, Antibiotic Medium...

  3. Isavuconazole, micafungin, and 8 comparator antifungal agents' susceptibility profiles for common and uncommon opportunistic fungi collected in 2013: temporal analysis of antifungal drug resistance using CLSI species-specific clinical breakpoints and proposed epidemiological cutoff values.

    Science.gov (United States)

    Pfaller, M A; Rhomberg, P R; Messer, S A; Jones, R N; Castanheira, M

    2015-08-01

    The in vitro activities of isavuconazole, micafungin, and 8 comparator antifungal agents were determined for 1613 clinical isolates of fungi (1320 isolates of Candida spp., 155 of Aspergillus spp., 103 of non-Candida yeasts, and 35 non-Aspergillus molds) collected during a global survey conducted in 2013. The vast majority of the isolates of the 21 different species of Candida, with the exception of Candida glabrata (MIC90, 2 μg/mL), Candida krusei (MIC90, 1 μg/mL), and Candida guilliermondii (MIC90, 8 μg/mL), were inhibited by ≤0.25 μg/mL of isavuconazole. C. glabrata and C. krusei were largely inhibited by ≤1 μg/mL of isavuconazole. Resistance to fluconazole was seen in 0.5% of Candida albicans isolates, 11.1% of C. glabrata isolates, 2.5% of Candida parapsilosis isolates, 4.5% of Candida tropicalis isolates, and 20.0% of C. guilliermondii isolates. Resistance to the echinocandins was restricted to C. glabrata (1.3-2.1%) and C. tropicalis (0.9-1.8%). All agents except for the echinocandins were active against 69 Cryptococcus neoformans isolates, and the triazoles, including isavuconazole, were active against the other yeasts. Both the mold active triazoles as well as the echinocandins were active against 155 Aspergillus spp. isolates belonging to 10 species/species complex. In general, there was low resistance levels to the available systemically active antifungal agents in a large, contemporary (2013), global collection of molecularly characterized yeasts and molds. Resistance to azoles and echinocandins was most prominent among isolates of C. glabrata, C. tropicalis, and C. guilliermondii.

  4. Augmenting the Antifungal Activity of an Oxidizing Agent with Kojic Acid: Control of Penicillium Strains Infecting Crops

    Directory of Open Access Journals (Sweden)

    Jong H. Kim

    2014-11-01

    Full Text Available Oxidative treatment is one of the strategies for preventing Penicillium contamination in crops/foods. The antifungal efficacy of hydrogen peroxide (H2O2; oxidant was investigated in Penicillium strains by using kojic acid (KA as a chemosensitizing agent, which can enhance the susceptibility of pathogens to antifungal agents. Co-application of KA with H2O2 (chemosensitization resulted in the enhancement of antifungal activity of either compound, when compared to the independent application of each agent alone. Of note, heat enhanced the activity of H2O2 to a greater extent during chemosensitization, whereby the minimum inhibitory or minimum fungicidal concentrations of H2O2 was decreased up to 4 or 13 fold, respectively, at 35–45 °C (heat, when compared to that at 28 °C (normal growth temperature. However, heat didn’t increase the antifungal activity of KA, indicating specificity exists between heat and types of antifungals applied. The effect of chemosensitization was also strain-specific, where P. expansum (both parental and fludioxonil-resistant mutants or P. italicum 983 exhibited relatively higher susceptibility to the chemosensitization, comparing to other Penicillium strains tested. Collectively, chemosensitization can serve as a potent antifungal strategy to lower effective dosages of toxic antifungal substances, such as H2O2. This can lead to coincidental lowering of environmental and health risks.

  5. Comparing the disk-diffusion and agar dilution tests for Neisseria gonorrhoeae antimicrobial susceptibility testing

    Directory of Open Access Journals (Sweden)

    Hsi Liu

    2016-11-01

    Full Text Available Abstract Background We assessed the validity of testing for antimicrobial susceptibility of clinical and mutant Neisseria gonorrhoeae (GC isolates by disk diffusion in comparison to agar dilution, and Etest® (bioMerieux, France, respectively, for three third generation extended spectrum cephalosporins (ESC: ceftriaxone (CRO, cefixime (CFX, and cefpodoxime (CPD. Methods One hundred and five clinical isolates and ten laboratory-mutants were tested following Clinical Laboratory Standard Institute (CLSI and manufacturer’s standards for each of the three methods. The measured diameters by the disk diffusion method were tested for correlation with the MIC values by agar dilution. In addition, comparisons with the Etest® were made. Categorical results for concordance, based on standard CLSI cutoffs, between the disk diffusion and the other two methods, respectively, were tested using the Chi-square statistics. Reproducibility was tested for CFX across a 6-month interval by repeated disk tests. Results Across all 115 specimens, the disk diffusion tests produced good categorical agreements, exhibiting concordance of 93.1%, 92.1%, and 90.4% with agar dilution and 93.0%, 92.1%, and 90.4% with Etest®, for CRO, CFX, and CPD, respectively. Pearson correlations between disk-diffusion diameters and agar dilution MIC’s were -0.59, -0.67, and -0.81 for CRO, CFX, and CPD, respectively. The correlations between disk diffusion and Etest® were -0.58, -0.73, and -0.49. Pearson correlation between the CFX disk readings over a 6-month interval was 91%. Conclusions Disk diffusion tests remain to be a useful, reliable and fast screening method for qualitative antimicrobial susceptibility testing for ceftriaxone, cefixime, and cefpodoxime.

  6. Routine disc diffusion antimicrobial susceptibility testing of Clostridium difficile and association with PCR ribotype 027

    DEFF Research Database (Denmark)

    Holt, H M; Danielsen, T K; Justesen, U S

    2015-01-01

    Reduced susceptibility to metronidazole and vancomycin in Clostridium difficile has been reported, which emphasises the need for simple antimicrobial susceptibility testing methods. The aim of this study was to apply a published disc diffusion method and zone diameter breakpoint correlates...... the published breakpoint (difficile PCR ribotype 027 isolates had smaller zone...... diameters than non-027 isolates. The disc diffusion method is very simple and inexpensive, and the published zone diameter breakpoints will detect C. difficile isolates with reduced susceptibility to metronidazole and vancomycin....

  7. Interlaboratory Study of Quality Control Isolates for a Broth Microdilution Method (Modified CLSI M38-A) for Testing Susceptibilities of Dermatophytes to Antifungals▿

    Science.gov (United States)

    Ghannoum, M. A.; Arthington-Skaggs, B.; Chaturvedi, V.; Espinel-Ingroff, A.; Pfaller, M. A.; Rennie, R.; Rinaldi, M. G.; Walsh, T. J.

    2006-01-01

    The Clinical and Laboratory Standards Institute (CLSI; formerly National Committee for Clinical Laboratory Standards, or NCCLS) M38-A standard for the susceptibility testing of filamentous fungi does not specifically address the testing of dermatophytes. In 2003, a multicenter study investigated the reproducibility of the microdilution method developed at the Center for Medical Mycology, Cleveland, Ohio, for testing the susceptibility of dermatophytes. Data from that study supported the introduction of this method for testing dermatophytes in the future version of the CLSI M38-A standard. In order for the method to be accepted by CLSI, appropriate quality control isolates needed to be identified. To that end, an interlaboratory study, involving the original six laboratories plus two additional sites, was conducted to evaluate potential candidates for quality control isolates. These candidate strains included five Trichophyton rubrum strains known to have elevated MICs to terbinafine and five Trichophyton mentagrophytes strains. Antifungal agents tested included ciclopirox, fluconazole, griseofulvin, itraconazole, posaconazole, terbinafine, and voriconazole. Based on the data generated, two quality control isolates, one T. rubrum isolate and one T. mentagrophytes isolate, were identified and submitted to the American Type Culture Collection (ATCC) for inclusion as reference strains. Ranges encompassing 95.2 to 97.9% of all data points for all seven drugs were established. PMID:17050812

  8. Chalcone derivatives as potential antifungal agents: Synthesis, and antifungal activity

    Directory of Open Access Journals (Sweden)

    Deepa Gupta

    2015-01-01

    Full Text Available Much research has been carried out with the aim to discover the therapeutic values of chalcone derivatives. Chalcones possess wide range of pharmacological activity such as antibacterial, antimalarial, antiprotozoal, antitubercular, anticancer, and antifungal agents etc. The presence of reactive α,β-unsaturated keto group in chalcones is found to be responsible for their biological activity. The rapid developments of resistance to antifungal agents, led to design, and synthesize the new antifungal agents. The derivatives of chalcones were prepared using Claisen-Schmidt condensation scheme with appropriate tetralone and aldehyde derivatives. Ten derivatives were synthesized and were biologically screened for antifungal activity. The newly synthesized derivatives of chalcone showed antifungal activity against fungal species, Microsporum gypseum. The results so obtained were superior or comparable to ketoconazole. It was observed that none of the compounds tested showed positive results for fungi Candida albicans nor against fungi Aspergillus niger. Chalcone derivatives showed inhibitory effect against M. gypseum species of fungus. It was found that among the chalcone derivatives so synthesized, two of them, that is, 4-chloro derivative, and unsubstituted derivative of chalcone showed antifungal activity superior to ketoconazole. Thus, these can be the potential new molecule as antifungal agent.

  9. A Novel Hypoxia Challenge Test Demonstrates Cardiovascular and Pulmonary Susceptibility to Acrolein Gas in Hypertensive Rats.

    Science.gov (United States)

    High levels of air pollution increase the risk of cardiovascular morbidity and mortality, especially in susceptible populations including those with hypertension. Stress tests are useful for manifesting latent effects of exposure, particularly at low concentrations, often when no...

  10. Interlaboratory comparison of results of susceptibility testing with caspofungin against Candida and Aspergillus species.

    NARCIS (Netherlands)

    Odds, F.C.; Motyl, M.; Andrade, R.; Bille, J.; Canton, E.; Cuenca-Estrella, M.; Davidson, A.; Durussel, C.; Ellis, D.; Foraker, E.; Fothergill, A.; Ghannoum, M.A.; Giacobbe, R.A.; Gobernado, M.; Handke, R.; Laverdiere, M.; Lee-Yang, W.; Merz, W.G.; Ostrosky-Zeichner, L.; Peman, J.; Perea, S.; Perfect, J.R.; Pfaller, M.A.; Proia, L.; Rex, J.H.; Rinaldi, M.; Rodriguez-Tudela, J.L.; Schell, W.A.; Shields, C.; Sutton, D.A.; Verweij, P.E.; Warnock, D.W.

    2004-01-01

    Seventeen laboratories participated in a study of interlaboratory reproducibility with caspofungin microdilution susceptibility testing against panels comprising 30 isolates of Candida spp. and 20 isolates of Aspergillus spp. The laboratories used materials supplied from a single source to determine

  11. Family system characteristics and psychological adjustment to cancer susceptibility genetic testing : a prospective study

    NARCIS (Netherlands)

    van Oostrom, I.; Meijers-Heijboer, H.; Duivenvoorden, H. J.; Brocker-Vriends, A. H. J. T.; van Asperen, C. J.; Sijmons, R. H.; Seynaeve, C.; Van Gool, A. R.; Klijn, J. G. M.; Tibben, A.

    2007-01-01

    This study examined prospectively the contribution of family functioning, differentiation to parents, family communication and support from relatives to psychological distress in individuals undergoing genetic susceptibility testing for a known familial pathogenic BRCA1/2 or Hereditary nonpolyposis

  12. Oral Isolates of Saccharomyces in Patients with Oral Fungal Infection and Their Susceptibility to Antifungal%口腔念珠菌感染患者口内菌株检出情况和药敏性观察

    Institute of Scientific and Technical Information of China (English)

    周小武; 姜巍巍; 毛利艳; 梅丽琴

    2012-01-01

    目的 探讨口腔念珠菌病患者口内念珠菌珠菌种的分布和体外药物敏感性.方法 收集患者标本采用念珠菌显色培养基对其进行分离鉴定,采用NCCLS M27-A2微量稀释法测定念珠菌株对特比萘芬、氟康唑、伊曲康唑、制霉菌素四种抗真菌药物以及联合用药的体外药敏情况.结果 试验组和对照组口腔念珠菌检出率分别为74.65%%和35.42%,其中白色念珠菌分别占62.26%和58.82%.特比萘芬、氟康唑、伊曲康唑、制霉菌素对口腔念珠菌的体外试验敏感性总有效率分别为57.89%、86.84%、84.21%和100.00%.特比萘芬与氟康唑联用时,念珠菌对药物敏感性增高.结论 口腔念珠菌病仍以白色念珠菌为主,治疗时应进行真菌常规菌种鉴定及药敏试验.对耐药菌株的治疗,联合用药也是重要方法.%Objective To explore the distribution of strains and the invitro antifungal susceptibilities of Candidaspe-cies. Methods The researchers collected the specimens of patients and had Candida isolated and identified by using Candida color culture medium. The invitro antifungal susceptibilities to Terbinafine, Fluconazole, Nystatine and Itraconazole as well as the drug combination of them were detected with checkerboard micro dilution method based on M27 - A2 ( 2002 ) of NCCLS. Result The incidence of Saccharomyces was 74. 65%% and 35. 42% in experimental group and control group respectively. The proportion of Saccharomyces albicans was 62. 26% ( experimental group ) and 58. 82% ( control group ). The overall effective rate of the invitro antifungal susceptibilities of Candida albicans isolates to Terbinafine, Fluconazole, Nystatine and Itraconazole were 57. 89% , 86. 84% , 84. 21% and 100.00% respectively. When Terbinafine being combinated with Fluconazole, susceptibilities of the isolates were enhanced accordingly. Conclusion Saccharomyces albicans is the main infection strain. It is necessary to make strain identification and

  13. Development of a rapid ATP bioluminescence assay for biocidal susceptibility testing of rapidly growing mycobacteria.

    Science.gov (United States)

    Kapoor, Renuka; Yadav, Jagjit S

    2010-10-01

    An ATP-based biocide susceptibility assay for mycobacteria was developed by optimizing the cell lysis and assay conditions. Compared to the conventional agar plating method, the assay was rapid (1.5 h) and showed high sensitivity and specificity as determined by receiver operating characteristic (ROC) analysis. The test species, Mycobacterium immunogenum, M. chelonae, and M. abscessus, showed various susceptibilities to the glutaraldehyde- and isothiazolone-based test biocides.

  14. Susceptibility of Dermatophytes to Thiabendazole Using CLSI Broth Macrodilution

    Science.gov (United States)

    Robledo-Leal, Efrén; Elizondo-Zertuche, Mariana; González, Gloria M.

    2012-01-01

    Objective. To evaluate in vitro antifungal activity of thiabendazole against strains of dermatophytes using a reference method for filamentous fungi. Materials and Methods. Dermatophytes' susceptibility to thiabendazole (TBZ) and fluconazole (FCZ) was evaluated using macrodilution method of protocol M38-A2 of the Clinical and Laboratory Standards Institute (CLSI). Results. MIC ranges of TBZ for all strains were narrower and/or smaller than those of FCZ. TBZ showed a significantly greater potency than FCZ (P = 0.05) against all isolates. Discussion. Although there have been approaches to evaluate the antifungal activity of TBZ in human mycoses, no tests had been made with a standardized protocol. Susceptibility data resulted from this study shows that although TBZ is not a particularly strong inhibitor of dermatophytes, it displays a stable and constant effect against all isolates tested. Conclusion. Results show that TBZ is more effective against strains of dermatophytes than FCZ. We acknowledge the antifungal effect of TBZ against dermatophyte isolates. PMID:23008781

  15. 93例泌尿系真菌感染菌群分布及药物敏感试验结果%An analysis of species distribution and antifungal susceptibility of 93 strains of infecting urinary system epiphytes

    Institute of Scientific and Technical Information of China (English)

    夏玲玲; 周仲松; 刘贵育; 李家斌; 余鑫之

    2009-01-01

    Objective To study and analyze the detected rate, distribution of fungal species and antifungal susceptibility of 93 strains of infecting urinary system epiphytes in the last three years to provide foundation for the reasonable clinical use of anti-epiphyte medicines. Methods The epiphytes isolated from epiphyte culture of midstream urine were tested and their drug sensitivity was tested with ATB Fungus 3 diffusion method. Results Of 93 strains isolated in the experiment, 49 were Candida albicans, accounted for 52.7%; 27 were C. glabrata, ac-counted for 29.0%; and 13 were C.tropiclis, accounted for 13.9%. And their drug resistances to fluconazole were 16.7%, 3.8% and 23.1%, re-spectively, itraconazole were 21.1%, 39.1% and 33.3%, respectively, flucytosine were 37.8%, 20.0% and 15.4%, respectively. C.tropiclis resis-tances to amphotericin B were 23.1%, other tested epiphytes were sensitive to amphotericin B. Conclusion The majority of tested epiphytes are C. albicans, C. glabrata and C.tropiclis in our hospital epiphytic infections of urine system, which are comparatively sensitive to zole medicines. Some C.tropiclis is resistances to amphotericin B.%目的 了解近3年泌尿系统感染真菌的检出率、菌种分布及药物敏感试验结果,为临床合理使用抗真菌药物提供依据.方法 中段尿真菌培养分离的菌株鉴定,采用ATB Fungus 3纸片扩散法测定药敏性.结果 共分离到真菌93株,其中白色念珠菌49株(52.7%)、光滑念珠菌27株(29.0%)、热带念珠菌13株(13.9%),它们对氟康唑的耐药率分别为16.7%、3.8%及23.1%;对伊曲康唑耐药率分别为21.1%、39.1%及33.3%;对氟胞嘧啶耐药率分别为37.8%、20.0%及15.4%;热带念珠菌对两性霉素B耐药率为23.1%,其他真菌对两性霉素B敏感.结论 泌尿系感染真菌以白色念珠菌、光滑念珠菌及热带念珠菌为主,对三唑类药物敏感性较好,部分热带念珠菌对两性霉素B耐药.

  16. Rapid identification and antibiotic susceptibility testing of Yersinia pestis using bioluminescent reporter phage.

    Science.gov (United States)

    Schofield, David A; Molineux, Ian J; Westwater, Caroline

    2012-08-01

    The rapid identification and antibiotic susceptibility testing of Yersinia pestis is paramount for a positive prognosis. We previously engineered a Y. pestis-specific 'bioluminescent' reporter phage for the identification of Y. pestis. In this study, we generated an improved reporter phage and evaluated the ability of this phage to provide direct and rapid susceptibility testing. Compared to the first generation reporter, the second generation reporter exhibited a 100-fold increase in signal strength, leading to a 10-fold increase in assay sensitivity. Y. pestis antimicrobial testing in the presence of the reporter elicited bioluminescent signals that were drug concentration-dependent, and produced susceptibility profiles that mirrored the standard CLSI method. The phage-generated susceptibility profiles, however, were obtained within hours in contrast to days with the conventional method.

  17. Rapid identification and antibiotic susceptibility testing of Yersinia pestis using bioluminescent reporter phage

    Science.gov (United States)

    Schofield, David A.; Molineux, Ian J.; Westwater, Caroline

    2012-01-01

    The rapid identification and antibiotic susceptibility testing of Yersinia pestis is paramount for a positive prognosis. We previously engineered a Y. pestis-specific ‘bioluminescent’ reporter phage for the identification of Y. pestis. In this study, we generated an improved reporter phage and evaluated the ability of this phage to provide direct and rapid susceptibility testing. Compared to the first generation reporter, the second generation reporter exhibited a 100-fold increase in signal strength, leading to a 10-fold increase in assay sensitivity. Y. pestis antimicrobial testing in the presence of the reporter elicited bioluminescent signals that were drug concentration-dependent, and produced susceptibility profiles that mirrored the standard CLSI method. The phage-generated susceptibility profiles, however, were obtained within hours in contrast to days with the conventional method. PMID:22579583

  18. Susceptibility to heat and antifungal agents of Cryptococcus neoformans var. neoformans (serotype D isolated from Eucalyptus spp in Rio Grande do Sul, Brazil Susceptibilidade de Cryptococcus neoformans var. neoformans (sorotipo D isolados de Eucalyptus spp., no Rio Grande do Sul (Brasil, frente ao calor e a agentes antifúngicos

    Directory of Open Access Journals (Sweden)

    Jorge A Horta

    2005-03-01

    Full Text Available In this work we studied the susceptibility to heat and antifungal agents of the first strains of environmental Cryptococcus neoformans var neoformans (serotype D isolated in the state of Rio Grande do Sul, Brazil. In order to achieve a rigorous analysis, we employed the methodology recommended by NCCLS, Yeast Nitrogen Base (YNB proposed by Ghannoum et al (YNB-1, Antibiotic medium 3 (AM3 indicated by others, YNB adjusted to the NCCLS methodology (YNB-2 and Etest. Our results indicate that all strains were susceptible to amphotericin B (0.0625-0.5 µg/mL, fluconazole (0.125-8.0 µg/mL, itraconazole (0.031-0.25 µg/mL and flucytosine (0.125-4.0 µg/mL. The C. neoformans serotype D strains were more susceptible to heat (47ºC/30 min than C. neoformans serotype A.Este estudo foi realizado com os primeiros isolados ambientais de C. neoformans sorotipo D, obtidos no Rio Grande do Sul. Objetivando-se avaliar a susceptibilidade a agentes antifúngicos de forma mais rigorosa, utilizou-se a técnica de referência proposta pelo NCCLS, Caldo Yeast Nitrogen Base (YNB proposto por Ghannoum et al., Antibiotic medium 3, caldo YNB adequado à metodologia do NCCLS e o E-test. Os resultados indicaram que todos os isolados foram sensíveis à anfotericina B (0,0625-0,5 µg/mL, fluconazol (0,125-4,0 µg/mL, itraconazol (0,031-0,25 µg/ml e fluorocitosina (0,125-4,0 µg/mL através das técnicas empregadas. Nos testes de termotolerância (47ºC/30 min, observou-se que as culturas de C. neoformans sorotipo D são mais sensíveis do que as de C. neoformans sorotipo A.

  19. Bacterial Cytological Profiling (BCP as a Rapid and Accurate Antimicrobial Susceptibility Testing Method for Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    D.T. Quach

    2016-02-01

    Full Text Available Successful treatment of bacterial infections requires the timely administration of appropriate antimicrobial therapy. The failure to initiate the correct therapy in a timely fashion results in poor clinical outcomes, longer hospital stays, and higher medical costs. Current approaches to antibiotic susceptibility testing of cultured pathogens have key limitations ranging from long run times to dependence on prior knowledge of genetic mechanisms of resistance. We have developed a rapid antimicrobial susceptibility assay for Staphylococcus aureus based on bacterial cytological profiling (BCP, which uses quantitative fluorescence microscopy to measure antibiotic induced changes in cellular architecture. BCP discriminated between methicillin-susceptible (MSSA and -resistant (MRSA clinical isolates of S. aureus (n = 71 within 1–2 h with 100% accuracy. Similarly, BCP correctly distinguished daptomycin susceptible (DS from daptomycin non-susceptible (DNS S. aureus strains (n = 20 within 30 min. Among MRSA isolates, BCP further identified two classes of strains that differ in their susceptibility to specific combinations of beta-lactam antibiotics. BCP provides a rapid and flexible alternative to gene-based susceptibility testing methods for S. aureus, and should be readily adaptable to different antibiotics and bacterial species as new mechanisms of resistance or multidrug-resistant pathogens evolve and appear in mainstream clinical practice.

  20. Effect of physiological heterogeneity of E. coli population on antibiotic susceptivity test

    Institute of Scientific and Technical Information of China (English)

    LIU; YuQing; ZHANG; HuaiQiang; SHEN; JianZhong; GAO; PeiJi

    2007-01-01

    According to the instantaneous growth rate (dN/dt) of E. coli CVCC249 growing in batch culture, the entire growth progress was distinguished into four phases: accelerating growth phase, constant growth phase, decelerating growth phase and declining phase, in each of which obvious variation in physiological and biochemical properties was detected, including total DNA, total protein, and MTT-dehydrogenase activity, etc., that led to difference in their antibiotic susceptivity. Antibiotic susceptivity of the population sampled from each phase was tested by Concentration-killing Curve (CKC) approach following the formula N=N0/{1+exp[r((x-BC50)]}, showing as normal distribution at the individual cell level for an internal population, in which the median bactericidal concentration BC50 represents the mean level of susceptivity, while the bactericidal span BC1-99=(2lnN0)/r indicates the variation degree of the antibiotic susceptivity. Furthermore, tested by CKC approach, the antibiotic susceptivity of E. coli CVCC249 population in each physiological phase to gentamicin or enoxacin was various: susceptivity of the population in the constant growth phase and declining phase all increased compared with that in the accelerating growth phase for gentamicin but declined for enoxacin. The primary investigations revealed that the physiological phase should be taken into account in the context of antibiotic susceptivity and research into antimicrobial mechanism. However there are few reports concerned with this study. Further research using different kinds of antibiotics with synchronized continuous culture of different bacterial strains is required.

  1. Direct Susceptibility testing of Urine samples after 6, 9 and 16 hours incubation

    DEFF Research Database (Denmark)

    Ghatian, Khaled; Littauer, Pia; Llado, Minna Fyhn Lykke

    2015-01-01

    Objectives: At present, susceptibility testing of infective bacteria from urine samples, according to EUCAST, is interpretation of zones after 16 hours of incubation. Using a semi-automated sample processing, the Kiestra (BD) system, to incubate of the agar plates and digital image processing...... testing on urine samples. Methods: In total 675 consecutive urine samples from out-patients, were inoculated directly, according to our laboratory standard, as 10µL on each part of a bi-plate (oxoid chomogene agar and 5% horse blood-agar) and 10µL on a MH plate for resistance testing, using an Inoqula...... (Kiestra-BD). The plates were photographed at 6, 9 and 16 hours after incubation. As a gold standard, the susceptibility test was performed according to EUCAST, as a secondary susceptibility testing, for colonies identified on the biplates, and read after 16 hours incubation.The disks used (Oxoid, Termo...

  2. 抗真菌药物治疗外阴阴道念珠菌病敏感性的临床研究%Clinical Research of Susceptibility to Antifungal Agents in the Treatment of Vulvovaginal Candidiasis

    Institute of Scientific and Technical Information of China (English)

    郭爱芹

    2012-01-01

    目的 探讨外阴阴道念珠菌病(VVC)的临床特征及抗真菌药物治疗VVC的敏感性.方法 对我院自2008年1月至2011年8月收治的1000例妇科门诊患者的VVC发病情况进行回顾性分析,观察念珠菌对抗真菌药物的敏感性.结果 通过分离1000株念珠菌,白念珠菌为610例(61%),非白念珠菌为390例(39%).白念珠菌及非白念珠菌对氟康唑和伊曲康唑的敏感度比较差异有统计学意义(P<0.05);对酮康唑和两性霉素B的敏感度比较,差异无统计学意义(P>0.05).结论 白念珠菌是VVC的主要致病菌,非白念珠菌的药物敏感性较白念珠菌低,白念珠菌对抗真菌药物的耐药性不常见.%Objective To discuss the clinical features of vulvovaginal candidiasis( VVC )and the susceptibility to antifungal agents in the treatment. Methods Retrospective analysis of VVC incidence of 1000 gynecological outpatients of our hospital from January 2008 to August 2011 was done, Candida albicans sensitivity to antifungal agents was observed. Results Among the 1000 cases,610 patients( 61% )had Candida albicans,and 390 patients( 39% )had non-Candida albicans. Sensitivity of Candida albicans and non-Candida albicans to fluconazole and itraconazole werestatistically significantly different^ P 0. 05 ). Conclusion The main pathogen of VCC is Candida albicans, non-Candida albicans drug sensitivity is lower than Candida albicans, antifungal drug resistance is not common in Candida albicans.

  3. Pyrazinamide susceptibility testing: proposed new standard with the BACTECTM MGITTM 960 system.

    Science.gov (United States)

    Piersimoni, C; Mustazzolu, A; Iacobino, A; Giannoni, F; Santoro, G; Gherardi, G; Del Giudice, A; Perna, R; Fattorini, L

    2016-12-01

    The susceptibility of 253 Mycobacterium tuberculosis complex isolates to pyrazinamide (PZA) was assessed using the BACTECTM MGITTM 960 (M960) system. Resistant strains underwent paired repeat testing using 1) a critical concentration of 200 g/ml (PZA-200), and 2) a reduced inoculum of 0.25 ml. They were also examined using the BACTEC 460 (B460) reference method and investigated for pncA mutations. On M960, 37 isolates were resistant. In the PZA-200 assay, 20 of these were resistant and 17 susceptible, while 18 were resistant and 19 susceptible with reduced inoculum. The B460 assay and pncA sequencing confirmed results with reduced inoculum.

  4. Genetic susceptibility testing from a stress and coping perspective.

    Science.gov (United States)

    Gooding, Holly C; Organista, Kurt; Burack, Jeffrey; Biesecker, Barbara Bowles

    2006-04-01

    Four theories of health behavior and of stress and coping are reviewed for their ability to illuminate interest in uptake and outcomes of genetic testing for adult-onset diseases. These theories are the Health Belief Model, the Theory of Planned Behavior (TPB), the Common Sense Model of Self-regulation (CSM), and the Transactional Model of Stress and Coping (TMSC). Basic concepts of each theory are discussed, followed by evidence from the literature supporting the relevance of these concepts to the understanding of genetic testing for four adult-onset diseases: Huntington's disease, Alzheimer's disease, hereditary breast/ovarian cancer, and hereditary colorectal cancer. Emphasis is placed on the finding that a decision to undergo genetic testing may be considered as a way to cope with both the cognitive and affective concerns that arise from living at increased risk of developing a disease in the future. The potential value of genetic testing for reducing uncertainty about and gaining a sense of control over one's risk of developing a chronic disease is highlighted. We argue that theories which focus on stress and coping provide a useful framework for future studies of genetic testing decisions for adult-onset disease risk.

  5. What's New in Genetic Testing for Cancer Susceptibility?

    Science.gov (United States)

    Plichta, Jennifer K; Griffin, Molly; Thakuria, Joseph; Hughes, Kevin S

    2016-09-15

    The advent of next-generation sequencing, and its transition further into the clinic with the US Food and Drug Administration approval of a cystic fibrosis assay in 2013, have increased the speed and reduced the cost of DNA sequencing. Coupled with a historic ruling by the Supreme Court of the United States that human genes are not patentable, these events have caused a seismic shift in genetic testing in clinical medicine. More labs are offering genetic testing services; more multigene panels are available for gene testing; more genes and gene mutations are being identified; and more variants of uncertain significance, which may or may not be clinically actionable, have been found. All these factors, taken together, are increasing the complexity of clinical management. While these developments have led to a greater interest in genetic testing, risk assessment, and large-scale population screening, they also present unique challenges. The dilemma for clinicians is how best to understand and manage this rapidly growing body of information to improve patient care. With millions of genetic variants of potential clinical significance and thousands of genes associated with rare but well-established genetic conditions, the complexities of genetic data management clearly will require improved computerized clinical decision support tools, as opposed to continued reliance on traditional rote, memory-based medicine.

  6. A Simplified Test for Blanching Susceptibility of Copper Alloys

    Science.gov (United States)

    Thomas-Ogbuji, Linus U.; Humphrey, Donald; Setlock, John

    2003-01-01

    GRCop-84 (Cu-8Cr-4Nb) is a dispersion-strengthened alloy developed for space-launch rocket engine applications, as a liner for the combustion chamber and nozzle ramp. Its main advantage over rival alloys, particularly NARloy-Z (Cu-Ag-Zr), the current liner alloy, is in high temperature mechanical properties. Further validation required that the two alloys be compared with respect to service performance and durability. This has been done, under conditions resembling those expected in reusable launch engine applications. GRCop-84 was found to have a superior resistance to static and cyclic oxidation up to approx. 700 C. In order to improve its performance above 700 C, Cu-Cr coatings have also been developed and evaluated. The major oxidative issue with Cu alloys is blanching, a mode of degradation induced by oxidation-reduction fluctuations in hydrogen-fueled engines. That fluctuation cannot be addressed with conventional static or cyclic oxidation testing. Hence, a further evaluation of the alloy substrates and Cu-Cr coating material necessitated our devising a test protocol that involves oxidaton-reduction cycles. This paper describes the test protocols used and the results obtained.

  7. In vitro antibiotic susceptibilities of Burkholderia mallei (causative agent of glanders) determined by broth microdilution and E-test.

    Science.gov (United States)

    Heine, H S; England, M J; Waag, D M; Byrne, W R

    2001-07-01

    In vitro susceptibilities to 28 antibiotics were determined for 11 strains of Burkholderia mallei by the broth microdilution method. The B. mallei strains demonstrated susceptibility to aminoglycosides, macrolides, quinolones, doxycycline, piperacillin, ceftazidime, and imipenem. For comparison and evaluation, 17 antibiotic susceptibilities were also determined by the E-test. E-test values were always lower than the broth dilution values. Establishing and comparing antibiotic susceptibilities of specific B. mallei strains will provide reference information for assessing new antibiotic agents.

  8. Comparison of CLSI broth macrodilution and microdilution methods for echinocandin susceptibility testing of 5 Candida species.

    Science.gov (United States)

    Bopp, Lawrence H; Baltch, Aldona L; Ritz, William J; Smith, Raymond P

    2011-11-01

    In order to compare the Clinical and Laboratory Standards Institute (CLSI) broth macrodilution and microdilution methods of susceptibility testing for echinocandins and yeast, 55 strains of Candida representing 5 species were tested using the CLSI-recommended broth macro- and microdilution methods. Small (1-3 log(2)) but potentially important method-, species-, and drug-dependent differences in MICs were observed.

  9. Comparison and Development of Pyrazinamide Susceptibility Testing Methods for Tuberculosis in Thailand

    Science.gov (United States)

    Foongladda, Suporn; Klayut, Wiphat; Pholwat, Suporn; Houpt, Eric

    2015-01-01

    Pyrazinamide (PZA) plays a critical role in shortening tuberculosis treatment duration and in treating MDR-TB. The standard phenotypic MGIT PZA susceptibility testing method is imperfect because it is slow and has potential for false resistance. In this study we evaluated two different phenotypic based methods, qPCR phage assay and MTT assay, as well as genotypic sequencing. The assay was evaluated on 71 clinical M. tuberculosis isolates (37 MGIT PZA susceptible, 34 MGIT PZA resistant) and compared to the MGIT result. Of these methods the qPCR phage assay yielded an accuracy of 89% versus standard MGIT while MTT yielded 83%. The genotypic sequencing method yielded 90% accuracy. We conclude that any of these faster PZA susceptibility methods perform reasonably well against a MGIT PZA susceptibility standard. PMID:26298819

  10. Rapid antimicrobial susceptibility testing of clinical isolates by digital time-lapse microscopy

    DEFF Research Database (Denmark)

    Fredborg, M; Rosenvinge, F S; Spillum, E;

    2015-01-01

    Rapid antimicrobial susceptibility testing (AST) is essential for early and appropriate therapy. Methods with short detection time enabling same-day treatment optimisation are highly favourable. In this study, we evaluated the potential of a digital time-lapse microscope system, the oCelloScope s......Rapid antimicrobial susceptibility testing (AST) is essential for early and appropriate therapy. Methods with short detection time enabling same-day treatment optimisation are highly favourable. In this study, we evaluated the potential of a digital time-lapse microscope system, the o...... (168 antimicrobial agent-organism combinations) demonstrated 3.6 % minor, no major and 1.2 % very major errors of the oCelloScope system compared to conventional susceptibility testing, as well as a rapid and correct phenotypic detection of strains with methicillin-resistant Staphylococcus aureus (MRSA...

  11. Harmonization of antimicrobial susceptibility testing among veterinary diagnostic laboratories in the five Nordic countries

    DEFF Research Database (Denmark)

    Petersen, A.; Aarestrup, Frank Møller; Hofshagen, Merete

    2003-01-01

    , whereas for Enterococcus spp., two laboratories had less than 90 % concordant results. Susceptibility testing of Salmonella to fluoroquinolones gave rise to almost 0.5% nonconcordant results and susceptibility testing of S. aureus to vancomycin resulted in that 1.8% of the strains were incorrectly......A total of 100 bacterial strains (25 Escherichia coli, 25 Salmonella enterica, 25 Staphylococcus aureus, and 25 Enterococcus strains) and four reference strains were tested for susceptibility toward 8-12 antimicrobial agents in 12 veterinary diagnostic laboratories in the five Nordic countries...... Committee of Clinical Laboratory Standards breakpoints, the percentage of concordant results increased to 98.4% and the performance between laboratories varied between 94.2 and 99.4% concordant results. For E. coli, S., aureus, and Salmonella, all laboratories except one had more than 97% concordant results...

  12. A two-hour antibiotic susceptibility test by ATP-bioluminescence.

    Science.gov (United States)

    March Rosselló, Gabriel Alberto; García-Loygorri Jordán de Urries, María Cristina; Gutiérrez Rodríguez, María Purificación; Simarro Grande, María; Orduña Domingo, Antonio; Bratos Pérez, Miguel Ángel

    2016-01-01

    The antibiotic susceptibility test (AST) in Clinical Microbiology laboratories is still time-consuming, and most procedures take 24h to yield results. In this study, a rapid antimicrobial susceptibility test using ATP-bioluminescence has been developed. The design of method was performed using five ATCC collection strains of known susceptibility. This procedure was then validated against standard commercial methods on 10 strains of enterococci, 10 staphylococci, 10 non-fermenting gram negative bacilli, and 13 Enterobacteriaceae from patients. The agreement obtained in the sensitivity between the ATP-bioluminescence method and commercial methods (E-test, MicroScan and VITEK2) was 100%. In summary, the preliminary results obtained in this work show that the ATP-bioluminescence method could provide a fast and reliable AST in two hours.

  13. Comparison of agar dilution and antibiotic gradient strip test with broth microdilution for susceptibility testing of swine Brachyspira species.

    Science.gov (United States)

    Mirajkar, Nandita S; Gebhart, Connie J

    2016-03-01

    Production-limiting diseases in swine caused by Brachyspira are characterized by mucohemorrhagic diarrhea (B. hyodysenteriae and "B. hampsonii") or mild colitis (B. pilosicoli), while B. murdochii is often isolated from healthy pigs. Emergence of novel pathogenic Brachyspira species and strains with reduced susceptibility to commonly used antimicrobials has reinforced the need for standardized susceptibility testing. Two methods are currently used for Brachyspira susceptibility testing: agar dilution (AD) and broth microdilution (BMD). However, these tests have primarily been used for B. hyodysenteriae and rarely for B. pilosicoli. Information on the use of commercial susceptibility testing products such as antibiotic gradient strips is lacking. Our main objective was to validate and compare the susceptibility results, measured as the minimum inhibitory concentration (MIC), of 6 antimicrobials for 4 Brachyspira species (B. hyodysenteriae, "B. hampsonii", B. pilosicoli, and B. murdochii) by BMD and AD (tiamulin, valnemulin, lincomycin, tylosin, and carbadox) or antibiotic gradient strip (doxycycline) methods. In general, the results of a high percentage of all 4 Brachyspira species differed by ±1 log2 dilution or less by BMD and AD for tiamulin, valnemulin, lincomycin, and tylosin, and by BMD and antibiotic gradient strip for doxycycline. The carbadox MICs obtained by BMD were 1-5 doubling dilutions different than those obtained by AD. BMD for Brachyspira was quicker to perform with less ambiguous interpretation of results when compared with AD and antibiotic gradient strip methods, and the results confirm the utility of BMD in routine diagnostics.

  14. Candida colonization in urine samples of ICU patients: determination of etiology, antifungal susceptibility testing and evaluation of associated risk factors.

    Science.gov (United States)

    Singla, Nidhi; Gulati, Neelam; Kaistha, Neelam; Chander, Jagdish

    2012-08-01

    The presence of Candida in urine presents a therapeutic challenge for the physician as it is often asymptomatic, and management guidelines have not been clearly laid down on this issue. The presence of Candida in urine may represent contamination of clinical sample, actual colonization of the lower urinary tract or may be a true indicator of invasive infection of lower and/or upper urinary tract. In a clinical setting like the ICU, multiple risk factors for Candida colonization may be present in the same patient, thereby increasing the chances of candiduria, manifold. In the present study on 80 patients in ICU, high rate of Candida colonization (57.5%) was found in urine samples of ICU patients with C. tropicalis (57.3%) being the predominant species. We also isolated 8 strains of Trichosporon species, all of these presented as a mixed infection along with Candida species. Among the various risk factors studied, urinary catheterization and previous antibiotic therapy were identified as statistically significant (P value albicans, C. glabrata and Trichosporon species, respectively.

  15. Suscetibilidade a antifúngicos de cepas de Candida albicans isoladas de pacientes com estomatite protética Susceptibility to antifungal drugs of Candida albicans strains isolated from patients with denture stomatitis

    Directory of Open Access Journals (Sweden)

    Jéssica Moreira BATISTA

    1999-12-01

    Full Text Available Pacientes portadores de próteses totais, apresentam, com freqüência a chamada estomatite protética, com a qual associa-se Candida albicans determinando a chamada candidíase eritematosa. Assim, procuramos avaliar a suscetibilidade dessa levedura a agentes antifúngicos. A suscetibilidade de dezenove cepas de Candida albicans isoladas de pacientes apresentando estomatite protética foi estudada frente a: um derivado poliênico representado, pela anfotericina B (AnB, e dois derivados azóicos, cetoconazol e miconazol. A atividade antifúngica foi estudada a partir da determinação da concentração inibitória mínima (CIM e da concentração fungicida mínima (CFM, pela técnica de diluição em ágar. Os resultados obtidos, mostraram baixos valores de CIMs e CFMs (mg/ml para a AnB frente a todas as leveduras. Para o miconazol e o cetoconazol, foram observadas CIMs invariavelmente £ 4,00 mg/ml; para as CFMs, foram obtidos valores ³ 16,00 mg/ml frente a maioria das cepas. Conclui-se que a AnB apresentou maior ação fungicida in vitro enquanto os azóis demonstraram ação fungistática mas não fungicida. Acreditamos que a pesquisa de novas drogas, principalmente de uso tópico ainda é necessária, a fim de se tratar, com sucesso, a candidíase eritematosa, comumente observada nas chamadas estomatites protéticas.Users of total prosthesis present in general a high frequency of the so called denture stomatitis, associated to erythematous candidiasis. So, we evaluated the susceptibility of oral yeast to three antifungal agents. Strains of Candida albicans isolated from patients with denture stomatitis were evaluated in relation to the susceptibility of the antifungal drugs as amphotericin B (polyenic derivatives, and azole agents as ketoconazole and miconazole. The antifungal activity was evaluated, and the minimal inhibitory concentration (MIC and minimal fungicide concentration (MFC were determined utilizing the agar dilution method. The

  16. Rapid antibiotic susceptibility testing of Mycobacterium tuberculosis : Its utility in resource poor settings

    Directory of Open Access Journals (Sweden)

    Poojary A

    2006-01-01

    Full Text Available Purpose: To compare the rapid colorimetric nitrate reductase based antibiotic susceptibility (CONRAS test performed on Mycobacterium tuberculosis isolates with the conventional method i.e., the proportion method. Methods: One hundred clinical isolates of M. tuberculosis were tested for susceptibility to isoniazid (INH and rifampicin (RIF by the conventional proportion method and CONRAS in Middlebrook 7H9 liquid medium enriched with growth supplements (MB7H9S. Results: The performance of the CONRAS test was evaluated using proportion method as the gold standard. The sensitivity (ability to detect true drug resistance and specificity (ability to detect true drug susceptibility of the CONRAS test to INH was 93.75 and 98.52% and for RIF it was 96.10 and 100% respectively. The mean time for reporting was 6.3 days and the test showed excellent reproducibility. The kappa (k value for INH was 0.92 and for RIF was 0.99, indicating excellent agreement between the two methods. Conclusions: CONRAS test is a rapid and reliable method of drug susceptibility for M. tuberculosis.

  17. Antibiotic susceptibility testing of the Gram-negative bacteria based on flow cytometry

    Directory of Open Access Journals (Sweden)

    Claude Saint-Ruf

    2016-07-01

    Full Text Available Rapidly treating infections with adequate antibiotics is of major importance. This requires a fast and accurate determination of the antibiotic susceptibility of bacterial pathogens. The most frequently used methods are slow because they are based on the measurement of growth inhibition. Faster methods, such as PCR-based detection of determinants of antibiotic resistance, do not always provide relevant information on susceptibility, particularly that which is not genetically based. Consequently, new methods, such as the detection of changes in bacterial physiology caused by antibiotics using flow cytometry and fluorescent viability markers, are being explored. In this study, we assessed whether Alexa Fluor® 633 Hydrazide (AFH, which targets carbonyl groups, can be used for antibiotic susceptibility testing. Carbonylation of cellular macromolecules, which increases in antibiotic-treated cells, is a particularly appropriate to assess for this purpose because it is irreversible. We tested the susceptibility of clinical isolates of Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa, to antibiotics from the three classes: β-lactams, aminoglycosides and fluoroquinolones. In addition to AFH, we used TO-PRO®-3, which enters cells with damaged membranes and binds to DNA, and DiBAC4 (3, which enters cells with depolarized membranes. We also monitored antibiotic-induced morphological alterations of bacterial cells by analyzing light scattering signals. Although all tested dyes and light scattering signals allowed for the detection of antibiotic-sensitive cells, AFH proved to be the most suitable for the fast and reliable detection of antibiotic susceptibility.

  18. Antibiotic Susceptibility Testing of the Gram-Negative Bacteria Based on Flow Cytometry.

    Science.gov (United States)

    Saint-Ruf, Claude; Crussard, Steve; Franceschi, Christine; Orenga, Sylvain; Ouattara, Jasmine; Ramjeet, Mahendrasingh; Surre, Jérémy; Matic, Ivan

    2016-01-01

    Rapidly treating infections with adequate antibiotics is of major importance. This requires a fast and accurate determination of the antibiotic susceptibility of bacterial pathogens. The most frequently used methods are slow because they are based on the measurement of growth inhibition. Faster methods, such as PCR-based detection of determinants of antibiotic resistance, do not always provide relevant information on susceptibility, particularly that which is not genetically based. Consequently, new methods, such as the detection of changes in bacterial physiology caused by antibiotics using flow cytometry and fluorescent viability markers, are being explored. In this study, we assessed whether Alexa Fluor® 633 Hydrazide (AFH), which targets carbonyl groups, can be used for antibiotic susceptibility testing. Carbonylation of cellular macromolecules, which increases in antibiotic-treated cells, is a particularly appropriate to assess for this purpose because it is irreversible. We tested the susceptibility of clinical isolates of Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa, to antibiotics from the three classes: β-lactams, aminoglycosides, and fluoroquinolones. In addition to AFH, we used TO-PRO®-3, which enters cells with damaged membranes and binds to DNA, and DiBAC4 (3), which enters cells with depolarized membranes. We also monitored antibiotic-induced morphological alterations of bacterial cells by analyzing light scattering signals. Although all tested dyes and light scattering signals allowed for the detection of antibiotic-sensitive cells, AFH proved to be the most suitable for the fast and reliable detection of antibiotic susceptibility.

  19. Clinically significant anaerobic bacteria isolated from patients in a South African academic hospital: antimicrobial susceptibility testing.

    Science.gov (United States)

    Naidoo, S; Perovic, O; Richards, G A; Duse, A G

    2011-09-27

    BACKGROUND. Increasing resistance to some antimicrobial agents among anaerobic bacteria has made susceptibility patterns less predictable. METHOD. This was a prospective study of the susceptibility data of anaerobic organisms isolated from clinical specimens from patients with suspected anaerobic infections from June 2005 until February 2007. Specimens were submitted to the microbiology laboratory at Charlotte Maxeke Johannesburg Academic Hospital, where microscopy, culture and susceptibility testing were performed the using E test® strip minimum inhibitory concentration method. Results were interpreted with reference to Clinical and Laboratory Standards Institute guidelines for amoxicillin-clavulanate, clindamycin, metronidazole, penicillin, ertapenem, cefoxitin, ceftriaxone, chloramphenicol and piperacillin-tazobactam. RESULTS. One hundred and eighty anaerobic isolates were submitted from 165 patients. The most active antimicrobial agents were chloramphenicol (100% susceptible), ertapenem (97.2%), piperacillin-tazobactam (99.4%) and amoxicillin-clavulanic acid (96.7%). Less active were metronidazole (89.4%), cefoxitin (85%), clindamycin (81.7%), ceftriaxone (68.3%) and penicillin (33.3%). CONCLUSION. Susceptibility testing should be performed periodically to identify emerging trends in resistance and to modify empirical treatment of anaerobic infections.

  20. Evaluation of susceptibility of high strength steels to delayed fracture by using cyclic corrosion test and slow strain rate test

    Energy Technology Data Exchange (ETDEWEB)

    Li Songjie [School of Materials Science and Engineering, University of Science and Technology Beijing, No. 30 Xueyuan Road, Hidian Zone, Beijing 100083 (China); Structural Metals Center, National Institute for Materials Science, 1-2-1 Sengen, Tsukuba, Ibaraki 305-0047 (Japan); Zhang Zuogui [Structural Metals Center, National Institute for Materials Science, 1-2-1 Sengen, Tsukuba, Ibaraki 305-0047 (Japan); Akiyama, Eiji [Structural Metals Center, National Institute for Materials Science, 1-2-1 Sengen, Tsukuba, Ibaraki 305-0047 (Japan)], E-mail: AKIYAMA.Eiji@nims.go.jp; Tsuzaki, Kaneaki [Structural Metals Center, National Institute for Materials Science, 1-2-1 Sengen, Tsukuba, Ibaraki 305-0047 (Japan); Zhang Boping [School of Materials Science and Engineering, University of Science and Technology Beijing, No. 30 Xueyuan Road, Hidian Zone, Beijing 100083 (China)

    2010-05-15

    To evaluate susceptibilities of high strength steels to delayed fracture, slow strain rate tests (SSRT) of notched bar specimens of AISI 4135 with tensile strengths of 1300 and 1500 MPa and boron-bearing steel with 1300 MPa have been performed after cyclic corrosion test (CCT). During SSRT the humidity around the specimen was kept high to keep absorbed diffusible hydrogen. The fracture stresses of AISI 4135 steels decreased with increment of diffusible hydrogen content which increased with CCT cycles. Their delayed fracture susceptibilities could be successfully evaluated in consideration of both influence of hydrogen content on mechanical property and hydrogen entry.

  1. Change of antibiotic susceptibility testing guidelines from CLSI to EUCAST: influence on cumulative hospital antibiograms.

    Directory of Open Access Journals (Sweden)

    Aline Wolfensberger

    Full Text Available OBJECTIVE: We studied whether the change in antibiotic susceptibility testing (AST guidelines from CLSI to EUCAST influenced cumulative antibiograms in a tertiary care hospital in Switzerland. METHODS: Antibiotic susceptibilities of non-duplicate isolates collected within a one-year period before (period A and after (period B changing AST interpretation from CLSI 2009 to EUCAST 1.3 (2011 guidelines were analysed. In addition, period B isolates were reinterpreted according to the CLSI 2009, CLSI 2013 and EUCAST 3.1 (2013 guidelines. RESULTS: The majority of species/drug combinations showed no differences in susceptibility rates comparing periods A and B. However, in some gram-negative bacilli, decreased susceptibility rates were observed when comparing CLSI 2009 with EUCAST 1.3 within period B: Escherichia coli / cefepime, 95.8% (CLSI 2009 vs. 93.1% (EUCAST 1.3, P=0.005; Enterobacter cloacae / cefepime, 97.0 (CLSI 2009 vs. 90.5% (EUCAST 1.3, P=0.012; Pseudomonas aeruginosa / meropenem, 88.1% (CLSI 2009 vs. 78.3% (EUCAST 1.3, P=0.002. These differences were still evident when comparing susceptibility rates according to the CLSI 2013 guideline with EUCAST 3.1 guideline. For P. aeruginosa and imipenem, a trend towards a lower antibiotic susceptibility rate in ICUs compared to general wards turned into a significant difference after the change to EUCAST: 87.9% vs. 79.8%, P=0.08 (CLSI 2009 and 86.3% vs. 76.8%, P=0.048 (EUCAST 1.3. CONCLUSIONS: The change of AST guidelines from CLSI to EUCAST led to a clinically relevant decrease of susceptibility rates in cumulative antibiograms for defined species/drug combinations, particularly in those with considerable differences in clinical susceptibility breakpoints between the two guidelines.

  2. EUCAST recommendations for antimicrobial susceptibility testing applied to the three main Campylobacter species isolated in humans.

    Science.gov (United States)

    Sifré, Elodie; Salha, Ben Amor; Ducournau, Astrid; Floch, Pauline; Chardon, Hubert; Mégraud, Francis; Lehours, Philippe

    2015-12-01

    Antimicrobial susceptibility testing of Campylobacter isolates is of great importance for treatment options especially in systemic diseases. The European Committee for Antimicrobial Susceptibility Testing (EUCAST) recently proposed epidemiological cut-offs (ECOFFs) for a limited number of antimicrobial compounds and for Campylobacter jejuni and Campylobacter coli only. In the present study, the EUCAST method was used after minor modifications to define antimicrobial susceptibility patterns for, 1997 C. jejuni, 419 C. coli and 100 Campylobacter fetus strains received at the French National Reference Center for Campylobacters and Helicobacters. Our results show that the ECOFFs defined by EUCAST for tetracycline and ciprofloxacin can be used for C. jejuni and C. coli. The same ECOFF can be used for erythromycin for the three species. The C. jejuni and C. coli ECOFFs for ciprofloxacin however cannot be applied to C. fetus. We also provide data to categorise two 2 β-lactams of interest for systemic diseases, ampicillin and amoxicillin+clavulanate, for the three species.

  3. Pinhole test for identifying susceptibility of soils to piping erosion: effect water quality and hydraulic head

    Energy Technology Data Exchange (ETDEWEB)

    Nadal Romero, E.; Verachtert, E.; Poesen, J.

    2009-07-01

    Piping has been observed in both natural and soils, as well as under different types of land uses and vegetation covers. Despite its importance, no standard widely-applied methodology exists to assess susceptibility of soils to piping. This study aims at evaluating the pinhole test for assessing the susceptibility of soils to piping under different conditions. More precisely, the effects of hydraulic head and water quality are being assessed. Topsoil samples (remoulded specimens) with a small range of water contents were taken in Central Belgium (Heverlee) and the susceptibility of these soil samples are investigated under standardized laboratory conditions with a pinhole test device. Three hydraulic heads (50,180 and 380 mm) and two water qualities (tap and distilled water) were used, reflecting dominant field conditions. (Author) 6 refs.

  4. Serogenotyping and antimicrobial susceptibility testing of Salmonella spp. isolated from retail meat samples in Lagos, Nigeria.

    Science.gov (United States)

    Smith, Stella; Braun, Sascha; Akintimehin, Faith; Fesobi, Toun; Bamidele, Moses; Coker, Akitoye; Monecke, Stefan; Ehricht, Ralf

    2016-08-01

    Microarray-based serogenotyping, antimicrobial susceptibility tests and the detection of relevant resistance genes were performed on isolates of Salmonella spp. from retail meat samples obtained in Lagos, Nigeria. Out of 151 meat samples, 33 Salmonella isolates were obtained. Nine different Salmonella serovars (S. Amoutive, S. Bargny, S. Drac, S. Ealing, S. Urbana, S. Hadar, S. Nyborg, S. Anatum and S. Havana) were identified by microarray-based serogenotyping and confirmed afterwards using classical serotyping. Antibiotic susceptibility tests with 17 antibiotics showed that almost all isolates were fully susceptible to this panel. The results of this study indicated a high prevalence of Salmonella in retail meat, the presence of some previously rather rarely described Serovars in retail meat samples from Lagos, and a need to monitor for Salmonella and their antibiotic resistance determinants. The microarray-based system used herein proved to be perfectly suited as epidemiological tool to replace classical serotyping.

  5. Antimicrobial susceptibility testing of bovine digital dermatitis treponemes identifies macrolides for in vivo efficacy testing.

    Science.gov (United States)

    Evans, N J; Brown, J M; Hartley, C; Smith, R F; Carter, S D

    2012-12-07

    Digital dermatitis (DD) is a major infectious lameness of dairy cattle and sheep considered to be caused by treponemes. The aim of this study was to identify antibiotics effective against DD treponemes that might be useful in the treatment of ruminant DD in the future or to identify antibiotics useful in isolation studies. Here, a microdilution method was used to identify in vitro antimicrobial susceptibility of treponemes cultured from DD lesions to eight relevant antibiotics. DD treponemes exhibited highest susceptibility to amoxicillin, azithromycin and gamithromycin. Unfortunately, amoxicillin whilst having potential for DD treatment in other animals (e.g. sheep) would require milk withhold periods in dairy cattle. DD treponemes were not particularly susceptible to two cephalosporins: cefalexin and ceftiofur, which do not require milk withhold. The bacteria demonstrated low susceptibility to trimethoprim and especially colistin suggesting these antimicrobials may be particularly useful in isolation of DD treponemes. The most promising high susceptibility results for macrolides indicate a rationale to consider veterinary licensed macrolides as DD treatments. Furthermore, given the DD treponeme antibiotic susceptibility similarities to established treatments for human treponematoses, identification of treponemacidal, long acting β-lactam analogues not requiring milk withhold may allow for development of a successful treatment for dairy cattle DD.

  6. Rapid drug susceptibility testing of mycobacteria by culture on a highly porous ceramic support

    NARCIS (Netherlands)

    Ingham, C.J.; Ayad, A.B.; Nolsen, K.; Mulder, B.

    2008-01-01

    BACKGROUND: Phenotypic, culture-based methods for drug susceptibility testing (DST) of Mycobacterium tuberculosis are relatively simple and may be particularly appropriate for resource-limited settings where tuberculosis (TB) is most prevalent. However, these methods can be slow and generate signifi

  7. Molecular drug susceptibility testing in the Netherlands: performance of the MTBDRplus and MTBDRsl assays

    NARCIS (Netherlands)

    Simons, S.O.; Laan, T. van der; Zwaan, R. de; Kamst, M.; Ingen, J. van; Dekhuijzen, P.N.; Boeree, M.J.; Soolingen, D. van

    2015-01-01

    BACKGROUND: The performance of molecular drug susceptibility testing in countries with a low prevalence of drug resistance, such as the Netherlands, has not been adequately studied. OBJECTIVE: To evaluate the diagnostic accuracy of the GenoType((R)) MTBDRplus and MTBDRsl assays to detect resistance

  8. Mixed-morphotype broth microdilution susceptibility testing of Pseudomonas aeruginosa from cystic fibrosis patients.

    OpenAIRE

    1993-01-01

    Multiple morphotypes of Pseudomonas aeruginosa isolated from 50 respiratory specimens of cystic fibrosis patients were tested for correlation of broth microdilution susceptibility results of a mixed-morphotype inoculum with a predicted antibiogram of the individual isolates. The overall correlation was 96.0%, with only 1.6% very major or major errors.

  9. Evaluation of Ciprofloxacin as a Representative of Veterinary Fluoroquinolones in Susceptibility Testing

    OpenAIRE

    Riddle, Christa; Lemons, Carol L.; Papich, Mark G.; Altier, Craig

    2000-01-01

    Currently in veterinary medicine, ciprofloxacin is often used in susceptibility testing to represent the entire class of fluoroquinolone antimicrobials. Using quality control organisms as well as clinical isolates, we compared the MIC of ciprofloxacin to those of three other fluoroquinolones used in animals and found that ciprofloxacin is not an adequate representative of other members of this class.

  10. Interpretive criteria for mupirocin susceptibility testing of Staphylococcus spp. using CLSI guidelines.

    LENUS (Irish Health Repository)

    Creagh, S

    2012-02-03

    Mupirocin is an antimicrobial agent commonly used to treat staphylococcal infection or to eliminate persistent carriage. To date, interpretive criteria have not been established to define susceptibility or resistance when performing mupirocin susceptibility testing. In this evaluation, using CLSI guidelines, a total of 502 staphylococci comprising 219 methicillin-sensitive Staphylococcus aureus, 222 methicillin-resistant S. aureus and 61 coagulase-negative staphylococci are tested by broth microdilution, disc diffusion and E-test. Disc diffusion using 5 microg mupirocin discs was found to be a reliable method to distinguish susceptible and resistant strains. Minimum inhibitory concentration (MIC) determination was required to differentiate low-level and high-level resistance to mupirocin. E-test was found to be an accurate alternative to broth microdilution for the routine determination of MIC values of staphylococci to mupirocin. Broth microdilution and disc-diffusion results were plotted on a scattergram, and error rates were calculated. No errors were found using susceptibility criteria of < 4 microg\\/mL (MIC) and > 19 mm (zone diameter).

  11. Screening of pharmacologically active small molecule compounds identifies antifungal agents against Candida biofilms

    Directory of Open Access Journals (Sweden)

    Takao eWatamoto

    2015-12-01

    Full Text Available Candida species have emerged as important and common opportunistic human pathogens, particularly in immunocompromised individuals. The current antifungal therapies either have toxic side effects or are insufficiently effect. The aim of this study is develop new small-molecule antifungal compounds by library screening methods using C. albicans, and to evaluate their antifungal effects on Candida biofilms and cytotoxic effects on human cells. Wild-type C. albicans strain SC5314 was used in library screening. To identify antifungal compounds, we screened a small-molecule library of 1,280 pharmacologically active compounds (LOPAC1280TM using an antifungal susceptibility test (AST. To investigate the antifungal effects of the hit compounds, ASTs were conducted using Candida strains in various growth modes, including biofilms. We tested the cytotoxicity of the hit compounds using human gingival fibroblast (hGF cells to evaluate their clinical safety. Only 35 compounds were identified by screening, which inhibited the metabolic activity of C. albicans by >50%. Of these, 26 compounds had fungistatic effects and 9 compounds had fungicidal effects on C. albicans. Five compounds, BAY11-7082, BAY11-7085, sanguinarine chloride hydrate, ellipticine and CV-3988, had strong fungicidal effects and could inhibit the metabolic activity of Candida biofilms. However, BAY11-7082, BAY11-7085, sanguinarine chloride hydrate and ellipticine were cytotoxic to hGF cells at low concentrations. CV-3988 showed no cytotoxicity at a fungicidal concentration.Four of the compounds identified, BAY11-7082, BAY11-7085, sanguinarine chloride hydrate and ellipticine, had toxic effects on Candida strains and hGF cells. In contrast, CV-3988 had fungicidal effects on Candida strains, but low cytotoxic effects on hGF cells. Therefore, this screening reveals agent, CV-3988 that was previously unknown to be antifungal agent, which could be a novel therapies for superficial mucosal

  12. Contemporary tetracycline susceptibility testing: doxycycline MIC methods and interpretive criteria (CLSI and EUCAST) performance when testing Gram-positive pathogens.

    Science.gov (United States)

    Jones, Ronald N; Stilwell, Matthew G; Wilson, Michael L; Mendes, Rodrigo E

    2013-05-01

    International susceptibility testing breakpoint organizations and regulatory agencies have markedly differing interpretive criteria for the tetracycline class. Here we examined the magnitude of these differences for doxycycline and tetracycline hydrochloride (HCL) when tested against a collection of 13,176 Gram-positive cocci from a worldwide surveillance network (SENTRY Antimicrobial Surveillance Program, 2010). Clinical and Laboratory Standards Institute (CLSI) breakpoints are routinely higher, usually 4-fold, compared to those of the European Committee on Antimicrobial Susceptibility Testing (EUCAST); however, CLSI recently (2013) modified Streptococcus pneumoniae breakpoints (≤ 2 μg/mL in 2012) to ≤ 0.25 and ≤ 1 μg/mL for doxycycline and tetracycline HCL, respectively. We report that these changes are a promising step toward international breakpoint harmonization, but lack a comprehensive approach needed for testing tetracyclines against all Gram-positive cocci. Generally, EUCAST breakpoint criteria showed i) lower spectrums (reduced susceptibility rates) for the tetracyclines, but highest for doxycycline versus all species examined; ii) greater test accuracy (lower predictive categorical errors), especially for tetracycline to predict doxycycline susceptibility (99.91%); and iii) zone diameter correlate breakpoints which are generally available online. Molecular tests for tet resistance genes demonstrate that tet (K) and tet (M) containing strains can occur in the susceptible population of MIC results by both CLSI and EUCAST breakpoint criteria. In summary, doxycycline continues to show greater comparative potency versus tetracycline HCL against all monitored Gram-positive species and the international harmonization of tetracycline breakpoints should be a priority, as the most recent CLSI update only addressed 1 streptococcal species and 2 tetracycline agents.

  13. Levofloxacin susceptibility testing against Helicobacter pylori: evaluation of a modified disk diffusion method compared to E test.

    Science.gov (United States)

    Boyanova, Lyudmila; Ilieva, Juliana; Gergova, Galina; Mitov, Ivan

    2016-01-01

    We compared levofloxacin (1 μg/disk) disk diffusion method to E test against 212 Helicobacter pylori strains. Using diameter breakpoints for susceptibility (≥15 mm) and resistance (≤9 mm), very major error, major error rate, and categoric agreement were 0.0%, 0.6%, and 93.9%, respectively. The method may be useful in low-resource laboratories.

  14. Computerised emission and susceptibility MIL.STD testing with automated NB/BB detection

    Science.gov (United States)

    Vanessen, J. C.

    1990-09-01

    Automation of Electromagnetic Compatibility (EMC) testing is becoming common at many EMC test facilities. Commercial automated systems have become available in the past few years. The test and operations section has developed its own EMC automation to enhance and aid in testing. A complete overview of the automated EMC test facility in operation for emission and susceptibility measurements is presented. It includes a hardware description, the program structure and some of the methods required to complete such a program on the equipment chosen, including the Narrow Band (NB) and Broad Band (BB).

  15. A fatal case of AIDS-defining meningoencephalitis by C. Neoformans, sensitive to antifungal therapy

    Directory of Open Access Journals (Sweden)

    Mohammad-Khani S

    2010-11-01

    Full Text Available Abstract Cryptococcus neoformans is the most common cause of life threatening meningoencephalitis in HIV-infected patients. Diagnosis is based on tests for cryptoccocal antigen in serum and cerebrospinal fluid, and on culture of the organism. We present a case of AIDS-related cryptococcal meningoencephalitis unresponsive to antifungal combination therapy, despite of evidence of fungal susceptibility in vitro. Significant decreases in cryptococcal antigen titers in serum and cerebrospinal fluid did not correlate with progress in disease and fatal outcome.

  16. [Performance evaluation of VITEK 2 system in meropenem susceptibility testing of clinical Pseudomonas aeruginosa isolates].

    Science.gov (United States)

    Acuner, Ibrahim Cağatay; Bayramoğlu, Gülçin; Birinci, Asuman; Cekiç Cihan, Ciğdem; Bek, Yüksel; Durupınar, Belma

    2011-07-01

    Pseudomonas aeruginosa is an important opportunistic pathogen associated with various community-acquired or nosocomial infections. Multi-drug resistant P.aeruginosa strains increasingly cause epidemics and spread in various hospital wards and geographic regions. Carbapenems are among the most effective antimicrobials in the treatment of multi-drug resistant P.aeruginosa infections, and meropenem is the most successful among alternatives in initial therapy. Particularly in severe infections, inappropriate or inadequate initial antimicrobial therapy is independently associated with adverse clinical and economic outcomes. Availability of accurate and rapid susceptibility testing is a priority. Most of the automated microbiology systems can provide rapid results within 8 to 12 hours. In comparison to standard methods, problems in the antimicrobial susceptibility testing of particular microorganisms and antimicrobial agents have been reported for automated microbiology systems. Failures have been reported previously especially in the susceptibility testing of P.aeruginosa versus carbapenem. Most of these studies are designed according to the Food and Drug Administration (FDA, USA) performance analysis scheme (Class II Special Controls Guidance Document: Antimicrobial Susceptibility Test Systems) in a simplified form. However, there are many lacking issues in the design of most of these studies. Among these, insufficient sample size, use of inappropriate reference method, lack of reproducibility testing, and inadequate distribution of study isolates in interpretative categories are of notice. There are only few studies in the literature that evaluate the performance of automated systems in antimicrobial susceptibility testing of carbapenems in clinical P.aeruginosa isolates with a sufficient sample size (n ? 100). However, most of these studies still have one or more major deficiencies in the study design. Furthermore, none of these studies evaluate the performance of

  17. Comparison of the CLSI guideline and ISO/IDF standard for antimicrobial susceptibility testing of Lactobacilli.

    Science.gov (United States)

    Mayrhofer, Sigrid; Zitz, Ulrike; Birru, Firew H; Gollan, Dagmar; Gołoś, Aleksandra K; Kneifel, Wolfgang; Domig, Konrad J

    2014-12-01

    Lactobacilli play a crucial role as probiotics and as starter cultures in the production of fermented foods. Although lactobacilli are a technologically useful and beneficial group of bacteria, a few members of them have been rarely correlated with bacterial infections. Correspondingly, clinicians are interested in the antimicrobial susceptibility of lactobacilli. In addition, antimicrobial susceptibility testing (AST) is also relevant for commercially applied lactobacilli as bacterial strains harboring transferable antibiotic resistance genes should not be used in fermented and probiotic foods. Therefore, two methods were developed by different organizations, which were compared within this study. For this purpose, 22 Lactobacillus-type strains were tested for their antimicrobial susceptibility to 16 antibiotics following the procedures of the Clinical and Laboratory Standards Institute (CLSI) and the International Organization of Standardization (ISO)/International Dairy Federation (IDF). Crucial discrepancies between both procedures were detected mainly due to the different AST media. Hence, half of the strains tested did not consistently grow in the CLSI medium, whereas all showed evaluable growth in the ISO/IDF medium. However, some antibiotics were influenced by the latter medium. In particular, low levels of essential agreement between both methods were obtained with seven antibiotics. Accordingly, different interpretative criteria are needed for both procedures to distinguish resistant from susceptible strains.

  18. Sporothrix schenckii associated with armadillo hunting in Southern Brazil: epidemiological and antifungal susceptibility profiles Sporothrix schenckii relacionado à caça ao tatu no Sul do Brasil: aspectos epidemiológicos e suscetibilidade dos isolados aos antifúngicos

    Directory of Open Access Journals (Sweden)

    Sydney Hartz Alves

    2010-10-01

    Full Text Available INTRODUCTION: Sporotrichosis is the most common subcutaneous mycosis observed in Brazil and it is generally consequent to a little trauma caused by vegetal particles or spines which inoculate the fungi in the subcutaneous area. Although sporotrichosis had been frequently mentioned with armadillo hunting this form has not been widely reported in Brazil until now. In this study we report ten cases of sporotrichosis evolving the armadillo's hunting diagnosed in some towns located in the central and west regions of Rio Grande do Sul State. METHODS: The cases were established based on clinical and classic mycological laboratorial techniques. The susceptibility tests were conducted by microdilution technique according to M38-A2 CLSI documents. RESULTS: Ten cases of sporotrichosis associated with armadillo hunting detected in the State of Rio Grande do Sul were diagnosed by mycological methods. The susceptibility tests of Sporothrix schenckii isolates to antifungal agents itraconazole, ketoconazole and terbinafine showed that all the isolates were susceptible. CONCLUSIONS: The paper discusses some cultural aspects related to hunting of this wild animal as well as possible causes of this unexpected occurrence in southern Brazil.INTRODUÇÃO: A esporotricose constitui-se na micose subcutânea mais frequentemente observada e, na maioria dos casos, a infecção é decorrente de pequenos traumas envolvendo fragmentos vegetais ou espinhos que inoculam o fungo no tecido subcutâneo. Embora frequentemente relacionada a caça a tatus, esta ocorrência tem sido raramente relatada no Brasil. Neste estudo relatamos dez casos envolvendo esta prática, observados em várias cidades das regiões centro e oeste do Estado do Rio Grande do Sul. MÉTODOS: o diagnóstico clínico foi confirmado pelos métodos clássicos de cultura em ágar Mycobiotic, identificação micromorfológica seguida de reversão a fase leveduriforme em ágar BHI. Os testes de suscetibilidade

  19. In Vitro Antifungal Susceptibility of Yeast Phase of Sporothrix schenckii by Microdilution Broth Method%微量稀释法检测申克孢子丝菌酵母相体外抗真菌药物敏感性

    Institute of Scientific and Technical Information of China (English)

    蔡晴; 李珊山; 宋洋; 沈永年; 吕桂霞; 陈伟; 刘维达

    2011-01-01

    Objective To evaluate the activities of potassium iodide, fluconazole, itraconazole, terbinafine on yeast phase of Sporothrix shenckii, and investigate the effect of potassium iodide on itraconazole and terbinafine by microdilution broth method in vitro. Methods Fifty-four isolates of Sporothrix shenckii were converted to the yeast form successive passages on Brain Heart Infusion Agar medium at 35℃. Then according to a modified CLSI microdilution method M27-A2, tested the susceptibility of these isolates to potassium iodide, fluconazole, itraconazole, terbinafine. We added potassium iodide in RPMI-1640, then compared the susceptibility of itraconazole and terbinafine with the former in vitro. Candida krusei ATCC 6258 was used for quality control. Results Potassium iodide showed no antifungal activity in vitro. Terbinafine had the best susceptibility, with geometric mean of 0. 17μg/mL, MIC range is 0.03 ~ 1 μg/mL. The MIC of itraconazole is 0.03 ~8μg/mL(0. 98μg/mL). Fluconazole has a high MIC( >64μg/mL). With potassium iodide in RPMI1640, the MICs of terbinafine and itraconazole were significantly different from the former results ( P <0. 05). But there were no significant difference in MIC between isolates from cutaneous sporotrichosis and lymphocutaneous sporotrichosis (P > 0.05). Conclusion A modified M27-A2 is an alternative method for testing yeast phase of Sporothrix schenckii. Potassium iodide can improve the antifungal susceptibility of terbinafine and itraconazole to yeast phase in vitro.%目的 用液基微量稀释法观察双相真菌申克孢子丝菌酵母相体外抗真菌药物敏感性.方法 将54株申克孢子丝菌临床株于脑心浸液琼脂培养基连续传代获得酵母相,参考美国临床实验室标准化委员会(CLSI)的微量稀释法M27-A2检测菌株酵母相对碘化钾、氟康唑、伊曲康唑和特比萘芬的体外敏感性,并观察碘化钾对伊曲康唑和特比萘芬体外抑菌作用的影响.

  20. 呼吸道白色假丝酵母菌分离株生物膜形成及药物敏感性检测%Biofilm formation and antifungal susceptibility of Candida albicans isolated from respiratory tract

    Institute of Scientific and Technical Information of China (English)

    阳隽; 张天托; 朱家馨; 黄静

    2011-01-01

    OBJECTIVE To monitor the biofilm formation and antifungal susceptibility of Candida albicans isolated from lower respiratory tract of critically ill patients.METHODS By forming biofilm in cell culture plate in vitro,based on the amount of light blocked passing through the wells, C.albicans isolates were divided into two groups:biofilm-negative isolates and biofilm-positive isolates.The MICs of antifungal drugs against planktonic cells and biofilm-associated adherent cells of 10 isolates were determined respectively.RESULTS Totally 14(26.92%)of 52 isolates were classified as biofilm producer, the other 38(73.08 %)isolates were classified as nonbiofilm producer.The MICs of FLU, CASPO and AMB for biofilm-associated adherent cells were much higher than that for planktonic cells.All biofilm-associated adherent ceils were resistant to FLU and CASPO(SMIC80 >256 μg/ml;>16 μg/ml).The MICs of AMB for biofilms of 4 strains were more than 8 μg/ml.CONCLUSION Biofilm formation varies greatly among individual C.albicans isolates.C.albicans biofilm is highly resistant to antifungal agents.%目的 监测危重病患者下呼吸道分离的白色假丝酵母菌(CAL)体外生物膜形成及对抗真菌药物的敏感性,为临床诊治提供依据.方法 接种CAL于96孔培养板黏附生长形成生物膜,根据相对于空白对照透光度下降的程度将CAL分为生物膜阳性和生物膜阴性菌株,并测定抗真菌药物对10株生物膜阳性CAL游离态和生物膜的MIC值.结果 52株CAL中有14株为生物膜阳性菌株,占26.92%;38株为生物膜阴性菌株,占73.08%;氟康唑、卡泊芬净及两性霉素B对生物膜CAL的MIC值明显高于其游离态MIC值,10株生物膜CAL对氟康唑、卡泊芬净均耐药(SMIC80>256μg/ml及>16μg/ml),而两性霉素B对其中4株生物膜CAL的SMIC80>8μg/ml.结论 呼吸道CAL分离株生物膜形成存在表型差异,生物膜CAL对抗真菌药物的耐药性增高.

  1. Quality Control and Reference Guidelines for CLSI Broth Microdilution Method (M38-A Document) for Susceptibility Testing of Anidulafungin against Molds▿

    Science.gov (United States)

    Espinel-Ingroff, A.; Fothergill, A.; Ghannoum, M.; Manavathu, E.; Ostrosky-Zeichner, L.; Pfaller, M. A.; Rinaldi, M. G.; Schell, W.; Walsh, T. J.

    2007-01-01

    The CLSI (formerly NCCLS) M38-A document for antifungal susceptibility testing of filamentous fungi does not describe guidelines for echinocandins. A multicenter study (eight centers) evaluated inter- and intralaboratory reproducibilities of two reading times (24 and 48 h or 48 and 72 h) and two end points (MICs and minimum effective concentrations [MECs]) for evaluating anidulafungin against molds. Anidulafungin MICs (≥50% inhibition) and MECs (morphological hyphal changes) were determined for seven Aspergillus isolates (four species) and one isolate each of Fusarium moniliforme, Fusarium solani, and Paecilomyces variotii and for two Scedosporium apiospermum isolates. The inter- and intralaboratory reproducibilities of 10 replicate tests performed in each laboratory on 10 different days for each isolate was 100% at 24 h (MECs, ≤0.015 μg/ml) for six Aspergillus and P. variotii isolates. The reproducibility was 94 to 96.7% at 72 h (MECs, 1 to 8 μg/ml) for S. apiospermum and 96.7 to 97.5% at 48 h (MICs, ≥32 μg/ml) for both Fusarium isolates. Introduction of these identified optimum testing conditions for anidulafungin into future versions of the M38 document is warranted. PMID:17475760

  2. Genetic testing for TMEM154 mutations associated with lentivirus susceptibility in sheep.

    Directory of Open Access Journals (Sweden)

    Michael P Heaton

    Full Text Available In sheep, small ruminant lentiviruses cause an incurable, progressive, lymphoproliferative disease that affects millions of animals worldwide. Known as ovine progressive pneumonia virus (OPPV in the U.S., and Visna/Maedi virus (VMV elsewhere, these viruses reduce an animal's health, productivity, and lifespan. Genetic variation in the ovine transmembrane protein 154 gene (TMEM154 has been previously associated with OPPV infection in U.S. sheep. Sheep with the ancestral TMEM154 haplotype encoding glutamate (E at position 35, and either form of an N70I variant, were highly-susceptible compared to sheep homozygous for the K35 missense mutation. Our current overall aim was to characterize TMEM154 in sheep from around the world to develop an efficient genetic test for reduced susceptibility. The average frequency of TMEM154 E35 among 74 breeds was 0.51 and indicated that highly-susceptible alleles were present in most breeds around the world. Analysis of whole genome sequences from an international panel of 75 sheep revealed more than 1,300 previously unreported polymorphisms in a 62 kb region containing TMEM154 and confirmed that the most susceptible haplotypes were distributed worldwide. Novel missense mutations were discovered in the signal peptide (A13V and the extracellular domains (E31Q, I74F, and I102T of TMEM154. A matrix-assisted laser desorption/ionization-time-of flight mass spectrometry (MALDI-TOF MS assay was developed to detect these and six previously reported missense and two deletion mutations in TMEM154. In blinded trials, the call rate for the eight most common coding polymorphisms was 99.4% for 499 sheep tested and 96.0% of the animals were assigned paired TMEM154 haplotypes (i.e., diplotypes. The widespread distribution of highly-susceptible TMEM154 alleles suggests that genetic testing and selection may improve the health and productivity of infected flocks.

  3. Suscetibilidade a antifúngicos in vitro de Candida spp. em pacientes do Hospital Universitário Regional de Maringá-PR Antifungal susceptibility of Candida spp. in vitro among patients from Regional University Hospital of Maringá-PR

    Directory of Open Access Journals (Sweden)

    Fernanda de Oliveira Demitto

    2012-10-01

    Full Text Available INTRODUÇÃO: No ambiente hospitalar, são frequentes as infecções por leveduras do gênero Candida spp., o que torna esse assunto um importante alvo de estudos. OBJETIVO: Avaliar o perfil de suscetibilidade aos antifúngicos de espécies de Candida de pacientes internados no Hospital Universitário Regional de Maringá-PR (HURM. MATERIAL E MÉTODOS: As amostras foram submetidas ao teste de microdiluição em caldo (MD, segundo o documento M27-A3 para determinação da concentração inibitória mínima (CIM, e ao teste de difusão em disco de acordo com o documento M44-A2, ambos do Clinical Laboratory Standarts Institute (CLSI. RESULTADOS E DISCUSSÃO: Foram obtidos 91 isolados provenientes de amostras de urina, hemocultura, ponta de cateter, secreção orotraqueal, entre outros, sendo 38 Candida albicans, 23 C. tropicalis, 16 C. gabrata, 10 C. parapsilosis e quatro C. krusei. Dos antifúngicos testados, anfotericina B, voriconazol e anidulafungina foram os mais eficazes. CONCLUSÃO: A comparação entre as metodologias de microdiluição em caldo e disco difusão (DD mostrou boa correlação para fluconazol para a maioria das espécies de Candida spp., sendo possível destacar que a DD é útil para triagem dos principais antifúngicos usados na prática clínica. No entanto, casos de resistência detectados por DD devem ser confirmados por meio do método de MD, evitando, assim, resultados falsos resistentes, melhorando a eficácia e a segurança do tratamento.INTRODUCTION: Due to the ubiquity of nosocomial yeast infections of the genus Candida spp., studies in this area have become increasingly relevant. OBJECTIVE: To evaluate the antifungal Candida spp. susceptibility profile of hospitalized patients from the Regional University Hospital of Maringá-PR (HURM. MATERIAL AND METHODS: The samples were submitted to broth microdilution test (MD according to protocol M27-A3 in order to determine the minimum inhibitory concentration (MIC and disk

  4. Multicenter evaluation of the MB/BACT system for susceptibility testing of Mycobacterium tuberculosis.

    Science.gov (United States)

    Bemer, Pascale; Bodmer, Thomas; Munzinger, Juerg; Perrin, Monique; Vincent, Véronique; Drugeon, Henri

    2004-03-01

    The reliability of the MB/BACT system for susceptibility testing of Mycobacterium tuberculosis to pyrazinamide, rifampin, isoniazid, streptomycin, and ethambutol was compared to the BACTEC 460TB system. The proportion method was used to resolve discrepant results by an independent arbiter. Two interpretative methods were used, with an undiluted control (direct control) and a diluted control (10(-1) control). As no significant difference was observed between the two controls, the method with the direct control was adopted as the most accurate one. One hundred sixty-six strains were tested, with an overall agreement of 98.3%. After resolution of the 18 discrepant results by the proportion method, the sensitivity and specificity of the MB/BACT system were 100% for rifampin, isoniazid, and pyrazinamide. For ethambutol, sensitivity was 92.3% at the critical concentration and 33% at the high concentration, and specificity was 100% at both concentrations. For streptomycin, sensitivity was 100% at the critical concentration and 80% at the high concentration, and specificity was 98.6% at the critical concentration and 100% at the high concentration. The rifampin, isoniazid, streptomycin, and ethambutol susceptibility test results were obtained in 6.6 days with the MB/BACT versus 5 days with the BACTEC 460TB. The pyrazinamide susceptibility test results were obtained in 7.8 days with the MB/BACT, versus 6.7 days with the BACTEC 460TB. These data demonstrate that the fully automated MB/BACT system is a very reliable method for rapid susceptibility testing of M. tuberculosis against rifampin, isoniazid, and pyrazinamide. Sensitivity results have to be improved for ethambutol and streptomycin, especially at the high concentration.

  5. Electromagnetic Compatibility Testing for Conducted Susceptibility Along Interconnecting Signal Lines. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Ewing, P. D. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Wood, R. T. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Korsah, K. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Shourbaji, A. A. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Wilson, T. L. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Beets, B. M. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

    2002-07-31

    This document presents recommendations and the associated technical basis for addressing the effects of conducted electromagnetic interference (EMI) and radio-frequency interference (RFI) along interconnecting signal lines in safety-related instrumentation and control (I&C) systems. Oak Ridge National Laboratory has been engaged in assisting the U.S. Nuclear Regulatory Commission Office of Nuclear Regulatory Research in developing the technical basis for regulatory guidance on EMIIRFI immunity and power surge withstand capability (SWC). Previous research efforts have provided recommendations on (1) electromagnetic compatibility design and installation practices, (2) the endorsement of EMI/RFI and SWC test criteria and test methods, (3) the determination of ambient electromagnetic conditions at nuclear power plants, and (4) the development of recommended electromagnetic operating envelopes applicable to locations where safety-related I&C systems will be installed. The current research focuses on the susceptibility of l&C systems to conducted EMIIRFI along interconnecting signal lines. Coverage of signal line susceptibility was identified as an open issue in previous research on establishing the technical basis for EMIIRFI and SWC in safety-related I&C systems. Research results provided in this report will be used to establish the technical basis for endorsing U.S. Department of Defense and European Committee for Electrotechnical Standardization test criteria and test methods that address signal-line susceptibility. In addition, recommendations on operating envelopes are presented based on available technical information.

  6. Comparative erythromycin and tylosin susceptibility testing of streptococci from bovine mastitis.

    Science.gov (United States)

    Entorf, Monika; Feßler, Andrea T; Kaspar, Heike; Kadlec, Kristina; Peters, Thomas; Schwarz, Stefan

    2016-10-15

    Tylosin, a 16-membered macrolide, is - besides other indications - used for the treatment of bovine mastitis. So far, there is only limited information available on the tylosin susceptibility of streptococci isolated from mastitis. The aim of the present study was to comparatively investigate 303 streptococci from bovine mastitis, including 101 Streptococcus agalactiae, 100 Streptococcus dysgalactiae and 102 Streptococcus uberis, for their tylosin and erythromycin susceptibility by broth microdilution and agar disk diffusion. Both tests followed the recommendations of the Clinical and Laboratory Standards Institute (CLSI). For erythromycin, the results were interpreted using the CLSI-approved clinical breakpoints. Moreover, erythromycin-resistant isolates were tested for the presence of macrolide resistance genes and for inducible macrolide resistance. In general, both testing methods showed a good correlation for the three streptococcal species, although for the erythromycin susceptibility testing 11 S. uberis isolates fell into the very major error category. All but one of the erythromycin-resistant isolates harbored at least one macrolide resistance gene, with the erm(B) gene being most common. Moreover, single isolates of S. agalactiae and S. dysgalactiae proved to be inducibly macrolide-resistant. Since inducible macrolide resistance can easily switch to constitutive resistance, tylosin should not be used for the treatment of infections caused by inducibly resistant streptococci.

  7. American Society of Clinical Oncology Policy Statement Update: Genetic and Genomic Testing for Cancer Susceptibility.

    Science.gov (United States)

    Robson, Mark E; Bradbury, Angela R; Arun, Banu; Domchek, Susan M; Ford, James M; Hampel, Heather L; Lipkin, Stephen M; Syngal, Sapna; Wollins, Dana S; Lindor, Noralane M

    2015-11-01

    The American Society of Clinical Oncology (ASCO) has long affirmed that the recognition and management of individuals with an inherited susceptibility to cancer are core elements of oncology care. ASCO released its first statement on genetic testing in 1996 and updated that statement in 2003 and 2010 in response to developments in the field. In 2014, the Cancer Prevention and Ethics Committees of ASCO commissioned another update to reflect the impact of advances in this area on oncology practice. In particular, there was an interest in addressing the opportunities and challenges arising from the application of massively parallel sequencing-also known as next-generation sequencing-to cancer susceptibility testing. This technology introduces a new level of complexity into the practice of cancer risk assessment and management, requiring renewed effort on the part of ASCO to ensure that those providing care to patients with cancer receive the necessary education to use this new technology in the most effective, beneficial manner. The purpose of this statement is to explore the challenges of new and emerging technologies in cancer genetics and provide recommendations to ensure their optimal deployment in oncology practice. Specifically, the statement makes recommendations in the following areas: germline implications of somatic mutation profiling, multigene panel testing for cancer susceptibility, quality assurance in genetic testing, education of oncology professionals, and access to cancer genetic services.

  8. In vitro susceptibility of 188 clinical and environmental isolates of Aspergillus flavus for the new triazole isavuconazole and seven other antifungal drugs

    NARCIS (Netherlands)

    Shivaprakash, M.R.; Geertsen, E.; Chakrabarti, A.; Mouton, J.W.; Meis, J.F.G.M.

    2011-01-01

    Recently isavuconazole, an experimental triazole agent, was found to be active against Aspergillus species. As Aspergillus flavus is the second-most common Aspergillus species isolated from human infection and the fungus has not been widely tested against the drug, we studied a large collection of c

  9. Emergence of non-albicans Candida species and antifungal resistance in intensive care unit patients

    Institute of Scientific and Technical Information of China (English)

    Ravinder Kaur; Megh Singh Dhakad; Ritu Goyal; Rakesh Kumar

    2016-01-01

    Objective: To evaluate the epidemiology of candidiasis and the antifungal susceptibility profile of Candida species isolated from the intensive care unit (ICU) patients. Methods: The study used a qualitative descriptive design. Relevant samples depending on organ system involvement from 100 ICU patients were collected and processed. Identification and speciation of the isolates was conducted by the biochemical tests. Antifungal susceptibility testing was carried out as per CLSI-M27-A3 document. Results: Ninety Candida isolates were isolated from the different clinical samples:urine (43.3%), tracheal aspirate (31.1%), urinary catheter (12.2%), endotracheal tube (7.8%), abdominal drains (3.3%), sputum (2.2%). The incidence of candidiasis caused by non-albicans Candida (NAC) species (63.3%) was higher than Candida albicans (36.7%). The various NAC species were isolated as: Candida tropicalis (41.1%), Candida glab-rata (10%), Candida parapsilosis (6.7%), Candida krusei (3.3%) and Candida kefyr (2.2%). The overall isolation rate of Candida species from samples was 53.3%. Anti-fungal susceptibility indicated that 37.8%and 7.8%of the Candida isolates were resistant to fluconazole and amphotericin B, respectively. Conclusions: Predominance of NAC species in ICU patients along with the increasing resistance being recorded to fluconazole which has a major bearing on the morbidity and management of these patients and needs to be further worked upon.

  10. Emergence of non-albicans Candida species and antifungal resistance in intensive care unit patients

    Institute of Scientific and Technical Information of China (English)

    Ravinder Kaur; Megh Singh Dhakad; Ritu Goyal; Rakesh Kumar

    2016-01-01

    Objective: To evaluate the epidemiology of candidiasis and the antifungal susceptibility profile of Candida species isolated from the intensive care unit(ICU) patients.Methods: The study used a qualitative descriptive design. Relevant samples depending on organ system involvement from 100 ICU patients were collected and processed.Identification and speciation of the isolates was conducted by the biochemical tests.Antifungal susceptibility testing was carried out as per CLSI-M27-A3 document.Results: Ninety Candida isolates were isolated from the different clinical samples: urine(43.3%), tracheal aspirate(31.1%), urinary catheter(12.2%), endotracheal tube(7.8%),abdominal drains(3.3%), sputum(2.2%). The incidence of candidiasis caused by nonalbicans Candida(NAC) species(63.3%) was higher than Candida albicans(36.7%).The various NAC species were isolated as: Candida tropicalis(41.1%), Candida glabrata(10%), Candida parapsilosis(6.7%), Candida krusei(3.3%) and Candida kefyr(2.2%). The overall isolation rate of Candida species from samples was 53.3%. Antifungal susceptibility indicated that 37.8% and 7.8% of the Candida isolates were resistant to fluconazole and amphotericin B, respectively.Conclusions: Predominance of NAC species in ICU patients along with the increasing resistance being recorded to fluconazole which has a major bearing on the morbidity and management of these patients and needs to be further worked upon.

  11. Comparison of the Etest and a rapid flow cytometry-based method with the reference CLSI broth microdilution protocol M27-A3 for the echinocandin susceptibility testing of Candida spp.

    Science.gov (United States)

    Vale-Silva, L A; Pinto, P; Lopes, V; Ramos, H; Pinto, E

    2012-06-01

    Reference broth microdilution protocols for the antifungal susceptibility testing (AST) of yeasts are available, but routine AST relies more on simpler alternatives. In this work, the Etest and a novel flow cytometry (FC) method were compared to the Clinical Laboratory Standards Institute (CLSI) protocol M27-A3. Caspofungin and anidulafungin were tested against 60 clinical isolates of Candida glabrata, C. krusei, and C. parapsilosis. There were two intermediate susceptibility results and 19 out of 20 tested C. krusei strains were categorized as resistant to caspofungin (minimum inhibitory concentration [MIC] of 1.0 mg/L). There was a generally excellent essential agreement with the reference method, only interrupted by Etest results for the anidulafungin susceptibility of C. glabrata (80%) and the FC method's results for caspofungin with C. krusei (40%). Categorical agreements were excellent, with the notable exception of the caspofungin-resistant C. krusei, with 19 very major errors for the FC method and 19 minor errors plus one very major error for the Etest (5% and 0% categorical agreements, respectively). Two additional minor errors were registered for the FC method when testing C. parapsilosis with anidulafungin and caspofungin. Overall, these data come to question the suitability of recently approved clinical breakpoints in the case of C. krusei. Further studies including fks mutants are now required.

  12. Effect of Susceptibility Testing Conditions on the In Vitro Antibacterial Activity of ETX0914.

    Science.gov (United States)

    Giacobbe, Robert A; Huband, Michael D; deJonge, Boudewijn L M; Bradford, Patricia A

    2017-02-01

    The effect of various conditions including pH, inoculum, temperature, atmosphere, divalent cations, and several body fluids on the in vitro activity of the novel antibacterial spiropyrimidinetrione ETX0914 in standard susceptibility tests was investigated against several species. None of the parameters investigated affected the activity of ETX0914, with the exception of pH. Whereas the MIC values for ETX0914 with S. aureus, E. faecalis, and E. coli did not change when the pH of the growth medium was varied from 5 to 8, they did increase at least 8-fold at pH values above 8. This loss of activity can be attributed to the deprotonation of the molecule at elevated pH. The data suggest that routine susceptibility testing with ETX0914 should result in reproducible MIC values.

  13. What's in a Name? The Impact of Accurate Staphylococcus pseudintermedius Identification on Appropriate Antimicrobial Susceptibility Testing.

    Science.gov (United States)

    Limbago, Brandi M

    2016-03-01

    Bacteria in the Staphylococcus intermedius group, including Staphylococcus pseudintermedius, often encode mecA-mediated methicillin resistance. Reliable detection of this phenotype for proper treatment and infection control decisions requires that these coagulase-positive staphylococci are accurately identified and specifically that they are not misidentified as S. aureus. As correct species level bacterial identification becomes more commonplace in clinical laboratories, one can expect to see changes in guidance for antimicrobial susceptibility testing and interpretation. The study by Wu et al. in this issue (M. T. Wu, C.-A. D. Burnham, L. F. Westblade, J. Dien Bard, S. D. Lawhon, M. A. Wallace, T. Stanley, E. Burd, J. Hindler, R. M. Humphries, J Clin Microbiol 54:535-542, 2016, http://dx.doi.org/10.1128/JCM.02864-15) highlights the impact of robust identification of S. intermedius group organisms on the selection of appropriate antimicrobial susceptibility testing methods and interpretation.

  14. Isolation,Identification and Antibiotics Susceptibility Test of Citrobacter freundii from Procambarus clarkia

    Institute of Scientific and Technical Information of China (English)

    Chen; Honglian; Song; Guangtong; He; Jixiang; Hou; Guanjun; Wang; Yongjie

    2014-01-01

    This experiment was conducted to clarify species and drug resistance of pathogen from the diseased Procambarus clarkia. Pathogenic bacteria from hepatopancreas of the diseased P. clarkia were examined using conventional methods,and then were isolated. The further tests and analysis of the isolated strain were developed,including the regression experiment to P. clarkia,the morphology,physiological and biochemical characteristics,sequence analysis of their 16 S rRNA and gyr B genes,and the susceptibility test to antibiotics. Large colonies with similar morphology and color were obtained. Strain X120523 was identified as Citrobacter freundii,proved to have strong pathogenicity,and was susceptible to quinolones and aminoglycosides.

  15. Antimicrobial susceptibility testing of 230 Helicobacter pylori strains: importance of medium, inoculum, and incubation time

    DEFF Research Database (Denmark)

    Hartzen, S H; Andersen, L P; Bremmelgaard, A;

    1997-01-01

    No standardized method of susceptibility testing for Helicobacter pylori is currently available, so before a large agar dilution study comprising 230 H. pylori strains belonging to more than 80 genetically different groups was initiated, we performed a relatively small preliminary study....../spot) as the inoculum and 72 h as the incubation time. A modest inoculum effect was noted for amoxicillin and metronidazole. By the methodology derived from our preliminary study, the susceptibilities of 230 H. pylori strains to six antibiotics were subsequently determined. The results were generally in accord...... with those of others, and apart from metronidazole, the MIC of which for approximately 25% of the strains tested was >8 microg/ml, resistance was low in Denmark. The situation might, however, quickly change when and if the number of indications for antibiotic therapy for H. pylori infections increase...

  16. Prevalence rates and antifungal susceptibility profiles of the Candida parapsilosis species complex: results from a nationwide surveillance of candidaemia in Brazil

    OpenAIRE

    Goncalves, S. S. [UNIFESP; Amorim, C. S.; M. Nucci; Padovan, Ana Carolina Barbosa [UNIFESP; Briones, Marcelo Ribeiro da Silva; Melo, A. S. A.; Colombo, Arnaldo Lopes

    2010-01-01

    P>The genetically heterogeneous taxon Candida parapsilosis was recently reclassified into three species: Candida parapsilosis, Candida orthopsilosis and Candida metapsilosis. the prevalences of these species among 141 bloodstream isolates tested in Brazil were 88% for C. parapsilosis, 9% for C. orthopsilosis, and 3% for C. metapsilosis. Except for three C. orthopsilosis isolates that were considered resistant to 5-flucytosine, all isolates representing the different species of this complex we...

  17. Antimicrobial susceptibility testing of newer quinolones against gram positive and gram negative clinical isolates.

    Science.gov (United States)

    Iffat, Wajiha; Shoaib, Muhammad Harris; Muhammad, Iyad Naeem; Rehana; Tasleem, Samiah; Gauhar, Shahnaz

    2010-07-01

    Antibiotic resistance development is an ongoing process associated with irrational antibiotic use. WHO recommends regular surveillance programs for monitoring of antibiotic resistance. The present study is a step in this direction. A total of 124 clinical isolates of Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae and Pseudomonas aeruginosa were collected from different hospitals in Karachi. In vitro antimicrobial susceptibility studies were carried out by agar dilution method using newer quinolones that included Gatifloxacin and Levofloxacin. It was observed that 50% (n=30) isolates of Staphylococcus aureus were resistant to gatifloxacin. Gatifloxacin was more active against Pseudomonas aeruginosa (n=23) and showing complete susceptibility with MIC 1mg/L except for three very resistant strains that shown resistance at even higher concentrations. Escherichia coli (n=45) has shown 15.5% and Klebsiella pneumoniae (n=26) 34.61% resistance to gatifloxacin. Levofloxacin was more active against Staphylococcus aureus and Escherichia coli showing complete susceptibility at 0.5 mg /L concentration. Pseudomonas aeruginosa and Klebsiella pneumoniae were found to be resistant to Levofloxacin showing 36.36% and 23.08% resistance respectively. The study strongly recommends the adherence to the antibiotic policy and regular susceptibility testing to overcome the problem associated with antimicrobial resistance.

  18. New method for rapid Susceptibility Testing on blood culture with HB&L system: preliminary data

    Directory of Open Access Journals (Sweden)

    Vincenzo Rondinelli

    2010-12-01

    Full Text Available Blood culture, although represents the gold standard in detecting the ethiological agent of sepsis, is rather rarely required in relation to the real diagnostic importance. The result of this test depends in fact on many factors (sample volume, time of collection, accuracy, antibiotic therapy, contamination, number of drawings, drawing site, interpretation difficulties, etc. that are often considered by many clinicians so limited as to doubt about their actual value. The disadvantages are therefore represented by the lack of standardization but also by the low sensitivity and above all by the technical times too long for the clinical needs. Blood culture begins with the drawing of samples from the “septic” patient followed incubation of the bottles in automatic thermostated systems. In case of positive result (36 hours, the culture is Gram stained and streaked on solid media in order to obtain isolated colonies for the identification and the susceptibility testing (48 hours from positive result. The long time required for pathogen identification and susceptibility testing involves empirical broad spectrum antibiotic therapy that can promote the increase of bacterial resistance but also patient management costs. A clinically useful report should be available on short notice in order to guide the clinician to choose the most appropriate antibiotic. The microbiologist has therefore the hard work of reviewing the organization and the management of the procedures.We have therefore started to consider the possibility of treating the blood as an biological liquid in order to quickly determine the susceptibility of bacteria to antibiotics.

  19. Study on hydrogen assisted cracking susceptibility of HSLA steel by implant test

    Directory of Open Access Journals (Sweden)

    Gopa Chakraborty

    2016-12-01

    Full Text Available DMR-249A is an indigenously developed high strength low alloy steel for Indian ship building industry for making ship-hull and is extensively used in the construction of war ships and submarines. Welding electrodes conforming to SFA 5.5 AWS E8018 C1 has been indigenously developed for welding of this steel using shielded metal arc welding process. In the present study, susceptibility to hydrogen assisted cracking of DMR-249A steel welds made using this electrode has been assessed using implant test. Implant tests were conducted using this electrode at two different levels of diffusible hydrogen, measured using gas chromatography technique. It is observed that both the steel and the welding consumable are not susceptible to hydrogen assisted cracking even with a high diffusible hydrogen level of 9 mL/100g of weld metal. In implant tests, specimen did not fracture even after loading to stress levels higher than the yield strength of the base metal. The good resistance of this steel and the welding consumable, even with high levels of diffusible hydrogen, is attributed to absence of a susceptible microstructure in both the weld metal and heat affected zone. Hence, this study shows that, in the absence of a susceptible microstructure, hydrogen assisted cracking is unlikely to occur even if hydrogen level is high. It also confirms that in welding of DMR-249A with indigenously developed E8018 C1 electrode, hydrogen assisted cracking is not a concern and no preheating is required to avoid it during welding.

  20. Study on hydrogen assisted cracking susceptibility of HSLA steel by implant test

    Institute of Scientific and Technical Information of China (English)

    Gopa CHAKRABORTY; R. REJEESH; S.K. ALBERT

    2016-01-01

    DMR-249A is an indigenously developed high strength low alloy steel for Indian ship building industry for making ship-hull and is extensively used in the construction of war ships and submarines. Welding electrodes conforming to SFA 5.5 AWS E8018 C1 has been indigenously developed for welding of this steel using shielded metal arc welding process. In the present study, susceptibility to hydrogen assisted cracking of DMR-249A steel welds made using this electrode has been assessed using implant test. Implant tests were conducted using this electrode at two different levels of diffusible hydrogen, measured using gas chromatography technique. It is observed that both the steel and the welding consumable are not susceptible to hydrogen assisted cracking even with a high diffusible hydrogen level of 9 mL/100g of weld metal. In implant tests, specimen did not fracture even after loading to stress levels higher than the yield strength of the base metal. The good resistance of this steel and the welding consumable, even with high levels of diffusible hydrogen, is attributed to absence of a susceptible microstructure in both the weld metal and heat affected zone. Hence, this study shows that, in the absence of a susceptible microstructure, hydrogen assisted cracking is unlikely to occur even if hydrogen level is high. It also confirms that in welding of DMR-249A with indigenously developed E8018 C1 electrode, hydrogen assisted cracking is not a concern and no preheating is required to avoid it during welding.

  1. Self-assembled magnetic bead biosensor for measuring bacterial growth and antimicrobial susceptibility testing.

    Science.gov (United States)

    Kinnunen, Paivo; McNaughton, Brandon H; Albertson, Theodore; Sinn, Irene; Mofakham, Sima; Elbez, Remy; Newton, Duane W; Hunt, Alan; Kopelman, Raoul

    2012-08-20

    Bacterial antibiotic resistance is one of the major concerns of modern healthcare worldwide, and the development of rapid, growth-based, antimicrobial susceptibility tests is key for addressing it. The cover image shows a self-assembled asynchronous magnetic bead rotation (AMBR) biosensor developed for rapid detection of bacterial growth. Using the biosensors, the minimum inhibitory concentration of a clinical E. coli isolate can be measured within two hours, where currently tests take 6-24 hours. A 16-well prototype is also constructed for simple and robust observation of the self-assembled AMBR biosensors.

  2. Antimicrobial Susceptibility Testing for Escherichia coli Strains to Fluoroquinolones, in Urinary Tract Infections

    Directory of Open Access Journals (Sweden)

    FA Nakhjavani

    2007-05-01

    Full Text Available Background: Urinary Tract Infections (UTIs are one of the most common infectious diseases diagnosed all over the world. Meanwhile most episode of UTIs are caused by Escherichia coli (up to 85% and frequently fluoroquinolones are preferred as initial agents for empiric therapy of UTIs. Widespread use of fluoroquinolones has resulted in an increasing incidence of resistance these agents all over the world. The aim of this study was to assess, susceptibility of Escherichia coli strains from UTI patients against common fluoroquinolones. Methods: Antimicrobial susceptibility testing was determined by disk agar diffusion (DAD and Minimal Inhibitory Concentration methods as described by the National Committee for Clinical Laboratory Standards (NCCLS. Results: One hundred sixty four clinical isolates of E. coli were collected by urine cultures from patients with UTI. The extent of resistant to nalidixic acid, ofloxacin, norfloxacin and ciprofloxacin, by disk diffusion method was 49.3%, 44.5%, 41.4% and 40.2%, respectively. Resistance to ciprofloxacin by MIC method was 4.9%. Conclusion: This study represents high level resistant of E. coli isolates from UTI patients. It is because of inappropriate and incorrect administration of antimicrobial agents in blind cases. This problem remarks significance of performing antimicrobial susceptibility testing before empiric antibiotic therapy. To overcome this problem use of unnecessary antibiotics therapy should be limited.

  3. Plant latex: a promising antifungal agent for post harvest disease control.

    Science.gov (United States)

    Sibi, G; Wadhavan, Rashmi; Singh, Sneha; Shukla, Abhilasha; Dhananjaya, K; Ravikumar, K R; Mallesha, H

    2013-12-01

    Bioactive compounds from plant latex are potential source of antifungic against post harvest pathogens. Latex from a total of seven plant species was investigated for its phytochemical and antifungal properties. Six fungi namely Aspergillus fumigatus, A. niger, A. terreus, F. solani, P. digitatum and R. arrhizus were isolated from infected fruits and vegetables and tested against various solvent extracts of latex. Analysis of latex extracts with phytochemical tests showed the presence of alkaloids, flavonoids, glycosides, phenols, saponins, steroids, tannins and terpenoids. Antifungal assay revealed the potential inhibitory activity of petroleum ether extracts against the postharvest fungal isolates. Various degree of sensitivity was observed irrespective of plant species studied with A. terreus and P. digitatum as the most susceptible ones. F. solani and A. fumigatus were moderately sensitive to the latex extracts tested. Among the plants, latex of Thevetia peruviana (75.2%) and Artocarpus heterophyllus (64.8%) were having potential antifungal activity against the isolates followed by Manilkara zapota (51.1%). In conclusion, use of plant latex makes interest to control postharvest fungal diseases and is fitting well with the concept of safety for human health and environment.

  4. Candidemia em um hospital terciário brasileiro: incidência, freqüência das diferentes espécies, fatores de risco e suscetibilidade aos antifúngicos Candidemia in a Brazilian tertiary care hospital: incidence, frequency of different species, risk factors and antifungal susceptibility

    Directory of Open Access Journals (Sweden)

    João Cesar Beenke França

    2008-02-01

    Full Text Available São apresentados os resultados de estudo transversal e observacional sobre candidemia realizado no Hospital de Clínicas da Universidade Federal do Paraná. No período de janeiro de 2001 a dezembro de 2004, foram analisados 100 episódios de candidemia. A incidência foi de 1,27 por 1.000 internações, sendo Candida spp o oitavo agente mais isolado nas infecções da corrente sanguínea. A idade variou de 5 dias a 89 anos com uma média de 32 anos, 60% dos casos ocorreram em adultos (66% > 50 anos e 40% em crianças (52% Results from a cross-sectional observational study on candidemia conducted at Hospital de Clínicas, Federal University of Paraná, are presented. From January 2001 to December 2004, one hundred candidemia cases were evaluated. The incidence was 1.27 episodes per 1,000 hospitalizations and Candida was the eighth most frequently isolated agent from bloodstream infections. The patients’ ages ranged from five days to 89 years, with a mean of 32 years. Sixty percent of the cases occurred in adults (66% > 50 years old and 40% in children (52% < one year old. Fifty-nine percent had been admitted to medical wards and 41% to the intensive care unit. Candida albicans was the most (59% frequent species followed by Candida tropicalis (15% and Candida parapsilosis (9%. The most (97% prevalent coexisting conditions were the use of antibacterial drugs, central venous catheter (77%, H2 blockers (57%, total parenteral nutrition (49% and admission to the intensive care unit (41%. Out of the 51 isolates tested, three of Candida glabrata presented dose-dependent susceptibility to fluconazole and resistance to itraconazole. One sample of Candida krusei presented dose-dependent susceptibility to fluconazole and one of Candida pelliculosa presented dose-dependent susceptibility to itraconazole. Among the study population, 68% received antifungal therapy, but the mortality was 56%.

  5. Preliminary indications for antibiotic susceptibility tests in less than six hour in positive blood cultures

    Directory of Open Access Journals (Sweden)

    Vesselina Kroumova

    2010-03-01

    Full Text Available A rapid determination of the antibiotic susceptibility patterns of pathogens responsible for sepsis represents a significant milestone for a timely correct antibiotic therapy.The system HB&L® (ALIFAX allows reduced time in the detection of bacterial growth and consequently is able to detect the growth or absence of certain microorganisms in the presence of a given antibiotic. In this study three system for rapid antibiotic susceptibility tests among bacteria isolated from blood were compared: HB&L® (ALIFAX,VITEK®2 (bioMérieux and essays Etest® (bioMérieux. Present findings indicate that HB&L® (ALIFAX is rapid reliable instrument that may support the clinician for a rapid and appropriate treatment, particularly in the critical patient.

  6. Rapid antimicrobial susceptibility testing with electrokinetics enhanced biosensors for diagnosis of acute bacterial infections.

    Science.gov (United States)

    Liu, Tingting; Lu, Yi; Gau, Vincent; Liao, Joseph C; Wong, Pak Kin

    2014-11-01

    Rapid pathogen detection and antimicrobial susceptibility testing (AST) are required in diagnosis of acute bacterial infections to determine the appropriate antibiotic treatment. Molecular approaches for AST are often based on the detection of known antibiotic resistance genes. Phenotypic culture analysis requires several days from sample collection to result reporting. Toward rapid diagnosis of bacterial infection in non-traditional healthcare settings, we have developed a rapid AST approach that combines phenotypic culture of bacterial pathogens in physiological samples and electrochemical sensing of bacterial 16S rRNA. The assay determines the susceptibility of pathogens by detecting bacterial growth under various antibiotic conditions. AC electrokinetic fluid motion and Joule heating induced temperature elevation are optimized to enhance the sensor signal and minimize the matrix effect, which improve the overall sensitivity of the assay. The electrokinetics enhanced biosensor directly detects the bacterial pathogens in blood culture without prior purification. Rapid determination of the antibiotic resistance profile of Escherichia coli clinical isolates is demonstrated.

  7. In vitro susceptibility testing of fluoroquinolone activity against Salmonella: recent changes to CLSI standards.

    Science.gov (United States)

    Humphries, Romney M; Fang, Ferric C; Aarestrup, Frank M; Hindler, Janet A

    2012-10-01

    Fluoroquinolone (FQ) resistance in Salmonella enterica is a significant clinical concern. Recognition of resistance by the clinical laboratory is complicated by the multiple FQ resistance mechanisms found in Salmonella. The Clinical Laboratory Standards Institute (CLSI) recently addressed this issue by revising the ciprofloxacin break points for Salmonella species. It is critical for clinicians and laboratory workers to be aware of the multiple technical issues surrounding these revised break points. In this article, we review FQ resistance mechanisms in Salmonella, their clinical significance, and data supporting the revised ciprofloxacin break points. We encourage clinical laboratories to adopt the revised CLSI ciprofloxacin break points for all Salmonella isolates in which susceptibility testing is indicated and discuss the technical issues for laboratories using commercial antimicrobial susceptibility systems.

  8. Patients' understanding of genetic susceptibility testing in mainstream medicine: qualitative study on thrombophilia

    Directory of Open Access Journals (Sweden)

    Shepherd Maggie H

    2007-06-01

    Full Text Available Abstract Background UK and US policy initiatives have suggested that, in the future, patients and clinicians in mainstream medicine could use genetic information to prevent common illnesses. There are no studies on patients' experience and understanding of the process of testing for common genetic susceptibilities in mainstream medicine. Methods Qualitative interviews with 42 individuals who had undergone testing for a genetic susceptibility for deep vein thrombosis in primary and secondary care in the UK. Results Some participants, often from higher social classes, had a good understanding of the test and its implications. They had often sought additional information on thrombophilia from relatives and from the Internet. Others, often from less privileged backgrounds, had a poorer understanding of the test – seven individuals were unaware of having had the genetic test. Features of genetic information led to misunderstandings: (i at referral, (ii when communicating results, and (iii when making sense of the implications of testing. Participants' accounts indicated that non-specialist doctors may feel obliged to refer a patient for a genetic test they know little about, because a patient requests it after a relative had tested positive. Sometimes a referral for a genetic test was lost under information overload when multiple tests and issues were considered. The inconsistent and informal ways of communicating test results – for example by phone – in mainstream medicine also led to confusion. Participants did not generally overestimate their risk, but some were uncertain about whether they were taking the right preventive actions and/or whether their children were at risk. Information about genetic susceptibilities was difficult to make sense of, as it related to ambiguous risks for participants and family members, complicated and unfamiliar terminology and multiple genes and preventive strategies. Conclusion Policy visions of clinicians

  9. Classical against molecular-genetic methods for susceptibility testing of antituberculotics.

    Science.gov (United States)

    Porvaznik, I; Mokry, J; Solovic, I

    2015-01-01

    Tuberculosis currently belongs to rare respiratory diseases in Slovakia. However, the emergence and spread of multi-drug resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) are major challenges for global tuberculosis control, since the treatment of resistant forms creates both medical and financial problems. Cultivation methods of diagnosis are time-consuming, many times exceeding the time of the initial phase of tuberculosis treatment. Therefore, in the presented study we compared the standard procedures, based on the cultivation of mycobacteria and subsequent drug susceptibility testing to antituberculotics, with molecular-genetic methods using PCR diagnostic kits. The molecular-genetic testing enables to obtain direct and fast evidence of Mycobacterium tuberculosis, with genomic verification of resistance to the most important anti-tuberculosis drugs - isoniazid and rifampicin in MDR-TB, and ethambutol, aminoglycosides, and fluoroquinolones in XDR-TB. In 2012-2013, we confirmed 19 cases of drug-resistant tuberculosis in Slovakia. The resistance to rifampicin was confirmed in all strains with both methods. In two cases, the molecular-genetic testing did not show resistance to isoniazid, as confirmed by conventional cultivation. Furthermore, two strains demonstrating susceptibility in conventional microbiological testing to ethambutol and five strains to fluoroquinolones were verified as actually being resistant using a PCR method. Rapid diagnosis and identification of MDR-TB or XDR-TB strains using molecular-genetic testing is an essential tool for the timely and appropriate drug treatment and prevention of spread of drug resistant strains.

  10. Results from the ARTEMIS DISK Global Antifungal Surveillance Study, 1997 to 2007: a 10.5-Year Analysis of Susceptibilities of Candida Species to Fluconazole and Voriconazole as Determined by CLSI Standardized Disk Diffusion ▿

    OpenAIRE

    Pfaller, M.A.; Diekema, D. J.; Gibbs, D. L.; Newell, V. A.; Ellis, D.; Tullio, V; Rodloff, A.; Fu, W; Ling, T. A.

    2010-01-01

    Fluconazole in vitro susceptibility test results for 256,882 isolates of Candida spp. were collected from 142 sites in 41 countries from June 1997 to December 2007. Data were collected for 197,619 isolates tested with voriconazole from 2001 to 2007. A total of 31 different species of Candida were isolated. Increased rates of isolation of the common non-albicans species C. glabrata (10.2% to 11.7%), C. tropicalis (5.4% to 8.0%), and C. parapsilosis (4.8% to 5.6%) were noted when the time perio...

  11. Assessment of Etest as an alternative to agar dilution for antimicrobial susceptibility testing of Neisseria gonorrhoeae.

    Science.gov (United States)

    Liu, Hsi; Taylor, Thomas H; Pettus, Kevin; Trees, David

    2014-05-01

    We studied whether the Etest can be used as an alternative to agar dilution to determine antimicrobial susceptibilities of ceftriaxone, cefixime, and cefpodoxime in Neisseria gonorrhoeae surveillance. One hundred fifteen clinical and laboratory isolates of N. gonorrhoeae were tested following the Clinical Laboratory Improvement Amendments (CLIA)-approved CLSI standard agar dilution method and, separately, by the Etest according to the manufacturer's recommendations. The MICs were determined and compared. Ten laboratory-generated mutants were used to simulate substantially nonsusceptible specimens. The Etest and agar dilution methods were well correlated. Statistical tests produced regression R2 values of 88%, 82%, and 85% and Pearson correlation coefficients of 92%, 91%, and 92% for ceftriaxone, cefixime, and cefpodoxime, respectively. When paired comparisons were made, the two tests were 88.7%, 80%, and 87% within 1 log2 dilution from each other for ceftriaxone, cefixime, and cefpodoxime, respectively. The within-2-log2 agreements were 99.1%, 98.3%, and 94.8% for ceftriaxone, cefixime, and cefpodoxime, respectively. Notwithstanding the good correlations and the within-2-log2 general agreement, the Etest results produced slightly lower MICs than the agar dilution results. In conclusion, we found that the Etest can be effectively used as an alternative to agar dilution testing to determine the susceptibility of N. gonorrhoeae to ceftriaxone, cefixime, and cefpodoxime, although we recommend further research into extremely resistant isolates. For isolates within the typical range of clinical MICs, reexamination of the Etest interpretation of susceptible and nonsusceptible categories would likely allow for successful transition from agar dilution to the Etest.

  12. Direct Susceptibility Testing of Mycobacterium tuberculosis for Pyrazinamide by Use of the Bactec MGIT 960 System.

    Science.gov (United States)

    Demers, Anne-Marie; Venter, Amour; Friedrich, Sven O; Rojas-Ponce, Gabriel; Mapamba, Daniel; Jugheli, Levan; Sasamalo, Mohammed; Almeida, Deepak; Dorasamy, Afton; Jentsch, Ute; Gibson, Mara; Everitt, Daniel; Eisenach, Kathleen D; Diacon, Andreas H

    2016-05-01

    Pyrazinamide (PZA) is a key antituberculosis drug, yet no rapid susceptibility test is commercially available. PZA drug susceptibility testing (DST) was performed directly on sputum samples from 327 patients and compared with the indirect method by using the Bactec MGIT 960 system in the context of patient screening for participation in a drug trial. Compared to standard indirect PZA DST, direct DST was successful in only 59% of cases, but results obtained were highly accurate and available faster. Agreement between the direct and indirect methods varied from 90 to 100% in each laboratory. The median times for obtaining PZA results from the time when the specimen was collected ranged from 11 to 16 days for the direct test and 18 to 95 days for the indirect test across laboratories. The direct method is accurate and reproducible across laboratories. It can be expected to accelerate results in >50% of cases, but it cannot replace indirect DST for PZA. Phenotypic methods remain the gold standard for DST in drug trials. If future studies can optimize the method to decrease the number of uninterpretable results, direct MGIT DST could be the new phenotypic DST standard for clinical trials, providing more rapid detection of resistance to new drugs in experimental regimens.

  13. Direct Susceptibility Testing of Mycobacterium tuberculosis for Pyrazinamide by Use of the Bactec MGIT 960 System

    Science.gov (United States)

    Demers, Anne-Marie; Venter, Amour; Friedrich, Sven O.; Rojas-Ponce, Gabriel; Mapamba, Daniel; Jugheli, Levan; Sasamalo, Mohammed; Almeida, Deepak; Dorasamy, Afton; Jentsch, Ute; Gibson, Mara; Everitt, Daniel; Diacon, Andreas H.

    2016-01-01

    Pyrazinamide (PZA) is a key antituberculosis drug, yet no rapid susceptibility test is commercially available. PZA drug susceptibility testing (DST) was performed directly on sputum samples from 327 patients and compared with the indirect method by using the Bactec MGIT 960 system in the context of patient screening for participation in a drug trial. Compared to standard indirect PZA DST, direct DST was successful in only 59% of cases, but results obtained were highly accurate and available faster. Agreement between the direct and indirect methods varied from 90 to 100% in each laboratory. The median times for obtaining PZA results from the time when the specimen was collected ranged from 11 to 16 days for the direct test and 18 to 95 days for the indirect test across laboratories. The direct method is accurate and reproducible across laboratories. It can be expected to accelerate results in >50% of cases, but it cannot replace indirect DST for PZA. Phenotypic methods remain the gold standard for DST in drug trials. If future studies can optimize the method to decrease the number of uninterpretable results, direct MGIT DST could be the new phenotypic DST standard for clinical trials, providing more rapid detection of resistance to new drugs in experimental regimens. PMID:26912751

  14. Rapid susceptibility testing of Mycobacterium tuberculosis by bioluminescence assay of mycobacterial ATP

    Energy Technology Data Exchange (ETDEWEB)

    Nilsson, L.E.; Hoffner, S.E.; Ansehn, S.

    1988-08-01

    Mycobacterial growth was monitored by bioluminescence assay of mycobacterial ATP. Cultures of Mycobacterium tuberculosis H37Rv and of 25 clinical isolates of the same species were exposed to serial dilutions of ethambutol, isoniazid, rifampin, and streptomycin. A suppression of ATP, indicating growth inhibition, occurred for susceptible but not resistant strains within 5 to 7 days of incubation. Breakpoint concentrations between susceptibility and resistance were determined by comparing these results with those obtained by reference techniques. Full agreement was found in 99% of the assays with the resistance ratio method on Lowenstein-Jensen medium, and 98% of the assays were in full agreement with the radiometric system (BACTEC). A main advantage of the bioluminescence method is its rapidity, with results available as fast as with the radiometric system but at a lower cost and without the need for radioactive culture medium. The method provides kinetic data concerning drug effects within available in vivo drug concentrations and has great potential for both rapid routine susceptibility testing and research applications in studies of drug effects on mycobacteria.

  15. Genetic antimicrobial susceptibility testing in Gram-negative sepsis - impact on time to results in a routine laboratory.

    Science.gov (United States)

    Kommedal, Øyvind; Aasen, Johanne Lind; Lindemann, Paul Christoffer

    2016-07-01

    Diagnostic testing of positive blood cultures is among the most critical tasks performed by clinical microbiology laboratories, and the total analysis time from sampling to results should be kept as short as possible. By providing identification of pelleted bacteria directly from positive blood-cultures, MALDI-TOF MS opens for relatively low-complex species-adjusted genetic susceptibility testing from the same bacterial pellet. In our lab routine, we prospectively evaluated a rapid in-house real-time PCR targeting the most common aminoglycoside and cephalosporin resistance genes in Escherichia coli and Klebsiella pneumoniae and measured time to preliminary susceptibility reporting for 138 samples. The results were compared to direct phenotypic susceptibility testing with interpretation after 6 h and overnight incubation respectively. Results from the genetic susceptibility testing were available for 69.5% (96/138) of the positive blood cultures within 24 h after sample collection. No phenotypic susceptibility results were available at this time. Compared to overnight direct susceptibility testing, the average time from sample collection to preliminary susceptibility reporting was reduced with 43%, from 45 h and 5 min to 25 h and 44 min, providing an earlier adjustment of antimicrobial therapy for 12 patients. Minor logistic adjustments have the potential to save yet another 4 h.

  16. Risk perceptions, worry, and attitudes about genetic testing for breast cancer susceptibility.

    Science.gov (United States)

    Cameron, Linda D; Reeve, Jeanne

    2006-01-01

    This study assessed the unique associations of risk perceptions and worry with attitudes about genetic testing for breast cancer susceptibility. Women (general practitioner clinic attenders, university students, and first-degree relatives of breast cancer survivors; N = 303) read information about genetic testing and completed measures assessing perceived cancer risk, cancer worry, and genetic testing attitudes and beliefs. Worry was associated with greater interest in genetic testing, stronger beliefs that testing has detrimental emotional consequences, and positive beliefs about benefits of testing and risk-reducing surgeries. Perceived risk was unrelated to interest and associated with more skeptical beliefs about emotional consequences and benefits of testing and risk-reducing surgeries. At low worry levels, testing interest increased with more positive beliefs about testing benefits; at high worry levels, interest was high regardless of benefits beliefs. The findings support Leventhal's Common-Sense Model of self-regulation delineating interactive influences of risk-related cognitions and emotions on information processing and behavior.

  17. Rapid, automated, nonradiometric susceptibility testing of Mycobacterium tuberculosis complex to four first-line antituberculous drugs used in standard short-course chemotherapy

    DEFF Research Database (Denmark)

    Johansen, Isik Somuncu; Thomsen, Vibeke Østergaard; Marjamäki, Merja;

    2004-01-01

    The increasing prevalence of drug-resistant tuberculosis necessitates rapid and accurate susceptibility testing. The nonradiometric BACTEC Mycobacteria Growth Indicator Tube 960 (MGIT) system for susceptibility testing was evaluated on 222 clinical Mycobacterium tuberculosis complex isolates for ...

  18. CLSI performance standards for antimicrobial susceptibility testing of bacteria isoloated from aquatic animals; second information supplement. CLSI document VET03/VET04-S2

    Science.gov (United States)

    The supplemental information presented in this document is intended for use with the antimicrobial susceptibility testing procedures published in the following Clinical and Laboratory Standards Institute (CLSI) approved documents VET03-A Methods for Antimicrobial Disk Susceptibility Testing of Bacte...

  19. Biochemical approaches to selective antifungal activity. Focus on azole antifungals.

    Science.gov (United States)

    Vanden Bossche, H; Marichal, P; Gorrens, J; Coene, M C; Willemsens, G; Bellens, D; Roels, I; Moereels, H; Janssen, P A

    1989-01-01

    Azole antifungals (e.g. the imidazoles: miconazole, clotrimazole, bifonazole, imazalil, ketoconazole, and the triazoles: diniconazole, triadimenol, propiconazole, fluconazole and itraconazole) inhibit in fungal cells the 14 alpha-demethylation of lanosterol or 24-methylenedihydrolanosterol. The consequent inhibition of ergosterol synthesis originates from binding of the unsubstituted nitrogen (N-3 or N-4) of their imidazole or triazole moiety to the heme iron and from binding of their N-1 substituent to the apoprotein of a cytochrome P-450 (P-450(14)DM) of the endoplasmic reticulum. Great differences in both potency and selectivity are found between the different azole antifungals. For example, after 16h of growth of Candida albicans in medium supplemented with [14C]-acetate and increasing concentrations of itraconazole, 100% inhibition of ergosterol synthesis is achieved at 3 x 10(-8) M. Complete inhibition of this synthesis by fluconazole is obtained at 10(-5) M only. The agrochemical imidazole derivative, imazalil, shows high selectivity, it has almost 80 and 98 times more affinity for the Candida P-450(s) than for those of the piglet testes microsomes and bovine adrenal mitochondria, respectively. However, the topically active imidazole antifungal, bifonazole, has the highest affinity for P-450(s) of the testicular microsomes. The triazole antifungal itraconazole inhibits at 10(-5) M the P-450-dependent aromatase by 17.9, whereas 50% inhibition of this enzyme is obtained at about 7.5 x 10(-6)M of the bistriazole derivative fluconazole. The overall results show that both the affinity for the fungal P-450(14)DM and the selectivity are determined by the nitrogen heterocycle and the hydrophobic N-1 substituent of the azole antifungals. The latter has certainly a greater impact. The presence of a triazole and a long hypdrophobic nonligating portion form the basis for itraconazole's potency and selectivity.

  20. Antimicrobial susceptibility testing for Helicobacter pylori in times of increasing antibiotic resistance.

    Science.gov (United States)

    Smith, Sinéad M; O'Morain, Colm; McNamara, Deirdre

    2014-08-07

    The gram-negative bacterium Helicobacter pylori (H. pylori) causes chronic gastritis, gastric and duodenal ulcers, gastric cancer and mucosa-associated lymphoid tissue lymphoma. Treatment is recommended in all symptomatic patients. The current treatment options for H. pylori infection are outlined in this review in light of the recent challenges in eradication success, largely due to the rapid emergence of antibiotic resistant strains of H. pylori. Antibiotic resistance is a constantly evolving process and numerous studies have shown that the prevalence of H. pylori antibiotic resistance varies significantly from country to country, and even between regions within the same country. In addition, recent data has shown that previous antibiotic use is associated with harbouring antibiotic resistant H. pylori. Local surveillance of antibiotic resistance is warranted to guide clinicians in their choice of therapy. Antimicrobial resistance is assessed by H. pylori culture and antimicrobial susceptibility testing. Recently developed molecular tests offer an attractive alternative to culture and allow for the rapid molecular genetic identification of H. pylori and resistance-associated mutations directly from biopsy samples or bacterial culture material. Accumulating evidence indicates that surveillance of antimicrobial resistance by susceptibility testing is feasible and necessary to inform clinicians in their choice of therapy for management of H. pylori infection.

  1. Screening of antifungal agents using ethanol precipitation and bioautography of medicinal and food plants.

    Science.gov (United States)

    Schmourlo, Gracilene; Mendonça-Filho, Ricardo R; Alviano, Celuta Sales; Costa, Sônia S

    2005-01-15

    In the search for bioactive compounds, bioautography and ethanol precipitation of macromolecules (proteins, polysaccharides, etc.) of plant aqueous extracts were associated in an antifungal screening. Thus, the supernatants, precipitates (obtained by ethanol precipitation) and aqueous extracts were investigated of medicinal and fruit bearing plants used against skin diseases by the Brazilian population. The agar diffusion and broth dilution methods were used to assess the activity against three fungi: Candida albicans, Trichophyton rubrum and Cryptococcus neoformans. The results, evaluated by the diameter of the inhibition zone of fungal growth, indicate that six plant species, among the 16 investigated, showed significant antifungal activity. The minimal inhibitory concentration (MIC) was determined on plant extracts that showed high efficacy against the tested microorganisms. The most susceptible yeast was Trichophyton rubrum and the best antifungal activity was shown by Xanthosoma sagittifolium supernatant. The bioautography was performed only for the aqueous extracts and supernatants of those plants that showed antifungal activity against Candida albicans and Cryptococcus neoformans, using n-butanol/acetic acid/water (BAW) 8:1:1 to develop silica gel TLC plates. Clear inhibition zones were observed for aqueous extracts of Schinus molle (R(f) 0.89) and Schinus terebinthifolius (R(f) 0.80) against Candida albicans, as for supernatant of Anacardium occidentale (R(f) 0.31) against Cryptococcus neoformans. The separation of macromolecules from metabolites, as in the case of Anacardium occidentale, Solanum sp. and Xanthosoma sagittifolium, enhances antifungal activity. In other cases, the antifungal activity is destroyed, as observed for Momordica charantia, Schinus molle and Schinus terebinthifolius.

  2. Calcium signaling mediates antifungal activity of triazole drugs in the Aspergilli.

    Science.gov (United States)

    Liu, Fei-fei; Pu, Li; Zheng, Qing-qing; Zhang, Yuan-wei; Gao, Rong-sui; Xu, Xu-shi; Zhang, Shi-zhu; Lu, Ling

    2015-08-01

    Azoles are widely applied and largely effective as antifungals; however, the increasing prevalence of clinically resistant isolates has yet to be matched by approaches to improve the efficacy of antimicrobial therapy. In this study, using the model fungus Aspergillus nidulans and one of the most common human pathogen Aspergillus fumigatus as research materials, we present the evidence that calcium signaling is involved in the azole-antifungals-induced stress-response reactions. In normal media, antifungal-itraconazole (ITZ) is able to induce the [Ca(2+)]c increased sharply but the addition of calcium chelator-EGTA or BAPTA almost blocks the calcium influx responses, resulted in the dramatically decreasing of [Ca(2+)]c transient. Real-time PCR analysis verified that six-tested Ca(2+)-inducible genes-two calcium channels (cchA/midA), a calmodulin-dependent phosphatase-calcineurin (cnaA), a transcription factor-crzA, and two calcium transporters (pmrA/pmcA)-could be transiently up-regulated by adding ITZ, indicating these components are involved in the azole stress-response reaction. Defect of cnaA or crzA caused more susceptibility to azole antifungals than did single mutants or double deletions of midA and cchA. Notably, EGTA may influence Rh123 accumulation as an azole-mimicking substrate through the process of the drug absorption. In vivo studies of a Galleria mellonella model identified that the calcium chelator works as an adjunct antifungal agent with azoles for invasive aspergillosis. Most importantly, combination of ITZ and EGTA or ITZ with calcium signaling inhibitor-FK506 greatly enhances the ITZ efficacy. Thus, our study provides potential clues that specific inhibitors of calcium signaling could be clinically useful adjuncts to conventional azole antifungals in the Aspergilli.

  3. Evaluating Constraints on Heavy-Ion SEE Susceptibility Imposed by Proton SEE Testing and Other Mixed Environments

    Science.gov (United States)

    Ladbury, R. L.; Lauenstein, J.-M.

    2016-01-01

    We develop metrics for assessing the effectiveness of proton SEE data for bounding heavy-ion SEE susceptibility. The metrics range from simple geometric criteria requiring no knowledge of the test articles to bounds of SEE rates.

  4. Correlation between bactericidal activity of fosfomycin trometamol in an in vitro model of the urinary bladder and susceptibility testing.

    Science.gov (United States)

    Pinasi, C; Albini, E; Marca, G

    1987-01-01

    The present study was undertaken to define an interpretative guideline for disk diffusion susceptibility testing with fosfomycin trometamol, a new antimicrobial agent which has been developed for the treatment of urinary tract infections. Two potencies of fosfomycin disk were used: 50 and 200 micrograms, prepared in the presence and absence of glucose-6-phosphate. To verify the reliability of the results obtained in susceptibility testing, we have also evaluated the bactericidal activity of fosfomycin trometamol versus sensitive and resistant strains in an 'in vitro' model simulating the hydrokinetic aspects involved in the treatment of bacterial cystitis. The data obtained evidenced the role of glucose-6-phosphate in antimicrobial susceptibility tests as well as the importance of the urinary antibiotic concentrations to define sensitive and resistant bacteria. On the basis of our results, we recommend that a 200-microgram disk of fosfomycin containing 50 micrograms of glucose-6-phosphate should be used in antimicrobial susceptibility testing with fosfomycin trometamol.

  5. Vancomycin-resistant Enterococcus spp.: validation of susceptibility testing and in vitro activity of vancomycin, linezolid, tigecycline and daptomycin

    DEFF Research Database (Denmark)

    Rathe, Mathias; Kristensen, Lise; Ellermann-Eriksen, Svend;

    2010-01-01

    Vancomycin-resistant enterococci (VRE) have emerged to become a significant nosocomial pathogen. However, detection may be challenging and treatment possibilities are limited. Reports of resistance to linezolide, daptomycin and tigecycline underline the need for reliable susceptibility testing wi...

  6. Determination of disk diffusion susceptibility testing interpretive criteria using model-based analysis: development and implementation.

    Science.gov (United States)

    DePalma, Glen; Turnidge, John; Craig, Bruce A

    2017-02-01

    The determination of diffusion test breakpoints has become a challenging issue due to the increasing resistance of microorganisms to antibiotics. Currently, the most commonly-used method for determining these breakpoints is the modified error-rate bounded method. Its use has remained widespread despite the introduction of several model-based methods that have been shown superior in terms of precision and accuracy. However, the computational complexities associated with these new approaches has been a significant barrier for clinicians. To remedy this, we developed and examine the utility of a free online software package designed for the determination of diffusion test breakpoints: dBETS (diffusion Breakpoint Estimation Testing Software). This software package allows clinicians to easily analyze data from susceptibility experiments through visualization, error-rate bounded, and model-based approaches. We analyze four publicly available data sets from the Clinical and Laboratory Standards Institute using dBETS.

  7. Physically-based landslide susceptibility modelling: geotechnical testing and model evaluation issues

    Science.gov (United States)

    Marchesini, Ivan; Mergili, Martin; Schneider-Muntau, Barbara; Alvioli, Massimiliano; Rossi, Mauro; Guzzetti, Fausto

    2015-04-01

    We used the software r.slope.stability for physically-based landslide susceptibility modelling in the 90 km² Collazzone area, Central Italy, exploiting a comprehensive set of lithological, geotechnical, and landslide inventory data. The model results were evaluated against the inventory. r.slope.stability is a GIS-supported tool for modelling shallow and deep-seated slope stability and slope failure probability at comparatively broad scales. Developed as a raster module of the GRASS GIS software, r.slope.stability evaluates the slope stability for a large number of randomly selected ellipsoidal potential sliding surfaces. The bottom of the soil (for shallow slope stability) or the bedding planes of lithological layers (for deep-seated slope stability) are taken as potential sliding surfaces by truncating the ellipsoids, allowing for the analysis of relatively complex geological structures. To take account for the uncertain geotechnical and geometric parameters, r.slope.stability computes the slope failure probability by testing multiple parameter combinations sampled deterministically or stochastically, and evaluating the ratio between the number of parameter combinations yielding a factor of safety below 1 and the total number of tested combinations. Any single raster cell may be intersected by multiple sliding surfaces, each associated with a slope failure probability. The most critical sliding surface is relevant for each pixel. Intensive use of r.slope.stability in the Collazzone Area has opened up two questions elaborated in the present work: (i) To what extent does a larger number of geotechnical tests help to better constrain the geotechnical characteristics of the study area and, consequently, to improve the model results? The ranges of values of cohesion and angle of internal friction obtained through 13 direct shear tests corresponds remarkably well to the range of values suggested by a geotechnical textbook. We elaborate how far an increased number of

  8. Psychological impact of genetic testing for cancer susceptibility: an update of the literature.

    Science.gov (United States)

    Meiser, Bettina

    2005-12-01

    This article presents an overview of the rapidly evolving body of literature on the psychological impact of genetic testing for hereditary breast/ovarian cancer susceptibility, hereditary non-polyposis colorectal cancer (HNPCC) and familial adenomatous polyposis (FAP). Uptake of genetic testing for BRCA1/2 and HNPCC-related mutations is more consistently related to psychological factors, rather than sociodemographic variables. Most studies on the psychological impact of genetic testing amongst individuals who have never been affected by cancer demonstrate that non-carriers derive significant psychological benefits from genetic testing, while no adverse effects have been observed amongst carriers. These benefits are more clear-cut for HNPCC, compared to hereditary breast/ovarian cancer, reflecting differences in risk management options. The few studies available on individuals affected with cancer indicate that the impact of genetic testing is mediated and amplified by their former experience of cancer. Future directions and challenges of research in this area are reviewed. In particular, more empirical data are needed on the broader impact of genetic testing on those with inconclusive results or results of uncertain significance. As genetic testing is becoming available for other types of familial cancer, additional investigations will be needed as there is evidence to suggest that the impact of genetic testing may be unique to each type of familial cancer.

  9. Validation of antibiotic susceptibility testing guidelines in a routine clinical microbiology laboratory exemplifies general key challenges in setting clinical breakpoints.

    Science.gov (United States)

    Hombach, Michael; Courvalin, Patrice; Böttger, Erik C

    2014-07-01

    This study critically evaluated the new European Committee for Antimicrobial Susceptibility Testing (EUCAST) antibiotic susceptibility testing guidelines on the basis of a large set of disk diffusion diameters determined for clinical isolates. We report several paradigmatic problems that illustrate key issues in the selection of clinical susceptibility breakpoints, which are of general importance not only for EUCAST but for all guidelines systems, i.e., (i) the need for species-specific determinations of clinical breakpoints/epidemiological cutoffs (ECOFFs), (ii) problems arising from pooling data from various sources, and (iii) the importance of the antibiotic disk content for separating non-wild-type and wild-type populations.

  10. Dermatophyte susceptibility varies towards antimicrobial textiles.

    Science.gov (United States)

    Hammer, Timo R; Mucha, Helmut; Hoefer, Dirk

    2012-07-01

    Dermatophytoses are a widespread problem worldwide. Textiles in contact with infected skin can serve as a carrier for fungus propagation. Hitherto, it is unknown, whether antifungal textiles could contribute in controlling dermatophytes e.g. by disrupting the chain of infection. Testing of antimicrobial fabrics for their antifungal activities therefore is a fundamental prerequisite to assess the putative clinical relevance of textiles for dermatophyte prevention. Fabrics finished with either didecyldimethylammonium chloride (DDAC), poly-hexamethylenbiguanide, copper and two silver chloride concentrations were tested for their antifungal activity against Trichophyton rubrum, Trichophyton mentagrophytes and Candida albicans. To prove dermatophyte susceptibility towards the textiles, swatches were subjected to DIN EN 14199 (Trichophyton sp.) or DIN EN ISO 20743 (C. albicans) respectively. In addition, samples were embedded, and semi-thin sections were analysed microscopically. While all samples showed a clear inhibition of C. albicans, activity against Trichophyton sp. varied significantly: For example, DDAC completely inhibited T. rubrum growth, whereas T. mentagrophytes growth remained unaffected even in direct contact to the fibres. The results favour to add T. mentagrophytes as a test organism in textile dermatophyte efficacy tests. Microscopic analysis of swatches allowed detailed evaluation of additional parameters like mycelium thickness, density and hyphae penetration depth into the fabric.

  11. ESBL Detection: Comparison of a Commercially Available Chromogenic Test for Third Generation Cephalosporine Resistance and Automated Susceptibility Testing in Enterobactericeae

    Science.gov (United States)

    El-Jade, Mohamed Ramadan; Parcina, Marijo; Schmithausen, Ricarda Maria; Stein, Christoph; Meilaender, Alina; Hoerauf, Achim; Molitor, Ernst

    2016-01-01

    Rapid detection and reporting of third generation cephalosporine resistance (3GC-R) and of extended spectrum betalactamases in Enterobacteriaceae (ESBL-E) is a diagnostic and therapeutic priority to avoid inefficacy of the initial antibiotic regimen. In this study we evaluated a commercially available chromogenic screen for 3GC-R as a predictive and/or confirmatory test for ESBL and AmpC activity in clinical and veterinary Enterobacteriaceae isolates. The test was highly reliable in the prediction of cefotaxime and cefpodoxime resistance, but there was no correlation with ceftazidime and piperacillin/tazobactam minimal inhibitory concentrations. All human and porcine ESBL-E tested were detected with exception of one genetically positive but phenotypically negative isolate. By contrast, AmpC detection rates lay below 30%. Notably, exclusion of piperacillin/tazobactam resistant, 3GC susceptible K1+ Klebsiella isolates increased the sensitivity and specificity of the test for ESBL detection. Our data further imply that in regions with low prevalence of AmpC and K1 positive E. coli strains chromogenic testing for 3GC-R can substitute for more time consuming ESBL confirmative testing in E. coli isolates tested positive by Phoenix or VITEK2 ESBL screen. We, therefore, suggest a diagnostic algorithm that distinguishes 3GC-R screening from primary culture and species-dependent confirmatory ESBL testing by βLACTATM and discuss the implications of MIC distribution results on the choice of antibiotic regimen. PMID:27494134

  12. ESBL Detection: Comparison of a Commercially Available Chromogenic Test for Third Generation Cephalosporine Resistance and Automated Susceptibility Testing in Enterobactericeae.

    Science.gov (United States)

    El-Jade, Mohamed Ramadan; Parcina, Marijo; Schmithausen, Ricarda Maria; Stein, Christoph; Meilaender, Alina; Hoerauf, Achim; Molitor, Ernst; Bekeredjian-Ding, Isabelle

    2016-01-01

    Rapid detection and reporting of third generation cephalosporine resistance (3GC-R) and of extended spectrum betalactamases in Enterobacteriaceae (ESBL-E) is a diagnostic and therapeutic priority to avoid inefficacy of the initial antibiotic regimen. In this study we evaluated a commercially available chromogenic screen for 3GC-R as a predictive and/or confirmatory test for ESBL and AmpC activity in clinical and veterinary Enterobacteriaceae isolates. The test was highly reliable in the prediction of cefotaxime and cefpodoxime resistance, but there was no correlation with ceftazidime and piperacillin/tazobactam minimal inhibitory concentrations. All human and porcine ESBL-E tested were detected with exception of one genetically positive but phenotypically negative isolate. By contrast, AmpC detection rates lay below 30%. Notably, exclusion of piperacillin/tazobactam resistant, 3GC susceptible K1+ Klebsiella isolates increased the sensitivity and specificity of the test for ESBL detection. Our data further imply that in regions with low prevalence of AmpC and K1 positive E. coli strains chromogenic testing for 3GC-R can substitute for more time consuming ESBL confirmative testing in E. coli isolates tested positive by Phoenix or VITEK2 ESBL screen. We, therefore, suggest a diagnostic algorithm that distinguishes 3GC-R screening from primary culture and species-dependent confirmatory ESBL testing by βLACTATM and discuss the implications of MIC distribution results on the choice of antibiotic regimen.

  13. STUDY OF ANTIBIOTIC SUSCEPTIBILITY TEST OF MODERN GENERATION OF DRUGS AGAINST UPPER RESPIRATORY TRACT PATHOGENS

    Directory of Open Access Journals (Sweden)

    Vinod Singh et al

    2012-10-01

    , nalidixic acid and ofloxacin, N. meningitidis 4 (100% were resistant to aztreonam, cefdinir, cefixime, cefotaxime, ceftazidime, nalidixic acid and ofloxacin whereas rest of gram negative strains showed moderate susceptible against amikacin, cefuroxime and ciprofloxacin. Hence we concluded that the efficacy of cefazolin, cephalexin, chloramphenical, tetracycline, amikacin and ciprofloxacin was recorded higher than other antibiotics tested against nasal infection causing pathogens.

  14. Comparison of Rosco Neo-Sensitabs with Oxoid paper disks in EUCAST disk diffusion antimicrobial susceptibility testing on Mueller-Hinton agar

    DEFF Research Database (Denmark)

    Justesen, U S; Acar, Ziyap; Olsson, K;

    2013-01-01

    This study compared Neo-Sensitabs with Oxoid paper disks using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) disk diffusion antimicrobial susceptibility test on Mueller-Hinton agar. The EUCAST-recommended quality control strains (Escherichia coli ATCC 25922, Pseudomonas...... paper disks for EUCAST disk diffusion antimicrobial susceptibility testing on Mueller-Hinton agar....

  15. In vitro and in vivo antifungal activity, liver profile test, and mutagenic activity of five plants used in traditional Mexican medicine

    Directory of Open Access Journals (Sweden)

    Jorge Cornejo-Garrido

    2015-02-01

    Full Text Available Berberis hartwegii Benth., Berberidaceae, Hamelia patens Jacq., Rubiaceae, Dendropanax arboreus (L. Decne & Planch., Araliaceae, Erythrina herbacea L., Fabaceae, and Zanthoxylum caribaeum Lam., Rutaceae, acetone extracts were selected on the basis of their use in traditional Mexican medicine to treat scabies or skin diseases. Anti-dermatophyte activity in vitro was evaluated using the agar dilution assay, and the therapeutic efficacy of B. hartwegii and Z. caribaeum were tested against experimental tinea pedis. The infected animals were treated intragastrically daily for seven days with 2.5 and 5 mg/kg of acetone extracts. The acetone extract of H. patens exhibited 100% growth inhibition against T. mentagrophytes and E. floccosum at 100.0 and 50.0 µg/ml, respectively, and B. hartwegii inhibited growth of M. canis and T. mentagrophytes at 100.0 µg/ml. Effective treatments with 2.5 mg/kg of Z. caribaeum and B. hartwegii extract were comparable with 1 mg/kg of clotrimazole in mice. Liver profile tests and histological analyses did not exhibit any signs of toxicity and the Ames test indicated that both extracts were safe when evaluated in strains TA98, TA100 and TA102. Our results suggest the potential for the future development of new antifungal drugs from B. hartwegii or Z. caribaeum.

  16. Antifungal activity of five species of Polygala

    Directory of Open Access Journals (Sweden)

    Susana Johann

    2011-09-01

    Full Text Available Crude extracts and fractions of five species of Polygala - P. campestris, P. cyparissias, P. paniculata, P. pulchella and P. sabulosa - were investigated for their in vitro antifungal activity against opportunistic Candida species, Cryptococcus gattii and Sporothrix schenckii with bioautographic and microdilution assays. In the bioautographic assays, the major extracts were active against the fungi tested. In the minimal concentration inhibitory (MIC assay, the hexane extract of P. paniculata and EtOAc fraction of P. sabulosa showed the best antifungal activity, with MIC values of 60 and 30 µg/mL, respectively, against C. tropicalis, C. gattii and S. schenckii. The compounds isolated from P. sabulosa prenyloxycoumarin and 1,2,3,4,5,6-hexanehexol displayed antifungal activity against S. schenckii (with MICs of 125 µg/mL and 250 µg/mL, respectively and C. gattii (both with MICs of 250 µg/mL. Rutin and aurapten isolated from P. paniculata showed antifungal activity against C. gattii with MIC values of 60 and 250 µg/mL, respectively. In the antifungal screening, few of the isolated compounds showed good antifungal inhibition. The compound α-spinasterol showed broad activity against the species tested, while rutin had the best activity with the lowest MIC values for the microorganisms tested. These two compounds may be chemically modified by the introduction of a substitute group that would alter several physico-chemical properties of the molecule, such as hydrophobicity, electronic density and steric strain.

  17. Standardization of a broth microdilution susceptibility testing method to determine minimum inhibitory concentrations of aquatic bacteria

    DEFF Research Database (Denmark)

    Miller, R.A.; Walker, R.D.; Carson, J.;

    2005-01-01

    A multiple laboratory study was conducted in accordance with the standards established by the Clinical and Laboratory Standards Institute (CLSI), formerly the National Committee for Clinical Laboratory Standards (NCCLS), for the development of quality control (QC) ranges using dilution antimicrob......A multiple laboratory study was conducted in accordance with the standards established by the Clinical and Laboratory Standards Institute (CLSI), formerly the National Committee for Clinical Laboratory Standards (NCCLS), for the development of quality control (QC) ranges using dilution......-Hinton broth. These QC ranges were accepted by the CLSI/NCCLS Subcommittee on Veterinary Antimicrobial Susceptibility Testing in January 2004. This broth microdilution testing method represents the first standardized method for determining MICs of bacterial isolates whose preferred growth temperatures...

  18. Contributions of culture and antimicrobial susceptibility tests to the retreatment of patients with pulmonary tuberculosis

    Directory of Open Access Journals (Sweden)

    Bruno Horta Andrade

    2013-08-01

    Full Text Available Introduction This study evaluated the efficacy of retreatment of pulmonary tuberculosis (TB with regard to treatment outcomes and antimicrobial susceptibility testing (ST profiles. Methods This retrospective cohort study analyzed 144 patients treated at a referral hospital in Brazil. All of them had undergone prior treatment, were smear-positive for TB and received a standardized retreatment regimen. Fisher's 2-tailed exact test and the χ2 test were used; RRs and 95% CIs were calculated using univariate and multivariate binary logistic regression. Results The patients were cured in 84 (58.3% cases. Failure was associated with relapsed treatment and abandonment (n=34. Culture tests were obtained for 103 (71.5% cases; 70 (48.6% had positive results. ST results were available for 67 (46.5% cases; the prevalence of acquired resistance was 53.7%. There were no significant differences between those who achieved or not therapeutic success (p=0.988, despite being sensitive or resistant to 1 or more drugs. Rifampicin resistance was independently associated with therapeutic failure (OR: 4.4, 95% CI:1.12-17.37, p=0.034. For those cases in which cultures were unavailable, a 2nd model without this information was built. In this, return after abandonment was significantly associated with retreatment failure (OR: 3.59, 95% CI:1.17-11.06, p=0.026. Conclusions In this cohort, the general resistance profile appeared to have no influence on treatment outcome, except in cases of rifampicin resistance. The form of reentry was another independent predictor of failure. The use of bacterial culture identification and ST in TB management must be re-evaluated. The recommendations for different susceptibility profiles must also be improved.

  19. Triazole antifungals: a review.

    Science.gov (United States)

    Peyton, L R; Gallagher, S; Hashemzadeh, M

    2015-12-01

    Invasive fungal infections and systemic mycosis, whether from nosocomial infection or immunodeficiency, have been on an upward trend for numerous years. Despite advancements in antifungal medication, treatment in certain patients can still be difficult for reasons such as impaired organ function, limited administration routes or poor safety profiles of the available antifungal medications. The growing number of invasive fungal species becoming resistant to current antifungal medications is of appreciable concern. Triazole compounds containing one or more 1,2,4-triazole rings have been shown to contain some of the most potent antifungal properties. Itracon-azole and fluconazole were some of the first triazoles synthesized, but had limitations associated with their use. Second-generation triazoles such as voriconazole, posa-conazole, albaconazole, efinaconazole, ravuconazole and isavuconazole are all derivatives of either itraconazole or fluconazole, and designed to overcome the deficiencies of their parent drugs. The goal of this manuscript is to review antifungal agents derived from triazole.

  20. 我院2009-2011年老年真菌感染菌种分布及其对抗真菌药敏感性分析%Analysis of Species Distribution and Drug Susceptibility to Antifungal Agents in the Aged with Fungal Infection of Our Hospital during 2009-2011

    Institute of Scientific and Technical Information of China (English)

    张慧儿; 裘莉佩

    2013-01-01

    目的:探讨临床老年患者标本中所分离的假丝酵母菌属的菌种分布及其对临床常用抗真菌药的敏感性,为老年人使用抗真菌药提供参考.方法:对我院2009-2011年从老年体内分离的213例假丝酵母菌进行回顾性分析,分析其菌种分布情况以及对5种抗真菌药的敏感性.结果:在213株假丝酵母菌中,白假丝酵母菌占42.3%(90/213),非白假丝酵母菌占57.7% (123/213);5-氟胞嘧啶、两性霉素B、氟康唑、伊曲康唑和伏立康唑的总敏感率分别为87.8%、100%、93.2%、85.9%和95.9%;90株白假丝酵母菌对上述5种抗真菌药的敏感率分别为93.3%、100%、95.4%、92.2%和96.3%,123株非白假丝酵母菌分别为83.7%、100%、91.6%、81.3%和95.6%.结论:在老年感染假丝酵母菌中最常见的菌种仍是白假丝酵母菌,但其比例较已报道的非老年感染者比例有所下降;白假丝酵母菌对常用抗真菌药仍有较高的敏感性,非白假丝酵母菌的耐药性则高于白假丝酵母菌.%OBJECTIVE: To discuss the species distribution and drug susceptibility to antifungal agent of isolated Candida, and to provide reference for the use of antifungal agent in the aged. METHODS: A total of 213 isolates were collected from our hospital during 2009 — 2011 were analyzed statistically, and the species distribution of Candida and drug resistance to 5 commonly used antifungal agents were analyzed. RESULTS: Among 213 Candida, Candida albicans accounted for 42.3% (90/213) and non-Candida albicans accounted for 57.7% (123/213). The overall percentage of strains susceptible to 5-flucytosine, amphotericin B, flucytosine, itraconazole and voriconazole were 87.8% , 100% , 93.2% , 85.9% and 95.9% , respectively. About 93.3% , 100%, 95.4% , 92.2% and 96.3% of 90 strains of Candida albicans were susceptible to these 5 antifungal agents. The susceptibility rates of 123 non-Candida albicans isolates were 83.7%, 100

  1. In Vitro Activity of Seven Systemically Active Antifungal Agents against a Large Global Collection of Rare Candida Species as Determined by CLSI Broth Microdilution Methods ▿

    Science.gov (United States)

    Diekema, D. J.; Messer, S. A.; Boyken, L. B.; Hollis, R. J.; Kroeger, J.; Tendolkar, S.; Pfaller, M. A.

    2009-01-01

    Five Candida species (C. albicans, C. glabrata, C. tropicalis, C. parapsilosis, and C. krusei) account for over 95% of invasive candidiasis cases. Some less common Candida species have emerged as causes of nosocomial candidiasis, but there is little information about their in vitro susceptibilities to antifungals. We determined the in vitro activities of fluconazole, voriconazole, posaconazole, amphotericin B, anidulafungin, caspofungin, and micafungin against invasive, unique patient isolates of Candida collected from 100 centers worldwide between January 2001 and December 2007. Antifungal susceptibility testing was performed by the CLSI M27-A3 method. CLSI breakpoints for susceptibility were used for fluconazole, voriconazole, anidulafungin, caspofungin, and micafungin, while a provisional susceptibility breakpoint of ≤1 μg/ml was used for amphotericin and posaconazole. Of 14,007 Candida isolates tested, 658 (4.7%) were among the less common species. Against all 658 isolates combined, the activity of each agent, expressed as the MIC50/MIC90 ratio (and the percentage of susceptible isolates) was as follows: fluconazole, 1/4 (94.8%); voriconazole, 0.03/0.12 (98.6%); posaconazole, 0.12/0.5 (95.9%); amphotericin, 0.5/2 (88.3%); anidulafungin, 0.5/2 (97.4%); caspofungin, 0.12/0.5 (98.0%); and micafungin, 0.25/1 (99.2%). Among the isolates not susceptible to one or more of the echinocandins, most (68%) were C. guilliermondii. All isolates of the less common species within the C. parapsilosis complex (C. orthopsilosis and C. metapsilosis) were susceptible to voriconazole, posaconazole, anidulafungin, caspofungin, and micafungin. Over 95% of clinical isolates of the rare Candida species were susceptible to the available antifungals. However, activity did vary by drug-species combination, with some species (e.g., C. rugosa and C. guilliermondii) demonstrating reduced susceptibilities to commonly used agents such as fluconazole and echinocandins. PMID:19710283

  2. The power of DNA double-strand break (DSB) repair testing to predict breast cancer susceptibility.

    Science.gov (United States)

    Keimling, Marlen; Deniz, Miriam; Varga, Dominic; Stahl, Andreea; Schrezenmeier, Hubert; Kreienberg, Rolf; Hoffmann, Isabell; König, Jochem; Wiesmüller, Lisa

    2012-05-01

    Most presently known breast cancer susceptibility genes have been linked to DSB repair. To identify novel markers that may serve as indicators for breast cancer risk, we performed DSB repair analyses using a case-control design. Thus, we examined 35 women with defined familial history of breast and/or ovarian cancer (first case group), 175 patients with breast cancer (second case group), and 245 healthy women without previous cancer or family history of breast cancer (control group). We analyzed DSB repair in peripheral blood lymphocytes (PBLs) by a GFP-based test system using 3 pathway-specific substrates. We found increases of microhomology-mediated nonhomologous end joining (mmNHEJ) and nonconservative single-strand annealing (SSA) in women with familial risk vs. controls (P=0.0001-0.0022) and patients with breast cancer vs. controls (P=0.0004-0.0042). Young age (DSB repair activities in PBLs as method to estimate breast cancer susceptibility beyond limitations of genotyping and to predict responsiveness to therapeutics targeting DSB repair-dysfunctional tumors.

  3. Child exposure to serious life events, COMT, and aggression: Testing differential susceptibility theory.

    Science.gov (United States)

    Hygen, Beate Wold; Belsky, Jay; Stenseng, Frode; Lydersen, Stian; Guzey, Ismail Cuneyt; Wichstrøm, Lars

    2015-08-01

    Both genetic and environmental factors contribute to individual differences in aggression. Catechol-O-methyltransferase Val158Met (COMT), a common, functional polymorphism, has been implicated in aggression and aggression traits, as have childhood experiences of adversity. It is unknown whether these effects are additive or interactional and, in the case of interaction, whether they conform to a diathesis-stress or differential susceptibility model. We examined Gene × Environment interactions between COMT and serious life events on measures of childhood aggression and contrasted these 2 models. The sample was composed of community children (N = 704); 355 were boys, and the mean age was 54.8 months (SD = 3.0). The children were genotyped for COMT rs4680 and assessed for serious life events and by teacher-rated aggression. Regression analysis showed no main effects of COMT and serious life events on aggression. However, a significant interactive effect of childhood serious life events and COMT genotype was observed: Children who had faced many serious life events and were Val homozygotes exhibited more aggression (p = .02) than did their Met-carrying counterparts. Notably, in the absence of serious life events, Val homozygotes displayed significantly lower aggression scores than did Met carriers (p = .03). When tested, this constellation of findings conformed to the differential susceptibility hypothesis: In this case, Val homozygotes are more malleable to the effect of serious life events on aggression and not simply more vulnerable to the negative effect of having experienced many serious life events.

  4. Drug susceptibility testing of clinical isolates of streptococci and enterococci by the Phoenix automated microbiology system

    Directory of Open Access Journals (Sweden)

    Sokeng Gertrude

    2007-05-01

    Full Text Available Abstract Background Drug resistance is an emerging problem among streptococcal and enterococcal species. Automated diagnostic systems for species identification and antimicrobial susceptibility testing (AST have become recently available. We evaluated drug susceptibility of clinical isolates of streptococci and enterococci using the recent Phoenix system (BD, Sparks, MD. Diagnostic tools included the new SMIC/ID-2 panel for streptococci, and the PMIC/ID-14 for enterococci. Two-hundred and fifty isolates have been investigated: β-hemolytic streptococci (n = 65, Streptococcus pneumoniae (n = 50, viridans group streptococci (n = 32, Enterococcus faecium (n = 40, Enterococcus faecalis (n = 43, other catalase-negative cocci (n = 20. When needed, species ID was determined using molecular methods. Test bacterial strains were chosen among those carrying clinically-relevant resistance determinants (penicillin, macrolides, fluoroquinolones, glycopeptides. AST results of the Phoenix system were compared to minimal inhibitory concentration (MIC values measured by the Etest method (AB Biodisk, Solna, Sweden. Results Streptococci: essential agreement (EA and categorical agreement (CA were 91.9% and 98.8%, respectively. Major (ME and minor errors (mE accounted for 0.1% and 1.1% of isolates, respectively. No very major errors (VME were produced. Enterococci: EA was 97%, CA 96%. Small numbers of VME (0.9%, ME (1.4% and mE (2.8% were obtained. Overall, EA and CA rates for most drugs were above 90% for both genera. A few VME were found: a teicoplanin and high-level streptomycin for E. faecalis, b high-level gentamicin for E. faecium. The mean time to results (± SD was 11.8 ± 0.9 h, with minor differences between streptococci and enterococci. Conclusion The Phoenix system emerged as an effective tool for quantitative AST. Panels based on dilution tests provided rapid and accurate MIC values with regard to clinically-relevant streptococcal and enterococcal

  5. ANTIFUNGAL ACTIVITY OF SOME COLEUS SPECIES GROWING IN NILGIRIS

    OpenAIRE

    Nilani, P.; Duraisamy, B.; Dhanabal, P.S.; khan, Saleemullah; Suresh, B.; Shankar, V.; Kavitha, K.Y.; Syamala, G.

    2006-01-01

    The in vitro antifungal activity of solvent extracts of Coleus forskohlii, Coleus blumei and Coleus barbatus were compared by testing against some pathogenic fungi like Aspergillus niger, Aspergillus fumigatus, Aspergillus ruantii, Proteus vulgaris and Candida albicans. The petroleum ether extract of Coleus forskohlii and Coleus barbatus exhibited significant antifungal activity against all the selected organisms. The extracts of Coleus blumei did not show any significant antifungal activity ...

  6. Antifungal activity of some coleus species growing in nilgiris.

    Science.gov (United States)

    Nilani, P; Duraisamy, B; Dhanabal, P S; Khan, Saleemullah; Suresh, B; Shankar, V; Kavitha, K Y; Syamala, G

    2006-07-01

    The in vitro antifungal activity of solvent extracts of Coleus forskohlii, Coleus blumei and Coleus barbatus were compared by testing against some pathogenic fungi like Aspergillus niger, Aspergillusfumigatus, Aspergillus ruantii, Proteus vulgaris and Candida albicans. The petroleum ether extract of Coleus forskohlii and Coleus barbatus exhibited significant antifungal activity against all the selected organisms. The extracts of Coleus blumei did not show any significant antifungal activity against the selected organisms.

  7. Rapid antimicrobial susceptibility test for identification of new therapeutics and drug combinations against multidrug-resistant bacteria

    OpenAIRE

    Sun, Wei; Weingarten, Rebecca A.; Xu, Miao; Southall, Noel; Dai, Sheng; Shinn, Paul; Sanderson, Philip E.; Williamson, Peter R.; Frank, Karen M.; Zheng,Wei

    2016-01-01

    Current antimicrobial susceptibility testing has limited screening capability for identifying empirical antibiotic combinations to treat severe bacterial infections with multidrug-resistant (MDR) organisms. We developed a new antimicrobial susceptibility assay using automated ultra-high-throughput screen technology in combination with a simple bacterial growth assay. A rapid screening of 5170 approved drugs and other compounds identified 25 compounds with activities against MDR Klebsiella pne...

  8. Antifungal Activity of 14-Helical β-Peptides against Planktonic Cells and Biofilms of Candida Species

    Directory of Open Access Journals (Sweden)

    Namrata Raman

    2015-08-01

    Full Text Available Candida albicans is the most prevalent cause of fungal infections and treatment is further complicated by the formation of drug resistant biofilms, often on the surfaces of implanted medical devices. In recent years, the incidence of fungal infections by other pathogenic Candida species such as C. glabrata, C. parapsilosis and C. tropicalis has increased. Amphiphilic, helical β-peptide structural mimetics of natural antimicrobial α-peptides have been shown to exhibit specific planktonic antifungal and anti-biofilm formation activity against C. albicans in vitro. Here, we demonstrate that β-peptides are also active against clinically isolated and drug resistant strains of C. albicans and against other opportunistic Candida spp. Different Candida species were susceptible to β-peptides to varying degrees, with C. tropicalis being the most and C. glabrata being the least susceptible. β-peptide hydrophobicity directly correlated with antifungal activity against all the Candida clinical strains and species tested. While β-peptides were largely ineffective at disrupting existing Candida biofilms, hydrophobic β-peptides were able to prevent the formation of C. albicans, C. glabrata, C. parapsilosis and C. tropicalis biofilms. The broad-spectrum antifungal activity of β-peptides against planktonic cells and in preventing biofilm formation suggests the promise of this class of molecules as therapeutics.

  9. Synthesis and biological evaluation of fluconazole analogs with triazole-modified scaffold as potent antifungal agents.

    Science.gov (United States)

    Hashemi, Seyedeh Mahdieh; Badali, Hamid; Irannejad, Hamid; Shokrzadeh, Mohammad; Emami, Saeed

    2015-04-01

    In order to find new azole antifungals, we have recently designed a series of triazole alcohols in which one of the 1,2,4-triazol-1-yl group in fluconazole structure has been replaced with 4-amino-5-aryl-3-mercapto-1,2,4-triazole motif. In this paper, we focused on the structural refinement of the primary lead, by removing the amino group from the structure to achieve 5-aryl-3-mercapto-1,2,4-triazole derivatives 10a-i and 11a-i. The in vitro antifungal susceptibility testing of title compounds demonstrated that most compounds had potent inhibitory activity against Candida species. Among them, 5-(2,4-dichlorophenyl)triazole analogs 10h and 11h with MIC values of fluconazole against Candida species.

  10. Antibacterial susceptibility of bovine-mastitis pathogens tested directly in milk from infected quarters.

    Science.gov (United States)

    Louhi-Lehtiö, M; Sandholm, M; Myllys, V; Honkanen-Buzalski, T

    1994-04-01

    Antibacterial susceptibilities of bovine-mastitis pathogens were analysed directly in 57 mastitic milk samples without inoculation with exogenous organisms. Aseptically collected milk was mixed with serial dilutions of antibacterials and the growth was observed using 2,3,5-triphenyltetrazolium chloride (TTC) reduction the following day. The results were compared with those obtained by using calibrated bacterial inocula in turbidimetric minimum-inhibitory-concentration (MIC) determination in broth cultures, and in TTC-broth culture-test and TTC-normal milk-test. The results of different methods all correlated positively when the entire data was used. However, taking the direct test in mastitic milk as the 'true' result, the total discrepancies varied from 34.7% to 48.8%. Antibacterial activities of the trimethoprim-sulphadoxine combination, and of spiramycin and ampicillin, decreased significantly when nutrient broth was replaced by milk as the test medium. The efficacy of trimethoprim-sulphadoxine as an antibacterial agent was also dependent on the source of milk.

  11. Comparison of methods for in vitro testing of susceptibility of porcine Mycoplasma species to antimicrobial agents.

    Science.gov (United States)

    Ter Laak, E A; Pijpers, A; Noordergraaf, J H; Schoevers, E C; Verheijden, J H

    1991-02-01

    The MICs of 18 antimicrobial agents used against strains of three porcine Mycoplasma species were determined by a serial broth dilution method. Twenty field strains of M. hyorhinis, ten field strains of M. hyopneumoniae, six field strains of M. flocculare, and the type strains of these species were tested. Twelve field strains and the type strain of M. hyorhinis were also tested by an agar dilution method. Tests were read at various time points. When the broth dilution method was used, the final MIC had to be read 2 days after color changes had stopped. MICs of tetracycline, oxytetracycline, doxycycline, and minocycline were low for the three Mycoplasma species tested. MICs of chlortetracycline were 8 to 16 times higher than MICs of the other tetracyclines. Spiramycin, tylosin, kitasamycin, spectinomycin, tiamulin, lincomycin, and clindamycin were effective against all strains of M. hyorhinis and M. hyopneumoniae. The quinolones were highly effective against M. hyopneumoniae but less effective against M. hyorhinis. The susceptibility patterns for M. hyopneumoniae and M. flocculare were similar.

  12. Miniaturized Antimicrobial Susceptibility Test by Combining Concentration Gradient Generation and Rapid Cell Culturing

    Directory of Open Access Journals (Sweden)

    Samuel C. Kim

    2015-10-01

    Full Text Available Effective treatment of bacterial infection relies on timely diagnosis and proper prescription of antibiotic drugs. The antimicrobial susceptibility test (AST is one of the most crucial experimental procedures, providing the baseline information for choosing effective antibiotic agents and their dosages. Conventional methods, however, require long incubation times or significant instrumentation costs to obtain test results. We propose a lab-on-a-chip approach to perform AST in a simple, economic, and rapid manner. Our assay platform miniaturizes the standard broth microdilution method on a microfluidic device (20 × 20 mm that generates an antibiotic concentration gradient and delivers antibiotic-containing culture media to eight 30-nL chambers for cell culture. When tested with 20 μL samples of a model bacterial strain (E. coli ATCC 25922 treated with ampicillin or streptomycin, our method allows for the determination of minimum inhibitory concentrations consistent with the microdilution test in three hours, which is almost a factor of ten more rapid than the standard method.

  13. CLSI broth microdilution method for testing susceptibility of Malassezia pachydermatis to thiabendazole

    OpenAIRE

    2009-01-01

    Thiabendazole, classified as antiparasitic and also used as an antifungal drug, can be found as otological solution indicated for treatment of parasitic and fungal external otitis in small animals. Malassezia pachydermatis is a yeast recognized as a normal inhabitant on the skin and mucous membranes of dogs and cats. However, it is considered an opportunistic agent that causes external otitis and dermatitis in these animals. The aim of this study was to evaluate the in vitro effect of thiaben...

  14. Antibacterial and antifungal activities of some Mexican medicinal plants.

    Science.gov (United States)

    Ruiz-Bustos, E; Velazquez, C; Garibay-Escobar, A; García, Z; Plascencia-Jatomea, M; Cortez-Rocha, M O; Hernandez-Martínez, J; Robles-Zepeda, R E

    2009-12-01

    In Mexico about 4,000 plant species have some medicinal use. The aim of this work was to evaluate the antimicrobial activity of six Mexican medicinal plants against fungi and Gram-positive and Gram-negative bacteria. Methanolic extracts were prepared from the Mexican medicinal plants Amphypteringium adstrigens, Castella tortuosa, Coutarea latiflora, Ibervillea sonorae, Jatropha cuneata, and Selaginella lepidophylla. The antibacterial and antifungal activities of the plants were determined by the broth microdilution method and the radial growth inhibition assay, respectively. All Mexican plants tested showed antimicrobial activity. Among the six plant extracts analyzed, J. cuneata showed the highest growth-inhibitory activity against fungi, Gram-positive and Gram-negative bacteria (J. cuneata > A. adstrigens > C. latiflora > C. tortuosa > I. sonorae approximately S. lepidophylla). Shigella flexneri and Staphylococcus aureus were the most susceptible bacteria to plant extracts. Complete inhibition of S. flexneri growth was observed with J. cuneata methanolic extract at 90 microg/mL. This plant extract also showed the strongest antifungal activity against Fusarium verticillioides and Aspergillus niger. Our data suggest that the medicinal plants tested have important antimicrobial properties. This is the first report describing the antimicrobial activities of several of the Mexican medicinal plants used in this study.

  15. In vitro susceptibilities of Zygomycota to polyenes.

    Science.gov (United States)

    Dannaoui, Eric; Meis, Jacques F G M; Mouton, Johan W; Verweij, Paul E

    2002-05-01

    The in vitro activity of polyenes was determined for 36 isolates of Zygomycota including Rhizopus spp. (15), Absidia corymbifera (10), Mucor spp. (6), Rhizomucor spp. (3), Cunninghamella bertholletiae (1) and Apophysomyces elegans (1). All isolates were tested with amphotericin B, heated amphotericin B and nystatin by a broth microdilution test. There were no significant differences between heated and unheated solutions of amphotericin B in terms of their antifungal activities. The MICs of amphotericin B for most of the strains were <2 mg/L. For all isolates, nystatin was significantly less active than amphotericin B (P < 0.001). The one C. bertholletiae and one A. elegans isolates were less susceptible to amphotericin B (MICs 2 mg/L) and were also less susceptible to nystatin.

  16. Bile Culture and Susceptibility Testing of Malignant Biliary Obstruction via PTBD

    Energy Technology Data Exchange (ETDEWEB)

    Yu Haipeng; Guo Zhi, E-mail: jieruke@yahoo.com.cn; Xing Wenge; Guo Xiuying; Liu Fang; Li Baoguo [Tinajin Medical University Cancer Institute and Hospital, Department of Interventional Therapy, Tianjin Key Cancer Prevention and Treatment Laboratory (China)

    2012-10-15

    Purpose: To assess the information obtained by bile culture and susceptibility testing for malignant biliary obstruction by a retrospective one-center study. Methods: A total of 694 patients with malignant biliary obstruction received percutaneous transhepatic biliary drainage during the period July 2003 to September 2010, and subsequently, bile specimens were collected during the procedure. Among the 694 patients, 485 were men and 209 were women, ranging in age from 38 to 78 years (mean age 62 years). Results: A total of 42.9% patients had a positive bile culture (298 of 694). Further, 57 species of microorganisms and 342 strains were identified; gram-positive bacteria accounted for 50.9% (174 of 342) and gram-negative bacteria accounted for 41.5% (142 of 342) of these strains. No anaerobes were obtained by culture during this study. The most common microorganisms were Enterococcus faecalis (41 of 342, 11.9%), Escherichia coli (34 of 342, 9.9%), Klebsiella pneumoniae (28 of 342, 8.2%), Staphylococcus epidermidis (19 of 342, 5.5%), Enterococcus (18 of 342, 5.3%), and Enterobacter cloacae (16 of 342, 4.7%). The percentage of {beta}-lactamase-producing gram-positive bacteria was 27.6% (48 of 174), and the percentage of gram-negative bacteria was 19.7% (28 of 142). The percentage of enzyme-producing Escherichia coli was 61.7% (21 of 34). Conclusion: The bile cultures in malignant biliary obstruction are different from those in the Tokyo Guidelines and other benign biliary obstruction researches, which indicates that a different antibacterial therapy should be applied. Thus, knowledge of the antimicrobial susceptibility data could aid in the better use of antibiotics for the empirical therapy of biliary infection combined with malignant biliary obstruction.

  17. Modified protocol for drug susceptibility testing of MGIT cultures of Mycobacterium tuberculosis by the MGIT 960.

    Science.gov (United States)

    Adami, Aline Gois; Gallo, Juliana Failde; Pinhata, Juliana Maira Watanabe; Martins, Maria Conceição; Giampaglia, Carmen Maria Saraiva; de Oliveira, Rosangela Siqueira

    2017-02-01

    A rapid detection of resistance in Mycobacterium tuberculosis is crucial for management and control of tuberculosis. This study evaluated a more rapid and cost-effective drug susceptibility testing (DST) protocol using primary isolates of M. tuberculosis in mycobacteria growth indicator tube (MGIT). Ninety-four M. tuberculosis isolates in MGIT were subjected to DST by the manufacturer's method, i.e., primary isolates were subcultured and DST was performed from positive cultures for a maximum of 5days; and by our modified method, i.e., DST was performed directly from primary MGIT cultures positive for more than 5days. Results were concordant for 76 (81%) isolates. Agreement between both methods was 92.0%, 98.9%, 97.7%, and 95.5% for streptomycin, isoniazid, rifampicin, and ethambutol, respectively. Six isolates failed to grow on the recommended method, including 3 resistant isolates. Not performing subculture of primary M. tuberculosis isolates yields reliable results, decreasing the turnaround time and the cost of the test.

  18. Rapid Real-Time Antimicrobial Susceptibility Testing with Electrical Sensing on Plastic Microchips with Printed Electrodes.

    Science.gov (United States)

    Safavieh, Mohammadali; Pandya, Hardik J; Venkataraman, Maanasa; Thirumalaraju, Prudhvi; Kanakasabapathy, Manoj Kumar; Singh, Anupriya; Prabhakar, Devbalaji; Chug, Manjyot Kaur; Shafiee, Hadi

    2017-03-30

    Rapid antimicrobial susceptibility testing is important for efficient and timely therapeutic decision making. Due to globally spread bacterial resistance, the efficacy of antibiotics is increasingly being impeded. Conventional antibiotic tests rely on bacterial culture, which is time-consuming and can lead to potentially inappropriate antibiotic prescription and up-front broad range of antibiotic use. There is an urgent need to develop point-of-care platform technologies to rapidly detect pathogens, identify the right antibiotics, and monitor mutations to help adjust therapy. Here, we report a biosensor for rapid (microchips with printed electrodes using antibodies (30 min), and its electrical response is monitored in the presence and absence of antibiotics over an hour of incubation time. We evaluated the microchip with Escherichia coli and methicillin-resistant Staphylococcus aureus (MRSA) as clinical models with ampicillin, ciprofloxacin, erythromycin, daptomycin, gentamicin, and methicillin antibiotics. The results are compared with the current standard methods, i.e. bacteria viability and conventional antibiogram assays. The technology presented here has the potential to provide precise and rapid bacteria screening and guidance in clinical therapies by identifying the correct antibiotics for pathogens.

  19. Evaluation of MGIT 960 System for the Second-Line Drugs Susceptibility Testing of Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Hyejin Kim

    2013-01-01

    Full Text Available Many laboratories validate DST of the second-line drugs by BACTEC MGIT 960 system. The objective of this study is to evaluate the critical concentration and perform DST for the 2nd line drugs. We evaluated 193 clinical strains of M. tuberculosis isolated from patients in South Korea. Testing the critical concentration of six second-line drugs was performed by MGIT 960 and compared with L-J proportion method. The critical concentration was determined to establish the most one that gave the difference between drug resistance and susceptibility in MGIT960 system. Good agreement of the following concentrations was found: Concordance was 95% for 0.5 μg/mL of moxifloxacin; 93.6%, 1.0 μg/mL of levofloxacin; 97.5%, 2.5 μg/mL of kanamycin; 90.6%, 2.5 μg/mL of capreomycin; 86.2%, 5.0 μg/mL of ethionamide; and 90.8%, 2.0 μg/mL of ρ-aminosalicylic acid. The critical concentrations of the four drugs, moxifloxacin, levofloxacin, kanamycin, and capreomycin, were concordant and reliable for testing 2nd line drug resistance. Further study of ethionamide and ρ-aminosalicylic acid is required.

  20. Multicenter evaluation of fully automated BACTEC Mycobacteria Growth Indicator Tube 960 system for susceptibility testing of Mycobacterium tuberculosis.

    Science.gov (United States)

    Bemer, Pascale; Palicova, Frantiska; Rüsch-Gerdes, Sabine; Drugeon, Henri B; Pfyffer, Gaby E

    2002-01-01

    The reliability of the BACTEC Mycobacteria Growth Indicator Tube (MGIT) 960 system for testing of Mycobacterium tuberculosis susceptibility to the three front-line drugs (isoniazid [INH], rifampin [RIF], and ethambutol [EMB]) plus streptomycin (STR) was compared to that of the BACTEC 460 TB system. The proportion method was used to resolve discrepant results by an independent arbiter. One hundred and ten strains were tested with an overall agreement of 93.5%. Discrepant results were obtained for seven strains (6.4%) with INH (resistant by BACTEC MGIT 960; susceptible by BACTEC 460 TB), for one strain (0.9%) with RIF (resistant by BACTEC MGIT 960; susceptible by BACTEC 460 TB), for seven strains (6.4%) with EMB (six resistant by BACTEC MGIT 960 and susceptible by BACTEC 460 TB; one susceptible by BACTEC MGIT 960 and resistant by BACTEC 460 TB), and for 19 strains (17.3%) with STR (resistant by BACTEC MGIT 960 and susceptible by BACTEC 460 TB). After resolution of discrepant results, the sensitivity of the BACTEC MGIT 960 system was 100% for all four drugs and specificity ranged from 89.8% for STR to 100% for RIF. Turnaround times were 4.6 to 11.7 days (median, 6.5 days) for BACTEC MGIT 960 and 4.0 to 10.0 days (median, 7.0 days) for BACTEC 460 TB. These data demonstrate that the fully automated and nonradiometric BACTEC MGIT 960 system is an accurate method for rapid susceptibility testing of M. tuberculosis.

  1. 209株临床分离的不同基因型白假丝酵母菌和丝状真菌的抗真菌药物敏感性及其耐药性趋势研究%Study on antifungal susceptibilities and resistance trends of 209 strains of C.albicans with various genotypes and filamentous fungi isolated from clinical specimens

    Institute of Scientific and Technical Information of China (English)

    王和; 康颖倩; 刘姝; 罗振华; 王丹霓

    2012-01-01

    目的 了解常见致病性真菌的药物敏感性及其耐药性趋势.方法 检测临床分离白假丝酵母菌和丝状菌的抗真菌药物敏感性.结果 176株白假丝酵母菌A型(55.1%)、B型(29.0%)和C型(15.9%)的FC和AP敏感率较高(88.7%~100%),FL、IT及VR 敏感率较低(小于26%).各基因型菌株在FC、AP、FL、IT、VR敏感性无明显差异(P>0.05),在FC与AP、FL、IT、VR,AP与FL、IT、VR之间存在显著的敏感率差异(P<0.001).33株丝状菌的MIC值,9株(27.3%)AP≥3μg/mL,7株(21.2 %)FC≥3μg/mL,32株(97%)FL>256μg/mL.各菌株的FC、AP、FL、IT、CS敏感率有差异,AP与FL、CS,IT与FL、CS,FC与FL、CS,FL与CS之间的敏感率差异显著(P<0.001).结论 白假丝酵母菌基因A型、B型和C型对所测试抗真菌药物的敏感性无明显差异.白假丝酵母菌和丝状菌多数菌株表现出较高的耐药率和明显的耐药性发展趋势.%Objective To investigate the antifungal susceptibilities and resistance trends of the pathogenic fungi. Methods With the method of minimum inhibitory concentration (MIC) test, the susceptibilities and the trend of drug-resistance to flucytosine, amphotericin B, fluconazole, itraconazole, voriconazole and/or caspofungin of 176 strains of C. albicans and 33 filamentous fungi which were isolated from patients with infectious diseases were detected. Results 176 strains of C. albicans, including 97 strains of genotype A (55.1%), 51 strains of genotype B (29.0%) and 28 strains of genotype C (15.9%), were sensitive to flucytosine and amphotericin B (sensitivity rate of 88.7% to 100%) but resistant to fluconazole, itraconazole and voriconazole (sensitivity rate<26%=. In various genotypes of C. albicans, no significant difference in susceptibility rates of flucytosine, amphotericin B, fluconazole, itraconazole, voriconazole can be found (P>0.05). However, the significant difference (P<0.001= in susceptibility rates of different antifungal

  2. Tolerance of yeast biofilm cells towards systemic antifungals

    DEFF Research Database (Denmark)

    Bojsen, Rasmus Kenneth

    was the only tested drug with activity against both growth arrested biofilm and planktonic cells but was found to only kill ~95 % of the cells. By using a collection of barcode tagged deletion mutants, we were identified that defects in protein synthesis, intracellular transport, cell cycle and lipid...... metabolism resulted in increased amphotericin B tolerance in both biofilm and planktonic cells. We furthermore observed that the tolerance level could be enhanced by nutrient starvation and inhibition of the TOR pathway. In conclusion, antifungal tolerance is the combined effect of the physiological state......Fungal infections have become a major problem in the hospital sector in the past decades due to the increased number of immune compromised patients susceptible to mycosis. Most human infections are believed to be associated with biofilm forming cells that are up to 1000-fold more tolerant...

  3. Sensitization of Candida albicans biofilms to various antifungal drugs by cyclosporine A

    Directory of Open Access Journals (Sweden)

    Shinde Ravikumar B

    2012-10-01

    Full Text Available Abstract Background Biofilms formed by Candida albicans are resistant towards most of the available antifungal drugs. Therefore, infections associated with Candida biofilms are considered as a threat to immunocompromised patients. Combinatorial drug therapy may be a good strategy to combat C. albicans biofilms. Methods Combinations of five antifungal drugs- fluconazole (FLC, voriconazole (VOR, caspofungin (CSP, amphotericin B (AmB and nystatin (NYT with cyclosporine A (CSA were tested in vitro against planktonic and biofilm growth of C. albicans. Standard broth micro dilution method was used to study planktonic growth, while biofilms were studied in an in vitro biofilm model. A chequerboard format was used to determine fractional inhibitory concentration indices (FICI of combination effects. Biofilm growth was analyzed using XTT-metabolic assay. Results MICs of various antifungal drugs for planktonic growth of C. albicans were lowered in combination with CSA by 2 to 16 fold. Activity against biofilm development with FIC indices of 0.26, 0.28, 0.31 and 0.25 indicated synergistic interactions between FLC-CSA, VOR-CSA, CSP-CSA and AmB-CSA, respectively. Increase in efficacy of the drugs FLC, VOR and CSP against mature biofilms after addition of 62.5 μg/ml of CSA was evident with FIC indices 0.06, 0.14 and 0.37, respectively. Conclusions The combinations with CSA resulted in increased susceptibility of biofilms to antifungal drugs. Combination of antifungal drugs with CSA would be an effective prophylactic and therapeutic strategy against biofilm associated C. albicans infections.

  4. Antifungal activity of selected Malaysian honeys: a comparison with Manuka honey

    Directory of Open Access Journals (Sweden)

    Siti Aisyah Sayadi

    2015-07-01

    Full Text Available Objective: To evaluate four selected Malaysian honey samples from different floral sources (Gelam, Tualang, Nenas and Acacia for their ability to inhibit the growth of fungi and yeast strains (Candida albicans, Aspergillus niger, Epidermophyton floccosum, Microsporum gypseum, Trichophyton rubrum and Trichophyton mentagrophytes. Methods: The broth microdilution method was used to assess the antifungal activity of honey against yeasts at different concentrations ranging from 0.01% to 70% (v/v. Minimum inhibitory concentration (MIC of the honeys were determined by visual inspection and spectrophotometric assay. Minimum fungicidal concentration test was performed by further sub-culturing from the plates which showed no visible growth in the MIC assay onto Sabroud dextrose agar. Results: All tested Malaysian honeys except Gelam showed antifungal activity against all species analysed, with the MIC ranging from 25% (v/v to 50% (v/v while MIC of Manuka honey ranged between 21% to 53% (v/v. Candida albicans was more susceptible to honey than other species tested. Conclusions: Locally produced honeys exhibited antifungal activity which is less than or equal to that of Manuka honey. Our data showed evidence in support of the therapeutic uses of Malaysian honeys.

  5. Antifungal activity of selected Malaysian honeys:a comparison with Manuka honey

    Institute of Scientific and Technical Information of China (English)

    Siti Aisyah Sayadi; Rozaini Mohd Zohdi; Nur Salme Suhana Shamshuddin; Muna Syairah Khairy; Nur Ashikin Hasan; Ahmad Syamil Yasin; Kalavathy Ramasamy

    2015-01-01

    Objective:To evaluate four selected Malaysian honey samples from different floral sources (Gelam,Tualang,Nenas and Acacia) for their ability to inhibit the growth of fungi and yeast strains (Candida albicans, Aspergillus niger, Epidermophyton floccosum, Microsporum gypseum, Trichophyton rubrum andTrichophyton mentagrophytes). Methods: The broth microdilution method was used to assess the antifungal activity of honey against yeasts at different concentrations ranging from 0.01% to 70% (v/v). Minimum inhibitory concentration (MIC) of the honeys were determined by visual inspection and spectrophotometric assay. Minimum fungicidal concentration test was performed by further sub-culturing from the plates which showed no visible growth in theMICassay onto Sabroud dextrose agar. Results: All tested Malaysian honeys except Gelam showed antifungal activity against all species analysed, with theMIC ranging from 25% (v/v) to 50% (v/v) whileMIC of Manuka honey ranged between 21% to 53%(v/v).Candida albicans was more susceptible to honey than other species tested. Conclusions: Locally produced honeys exhibited antifungal activity which is less than or equal to that of Manuka honey. Our data showed evidence in support of the therapeutic uses of Malaysian honeys.

  6. Nosocomial Candidiasis: Antifungal Stewardship and the Importance of Rapid Diagnosis.

    Science.gov (United States)

    Pfaller, Michael A; Castanheira, Mariana

    2016-01-01

    Candidemia and other forms of candidiasis are associated with considerable excess mortality and costs. Despite the addition of several new antifungal agents with improved spectrum and potency, the frequency of Candida infection and associated mortality have not decreased in the past two decades. The lack of rapid and sensitive diagnostic tests has led to considerable overuse of antifungal agents resulting in increased costs, selection pressure for resistance, unnecessary drug toxicity, and adverse drug interactions. Both the lack of timely diagnostic tests and emergence of antifungal resistance pose considerable problems for antifungal stewardship. Whereas antifungal stewardship with a focus on nosocomial candidiasis should be able to improve the administration of antifungal therapy in terms of drug selection, proper dose and duration, source control and de-escalation therapy, an important parameter, timeliness of antifungal therapy, remains a victim of slow and insensitive diagnostic tests. Fortunately, new proteomic and molecular diagnostic tools are improving the time to species identification and detection. In this review we will describe the potential impact that rapid diagnostic testing and antifungal stewardship can have on the management of nosocomial candidiasis.

  7. Methylxanthine Inhibit Fungal Chitinases and Exhibit Antifungal Activity

    Science.gov (United States)

    Tsirilakis, Kalliope; Kim, Christy; Vicencio, Alfin G.; Andrade, Christopher; Casadevall, Arturo; Goldman, David L.

    2015-01-01

    Chitinases are necessary for fungal cell wall remodeling and cell replication. Methylxanthines have been shown to competitively inhibit family 18 chitinases in vitro. We sought to determine the effects of methylxanthines on fungal chitinases. Fungi demonstrated variable chitinase activity and incubation with methylxanthines (0.5–10 mM) resulted in a dose-dependent decrease in this activity. All fungi tested, except for Candida spp., demonstrated growth inhibition in the presence of methylxanthines at a concentration of 10 mM. India ink staining demonstrated impaired budding and decreased cell size for methylxanthine-treated Cryptococcus neoformans. C. neoformans and Aspergillus fumigatus treated with pentoxifylline also exhibited abnormal cell morphology. In addition, pentoxifylline-treated C. neoformans exhibited increased susceptibility to calcofluor and a leaky melanin phenotype consistent with defective cell wall function. Our data suggest that a variety of fungi express chitinases and that methylxanthines have antifungal properties related to their inhibition of fungal chitinases. Our results highlight the potential utility of targeting chitinases in the development of novel antifungal therapies. PMID:21968902

  8. Cryptoccocal meningitis in Yaoundé (Cameroon) HIV infected patients: Diagnosis, frequency and Cryptococcus neoformans isolates susceptibility study to fluconazole.

    Science.gov (United States)

    Kammalac Ngouana, T; Dongtsa, J; Kouanfack, C; Tonfack, C; Fomena, S; Mallié, M; Delaporte, E; Boyom, F-Fekam; Bertout, S

    2015-03-01

    Cryptococcal meningitis is a mycosis encountered especially in patients with Acquired Immunodeficiency Syndrome and is fatal in the absence of treatment. Information on epidemiology, diagnosis and susceptibility profile to antifungal drugs, are scarce in Cameroon. Authors evaluated the diagnosis possibilities of the cryptococcal meningitis in Cameroon, and studied the antifungal susceptibility of isolated strains to fluconazole, used as first line treatment of the disease in Cameroon. Between December 2009 and July 2011, 146 cerebrospinal fluids obtained from HIV patients with suspicion of meningitis were analysed. The diagnosis procedure involved macroscopic and cyto-chemical analysis, India ink test, culture on Sabouraud chloramphenicol medium and antigen latex agglutination test. Antifungal susceptibility testing of isolated strains to fluconazole was done by the E-test(®) method. The diagnosis of cryptococcal meningitis gave 28.08% positive cases. Among these patients, 80% were at stages III and IV and 20% at stage I of the HIV infection, according to the WHO previous classification. Cyto-chemical analysis showed current findings in the case of cryptococcal meningitis. India ink test and latex agglutination test exhibited very high sensitivity and specificity (>94%). Fluconazole antifungal susceptibility testing gave MICs lower than 32μg/mL to 92.7% of isolated strains and MICs greater than this value to 7.3% of isolates. These results showed that cryptococcal meningitis remains a real problem among HIV infected patients in Yaoundé. The emergence of fluconazole reduced susceptibility strains is worrying. Nevertheless, efficacy of rapid detection tests is interesting because this will help in rapid diagnosis and treatment of patients.

  9. An Antifungal Combination Matrix Identifies a Rich Pool of Adjuvant Molecules that Enhance Drug Activity against Diverse Fungal Pathogens

    Directory of Open Access Journals (Sweden)

    Nicole Robbins

    2015-11-01

    Full Text Available There is an urgent need to identify new treatments for fungal infections. By combining sub-lethal concentrations of the known antifungals fluconazole, caspofungin, amphotericin B, terbinafine, benomyl, and cyprodinil with ∼3,600 compounds in diverse fungal species, we generated a deep reservoir of chemical-chemical interactions termed the Antifungal Combinations Matrix (ACM. Follow-up susceptibility testing against a fluconazole-resistant isolate of C. albicans unveiled ACM combinations capable of potentiating fluconazole in this clinical strain. We used chemical genetics to elucidate the mode of action of the antimycobacterial drug clofazimine, a compound with unreported antifungal activity that synergized with several antifungals. Clofazimine induces a cell membrane stress for which the Pkc1 signaling pathway is required for tolerance. Additional tests against additional fungal pathogens, including Aspergillus fumigatus, highlighted that clofazimine exhibits efficacy as a combination agent against multiple fungi. Thus, the ACM is a rich reservoir of chemical combinations with therapeutic potential against diverse fungal pathogens.

  10. 曲霉临床和环境分离株基因型和药敏表型分析%Genotyping and susceptibility testing of Aspergillus species from clinical settings and environmental sources

    Institute of Scientific and Technical Information of China (English)

    敖俊红; 杨燕妮; 郝飞

    2010-01-01

    Objective To investigate the genotype and antifungal susceptibility of AspergiIlus species isolated from clinical settings and environmental sources. Methods Random amplification of polymorphic DNA (RAPD) analysis was performed to profile the genotypes of 48 Aspergillus fumigatus strains, 59 Aspergillus flavus strains and 30 Aspergillus niger strains isolated from clinical settings and environmental sources. NCCLS M38-A protocol was carried out for antifungal susceptibility testing of these Aspergillus isolates. Results As RAPD analysis showed, the 48 Aspergillus fumigatus strains were classified into 8 genotypes, 59 Aspergillus flavus strains into 12 genotypes, and 30 Aspergillus niger strains into 5 genotypes. A significant difference was observed in the susceptibility to terbinafine among the 8 genotypes of Aspergillus fumigatus (χ2 = 33.092, P <0.01 ) as well as in that to amphotericin B among the 5 genotypes of Aspergillus niger (χ2 = 15.185, P< 0.05).No statistical difference was found in the susceptibility to amphoteriein B, itraconazole, fluconazole or flucytosine among the 8 genotypes of Aspergillus fumigatus or in that to terbinafine, itraconazole, fluconazole or flucytosine among the 5 genotypes of Aspergillus niger. Conclusion There is some difference in the susceptibility to some antifungal agents between different genotypes of Aspergillus fumigatus and Aspergillus niger.%目的 探讨临床和环境来源曲霉的基因分型情况以及不同基因型曲霉的药敏结果.方法 采用随机扩增DNA多态性(RAPD)分型方法对45株烟曲霉、59株黄曲霉和30株黑曲霉进行基因分型,采用M38-A方案分析曲霉对抗真菌药物的敏感性.结果 45株烟曲霉产生8种基因型,59株黄曲霉产生12种基因型,30株黑曲霉产生5种基因型.8种基因型烟曲霉对特比萘芬敏感性差异有统计学意义(χ2=33.092,P<0.01),而对其他抗真菌药物敏感性差异无统计学意义.5种基因型黑曲霉对两性

  11. Antimicrobial susceptibility testing for Helicobacter pylori isolates from Brazilian children and adolescents: comparing agar dilution, E-test, and disk diffusion.

    Science.gov (United States)

    Ogata, Silvio Kazuo; Gales, Ana Cristina; Kawakami, Elisabete

    2014-01-01

    Antimicrobial susceptibility testing for Helicobacter pylori is increasingly important due to resistance to the most used antimicrobials agents. Only agar dilution method is approved by CLSI, but it is difficult to perform routinely. We evaluated the reliability of E-test and disk diffusion comparing to agar dilution method on Helicobacter pylori antimicrobial susceptibility testing. Susceptibility testing was performed for amoxicillin, clarithromycin, furazolidone, metronidazole and tetracycline using E-test, disk-diffusion and agar dilution method in 77 consecutive Helicobacter pylori strains from dyspeptic children and adolescents. Resistance rates were: amoxicillin - 10.4%, 9% and 68.8%; clarithromycin - 19.5%, 20.8%, 36.3%; metronidazole - 40.2%33.7%, 38.9%, respectively by agar dilution, E-test and disk diffusion method. Furazolidone and tetracycline showed no resistance rates. Metronidazole presented strong correlation to E-test (r = 0.7992, p Helicobacter pylori susceptibility testing. Disk diffusion method presented high disagreement and high rates of major errors.

  12. Rapid identification and antimicrobial susceptibility testing of positive blood cultures using MALDI-TOF MS and a modification of the standardised disc diffusion test: a pilot study.

    LENUS (Irish Health Repository)

    Fitzgerald, C

    2016-04-27

    In an era when clinical microbiology laboratories are under increasing financial pressure, there is a need for inexpensive, yet effective, rapid microbiology tests. The aim of this study was to evaluate a novel modification of standard methodology for the identification and antimicrobial susceptibility testing (AST) of pathogens in positive blood cultures, reducing the turnaround time of laboratory results by 24 h.

  13. Design, characterization, teratogenicity testing, antibacterial, antifungal and DNA interaction of few high spin Fe(II) Schiff base amino acid complexes

    Science.gov (United States)

    Abdel-Rahman, Laila H.; El-Khatib, Rafat M.; Nassr, Lobna A. E.; Abu-Dief, Ahmed M.; Lashin, Fakhr El-Din

    2013-07-01

    In this study, new Fe(II) Schiff base amino acid chelates derived from the condensation of o-hydroxynaphthaldehyde with L-alanine, L-phenylalanine, L-aspartic acid, L-histidine and L-arginine were synthesized and characterized via elemental, thermogravimetric analysis, molar conductance, IR, electronic, mass spectra and magnetic moment measurements. The stoichiometry and the stability constants of the complexes were determined spectrophotometrically. Correlation of all spectroscopic data suggested that Schiff bases ligands exhibited tridentate with ONO sites coordinating to the metal ions via protonated phenolic-OH, azomethine-N and carboxylate-O with the general formulae [Fe(HL)2]·nH2O. But in case of L-histidine, the ligand acts as tetradentate via deprotonated phenolic-OH, azomethine-N, carboxylate-O and N-imidazole ring ([FeL(H2O)2]·2H2O), where HL = mono anion and L = dianion of the ligand. The structure of the prepared complexes is suggested to be octahedral. The prepared complexes were tested for their teratogenicity on chick embryos and found to be safe until a concentration of 100 μg/egg with full embryos formation. Moreover, the interaction between CT-DNA and the investigated complexes were followed by spectrophotometric and viscosity measurements. It was found that, the prepared complexes bind to DNA via classical intercalative mode and showed a different DNA activity with the sequence: nhi > nari > nali > nasi > nphali. Furthermore, the free ligands and their complexes are screened for their in vitro antibacterial and antifungal activity against three types of bacteria, Escherichia coli, Pseudomonas aeruginosa and Bacillus cereus and three types of anti fungal cultures, Penicillium purpurogenium, Aspergillus flavus and Trichotheium rosium in order to assess their antimicrobial potential. The results show that the metal complexes are more reactive with respect to their corresponding Schiff base amino acid ligands.

  14. In vitro activities of new and conventional antifungal agents against clinical Scedosporium isolates.

    NARCIS (Netherlands)

    Meletiadis, J.; Meis, J.F.G.M.; Mouton, J.W.; Rodriguez-Tudela, J.L.; Donnelly, J.P.; Verweij, P.E.

    2002-01-01

    The susceptibilities of 13 clinical isolates of Scedosporium apiospermum and 55 clinical isolates of S. prolificans to new and conventional drugs belonging to three different classes of antifungal agents, the azoles (miconazole, itraconazole, voriconazole, UR-9825, posaconazole), the polyenes (ampho

  15. Rapid colorimetric testing for pyrazinamide susceptibility of M. tuberculosis by a PCR-based in-vitro synthesized pyrazinamidase method.

    Science.gov (United States)

    Zhou, Man; Geng, Xuelei; Chen, Jun; Wang, Xude; Wang, Dianbing; Deng, Jiaoyu; Zhang, Zhiping; Wang, Weihua; Zhang, Xian-En; Wei, Hongping

    2011-01-01

    Pyrazinamide (PZA) is an important first-line anti-tuberculosis drug. But PZA susceptibility test is challenging because PZA activity is optimal only in an acid environment that inhibits the growth of M. tuberculosis. For current phenotypic methods, inconsistent results between different labs have been reported. Direct sequencing of pncA gene is being considered as an accurate predictor for PZA susceptibility, but this approach needs expensive sequencers and a mutation database to report the results. An in-vitro synthesized Pyrazinamidase (PZase) assay was developed based on PCR amplification of pncA gene and an in vitro wheat germ system to express the pncA gene into PZase. The activity of the synthesized PZase was used as an indicator for PZA susceptibility. Fifty-one clinical isolates were tested along with pncA sequencing and the BACTEC MGIT 960 methods. The in-vitro synthesized PZase assay was able to detect PZA susceptibility of M. tuberculosis within 24 h through observing the color difference either by a spectrometer or naked eyes. This method showed agreements of 100% (33/33) and 88% (14/16) with the pncA sequencing method, and agreements of 96% (27/28) and 65% (15/23) with the BACTEC MGIT 960 method, for susceptible and resistant strains, respectively. The novel in-vitro synthesized PZase assay has significant advantages over current methods, such as its fast speed, simplicity, no need for expensive equipment, and the potentials of being a direct test, predicting resistance level and easy reading results by naked eyes. After confirmation by more clinical tests, this method may provide a radical change to the current PZA susceptibility assays.

  16. Rapid colorimetric testing for pyrazinamide susceptibility of M. tuberculosis by a PCR-based in-vitro synthesized pyrazinamidase method.

    Directory of Open Access Journals (Sweden)

    Man Zhou

    Full Text Available Pyrazinamide (PZA is an important first-line anti-tuberculosis drug. But PZA susceptibility test is challenging because PZA activity is optimal only in an acid environment that inhibits the growth of M. tuberculosis. For current phenotypic methods, inconsistent results between different labs have been reported. Direct sequencing of pncA gene is being considered as an accurate predictor for PZA susceptibility, but this approach needs expensive sequencers and a mutation database to report the results. An in-vitro synthesized Pyrazinamidase (PZase assay was developed based on PCR amplification of pncA gene and an in vitro wheat germ system to express the pncA gene into PZase. The activity of the synthesized PZase was used as an indicator for PZA susceptibility. Fifty-one clinical isolates were tested along with pncA sequencing and the BACTEC MGIT 960 methods. The in-vitro synthesized PZase assay was able to detect PZA susceptibility of M. tuberculosis within 24 h through observing the color difference either by a spectrometer or naked eyes. This method showed agreements of 100% (33/33 and 88% (14/16 with the pncA sequencing method, and agreements of 96% (27/28 and 65% (15/23 with the BACTEC MGIT 960 method, for susceptible and resistant strains, respectively. The novel in-vitro synthesized PZase assay has significant advantages over current methods, such as its fast speed, simplicity, no need for expensive equipment, and the potentials of being a direct test, predicting resistance level and easy reading results by naked eyes. After confirmation by more clinical tests, this method may provide a radical change to the current PZA susceptibility assays.

  17. Evaluation of four colourimetric susceptibility tests for the rapid detection of multidrug-resistant Mycobacterium tuberculosisisolates

    Directory of Open Access Journals (Sweden)

    Ahmet Yilmaz Coban

    2015-08-01

    Full Text Available The purpose of this study is to evaluate four rapid colourimetric methods, including the resazurin microtitre assay (REMA, malachite green decolourisation assay (MGDA, microplate nitrate reductase assay (MNRA and crystal violet decolourisation assay (CVDA, for the rapid detection of multidrug-resistant (MDR tuberculosis. Fifty Mycobacterium tuberculosisisolates were used in this study. Eighteen isolates were MDR, two isolates were only resistant to isoniazid (INH and the remaining isolates were susceptible to both INH and rifampicin (RIF. INH and RIF were tested in 0.25 µg/mL and 0.5 µg/mL, respectively. The agar proportion method was used as a reference method. MNRA and REMA were performed with some modifications. MGDA and CVDA were performed as defined in the literature. The agreements of the MNRA for INH and RIF were 96% and 94%, respectively, while the agreement of the other assays for INH and RIF were 98%. In this study, while the specificities of the REMA, MGDA and CVDA were 100%, the specificity of the MNRA was lower than the others (93.3% for INH and 90.9% for RIF. In addition, while the sensitivity of the MNRA was 100%, the sensitivities of the others were lower than that of the MNRA (from 94.1-95%. The results were reported on the seventh-10th day of the incubation. All methods are reliable, easy to perform, inexpensive and easy to evaluate and do not require special equipment.

  18. A Low Cost/Low Power Open Source Sensor System for Automated Tuberculosis Drug Susceptibility Testing

    Directory of Open Access Journals (Sweden)

    Kyukwang Kim

    2016-06-01

    Full Text Available In this research an open source, low power sensor node was developed to check the growth of mycobacteria in a culture bottle with a nitrate reductase assay method for a drug susceptibility test. The sensor system reports the temperature and color sensor output frequency change of the culture bottle when the device is triggered. After the culture process is finished, a nitrite ion detecting solution based on a commercial nitrite ion detection kit is injected into the culture bottle by a syringe pump to check bacterial growth by the formation of a pigment by the reaction between the solution and the color sensor. Sensor status and NRA results are broadcasted via a Bluetooth low energy beacon. An Android application was developed to collect the broadcasted data, classify the status of cultured samples from multiple devices, and visualize the data for the end users, circumventing the need to examine each culture bottle manually during a long culture period. The authors expect that usage of the developed sensor will decrease the cost and required labor for handling large amounts of patient samples in local health centers in developing countries. All 3D-printerable hardware parts, a circuit diagram, and software are available online.

  19. A Low Cost/Low Power Open Source Sensor System for Automated Tuberculosis Drug Susceptibility Testing.

    Science.gov (United States)

    Kim, Kyukwang; Kim, Hyeong Keun; Lim, Hwijoon; Myung, Hyun

    2016-06-22

    In this research an open source, low power sensor node was developed to check the growth of mycobacteria in a culture bottle with a nitrate reductase assay method for a drug susceptibility test. The sensor system reports the temperature and color sensor output frequency change of the culture bottle when the device is triggered. After the culture process is finished, a nitrite ion detecting solution based on a commercial nitrite ion detection kit is injected into the culture bottle by a syringe pump to check bacterial growth by the formation of a pigment by the reaction between the solution and the color sensor. Sensor status and NRA results are broadcasted via a Bluetooth low energy beacon. An Android application was developed to collect the broadcasted data, classify the status of cultured samples from multiple devices, and visualize the data for the end users, circumventing the need to examine each culture bottle manually during a long culture period. The authors expect that usage of the developed sensor will decrease the cost and required labor for handling large amounts of patient samples in local health centers in developing countries. All 3D-printerable hardware parts, a circuit diagram, and software are available online.

  20. CLSI broth microdilution method for testing susceptibility of Malasseziapachydermatis to thiabendazole CLSI método de Microdiluição em Caldo para teste de suscetibilidade da Malassezia pachydermatis frente ao tiabendazol

    Directory of Open Access Journals (Sweden)

    Patrícia da Silva Nascente

    2009-06-01

    Full Text Available Thiabendazole, classified as antiparasitic and also used as an antifungal drug, can be found as otological solution indicated for treatment of parasitic and fungal external otitis in small animals. Malassezia pachydermatis is a yeast recognized as a normal inhabitant on the skin and mucous membranes of dogs and cats. However, it is considered an opportunistic agent that causes external otitis and dermatitis in these animals. The aim of this study was to evaluate the in vitro effect of thiabendazole against 51 isolates of M. pachydermatis using the CLSI Broth Microdilution method that has been adapted for this yeast species (NCCLS, 2002. Based on this test, the Minimum Inhibitory Concentrations (MIC of thiabendazol was calculated. Subsequently, the susceptibility of each isolate against this antifungal was determined. It was observed that the MIC of thiabendazole against M. pachydermatis ranged from 0.03 to > 4 μg/mL. A total of 13.7% of the isolates were found to be resistant, 47.1% were intermediate and 39.2% were sensitive to the drug. The rate of resistance of the yeasts against thiabendazole was similar to the results previously obtained with other antifungals, while the adapted broth microdilution technique used in this study proved to be efficient.Tiabendazol, um fármaco classificado como antiparasitário e também usado como antifúngico, pode ser encontrado como solução otologica indicada no tratamento da otite externa parasitária e fungica em pequenos animais. Malassezia pachydermatis é uma levedura considerada habitante normal da pele e das mucosas de cães e gatos. Entretanto, considera-se um agente do oportunista causador de otite externa e dermatite nestes animais. A finalidade deste estudo foi avaliar o efeito in vitro do tiabendazol frente a 51 amostras de M. pachydermatis através do método CLSI de Microdiluição em Caldo adaptado para esta espécie de levedura (NCCLS, 2002. Baseado neste teste calculou-se as Concentra

  1. Direct identification and susceptibility testing of positive blood cultures using high speed cold centrifugation and Vitek II system.

    Science.gov (United States)

    Bazzi, Ali M; Rabaan, Ali A; Fawarah, Mahmoud M; Al-Tawfiq, Jaffar A

    2016-06-13

    Compared to routine isolated colony-based methods, direct testing of bacterial pellets from positive blood cultures reduces turnaround time for reporting of antibiotic susceptibility. The aim of this study was to compare the accuracy, and precision, of a rapid method for direct identification and susceptibility testing of blood cultures with the routine method used in our laboratory, using Vitek 2. A total of 60 isolates were evaluated using the candidate and the routine method. The candidate method had 100% accuracy for the identification of Gram negative bacteria, Staphylococcus and Enterococcus, 50% for Streptococcus and 33.3% for Corynebacterium species. Susceptibility testing of Gram negative isolates yielded 98-100% essential agreement. For Staphylococcus and Enterococcus isolates, essential agreement was 100% for 17 antibiotics except for moxifloxacin. Direct testing of blood culture samples with Vitek 2 produced reliable identification and susceptibility results 18-24h sooner for aerobic/anaerobic facultative Gram-negative bacteria and Gram-positive Staphylococcus and Enterococcus strains.

  2. Revisiting susceptibility testing in MDR-TB by a standardized quantitative phenotypic assessment in a European multicentre study

    NARCIS (Netherlands)

    Cambau, E.; Viveiros, M.; Machado, D.; Raskine, L.; Ritter, C.; Tortoli, E.; Matthys, V.; Hoffner, S.; Richter, E.; Perez del Molino, M.L.; Cirillo, D.M.; Soolingen, D. van; Bottger, E.C.

    2015-01-01

    OBJECTIVES: Treatment outcome of MDR-TB is critically dependent on the proper use of second-line drugs as per the result of in vitro drug susceptibility testing (DST). We aimed to establish a standardized DST procedure based on quantitative determination of drug resistance and compared the results w

  3. Fusarium keratitis in Brazil: genotyping, in vitro susceptibilities, and clinical outcomes

    Science.gov (United States)

    Oechsler, Rafael A; Yamanaka, Tiago M; Bispo, Paulo JM; Sartori, Juliana; Yu, Maria Cecilia Zorat; Melo, Analy Salles A; Miller, Darlene; Hofling-Lima, Ana Luisa

    2013-01-01

    Background The purpose of this paper is to describe clinical characteristics and determine correlations between clinical outcomes and antifungal susceptibility among molecularly characterized ocular Fusarium isolates in Brazil. Methods Forty-one Fusarium isolates obtained from 41 eyes of 41 patients were retrieved from the ophthalmic microbiology laboratory at São Paulo Federal University and grown in pure culture. These isolates were genotyped and antifungal susceptibilities determined for each isolate using a broth microdilution method. The corresponding medical records were reviewed to determine clinical outcomes. Results The 41 isolates were genotypically classified as Fusarium solani species complex (36 isolates, 88%), Fusarium oxysporum species complex (two isolates, 5%), Fusarium dimerum species complex (one isolate, 2%) and two isolates that did not group into any of the species complexes. Final best corrected visual acuity varied from 20/20 to light perception and was on average 20/800 (logarithm of the minimum angle of resolution (LogMAR) 1.6). A history of trauma was the most common risk factor, being present in 21 patients (51%). Therapeutic penetrating keratoplasty was necessary in 22 patients (54%). Amphotericin B had the lowest minimum inhibitory concentration for 90% of isolates (MIC90) value (2 μg/mL) and voriconazole had the highest (16 μg/mL). There was an association between a higher natamycin MIC and need for therapeutic penetrating keratoplasty (Mann–Whitney test, P < 0.005). Conclusion Trauma was the main risk factor, and therapeutic penetrating keratoplasty was necessary in 54% of patients. Amphotericin B had the lowest MIC90 (2 μg/mL) of the three antifungal agents tested. There was an association between higher natamycin MIC levels and corneal perforation, emphasizing the need for antifungal susceptibility testing and tailoring of antifungal strategies. PMID:24039389

  4. Evaluation of quinupristin/dalfopristin (Synercid) and RPR 106972 stability in susceptibility testing media.

    Science.gov (United States)

    Marshall, S A; Kugler, K C; Jones, R N

    2000-08-01

    In response to conflicting reports on the chemical stability of quinupristin/dalfopristin, a study was designed to assess the in vitro longevity and effects of media and storage conditions on this streptogramin combination. Broth microdilution trays containing parenteral (quinupristin/dalfopristin) and oral (RPR 106972) streptogramin combinations as well as pristinomycin components (P-I and P-II) were preincubated at 35 degrees C for 12-72 h before inoculation with control strains (Streptococcus pneumoniae ATCC 49619, Haemophilus influenzae ATCC 49247, Enterococcus faecalis ATCC 29212, Staphylococcus aureus ATCC 29213) and five clinical isolates with various drug resistance phenotypes. Overall, the mean quinupristin/dalfopristin activity loss was 24%/12 h, 41%/18 h, 43%/24 h, 69%/48 h and 79%/72 h with no detected loss of potency when measured by E. faecalis until 18 h. RPR 106972 mean activity loss was 6%/12 h, 19%/18 h, 19%/24 h, 56%/48 h and 71%/72 h with no loss of potency as measured by S. aureus until 48 h. Overall, P-I components had greater stability as compared with P-II for both drug combinations. Bioassays showed similar trends in decreased activity. Bioassay differences among media types were only significant (> 3 mm; greater loss of potency) for haemophilus test media for both P-II components at 72 h. The presence of an organism in the medium had no effect on stability assay results. The effect of storage temperature (4, 25 degrees C) on quinupristin/dalfopristin and RPR 106972 stability was also detrimental to drug potency indicating the requirement for rigid quality assurance for streptogramin diagnostic reagents when determining activity by reference or standardized susceptibility tests.

  5. Preparation of a blood culture pellet for rapid bacterial identification and antibiotic susceptibility testing.

    Science.gov (United States)

    Croxatto, Antony; Prod'hom, Guy; Durussel, Christian; Greub, Gilbert

    2014-10-15

    Bloodstream infections and sepsis are a major cause of morbidity and mortality. The successful outcome of patients suffering from bacteremia depends on a rapid identification of the infectious agent to guide optimal antibiotic treatment. The analysis of Gram stains from positive blood culture can be rapidly conducted and already significantly impact the antibiotic regimen. However, the accurate identification of the infectious agent is still required to establish the optimal targeted treatment. We present here a simple and fast bacterial pellet preparation from a positive blood culture that can be used as a sample for several essential downstream applications such as identification by MALDI-TOF MS, antibiotic susceptibility testing (AST) by disc diffusion assay or automated AST systems and by automated PCR-based diagnostic testing. The performance of these different identification and AST systems applied directly on the blood culture bacterial pellets is very similar to the performance normally obtained from isolated colonies grown on agar plates. Compared to conventional approaches, the rapid acquisition of a bacterial pellet significantly reduces the time to report both identification and AST. Thus, following blood culture positivity, identification by MALDI-TOF can be reported within less than 1 hr whereas results of AST by automated AST systems or disc diffusion assays within 8 to 18 hr, respectively. Similarly, the results of a rapid PCR-based assay can be communicated to the clinicians less than 2 hr following the report of a bacteremia. Together, these results demonstrate that the rapid preparation of a blood culture bacterial pellet has a significant impact on the identification and AST turnaround time and thus on the successful outcome of patients suffering from bloodstream infections.

  6. Antifungal Activity of Brazilian Propolis Microparticles against Yeasts Isolated from Vulvovaginal Candidiasis

    Directory of Open Access Journals (Sweden)

    Kelen Fátima Dalben Dota

    2011-01-01

    Full Text Available Propolis, a resinous compound produced by Apis mellifera L. bees, is known to possess a variety of biological activities and is applied in the therapy of various infectious diseases. The aim of this study was to evaluate the in vitro antifungal activity of propolis ethanol extract (PE and propolis microparticles (PMs obtained from a sample of Brazilian propolis against clinical yeast isolates of importance in the vulvovaginal candidiasis (VVC. PE was used to prepare the microparticles. Yeast isolates (n=89, obtained from vaginal exudates of patients with VVC, were exposed to the PE and the PMs. Moreover, the main antifungal drugs used in the treatment of VVC (Fluconazole, Voriconazole, Itraconazole, Ketoconazole, Miconazole and Amphotericin B were also tested. Minimum inhibitory concentration (MIC was determined according to the standard broth microdilution method. Some Candida albicans isolates showed resistance or dose-dependent susceptibility for the azolic drugs and Amphotericin B. Non-C. albicans isolates showed more resistance and dose-dependent susceptibility for the azolic drugs than C. albicans. However, all of them were sensitive or dose-dependent susceptible for Amphotericin B. All yeasts were inhibited by PE and PMs, with small variation, independent of the species of yeast. The overall results provided important information for the potential application of PMs in the therapy of VVC and the possible prevention of the occurrence of new symptomatic episodes.

  7. Performance and clinical significance of direct antimicrobial susceptibility testing on urine from hospitalized patients

    NARCIS (Netherlands)

    Breteler, K.B.; Rentenaar, R.J.; Verkaart, G.; Sturm, P.D.J.

    2011-01-01

    BACKGROUND: Urinary tract infections (UTIs) are common infections in the community and the hospital. With increasing antimicrobial resistance, specifically in the Gram-negative uropathogens, reliable, rapid antimicrobial susceptibility data would be useful to guide antimicrobial treatment. Direct an

  8. ANTIFUNGAL ACTIVITY OF Cymbopogon nardus (L. Rendle (CITRONELLA AGAINST Microsporum canis FROM ANIMALS AND HOME ENVIRONMENT

    Directory of Open Access Journals (Sweden)

    Isis Regina Grenier CAPOCI

    2015-12-01

    Full Text Available Dermatophytosis is a common zoonosis in urban centers. Dogs and cats have played an important role as its disseminators. Environmental decontamination is essential for the prevention of its propagation to humans and animals. However, sanitizers or disinfectants with antifungal activity, currently available, have high toxicity. The present study evaluated the in vitro effects of an extract of citronella (Cymbopogon nardus on 31 Microsporum canis isolates from animals and home environments. Susceptibility tests were performed based on document M38-A2 (2008 of the Clinical and Laboratory Standards Institute with modifications for natural products. Although susceptibility variation was observed between the fungus tested, the concentrations that inhibited the growth of 50 and 90% of the microorganisms were low (19.5 and 78 µg/mL, respectively. Thus, this citronella extract showed potent fungistatic and fungicide activities against M. canis isolated from animals and home environments. Therefore, it could be an alternative for dermatophytosis prophylaxis in the home environment.

  9. ANTIFUNGAL ACTIVITY OF Cymbopogon nardus (L.) Rendle (CITRONELLA) AGAINST Microsporum canis FROM ANIMALS AND HOME ENVIRONMENT.

    Science.gov (United States)

    Capoci, Isis Regina Grenier; Cunha, Michele Milano da; Bonfim-Mendonça, Patricia de Souza; Ghiraldi-Lopes, Luciana Dias; Baeza, Lilian Cristiane; Kioshima, Erika Seki; Svidzinski, Terezinha Inez Estivalet

    2015-12-01

    Dermatophytosis is a common zoonosis in urban centers. Dogs and cats have played an important role as its disseminators. Environmental decontamination is essential for the prevention of its propagation to humans and animals. However, sanitizers or disinfectants with antifungal activity, currently available, have high toxicity. The present study evaluated the in vitro effects of an extract of citronella (Cymbopogon nardus) on 31 Microsporum canis isolates from animals and home environments. Susceptibility tests were performed based on document M38-A2 (2008) of the Clinical and Laboratory Standards Institute with modifications for natural products. Although susceptibility variation was observed between the fungus tested, the concentrations that inhibited the growth of 50 and 90% of the microorganisms were low (19.5 and 78 µg/mL, respectively). Thus, this citronella extract showed potent fungistatic and fungicide activities against M. canis isolated from animals and home environments. Therefore, it could be an alternative for dermatophytosis prophylaxis in the home environment.

  10. SUSCEPTIBILITY TESTS OF ANOPHELES STEPHENSI WITH SOME CHLORINE, PHOSPHORUS, CARBAMATE AND PYRETILROID INSECTICICDES IN SOUTH OF IRAN

    Directory of Open Access Journals (Sweden)

    M.Iranpour

    1993-12-01

    Full Text Available Susceptibility tests were carried out on Anopheles stephensi with D.D.T(4%, dieldrin (4%, malathion(5%, propoxus (0.1%, primphos-methyl (O.lmg/cm2, lambda-cyhalothrin (0.1%, permethrin (0.25% and deltamethrin (0.025% during 1990-94 in Minab county, soutern Iran, using W.H.O standard method. Results showed that An. stephensi was susceptible to malathion, propoxur, primphos - methyl, lambda- cyhalothrin permethrin and deltamethrin, also LT5O was 5,19.5, less than 1.less than 1,3 and less than 1 minute, respectively. Results indicated that An.stephensi was still resistant to D.D.T and dieldrin but susceptibility level has increased. Mortality rate was 36.1 and 80.6 when the species was exposed to D.D.T and dieldrin impregnated papers for 60 minutes, respectively.

  11. [Recommendations from MENSURA for selection of antimicrobial agents for susceptibility testing and criteria for the interpretation of antibiograms].

    Science.gov (United States)

    2000-03-01

    This document includes the recommendations from the Spanish antibiogram committee (The MENSURA group, Mesa Española de Normalización de la Sensibilidad y Resistencia a los Antimicrobianos, under the auspices of the Sociedad Española de Quimioterapia and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica) for the selection of antimicrobials for susceptibility testing. Separate tables for each group of organism with proposed susceptibility and resistance breakpoints are updated and comparatively presented with those of other groups, such us NCCLS, CA-SFM and BSAC. The susceptibility breakpoint tends to identify the fully susceptible population, which probably lacks any specific resistance mechanism. The analysis of MIC distributions for different homogeneous populations (same species) is used to define breakpoints for susceptibility. The resistance breakpoint is based on pharmacological and clinical data obtained when the corresponding antibiotic is administered with a conventional schedule. The primary objective of the Spanish MENSURA group is to contribute to the international consensus on the establishment of breakpoints.

  12. Identification of Streptococcus pneumoniae: Development of a Standardized Protocol for Optochin Susceptibility Testing Using Total Lab Automation

    Science.gov (United States)

    Panitz, Jessica; Burckhardt, Florian; Zimmermann, Stefan

    2017-01-01

    Purpose. Optochin susceptibility is one parameter used in the laboratory to identify Streptococcus pneumoniae. However, a single standardized procedure does not exist. Optochin is included neither in the current EUCAST breakpoint tables nor in the CLSI performance standards for antimicrobial susceptibility testing. We wanted to establish an evidence-based protocol for optochin testing for our Total Lab Automation. Methods. We tested seven different agars and four different reading time points (7 h, 12 h, 18 h, and 24 h). To accommodate for serotype diversity, all tests were done with 99 different strains covering 34 different serotypes of S. pneumoniae. We calculated a multivariable linear regression using data from 5544 inhibition zones. Results. Reading was possible for all strains at 12 h. Agar type and manufacturer influenced the size of the inhibition zones by up to 2 mm and they varied considerably depending on serotype (up to 3 mm for serotype 3). Depending on agar and reading time point, up to 38% of inhibition zones were smaller than the cut-off of 14 mm; that is, the result of the test was false-negative. Conclusions. Shortening incubation time from 24 h to 12 h for optochin susceptibility testing is feasible. Agar and incubation time have to be chosen carefully to avoid false-negative results.

  13. Genotyping using whole-genome sequencing is a realistic alternative to surveillance based on phenotypic antimicrobial susceptibility testing

    DEFF Research Database (Denmark)

    Zankari, Ea; Hasman, Henrik; Kaas, Rolf Sommer;

    2013-01-01

    genes and MLST types (where MLST stands for multilocus sequence typing). ResFinder results were compared with phenotypic antimicrobial susceptibility testing results using EUCAST epidemiological cut-off values and MLST types. Results: A total of 3051 different phenotypic tests were performed; 482 led......-genome sequencing (WGS) may soon be within reach even for routine surveillance and clinical diagnostics. The aim of this study was to evaluate WGS as a routine tool for surveillance of antimicrobial resistance compared with current phenotypic procedures. Methods: Antimicrobial susceptibility tests were performed...... on 200 isolates originating from Danish pigs, covering four bacterial species. Genomic DNA was purified from all isolates and sequenced as paired-end reads on the Illumina platform. The web servers ResFinder and MLST (www.genomicepidemiology.org) were used to identify acquired antimicrobial resistance...

  14. Quality Control Guidelines for Amphotericin B, Itraconazole, Posaconazole, and Voriconazole Disk Diffusion Susceptibility Tests with Nonsupplemented Mueller-Hinton Agar (CLSI M51-A Document) for Nondermatophyte Filamentous Fungi▿

    Science.gov (United States)

    Espinel-Ingroff, A.; Canton, E.; Fothergill, A.; Ghannoum, M.; Johnson, E.; Jones, R. N.; Ostrosky-Zeichner, L.; Schell, W.; Gibbs, D. L.; Wang, A.; Turnidge, J.

    2011-01-01

    Although Clinical and Laboratory Standards Institute (CLSI) disk diffusion assay standard conditions are available for susceptibility testing of filamentous fungi (molds) to antifungal agents, quality control (QC) disk diffusion zone diameter ranges have not been established. This multicenter study documented the reproducibility of tests for one isolate each of five molds (Paecilomyces variotii ATCC MYA-3630, Aspergillus fumigatus ATCC MYA-3626, A. flavus ATCC MYA-3631, A. terreus ATCC MYA-3633, and Fusarium verticillioides [moniliforme] ATCC MYA-3629) and Candida krusei ATCC 6258 by the CLSI disk diffusion method (M51-A document). The zone diameter ranges for selected QC isolates were as follows: P. variotii ATCC MYA-3630, amphotericin B (15 to 24 mm), itraconazole (20 to 31 mm), and posaconazole (33 to 43 mm); A. fumigatus ATCC MYA-3626, amphotericin B (18 to 25 mm), itraconazole (11 to 21 mm), posaconazole (28 to 35 mm), and voriconazole (25 to 33 mm); and C. krusei, amphotericin B (18 to 27 mm), itraconazole (18 to 26 mm), posaconazole (28 to 38 mm), and voriconazole (29 to 39 mm). Due to low testing reproducibility, zone diameter ranges were not proposed for the other three molds. PMID:21543581

  15. Clinical application of micronucleus test: a case-control study on the prediction of breast cancer risk/susceptibility.

    Science.gov (United States)

    Bolognesi, Claudia; Bruzzi, Paolo; Gismondi, Viviana; Volpi, Samantha; Viassolo, Valeria; Pedemonte, Simona; Varesco, Liliana

    2014-01-01

    The micronucleus test is a well-established DNA damage assay in human monitoring. The test was proposed as a promising marker of cancer risk/susceptibility mainly on the basis of studies on breast cancer. Our recent meta-analysis showed that the association between micronuclei frequency, either at baseline or after irradiation, and breast cancer risk or susceptibility, has been evaluated in few studies of small size, with inconsistent results. The aim of the present study is to investigate the role of micronucleus assay in evaluating individual breast cancer susceptibility. Two-hundred and twenty untreated breast cancer patients and 295 female controls were enrolled in the study. All women were characterized for cancer family history and 155 subjects were evaluated for the presence of BRCA mutations. Micronuclei frequency was evaluated at baseline and after irradiation with 1-Gy gamma rays from a 137Cs source. The results show a non significant increase of frequency of micronucleated binucleated lymphocytes in cancer patients compared with the controls at baseline (Mean (S.E.): 16.8 (0.7) vs 15.7 (0.5), but not after irradiation (Mean (S.E.): 145.8 (3.0) vs 154.0 (2.6)). Neither a family history of breast cancer nor the presence of a pathogenic mutation in BRCA1/2 genes were associated with an increased micronuclei frequency. Our results do not support a significant role of micronucleus frequency as a biomarker of breast cancer risk/susceptibility.

  16. Antifungal activity of diethyldithiocarbamate.

    Science.gov (United States)

    Allerberger, F; Reisinger, E C; Söldner, B; Dierich, M P

    1989-10-01

    Sodium diethyldithiocarbamate (DTC) was evaluated for its ability to combat four different species of fungi in vitro. Using a microtiter-broth-dilution method we were able to demonstrate an antifungal activity against Candida albicans, Cryptococcus neoformans, Aspergillus fumigatus and Mucor mucedo in doses achievable by intravenous administration in man.

  17. Typhoid outbreak in Songkhla, Thailand 2009-2011: clinical outcomes, susceptibility patterns, and reliability of serology tests.

    Directory of Open Access Journals (Sweden)

    Wannee Limpitikul

    Full Text Available OBJECTIVE: To determine the clinical manifestations and outcomes, the reliability of Salmonella enterica serotype Typhi (S ser. Typhi IgM and IgG rapid tests, and the susceptibility patterns and the response to treatment during the 2009-2011 typhoid outbreak in Songkhla province in Thailand. METHOD: The medical records of children aged <15 years with S ser. Typhi bacteremia were analysed. The efficacy of the typhoid IgM and IgG rapid tests and susceptibility of the S ser. Typhi to the current main antibiotics used for typhoid (amoxicillin, ampicillin, cefotaxime, ceftriaxone, co-trimoxazole, and ciprofloxacin, were evaluated. RESULTS: S ser. Typhi bacteremia was found in 368 patients, and all isolated strains were susceptible to all 6 antimicrobials tested. Most of the patients were treated with ciprofloxacin for 7-14 days. The median time (IQR of fever before treatment and duration of fever after treatment were 5 (4, 7 days and 4 (3, 5 days, respectively. Complications of ascites, lower respiratory symptoms, anemia (Hct <30%, and ileal perforation were found in 7, 7, 22, and 1 patients, respectively. None of the patients had recurrent infection or died. The sensitivities of the typhoid IgM and IgG tests were 58.3% and 25.6% respectively, and specificities were 74.1% and 50.5%, respectively. CONCLUSION: Most of the patients were diagnosed at an early stage and treated with a good outcome. All S ser. Typhi strains were susceptible to standard first line antibiotic typhoid treatment. The typhoid IgM and IgG rapid tests had low sensitivity and moderate specificity.

  18. Fluconazole exposure rather than clonal spreading is correlated with the emergence of Candida glabrata with cross-resistance to triazole antifungal agents.

    Science.gov (United States)

    Chen, Tun-Chieh; Chen, Yen-Hsu; Chen, Yee-Chun; Lu, Po-Liang

    2012-06-01

    The emergence of antifungal resistance in Candida species has raised concern in recent years, especially resistance toward triazole. Several newer triazole antifungal agents have been introduced which have a broader spectrum for fungal infections, such as voriconazole. However, cross-resistance among triazoles is a major concern with regard to their clinical application. Antifungal susceptibility was performed using E-test for 166 clinical isolates (29 blood and 137 nonblood isolates) in 2003 and 2004. We applied pulsed-field gel electrophoresis for genotyping. Ninety isolates of C. albicans, 47 isolates of C. tropicalis, 27 isolates of C. glabrata, and two isolates of C. krusei were included. All isolates were susceptible to amphotericin B. Eleven (40.7%) of the 27 C. glabrata had intermediate resistance to caspofungin. Forty-seven (28.3%) of the 166 isolates were not susceptible to fluconazole, including two C. albicans, 16 C. tropicalis, 27 C. glabrata, and two C. krusei isolates. All except seven of the C. glabrata isolates were susceptible to voriconazole. All the triazole drugs had a positive correlation among their minimum inhibitory concentrations (MICs). Fluconazole MIC was a good predictor for susceptibility to voriconazole, as determined using a receiver operating characteristic curve. Furthermore, a high diversity of pulsotypes for the 27 clinical isolates of C. glabrata was observed. Previous fluconazole exposure within 3 months was associated with reduced triazole susceptibility for C. glabrata. We demonstrated a significant positive correlation of MIC values among the four tested triazole drugs. No amphotericin B and caspofungin resistant isolates were found in this study. The cross-resistance to triazole among C. glabrata isolates was associated with previous fluconazole exposure as opposed to clonal spreading. Selection pressure due to fluconazole use may play a major role in triazole cross-resistance.

  19. In vitro antimicrobial susceptibility testing of human Brucella melitensis isolates from Qatar between 2014 - 2015

    NARCIS (Netherlands)

    Deshmukh, A.; Hagen, F.; Sharabasi, O.A.; Abraham, M.; Wilson, G.; Doiphode, S.; Maslamani, M.A.; Meis, J.F.G.M.

    2015-01-01

    BACKGROUND: Brucellosis is one of the most common zoonotic disease affecting humans and animals and is endemic in many parts of the world including the Gulf Cooperation Council region (GCC). The aim of this study was to identify the species and determine the antimicrobial susceptibility pattern of B

  20. Adolescent behavioral and neural reward sensitivity : a test of the differential susceptibility theory

    NARCIS (Netherlands)

    Richards, J. S.; Vasquez, A. Arias; von Rhein, D.; van der Meer, D.; Franke, B.; Hoekstra, P. J.; Heslenfeld, D. J.; Oosterlaan, J.; Faraone, S. V.; Buitelaar, J. K.; Hartman, C. A.

    2016-01-01

    Little is known about the causes of individual differences in reward sensitivity. We investigated gene-environment interactions (GxE) on behavioral and neural measures of reward sensitivity, in light of the differential susceptibility theory. This theory states that individuals carrying plasticity g

  1. Ten years of antimicrobial susceptibility testing of Salmonella from Danish pig farms

    DEFF Research Database (Denmark)

    Emborg, Hanne-Dorthe; Baggesen, Dorte Lau; Aarestrup, Frank Møller

    2008-01-01

    Objectives: This study analysed the trends in antimicrobial resistance in Salmonella serovars and phage types from pigs in Denmark from 1997 to 2006. Methods: Salmonella isolates collected through the Salmonella surveillance programme in pigs were serotyped and phage-typed, and susceptibilities t...

  2. A colorimetric and spectrophotometric method for in vitro susceptibility testing of Aspergillus species against caspofungin.

    NARCIS (Netherlands)

    Dorsthorst, D.T.A. te; Zwaaftink, R.B.; Rijs, A.J.M.M.; Meletiadis, J.; Verweij, P.E.

    2007-01-01

    The in vitro susceptibility of 45 Aspergillus fumigatus, Aspergillus flavus and Aspergillus terreus isolates against caspofungin (CAS) was assessed by the CLSI reference method with spectrophotometric reading and by a colorimetric method that employed the dye MTT. Perfect agreement was found between

  3. Molecular and Growth-Based Drug Susceptibility Testing of Mycobacterium tuberculosis Complex for Ethambutol Resistance in the United States

    Directory of Open Access Journals (Sweden)

    Mitchell A. Yakrus

    2016-01-01

    Full Text Available Ethambutol (EMB is used as a part of drug regimens for treatment of tuberculosis (TB. Susceptibility of Mycobacterium tuberculosis complex (MTBC isolates to EMB can be discerned by DNA sequencing to detect mutations in the embB gene associated with resistance. US Public Health Laboratories (PHL primarily use growth-based drug susceptibility test (DST methods to determine EMB resistance. The Centers for Disease Control and Prevention (CDC provides a service for molecular detection of drug resistance (MDDR by DNA sequencing and concurrent growth-based DST using agar proportion. PHL and CDC test results were compared for 211 MTBC samples submitted to CDC from September 2009 through February 2011. Concordance between growth-based DST results from PHL and CDC was 88.2%. A growth-based comparison of 39 samples, where an embB mutation associated with EMB resistance was detected, revealed a higher percentage of EMB resistance by CDC (84.6% than by PHL (59.0% which was significant (P value = 0.002. Discordance between all growth-based test results from PHL and CDC was also significant (P value = 0.003. Most discordance was linked to false susceptibility using the BACTEC™ MGIT™ 960 (MGIT growth-based system. Our analysis supports coalescing growth-based and molecular results for an informed interpretation of potential EMB resistance.

  4. Task 2 - Limits for High-Frequency Conducted Susceptibility Testing - CS114 (NRC-HQ-60-14-D-0015)

    Energy Technology Data Exchange (ETDEWEB)

    Wood, Richard Thomas [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Ewing, Paul D. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Moses, Rebecca J. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

    2015-09-01

    A principal focus of Task 2 under this project was for ORNL to evaluate the basis for susceptibility testing against high-frequency conducted interference and to establish recommendations to resolve concerns about the severity of test limits for the conducted susceptibility (CS) test, CS114, from MIL-STD-461. The primary concern about the test limit has been characterized by the EPRI EMI Working Group in the following terms: Demonstrating compliance with the CS114 test limits recommended in TR-102323 has proven to be problematic, even for components that have been tested to commercial standards and demonstrated proper operation in industrial applications [6]. Specifically, EPRI notes that the CS114 limits approved in regulatory documents are significantly higher than those invoked by the US military and similar commercial standards in the frequency range below 200 kHz. For this task, ORNL evaluated the original approach to establishing the test limit, EPRI technical findings from a review of the limit, and the regulatory basis through which the currently approved limits were accepted. Based on this analysis, strategies have been developed regarding changes to the CS114 limit that can resolve the technical concerns raised by the industry. Guided by the principles that reasonable assurance of safety must not be compromised but excessive conservatism should be reduced, recommendations on a suitable basis for a revised limit have been developed and can be incorporated into the planned Revision 2 of RG 1.180.

  5. Active packaging with antifungal activities.

    Science.gov (United States)

    Nguyen Van Long, N; Joly, Catherine; Dantigny, Philippe

    2016-03-01

    There have been many reviews concerned with antimicrobial food packaging, and with the use of antifungal compounds, but none provided an exhaustive picture of the applications of active packaging to control fungal spoilage. Very recently, many studies have been done in these fields, therefore it is timely to review this topic. This article examines the effects of essential oils, preservatives, natural products, chemical fungicides, nanoparticles coated to different films, and chitosan in vitro on the growth of moulds, but also in vivo on the mould free shelf-life of bread, cheese, and fresh fruits and vegetables. A short section is also dedicated to yeasts. All the applications are described from a microbiological point of view, and these were sorted depending on the name of the species. Methods and results obtained are discussed. Essential oils and preservatives were ranked by increased efficacy on mould growth. For all the tested molecules, Penicillium species were shown more sensitiv