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The treatment of invasive aspergillosis caused by cryptic species remains a challenge due to the lack of randomised clinical trials and investigation of the efficacy and safety of different therapeutic strategies. We aimed to evaluate the in vitro activity of 23 conventional and new antifungal drugs against 54 clinical and environmental Aspergillus oryzae isolates by using the Clinical and Laboratory Standards Institute (CLSI) standard M38‐A3. The lowest geometric mean MIC values were found for luliconazole and lanoconazole (0.001 μg/ml), followed by anidulafungin (0.104 μg/ml), posaconazole (0.15 μg/ml), itraconazole (0.37 μg/ml), efinaconazole (0.5 μg/ml), voriconazole (0.51 μg/ml), tavaborole (0.72 μg/ml), and amphotericin B (0.79 μg/ml). In contrast, ketoconazole, terbinafine, econazole, tioconazole, ravuconazole, miconazole, nystatin, clotrimazole, griseofulvin, sertaconazole, natamycin, tolnaftate, and fluconazole had no or low activity. Further studies are required to determine how well this in vitro activity translates into in vivo efficacy. [ABSTRACT FROM AUTHOR]

In the last 2 decades, renewed attention to neglected tropical diseases (NTDs) has spurred the development of antiparasitic agents, especially in light of emerging drug resistance. The need for new drugs has required in vitro screening methods using parasite culture. Furthermore, clinical laboratories sought to correlate in vitro susceptibility methods with treatment outcomes, most notably with malaria. Parasites with their various life cycles present greater complexity than bacteria, for which standardized susceptibility methods exist. This review catalogs the state-of-the-art methodologies used to evaluate the effects of drugs on key human parasites from the point of view of drug discovery as well as the need for laboratory methods that correlate with clinical outcomes.

Objectives: This study aimed to evaluate the susceptibility of clinical isolates of Enterobacterales and Pseudomonas aeruginosa to fosfomycin and to determine the concordance of disk diffusion (DD) and broth microdilution (BMD) with agar dilution (AD) for fosfomycin susceptibility testing. Methods: The activity of fosfomycin against 225 clinical isolates of Escherichia coli (n = 64), Klebsiella pneumoniae (n = 68), Enterobacter spp. (n = 28) and P. aeruginosa (n = 65) was tested by AD, DD and BMD. For DD, results were recorded considering and not considering colonies growing within the inhibition halo as recommended by the CLSI and EUCAST, respectively. Escherichia coli breakpoints were used for all Enterobacterales. Results were reported as categorical agreement (CA), major error (ME; false-resistant), very major error (VME; false-susceptible) and minor error (any other discrepancies). Results: Fosfomycin susceptibility of all tested species was >90% by AD. Following CLSI guidelines, DD was the only method reaching ≥90% CA with AD for E. coli and K. pneumoniae, albeit yielding 6% ME. Neither DD nor BMD achieved acceptable CA percentages for Enterobacter spp. Following EUCAST guidelines, none of the methods had CA ≥ 90%. For Enterobacterales, the best performance of DD is achieved when read as indicated by EUCAST but interpreted according the CLSI breakpoints (>97% CA; 0% VME; ≤2% ME). For P. aeruginosa, BMD yielded the best results (89% CA; 0% VME; 11% ME). Conclusion: Neither DD or BMD provide accurate results owing to unacceptable ME and VME percentages even when performed as intended by the guidelines.

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.

Background: Dermatophytosis is one of the most common infections affecting 3%–17% of the population. Resistance to antifungals so far was not of concern in the therapeutic management. However, recent reports of terbinafine‐resistant strains in several countries are worrisome making antifungal susceptibility testing inevitable. Objectives: We aimed to develop and evaluate an optimised broth microdilution assay for antifungal drug susceptibility testing of dermatophytes. Methods: We first studied the effect of different inocula, incubation temperatures and incubation times to establish an optimised assay. Subsequently, we tested 79 clinical strains of 11 dermatophyte species with 13 antifungals. Results: We found inoculating with 0.5–5 × 104 colony forming units (CFU) and incubating at 29°C ± 1°C for 4 days to be appropriate. Terbinafine was the most active antifungal agent with minimum inhibitory concentration (MIC) values ≤ 0.06 µg/mL, expect for one resistant T mentagrophytes strain, which was isolated from an Indian patient. Also, a majority of MICs of other antifungals that are commonly used to treat dermatophytosis were low, except those of fluconazole. Fluconazole MICs do not correlate with the good efficacy in the clinical management. Conclusions: Our assay enables fast and reliable susceptibility testing of dermatophytes with a large panel of different antifungals. This helps to improve the therapeutic management of dermatophytosis by detecting resistant strains. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the antifungal susceptibility of Sporothrix globosa isolated from Shandong, China, and compare the differences of antifungal activity in vitro between yeast and mycelial phases. Methods: The in vitro sensitivity of mycelium phase and yeast phase of Sporothrix globosa to anidulafungin, micafungin, caspofungin, 5‐flucytosine, posaconazole, voriconazole, itraconazole, fluconazole and amphotericin B was tested by Sensititre™ YeastOne™. The minimum inhibitory concentration (MIC) values of mycelium phase and yeast phase were calculated. SPSS 19.0 software was used to conduct non‐parametric rank sum test for MIC values, and P <.05 was considered statistically significant. Results: The mycelium phase and yeast phase were the most sensitive to itraconazole and the least sensitive to fluconazole. The yeast phase of the same strain was more sensitive to itraconazole, voriconazole, posaconazole, micafungin, anidulafungin, caspofungin and 5‐fluorouracil, compared with the mycelium (P <.05). However, fluconazole and amphotericin B had no significant difference in mycelium phase and yeast phase. Conclusions: Itraconazole is the most active antifungal agent in vitro against S globosa. The yeast phase of the same strain is more sensitive than that of the mycelium. [ABSTRACT FROM AUTHOR]

Malodour from the axilla is commonly caused by specific microbes, and may be inhibited by zinc oxide. The aim of this study was to determine the effects of zinc oxide on the axillary microbiota, odour and pH in a randomized, double-blind, placebo-controlled trial in 30 healthy volunteers. In each participant one axilla was treated with zinc oxide and the other with a placebo for 13 days. The microbiota and pH were analysed before and during treatment. At the final visit, the participants judged their own axillary odour for comparison. With zinc oxide treatment total bacterial growth and, specifically, that of odour-producing Corynebacterium spp. and Staphylococcus hominis, decreased (p < 0.05), despite an increase (p < 0.0005) in skin-surface pH. Compared with the placebo, zinc oxide treatment reduced (p = 0.005) self-perceived malodour. In vitro, Corynebacterium spp. (19 isolated strains) survival was reduced (p < 0.0005) at pH 5.0 compared with pH 6.0; growth inhibition by zinc oxide occurred at = 400 mg/l, and cell death occurred at = 10,000 mg/l for 12 (63%) of the strains. In conclusion, application of zinc oxide reduced malodour and the counts of causative bacteria, but increased the pH of the axilla. [ABSTRACT FROM AUTHOR]

In the present study, the structural characteristics that impart antibacterial activity to C16 alkynoic fatty acids (aFA) were further investigated. The syntheses of hexadecynoic acids (HDA) containing triple bonds at C‐3, C‐6, C‐8, C‐9, C‐10, and C‐12 were carried out in four steps and with an overall yield of 34–78%. In addition, HDA analogs containing a sulfur atom at either C‐4 or C‐5 were also prepared in 69–77% overall yields, respectively. Results from this study revealed that the triple bond at C‐2 is pivotal for the antibacterial activity displayed by 2‐HDA, while the farther the position of the triple bond from the carbonyl group, the lower its bactericidal activity against gram‐positive bacteria, including clinical isolates of methicillin‐resistant Staphylococcus aureus (CIMRSA) strains. The potential of 2‐HDA as an antibacterial agent was also assessed in five CIMRSA strains that were resistant to Ciprofloxacin (Cipro) demonstrating that 2‐HDA was the most effective treatment in inhibiting their growth when compared with either Cipro alone or equimolar combinations of Cipro and 2‐HDA. Moreover, it was proved that the inhibition of S. aureus DNA gyrase can be linked to the antibacterial activity displayed by 2‐HDA. Finally, it was determined that the ability of HDA analogs to form micelles can be linked to their decreased activity against gram‐positive bacteria, since critical micellar concentrations (CMC) between 50 and 300 μg/mL were obtained. [ABSTRACT FROM AUTHOR]

Aim: Assess performance of broth microdilution (BMD) as well as agar dilution methods for antimicrobial susceptibility testing of Staphylococci using tetrazolium salt. Methods and Results: Minimum inhibitory concentration (MIC) of eight antimicrobials; vancomycin (VA), linezolid, oxacillin, gentamicin (CN), tetracycline, ciprofloxacin, erythromycin and clindamycin was investigated for 80 isolates of Staphylococci by BMD with the addition of dimethyl thiazole diphenyl tetrazolium bromide (MTT), agar dilution with the addition of MTT and triphenyl tetrazolium chloride at the standard bacterial concentration together with addition of MTT at an experimental bacterial concentration. BMD (MTT) showed the highest agreement in comparison with the standard BMD. Conclusions: Colorimetric BMD was rapid and easy to interpret. Colorimetric agar dilution (MTT) was less tedious than BMD. Significance and Impact of the Study: Colorimetric antibiotic susceptibility is a good option to provide rapid reliable results for critically ill patients. In addition, agar dilution (MTT) helps to investigate outbreaks of methicillin‐resistant Staphylococcusaureus (MRSA), VISA or VRSA. BMD (MTT) can be performed routinely to detect VA MIC in MRSA blood stream infections and hospital acquired pneumonia, where, high VA MIC is associated with a higher mortality rate. [ABSTRACT FROM AUTHOR]

The present study proposes a discussion about the use of breakpoints when plant derivatives are used for investigating potential agents against Mycobacterium tuberculosis strains. A systematic review on these aspects was performed and supported that an arbitrary breakpoint may be considered inadequate in this kind of study. In addition, we propose that the adoption of this limiter should be done from the toxicity value found using the same plant derivative. [ABSTRACT FROM AUTHOR]

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.

Summary: Tinea pedis and onychomycosis are among the commonest fungal diseases in the world. Dermatophytes and, less frequently, non‐dermatophyte moulds are aetiological agents of foot mycosis and are capable of forming biofilms. Fungal biofilm has demonstrated increasing drug resistance. This work aims to evaluate, in vitro, the ability to form biofilm and the susceptibility to antifungal drugs of sessile dermatophytes and non‐dermatophyte moulds involved in foot mycosis. Thirty‐six dermatophytes and non‐dermatophyte moulds isolated from Tunisian patients with foot mycoses, and identified with MALDI‐TOF have been tested. MICs of fluconazole, econazole, itraconazole, terbinafine and griseofulvin were carried out using CLSI broth microdilution method. The ability to form biofilm and antifungal activities of drugs against fungal biofilm formation has been quantified by Crystal Violet and Safranin Red staining. Biomass quantification revealed that all species studied were able to form biofilms in vitro after 72 hours. Fluconazole, econazole, itraconazole and terbinafine inhibited fungal growth with MIC values ranging from 0.031 to >64 μg mL−1. The best antifungal activity has been obtained with terbinafine against Fusarium solani. Econazole showed the highest activity against fungal biofilm formation. These findings can help clinicians to develop the appropriate therapy of foot mycosis. [ABSTRACT FROM AUTHOR]

Purpose: To report data of samples collected from January 2000 to August 2013, in the Department of Diagnosis and Laboratory Analysis, Ophthalmic Hospital, Turin, Italy, from different types of ocular infections and their antibiotic susceptibility.Methods: Collected samples were cultured using both liquid and solid media. Then bacterial isolates were tested for antibiotic susceptibility using the Kirby-Bauer diffusion method and the National Committee for Clinical Laboratory Standards (NCCLS) serum standards.Results: Staphylococcus aureus is the most common bacteria isolated in ocular samples.Conclusions: In vitro susceptibility tests showed that levofloxacin and moxifloxacin (introduced only in 2010) had the highest efficacy against bacterial isolates. [ABSTRACT FROM AUTHOR]

Coagulase-negative Staphylococcus spp. was considered nonpathogenic until the emergence of multiresistance and the demonstration of their participation as infectious agents. In Brazil, oxacillin resistance may be present in over 80% of isolates, and the Clinical and Laboratory Standards Institute standardized a disk-diffusion method to predict this resistance in Staphylococcus. The aim of this study was to evaluate the variability among commercial disks of oxacillin (1 µg) and cefoxitin (30 µg) widely used in clinical laboratories of microbiology, compared with mecA gene and minimum inhibitory concentration (MIC) of oxacillin. The use of oxacillin and cefoxitin disks simultaneously allowed the detection of important differences, particularly, in less frequent species such as S. cohnii, S. haemolyticus, S. saprophyticus, and S. sciuri. Disks of cefoxitin of the brand 2 displayed good correlation with the mecA gene (98.7%) and oxacillin MIC (97.8%), while major discrepancies were observed using disks of brand 1. One of the critical points in the diffusion disk test is the quality of the disks: the use of better quality disks associated with molecular methods lead to better results to define the best antibiotic therapy.

Background: Pythium insidiosum is the etiological agent of pythiosis, an emerging life-threatening infectious disease in tropical and subtropical regions. The pathogen is a fungus-like organism resistant to antifungal therapy, for this reason, most cases need extensive surgical debridments as treatment, but depending on the size and anatomical region of the lesion, such approach is unfeasible. We investigate the fungicidal effect and toxicity of crude bark extract of Stryphnodendron adstringens and commercially available tannin on Pythium insidiosum both in vitro and in vivo. Methods: Standardized fragments of mycelia of fifteen isolates of P. insidiosum were tested with different concentrations of bark extract (10 to 30% v/v) and tannin (0.5, 1.0 and 1.5 mg/mL). For in vivo study, fifteen rabbits were experimentally infected with zoospores of P. insidiosum and treated by oral and intralesional applications of bark extract and tannin. Acute toxicity tests with both substances were also performed in rats. Results: In vitro studies showed fungicidal effect for both substances at different concentrations and the SEM showed alteration on the cell wall surface of the pathogen. All infected rabbits developed a firm nodular mass that reached around 90 mm2 ninety days after inoculation, but neither the intralesional inoculation of tannin, nor the oral administration of crude extract and tannin were able to promote remission of the lesions. Conclusions: Lesions developed by rabbits presented an encapsulated abscess being quite different of naturally acquired pythiosis, which is characterized by ulcerated lesions. Since no toxicity was observed in rats or rabbits inoculated with these products, while in vitro experiments showed direct antifungal effect, therapeutic activity of S. adstringens and tannin should be clinically tested as an alternative for healing wounds in naturally acquired pythiosis. [ABSTRACT FROM AUTHOR]

The naturally high minimum inhibitory concentration exhibited by echinocandins against Candida parapsilosis has been known since the first introduction of these antifungal agents. Despite this awareness, clinical failures have not been reported; consequently, the resistance of C. parapsilosis to echinocandins remains unexplored. We exposed 30 isolates of C. parapsilosis to echinocandins (caspofungin, micafungin, and anidulafungin) in vitro and studied the effects of this exposure. After 60 exposures, 80, 67, and 60 % of the isolates changed from susceptible to non-susceptible to caspofungin, micafungin, and anidulafungin, respectively. In addition, four strains exhibited cross-resistance to all three echinocandins. Based on the M27-A3 (CLSI, 2008) and M27-S4 (CLSI, 2012) techniques, the susceptibility of the resistant strains to other antifungal agents was assayed. All of the tested echinocandin-resistant strains were susceptible to amphotericin B, and the resistance rate to fluconazole, voriconazole, and flucytosine was 73.3, 43.3, and 20 %, respectively. The exposure of C. parapsilosis to the three echinocandins generated cross-resistant strains and an unexpected in vitro resistance to azoles and flucytosine. [ABSTRACT FROM AUTHOR]

Invasive fungal infections, usually Aspergillus and Candida, represent a major cause of morbidity and mortality in patients with malignant haematological diseases, but in the last years rare fungal infections have more frequently been reported. Here, we report the clinical history of three patients affected with haematological malignancies who developed an infection caused by Geotrichum ( G.) clavatum. Two out of three patients were affected by acute myeloid leukaemia (AML), and one by mantle cell lymphoma (MCL). All patients received cytarabine-based chemotherapeutic regimens and developed G. clavatum infection within 3 weeks from therapy initiation. In all cases, G. clavatum was isolated from central venous catheter and peripheral blood cultures. In vitro susceptibility test confirmed an intrinsic resistance to echinocandins and, in all cases, visceral localisations (spleen, liver and lung) were documented by total body computed tomography (CT) scan. A prolonged antifungal therapy with high doses liposomal amphotericin-B was necessary to obtain fever resolution. Only the patient with MCL died while the other two AML recovered, and one of them after received an allogeneic stem cell transplantation. We consecutively reviewed all published cases of infection caused by G. clavatum. Our experience and literature review indicate that G. clavatum can cause invasive infection in haematological patients, mainly in those with acute leukaemia. [ABSTRACT FROM AUTHOR]

Whole plant-parts of Bidens pilosa were powdered and extracted in concentrated hexane, acetone, ethanol, methanol and water. The extracts were tested for antimicrobial activity against Escherichia coli (25922), Bacillus subtilis (ATCC 6051), Enterococcus faecalis (51299) Staphylococcus aureus (ATCC 29213) and Pseudomonas aeruginosa (ATCC127853), using standard microbiological techniques. Active crude extracts were macerated in concentrated methanol and tested for secondary metabolites including tannins, saponins, alkaloids, cardiac glycosides, anthraquinones, steroids and flavonoids using standard phytochemical procedures. Hexane and methanol extracts demonstrated similar activity producing 8-17 mm and 11-18 mm inhibition zone-diameter ranges respectively. Further analysis for minimum inhibitory concentrations (MIC50) recorded 1.25-20mg/mL and 2.5-20mg/mL for hexane and methanol extracts respectively. The highest zones of inhibition diameters (22-36mm) and lowest MIC50 values (0.0002-0.0006mg/mL) were recorded for Gentamicin, the positive control. Minimum bactericidal concentration (MBC) ranges were between 10-80mg/mL and 0.001-0.005mg/mL for extracts and control antibiotic respectively. With the exception of anthraquinones, the plant crude extracts tested positive for all secondary metabolites analyzed. These results provide scientific basis for the use of B. pilosa in South African traditional medicine. The antibacterial activity reported herein may be attributed to one or more of the 6 secondary metabolites detected in the plant crude extracts. [ABSTRACT FROM AUTHOR]

The emergence of multidrug-resistant strains of Mycobacterium tuberculosis has increased the need for rapid drug susceptibility tests, which are needed for adequate patient treatment. The objective of the present study was to evaluate the mycobacteria growth indicator tube (MGIT) system to detect multidrug-resistant M. tuberculosis strains. The MGIT system was compared with two standard methods (proportion and resistance ratio methods). One hundred clinical M. tuberculosis isolates [25 susceptible to isoniazid (INH) and rifampicin (RIF), 20 resistant to INH, 30 resistant to INH-RIF, and 25 resistant to INH-RIF and other drugs] obtained in the State of São Paulo were tested for INH and RIF susceptibility. Full agreement among the tests was found for all sensitive and all INH-resistant strains. For RIF-resistant strains results among the tests agreed for 53 (96.4%) of 55 isolates. Results were obtained within 6 days (range, 5 to 8 days), 28 days and 12 days when using MGIT, the proportion method and the resistance ratio methods, respectively. The MGIT system presented an overall agreement of 96% when compared with two standard methods. These data show that the MGIT system is rapid, sensitive and efficient for the early detection of multidrug-resistant M. tuberculosis.

Background: Fungal endocarditis is a low-frequency disease with a challenging diagnosis, as it can be mistaken with bacterial endocarditis. Fungal endocarditis causes higher mortality rates in immunocompromised patients. In the clinical practice, the endocarditis caused by fungi represents up to 10% of all infectious endocarditis cases and has a mortality rate of nearly 50%., Case Report: Here we present the case of a 53-year-old woman under corticosteroid therapy with a history of rheumatic heart disease, aortic valve replacement, and rheumatoid arthritis, who presented with fungal endocarditis caused by Candida albicans. Even though the patient received 3 years of antifungal prophylaxis with fluconazole, had valve replacement surgery, and received intensive care, the patient finally worsened and died., Conclusions: Comorbidities and corticosteroid therapy predisposed the patient to acquire fungal endocarditis. This case highlights the importance of implementing procedures for the isolation and identification of fungi, and for carrying out antifungal-susceptibility testing, as well as establishing surveillance programs to identify infection-causing species and drug resistance patterns in hospitals. Moreover, designing and upgrading the algorithm for infectious endocarditis is the key to future improvements in diagnosis., (Copyright © 2023 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.)

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 Instituição. Susceptibilidade dose-dependente ao itraconazol ocorreu em 14,2% e ao fluconazol 1,6%. Faz-se necessário monitoramento epidemiológico.

The biological effects of the bacterial insecticides Bacilod, VectoLex and Spinosad as well as the insect growth regulators (IGRs) Baycidal, Sumilarv and Dudim against mosquito larvae of Aedes aegypti have been evaluated. According to LC50 values (concentration which to kill 50% of larvae), the bioinsecticide Spinosad (0.011 ppm) proved to be the most effective compound, followed by Bacilod (0.11 ppm) and VectoLex (0.38 ppm). Taking IC50 values (concentration which to inhibit the emergence of 50% of adults) into consideration, mosquito larvae of A. aegypti were more susceptible to the IGR Dudim (0.00056 ppm) than Baycidal (0.0007 ppm) and Sumilarv (0.0042 ppm) by about 1.25 and 7.5 folds, respectively. Variations in the susceptibility status of the present mosquito larvae may be attributed to the differential mode of action of the test compounds and its effective concentrations. On the other hand, larval treatments with sublethal concentrations of the above insecticides led to a reduction in the egg production and hatchability of eggs produced by mosquito females that developed from surviving larvae. [ABSTRACT FROM AUTHOR]

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Copyright of Revista Biociências is the property of Revista Biociencias and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background. The objective of this study was: i) to compare the results of urine culture with polymerase chain reaction (PCR) -based detection of microorganisms using two commercially available kits, ii) to assess antimicrobial susceptibility of urine isolates from cancer patients to chosen antimicrobial drugs and, if necessary, to update the recommendation of empirical therapy. Materials and methods. A one-year hospital-based prospective study has been conducted in Greater Poland Cancer Centre and Genetic Medicine Laboratory CBDNA Research Centre in 2011. Urine cultures and urine PCR assay from 72 patients were examined Results. Urine cultures and urine PCR assay from 72 patients were examined. Urine samples were positive for 128 strains from which 95 (74%) were identical in both tests. The most frequently isolated bacteria in both culture and PCR assay were coliform organisms and Enterococcus spp. The Gram negative bacilli were most resistant to cotrimoxazol. 77.2% of these bacilli and 100% of E. faecalis and S. agalactiae were sensitive to amoxicillin-clavulanic acid. 4.7% of Gram positive cocci were resistant to nitrofurantoin. Conclusions. The PCR method quickly finds the causative agent of urinary tract infection (UTI) and, therefore, it can help with making the choice of the proper antimicrobial therapy at an early stage. It appears to be a viable alternative to the recommendations made in general treatment guidelines, in cases where diversified sensitivity patterns of microorganisms have been found. [ABSTRACT FROM AUTHOR]

Abstract: External laboratory proficiency programs are an important requirement for test quality assurance (EQA) and compliance to regulatory guidelines (Clinical Laboratory Improvement Amendments and inspections). The American Proficiency Institute (API) regularly distributes EQA sample challenges (test events) including an Educational Sample (ES) for antimicrobial susceptibility testing. Beginning in 2007, API has sent 3 ES samples annually, each a well-characterized (molecular/phenotypic methods) strain having an interesting/emerging mechanism of resistance. Hundreds of USA laboratories, usually serving small- to medium-size hospitals and clinics, participate in the API ungraded ES test event. Analysis of responses is made and reported electronically as ES critiques addressing contemporary susceptibility testing issues that affect patient therapy. Seven Gram-positive (+) and 8 Gram-negative (−) ES strains were tested over the 5 years (2007–2011) with organism identification (graded) accuracy of 95.3% (range, 91.0–99.2%) for Gram (−) and 97.0% (range, 94.2–100.0%) for Gram (+) challenges. Susceptibility testing categorical accuracy was generally greatest for the disk diffusion test (91.0/97.0%) compared to the MIC methods (commercial automated or manual) combined (89.9/96.1%, for Gram [−]/Gram [+], respectively). The most worrisome observations of these ES samples were as follows: 1) poor recognition of ESBL- and serine carbapenemase-producing strains (various types including Klebsiella pneumoniae carbapanemase) due to delayed application of Clinical and Laboratory Standards Institute [CLSI] guidelines; 2) overcalling of ESBL in organisms having wild-type non-ESBL enzymes (OXA series; OXA, 1/30) due to commercial system or participant interpretive error; and 3) occasional drug-bug discords noted in nonfermentative Gram (−) bacilli. In conclusion, the API ES series of ungraded susceptibility testing challenges (accuracy was >90%) has been well received by subscribers and has provided detailed educational opportunities to improve laboratory testing performance. ES samples have delivered guidance to enable laboratories to rapidly comply with CLSI document changes of interpretive breakpoints such as those for β-lactams when testing Enterobacteriaceae and Pseudomonas aeruginosa; the program was sustained into 2012 and beyond to document quality of susceptibility tests in USA laboratories. [Copyright &y& Elsevier]

Abstract: Ceftaroline, the bio-active form of parenterally administered ceftaroline fosamil, is a unique broad-spectrum cephalosporin with in vitro and in vivo activity against methicillin-resistant Staphylococcus aureus and was approved for clinical use by the United States Food and Drug Administration in October 2010. In over a year since ceftaroline fosamil approval, no widely used commercial susceptibility test system has added this new compound to its product, therefore requiring use of alternative agar diffusion methods for clinical microbiology laboratories that want to test clinical isolates for ceftaroline susceptibility. An alternative strategy of applying a surrogate β-lactam class marker agent was assessed here, using results from 14,902 organisms (2008–2010) sampled in the USA. Very high and acceptable accuracy (≥99.75%) was observed for predicting ceftaroline susceptibility as follows: 1) use of imipenem or meropenem minimum inhibitory concentrations (MICs) at ≤8 μg/mL (susceptible and intermediate categories) when testing S. aureus; 2) use of ceftriaxone MIC at ≤2 μg/mL (susceptible and intermediate categories) when testing Streptococcus pneumoniae as well as other streptococci (S. pyogenes and S. agalactiae); and 3) use of ceftriaxone, or cefepime, or ceftazidime at ≤2 μg/mL (susceptible category) when testing Haemophilus influenzae. Only when testing indicated Enterobacteriaceae species using ceftriaxone susceptibility results did the ceftaroline-nonsusceptible errors increase (4.11%). These presented analyses offer a validated surrogate marker strategy for ceftaroline susceptibility testing, pending development and validation by the commonly used automated systems and agar diffusion commercial methods. [Copyright &y& Elsevier]

The increasing prevalence of ESBL-, AmpC- and carbapenemase-producing Enterobacteriaceae necessitates reliable phenotypic tests for detection and categorization. The main objective of this study was to evaluate ROSCO Neo-Sensitabs with different β-lactam-β-lactam inhibitor combinations for phenotypic detection and categorization of β-lactamases in Enterobacteriaceae. Using standard CLSI/EUCAST methodology, differences in zones of inhibitions between a β-lactam alone compared with the combination with a β-lactamase inhibitor as well as subjective synergy observations were determined for 172 well characterized Enterobacteriaceae strains with defined resistance mechanisms. The results showed that for all ESBL-positive strains (n = 66), combinations of clavulanic acid synergy with cefotaxime, ceftazidime or cefepime, were observed. All acquired AmpC β-lactamases (n = 17) were detected using cloxacillin combined with cefotaxime and/or ceftazidime (both combinations were required). Carbapenemase producers (n = 59) with the exception of one KPC-producer were correctly grouped using the combination of meropenem ± aminophenylboronic acid (APBA) or dipicolinic acid (DPA). Ethylene diamine tetraacetic acid (EDTA) also inhibited all metallo-β-lactamases, but as with DPA, one false positive result was observed. Based upon these data, we propose a tablet layout for 14 cm agar plates, which could be used as a whole or in a targeted approach for detection and categorizing of relevant acquired β-lactamases in Enterobacteriaceae. [ABSTRACT FROM AUTHOR]

The susceptibility of Sporothrix schenckii isolates from clinical cases of canine, feline and human sporotrichosis, and from the environment, was evaluated with 4% sodium hypochlorite and 6.6% chlorhexidine digluconate using the broth microdilution, agar diffusion and direct exposure techniques. The minimal inhibitory concentration was smaller than 0.8% for chlorhexidine digluconate and between 8% and 4% for sodium hypochlorite. Inhibition zones were not found in agar diffusion for sodium hypochlorite, and zones averaging 1.9 mm were found for chlorhexidine digluconate. In the direct exposure test, sodium hypochlorite demonstrated best performance at 20 min of contact, as chlorhexidine digluconate presented little antimicrobial activity. [ABSTRACT FROM AUTHOR]

Introduction: In Kenya, which ranks thirteenth of 27 high tuberculosis burden countries, diagnosis is based on Ziehl-Neelsen staining alone and patients are treated without information on sensitivity patterns. This study aimed to determine resistance patterns of Mycobacterium tuberculosis isolated from pulmonary samples. Methodology: Pulmonary tuberculosis patients in Nairobi were randomly sampled after informed consent and recruited into the study using a structured questionnaire. Specimens were cultured in liquid and solid media, and drug susceptibility tests were performed for first-line drugs including (isoniazid, rifampin, streptomycin, ethambutol and pyrazinamide). Results: Eighty-six (30%) of 286 isolates were resistant to at least one of five antibiotics tested. Thirty-seven (30.2%) isolates were resistant to isoniazid; 15 (11.6%) to streptomycin; 13 (4.5%) to ethambutol; four (1.4%) to rifampin ; and 30 (10.4%) to pyrazinamide. Double resistance was seen as follows: four (1.4%) isolates were resistant to both isoniazid and pyrazinamide; four (1.4%) to streptomycin and isoniazid; and one (0.3%) to rifampin and streptomycin. Two isolates (0.7%) were multidrug resistant, and one was triple resistant with an additional resistance to ethambutol. Results also showed 88.7% of patients were below the age of 40 years, while 26.3% were HIV positive. The majority of the patients (66.5%) were unemployed or self-employed in small businesses, with 79.4% earning less than 100 USD per month. Conclusion: The high resistance observed in isoniazid, which is a first-line drug, could result in an increase in multidrug resistance unless control programs are strengthened. Poverty should be addressed to reduce infection rates. [ABSTRACT FROM AUTHOR]

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Cryptococcus neoformans causes disseminated infection in 7–8% of HIV positive patients admitted to Hospital F. J. Muñiz in Buenos Aires. Meningoencephalitis is the most frequent clinical manifestation and is one of the main causes of death in those patients with AIDS. The standard treatment for this mycosis consists of amphotericin B followed by fluconazole until two successive cultures of CFS are negative. Although resistance to these drugs is infrequent, minimal inhibitory concentrations (MIC) of some antifungals can be high. Since it is important to know the susceptibility levels of this fungus to the antifungal drugs usually employed in our institution, we analyzed the susceptibility test results of C. neoformans with two diffusion methods (Etest and NeoSensitabs tablets) employing Mueller-Hinton agar with 2% glucose and 0.5 μg/ml methylene blue. These results were compared with MICs obtained through the use of the broth microdilution reference method (CLSI). Results showed good agreement with the reference method, with no very major errors and only two major errors for fluconazole using NeoSensitabs tablets. For all the above mentioned, we confirm the usefulness of Mueller-Hinton agar to evaluate C. neoformans susceptibility to amphotericin B and fluconazole with these two agar diffusion methods. [ABSTRACT FROM AUTHOR]

Objectives: Mycobacterium kansasii causes a variety of infections. Although previous reports on the prognosis of antimicrobial therapy have been mostly satisfactory, problems involving treatment failure or relapse have been encountered. The purpose of this study was to establish a relationship between the clinical treatment outcomes of M. kansasii infections and bacterial drug susceptibility, and their clonality. [ABSTRACT FROM PUBLISHER]

Dermatophytoses caused by the genus Microsporum require a long-duration therapy compared to infections caused by other genera. Treatment of these cutaneous infections includes topical and systemic antifungal agents. Tinea capitis and tinea unguium caused by M. canis and M. gypseum are the most difficult-to-treat dermatophytoses. There are few specific studies about corresponding antifungal susceptibility in vitro. Recently, the Clinical and Laboratory Standards Institute proposed the M38A document as standard to determine the minimum inhibitory concentrations (MICs) of several antifungal agents against conidium-forming filamentous fungi; however, dermatophytes were not included in this document. This study aimed to contribute to continuing investigations concerning the optimal antifungal susceptibility testing conditions of Microsporum spp. to terbinafine, ciclopiroxolamine and griseofulvin . The results pointed out potato dextrose agar as the best culture medium for inducing conidia sporulation, inoculum density amounting to 1 × 103 conidia ml−1, containing only microconidia, with an incubation time of 7 days at 28 °C and 100% growth inhibition serving as an endpoint. The minimum fungicidal concentration values were in accordance with the MICs values, showing a fungicidal activity of these drugs towards the tested strains. According to our results, in general, terbinafine was more active than griseofulvin and ciclopiroxolamine. [ABSTRACT FROM AUTHOR]

Aim: The emergence of multidrug-resistant tuberculosis (MDR-TB) is increasing, and the standard short-course regimen used for the treatment of TB is likely to be ineffective against MDR-TB, leading to the need for second-line drugs. In such situations, drug susceptibility testing is necessary to select an appropriate treatment regimen. Unfortunately, there are few studies showing the pattern of the second-line drug resistance in Turkey. We aimed to analyze the resistance to second-line anti-tuberculosis drugs of MDR strains of Mycobacterium tuberculosis isolated from the Aegean region of Turkey. Materials and Methods: In this study, drug susceptibility testing of 40 MDR-TB strains isolated from the Aegean region of Turkey was performed using the BACTEC 460 TB radiometric system. Capreomycin, ethionamide, kanamycin, amikacin, clofazimine and ofloxacin were tested in 1.25 μg/ml, 1.25 μg/ml, 5.0 μg/ml, 1.0 μg/ml, 0.5 μg/ml, and 2.0 μg/ml concentrations, respectively. Results: The results showed that 37.5% of the strains were resistant to ethionamide, 25% to capreomycin, 5% to kanamycin, amikacin and ofloxacin, and 2.5% to clofazimine. One (2.5%) of the 40 MDR-TB cases was defined as extensively drug-resistant tuberculosis (XDR-TB). Conclusions: The results of the study indicate that the high rates of resistance to ethionamide and capreomycin may be a problem in the treatment of patients with MDR-TB; XDR-TB is not yet a serious problem in our region. [ABSTRACT FROM AUTHOR]

Abstract: Treatment options are increasingly limited for carbapenem-resistant Acinetobacter baumannii. This study set out to determine the in vitro susceptibility of multiresistant Acinetobacter spp. to colistin, minocycline, and rifampicin using agar dilution, and to compare the accuracy of testing results obtained from an automated susceptibility testing system (Vitek 2). MICs for colistin, minocycline, and rifampicin were obtained by agar dilution and Vitek 2 for 44 unrelated strains of multiresistant Acinetobacter spp. All tested strains were susceptible to colistin, whereas 61% were susceptible to minocycline and 52% were susceptible to rifampicin. Results for colistin testing showed categoric agreement between Vitek 2 and agar dilution, with no false-resistance reported by Vitek 2. However, there was a poor agreement for minocycline and rifampicin when results obtained from Vitek 2 testing were compared with those obtained by agar dilution. [Copyright &y& Elsevier]

The availability of new antifungal agents has multiplied the demand for in vitro antifungal susceptibility testing for Aspergillus spp. The European Committee on Antimicrobial Susceptibility Testing (EUCAST) has charged its Antifungal Susceptibility Testing Subcommittee (AFST-EUCAST) with the preparation of new guidelines for in vitro susceptibility testing of antifungals against Aspergillus spp (EUCAST-AST-ASPERGILLUS). This committee has modified the reference method for broth dilution antifungal susceptibility testing of filamentous fungi (M38-A) as follows: (i) RPMI 1640 supplemented with 2% glucose (RPMI 2%G) as assay medium, (ii) inoculum preparation by conidium counting in a haemocytometer and (iii) an inoculum size of 2×105-5×105 CFU/ml. The incubation time is about 48 h at 35°C and MIC is read visually. The MIC value is a no-growth visual endpoint. The standard method described herein is intended to provide a valid and economic method for testing the susceptibility to antifungal agents of Aspergillus spp., to identify resistance, and to facilitate an acceptable degree of conformity, e.g. agreement within specified ranges and between laboratories in measuring the susceptibility. [ABSTRACT FROM AUTHOR]

We present the results of studies of the in vitro susceptibility of 52 isolates of Trichophyton rubrum and 40 of Trichophyton mentagrophytes to griseofulvin, terbinafine, itraconazole, ketoconazole, fluconazole and cyclopiroxolamine. All test strains were recovered from patients with toe nail onychomycosis and the minimum inhibitory concentration (MIC) of each antifungal against both species was individually assessed. In addition, we investigated the MIC of the combination of cyclopiroxolamine and itraconazole and cyclopiroxolamine and ketoconazole. The NCCLS approved procedure M38-A as modified by Santos and Hamdan was employed. The studies of the two drug combinations were conducted with a checkerboard design. Analysis of the data revealed that terbinafine was the most effective in vitro against all isolates, followed in order by itraconazole, cyclopiroxolamine, ketoconazole and fluconazole. We observed no significant difference in the in vitro susceptibility profiles between either species to any of the antifungals ( P [ABSTRACT FROM AUTHOR]

Abstract: Minimal inhibitory concentrations (MICs) of prulifloxacin were evaluated in comparison with ciprofloxacin, levofloxacin and moxifloxacin against a large collection (N =300) of Pseudomonas aeruginosa strains characterised according to the CLSI/NCCLS microdilution method. Additional in vitro tests (time–kill curves and mutant prevention concentration (MPC) determinations) were carried out. Assuming a susceptibility breakpoint for prulifloxacin identical to that of ciprofloxacin, the new fluoroquinolone emerged as the most potent antibiotic (72% of susceptible strains versus 65%, 61% and 23% for ciprofloxacin, levofloxacin and moxifloxacin, respectively). Time–kill tests at 4× MIC confirmed the pronounced bactericidal potency of the drug against P. aeruginosa. Amongst the members of the fluoroquinolone class assessed, prulifloxacin produced the lowest MPC values (≤4mg/L). Our in vitro results indicate that prulifloxacin represents the most powerful antipseudomonal drug available today. [Copyright &y& Elsevier]

Thirty-two Escherichia coli and 21 Klebsiella pneumoniae septicemia isolates with varying degrees of resistance to ciprofloxacin were analyzed for the presence of point mutations within the quinolone-resistance target genes. The number of mutations observed in the resistant isolates agreed with the level of ciprofloxacin resistance in both species. Such isolates were also resistant to nalidixic acid. Isolates with borderline susceptibility to ciprofloxacin, on the other hand, behaved differently in the two species. In E. coli all the isolates harbored at least one mutation and these isolates were also resistant to nalidixic acid, while no mutations were detected in the K. pneumoniae isolates, and susceptibility to nalidixic acid was unpredictable. Therefore, nalidixic acid cannot be used as a class representative. Time-kill curve studies on an isolate with borderline susceptibility from each species showed higher degrees of resistance to ciprofloxacin in comparison to that of the wild-type E. coli. A previously unreported parC mutation, S57→T, was detected in a resistant E. coli isolate and might expand the QRDR of this gene. Normalized resistance interpretations of histograms confirmed the setting of microbiological zone breakpoints for ciprofloxacin testing. [ABSTRACT FROM AUTHOR]

Summary 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 edn. 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. [ABSTRACT FROM AUTHOR]

Background: This review addresses the ability of a commercially available genetic susceptibility test to determine the risk of developing severe chronic periodontitis. The test is used to detect the simultaneous occurrence of allele 2 at the IL-1A+4845 and IL-1B+3954 loci. If both of these polymorphisms are present, patients are referred to as being genotype-positive and considered predisposed to becoming afflicted with severe chronic periodontitis. A basic premise of this Lest is the assumption that individuals who are genotype-positive produce increased amounts of IL-β in response to microbial lipopolysaccharides, which allegedly predisposes them la an exaggerated inflammatory response and an increased incidence of chronic periodontitis. Methods: Controlled clinical trials were selected that evaluated the ability of the genetic test to predict which patients were susceptible to bleeding upon probing, periodontitis, peri-implantitis, and tooth loss. Results: Comparison of results from test (genotype-positive) and control groups (genotype-negative) revealed that there is ambiguity with regard to predicting which patients will manifest elevated subgingival levels of IL-β. Similarly, it is questionable if the test is able to forecast which individuals will demonstrate an increased occurrence of bleeding upon probing, diminished clinical attachment, decreased osseous support, or loss of teeth. Conclusions: There are many unanswered questions concerning the utility of detecting allele 2 at the IL-1A+4845 and IL-1B+3954 loci to foretell which patients will develop severe chronic periodontitis. Therefore, clinicians must cautiously interpret results obtained with the commercially available genetic susceptibility test before they alter maintenance schedules or treatment regimens of symptomatic or asymptomatic patients. [ABSTRACT FROM AUTHOR]

Summary Northern Kwazulu/Natal (KZN) Province of South Africa borders on southern Mozambique, between Swaziland and the Indian Ocean. To control malaria vectors in KZN, houses were sprayed annually with residual DDT 2 g/m2 until 1996 when the treatment changed to deltamethrin 20–25 mg/m2. At Ndumu (27°02′ S, 32°19′ E) the recorded malaria incidence increased more than six-fold between 1995 and 1999. Entomological surveys during late 1999 found mosquitoes of the Anopheles funestus group (Diptera: Culicidae) resting in sprayed houses in some sectors of Ndumu area. This very endophilic vector of malaria had been eliminated from South Africa by DDT spraying in the 1950s, leaving the less endophilic An. arabiensis Patton as the only vector of known importance in KZN. Deltamethrin-sprayed houses at Ndumu were checked for insecticide efficacy by bioassay using susceptible An. arabiensis (laboratory-reared) that demonstrated 100% mortality. Members of the An. funestus group from Ndumu houses (29 males, 116 females) were identified by the rDNA PCR method and four species were found: 74 An. funestus Giles sensu stricto, 34 An. parensis Gillies, seven An. rivulorum Leeson and one An. leesoni Evans. Among An. funestus s.s. females, 5.4% (4/74) were positive for Plasmodium falciparum by ELISA and PCR tests. To test for pyrethroid resistance, mosquito adults were exposed to permethrin discriminating dosage and mortality scored 24 h post-exposure: survival rates of wild-caught healthy males were 5/10 An. funestus, 1/9 An. rivulorum and 0/2 An. parensis; survival rates of laboratory-reared adult progeny from 19 An. funestus females averaged 14% (after 1 h exposure to 1% permethrin 25 : 75 cis : trans on papers in WHO test kits) and 27% (after 30 min in a bottle with 25 μg permethrin 40 : 60 cis : trans). Anopheles funestus families showing > 20% survival in these two resistance test procedures numbered 5/19 and 12/19, respectively. Progeny from 15 of the families were tested on 4% DDT impregnated papers and gave 100% mortality. Finding these proportions of pyrethroid-resistant An. funestus, associated with a malaria upsurge at Ndumu, has serious implications for malaria vector control operations in southern Africa. [ABSTRACT FROM AUTHOR]

Antifungal drug susceptibility tests (AST) for Candida albicans are increasingly demanded for women with refractory or recurrent Candida vaginitis due to fluconazole resistance. Given reduced activity of azole drugs at pH levels found in women with Candida vaginitis, it is proposed that AST be performed at pH 4.5, since testing at only the recommended pH 7.0 is likely to miss a significant number of clinically relevant azole-resistant C. albicans vaginal isolates.