Fungal infections due to Candida species represent an im- portant cause of nosocomial bloodstream infections (18, 20). Candida is the fourth most prevalent cause of nosocomial bloodstream infections in Brazil, with an incidence of 2.49 cases per 1,000 hospital admissions, and a crude mortality of 54% (1). Candida guilliermondii is an infrequent, but not un- usual, agent of candidiasis and has been described as an emerg- ing pathogen. As reviewed by Krcmery and Barnes (7), over a 40-year period ending in 1990, only 10 cases of C. guilliermondii fungemia had been reported, whereas 33 cases were reported between 1990 and 2002. In that study, the rate of fungemia due to C. guilliermondii varied from 0 to 0.7% in the period be- tween 1950 and 1990 and from 0.7 to 5.5% from 1991 to 1998. Most cases of C. guilliermondii infection are associated with oncology patients (8, 16, 19). A review of the cases of fungemia in oncology patients reported from 1960 to 1990 showed that 10 cases (0.7%) were due to C. guilliermondii, with three deaths (19). In Brazil, a study conducted in six hospitals from Sa ˜o Paulo and Rio de Janeiro by Colombo et al. (2) showed three cases of C. guilliermondii infection among 145 Candida blood- stream infection patients. In another study involving 11 Bra- zilian institutions, C. guilliermondii accounted for 2.4% of 712 candidemias (1). Outbreak investigations involving these or- ganisms must distinguish reliably among the various species involved. Most strains of C. guilliermondii are morphologically
The Gram stain and acid-fast bacteria (AFB) stain showed no bacteria or tuberculosis. Candida guilliermondii was cultured in the joint fluid on hospital day 7 and was confirmed on hospital day 8. Amphotericin B (0.7 mg/kg/day) was given intravenously. Two weeks later, CRP had normalized. After 4 weeks, the amphotericin B was discontinued and oral fluconazole (400 mg/day) was prescribed for 6 months. The last culture of joint fluid was negative for C. guilliermondii,
The microbiological, clinical, and epidemiological features of most non-Candida albicans Candida species are well known, but much less is known about species such as Candida guilliermondii, an uncommon pathogen causing a variety of deep-seated infections in immunocompromised hosts. To characterize C. guilliermondii fungemia in patients with hematological malignancies and its susceptibility to antifungal drugs, all cases of C. guilliermondii fungemia diagnosed in our department between 1983 and 2005 were retrospectively analyzed and the literature was reviewed. C. guilliermondii caused 29/243 (11.7%) candidemia episodes diagnosed during the study period. Central venous catheters were the documented sources of candidemia in 19/29 episodes (65.5%), and invasive tissue infections were documented in 2 (6.9%). In the remaining eight, the catheter was not removed and the source of the fungemia remained obscure. Seven episodes ended in death, but only one could be attributed to invasive C. guilliermondii infection. Molecular typing data reveal no evidence of common infection sources. Isolates displayed high rates of in vitro susceptibility to amphotericin B (100%), voriconazole (95%), and fluconazole (90%) and lower rates of in vitro susceptibility to flucytosine (86%), itraconazole (76%), and caspofungin (33%). Our literature review confirms that C. guilliermondii is a significantly more frequent cause of candidemia among cancer patients compared with the general hospital population. It accounted for <1% of the total number of Candida bloodstream isolates reported in the articles we reviewed, with higher rates in Europe (1.4%) and Asia (1.8%) compared with North America (0.3%).
Methods: We performed a comparative study analyzing the cell wall composition and organiza- tion of Candida tropicalis, Candida guilliermondii, Candida krusei, and Candida auris, along with their ability to stimulate cytokine production and phagocytosis by human innate immune cells. Results: We found that the wall of these species had the basic components already described in C. albicans, with most of the chitin and b1,3-glucan located underneath the mannan layer. However, the walls of C. krusei and C. auris were rich in chitin and the former had a lower content of mannans. C. guilliermondii contained changes in the mannan and the b1,3-glucan levels. These species were differentially phagocytosed by human macrophages and stimulated cytokine production in a dectin-1-dependent pathway. C. krusei showed the most significant changes in the tested parameters, whereas C. auris behaved like C. albicans.
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Case presentation: A 5-year-old male Boxer, positive to Leishmania infantum, was referred to the Veterinary Teach- ing Hospital of the Department of Veterinary Medicine, University of Perugia, Italy for examination of a non-weight bearing left hind limb lameness of a duration of at least 3 months. During this period, treatment involved systemic anti-inflammatory medications and intra-articular corticosteroid administration. On presentation, clinical examination and radiographic findings were suggestive of cranial cruciate ligament deficiency. To support this diagnosis a stifle arthroscopy was performed: it confirmed a partial rupture of cranial cruciate ligament. Samples culture of synovial fluid and membrane was routinely collected as well, and revealed Candida guilliermondii joint infection. Treatment for the C. guilliermondii joint infection involved systemic anti-fungal therapy, joint lavage and intra-articular administration of antifungal drugs. Lameness improved markedly during this treatment, but lameness did not resolve completely, probably due to cranial cruciate ligament deficiency. Tibial tuberosity advancement (TTA) was chosen in order to treat stifle instability and was performed 4 weeks following cessation of treatment of the C. guilliermondii joint infection. Six month after TTA the dog showed a completely recovery with no lameness.
The emerging pathogens Candida palmioleophila, Candida fermentati, and Debaryomyces nepalensis are often misidentified as Candida guilliermondii or Candida famata in the clinical laboratory. Due to the significant differences in antifungal susceptibil- ities and epidemiologies among these closely related species, a lot of studies have focused on the identification of these emerging yeast species in clinical specimens. Nevertheless, limited tools are currently available for their discrimination. Here, two new molecular approaches were established to distinguish these closely related species. The first approach differentiates these species by use of restriction fragment length polymorphism analysis of partial internal transcribed spacer 2 (ITS2) and large subunit ribosomal DNA with the enzymes BsaHI and XbaI in a double digestion. The second method involves a multiplex PCR based on the intron size differences of RPL18, a gene coding for a protein component of the large (60S) ribosomal subunit, and species- specific amplification. These two methods worked well in differentiation of these closely related yeast species and have the poten- tial to serve as effective molecular tools suitable for laboratory diagnoses and epidemiological studies.
Although a rare cause of invasive candidiasis, Candida guilliermondii has been reported to exhibit decreased susceptibility to antifungal agents. Aside from case reports and small surveys, there is little information regarding the epidemiology and antifungal susceptibility profile of C. guilliermondii. We report geographic and temporal trends in the isolation and antifungal susceptibilities of 1,029 C. guilliermondii clinical isolates collected from 127 medical centers as part of the ARTEMIS DISK Antifungal Surveillance Program. In addition, we report the in vitro susceptibility of 132 bloodstream isolates of C. guilliermondii to caspofungin. C. guilliermondii represented 1.4% of the 75,761 isolates collected from 2001 to 2003 and was most common among isolates from Latin America (3.7% versus 0.6 to 1.1%). Decreased susceptibility to fluconazole was noted (75% susceptible; range, 68 to 77% across regions), and voriconazole was more active in vitro against C. guilliermondii than fluconazole (91% susceptible; range, 88 to 93% across regions). Fluconazole was least active against isolates from dermatology (58%) and surgical (69%) services and against isolates associated with skin and soft tissue infection (68%, compared to 85% susceptible for bloodstream isolates). There was no evidence of increasing azole resistance over time among C. guilliermondii isolates tested from 2001 to 2003. Of 132 bloodstream isolates of C. guilliermondii tested against caspofungin, most were inhibited by < 2 g/ml (96%; MIC 50 /MIC 90 , 0.5/1.0 g/ml). C. guilliermondii, a species that exhibits reduced susceptibility to fluconazole, is
Case presentation: An 87-year-old Swiss man with German ethnic origin suffered from symptoms of osteoarthritis of the knee. We present the first described case of periprosthetic joint infection after total knee arthroplasty by both Mycobacterium bovis and Candida guilliermondii in the context of a zoonosis with 14 months of follow-up. The infection was presumed to originate more than 55 years earlier, when these infectious agents were still present in cattle in Switzerland. After diagnosis of the pathogens, our patient was successfully treated with tuberculostatic and mycocide medication, and a two-stage revision knee arthroplasty was performed. The medication was given for 1 year.
 O. Bengtsson, B. Hahn-Hägerdal and M. F. Gorwa- Grauslund, “Xylose Reductase from Pichia stipitis with Altered Coenzyme Preference Improves Ethanolic Xylose Fermentation by Recombinant Saccharomyces cerevis- ae,” Biotechnology for Biofuels, Vol. 2, No. 9, 2009, p. 10.  D. B. Gurpilhares, A. Pessoa Jr. and I. C. Roberto, “Glu- cose-6-phosphate Dehydrogenase and Xylitol Production by Candida guilliermondii FTI 20037 Using Statistical Experimental Design,” Process Biochemistry, Vol. 41, No. 3, 2005, pp. 631-637.
ABSTRACT Candida guilliermondii was isolated from sterile specimens with increas- ing frequency over a several-month period despite a paucity of clinical evidence suggesting true Candida infections. However, a health care-associated outbreak was strongly considered due to growth patterns in the microbiology laboratory that were more consistent with true infection than environmental contamination. There- fore, an extensive investigation was performed to identify its cause. With the excep- tion of one case, patient clinical courses were not consistent with true invasive fun- gal infections. Furthermore, no epidemiologic link between patients was identiﬁed. Rather, extensive environmental sampling revealed C. guilliermondii in an anaerobic holding jar in the clinical microbiology laboratory, where anaerobic plates were prer- educed and held before inoculating specimens. C. guilliermondii grows poorly under anaerobic conditions. Thus, we postulate that anaerobic plates became intermit- tently contaminated. Passaging from intermittently contaminated anaerobic plates to primary quadrants of aerobic media during specimen planting yielded a colonial growth pattern typical for true specimen infection, thus obscuring laboratory con- tamination. A molecular evaluation of the C. guilliermondii isolates conﬁrmed a com- mon source for pseudo-outbreak cases but not for the one true infection. In line with Reason’s model of organizational accidents, active and latent errors coincided to contribute to the pseudo-outbreak. These included organism factors (lack of growth in anaerobic conditions obscuring plate contamination), human factors (lack of strict adherence to plating order, leading to only intermittent observation of aero- bic plate positivity), and laboratory factors (novel equipment). All of these variables should be considered when evaluating possible laboratory-based pseudo-outbreaks.
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mentati, described by Bai in 1996 (1). Changes in nomenclature could also explain additional mistakes such as the P. jadinii synonym of C. guilliermondii var. nitratophila. The teleomor- phic state of C. fermentati has been recently described by Vaughan-Martini in 2005 based on molecular data and is named Pichia caribbica (34). Since P. caribbica has been de- scribed, many analyses using molecular tools (RAPD [ran- dom(ly) amplified polymorphic DNA], electrophoretic karyo- typing, DNA composition, chromosomal DNA banding profiles, and D1/D2 sequences) reclassified clinical and envi- ronmental isolates phenotypically identified as P. guilliermondii into several species, including P. guilliermondii but also P. caribbica (2, 14, 30, 34). In our analyses, some carbon assimi- lation profiles were shared by the two species, one of the codes being a frequent code for both (see Table S2 in the supple- mental material), and the similarity of the ITS and 26S se- quences was greater than 98%.
An incorrect species identification (major error) occurred for two isolates (3.8%) with the MALDI Biotyper (one C. fermentati and one K. ohmeri misidentified as C. guilliermondii); however, the misidentification of the C. fermentati can be explained by the fact that this organism, a cryptic species within the C. guilliermon- dii species complex (26), was not in the current database. Gener- ally, MALDI has the advantage of having a “no reliable result” response rather than making an erroneous identification (Table 2). Among the 10 discrepancies or low-score results observed be- tween MALDI and the molecular reference method, six were due to a report of no reliable identification by the MALDI Biotyper, two of which were species not included in the MALDI database (C. fermentati and C. fabianii). C. metapsilosis and C. orthopsilosis, newly recognized as members of the C. parapsilosis species com- plex, are frequently misidentified by phenotypic methods as C. parapsilosis sensu stricto (27). Previous studies show that these two species can be identified by MALDI (28, 29); however, in two instances (2, 7, 11, 13, 28, 29) the original manufacturer spectral database library was secondarily modified to include these rare yeast species. Interestingly Hendrickx et al. (30) were able to iden- tify both of these cryptic species by MALDI-TOF MS without any database modifications. Overall, our results are comparable with those obtained in previously published studies where MALDI was compared to a reference molecular identification method (22, 29, 31).
The use of ribosomal DNA (rDNA) genes for identification of fungal species is based on the detection of conserved se- quences in 5.8S rDNA and 28S rDNA that enable the ampli- fication of the ITS2 region between these two genes. In this study, we PCR amplified, by using a fluorescent primer pair, the ITS2 regions of 56 fungal species of clinical significance. The amplicons were rapidly and accurately sized with an au- tomated capillary electrophoresis system, ABI PRISM 310 ge- netic analyzer. No intraspecies variability was observed among species for which more than one strain was tested. These spe- cies included Candida albicans, Candida guilliermondii, Can- dida tropicalis, Candida krusei, Candida glabrata, Candida parapsilosis, Rhodotorula rubra, and Trichophyton verrucosum. An exception was Trichosporon beigelii, of which a reference strain and a clinical strain were tested. Kemker et al. (12) previously demonstrated, by using restriction fragment length polymorphism of a segment of the ribosomal genes including the ITS2 region, that isolates identified as Trichosporon beigelii are genetically distinct from each other, depending on the source of the organism. The authors suggest that Trichosporon beigelii may represent several distinct entities and that, for the purpose of molecular diagnosis research, isolates from invasive
A more generally accurate method of assessment is the broth dilution technique. In this study, therefore, the broth dilution method was used in determining the activities measured as MIC. In using this method, higher degrees of differences in susceptibility among Candida species were not observed. The crude methanol extract was found to have minimum inhibitory concentration of 800 µ g/ml, 800 µg/ml and 1600µg/ml aganist Candida parapsilosis SN 1980, Candida guilliermondii SN 2006 and Candida glabrata SN 2266 respectively. The results revealed that the test extract showed more killing in case of Candida parapsilosis SN 1980 and Candida guilliermondii SN 2006 respectively It would appear that Candida glabrata SN 2266 is the less sensitive yeast to the test extract. The results obtained provide us obvious evidence that the test extract used in the study has a substantial level of antifungal activity.
oratory at Royal North Shore Hospital, Sydney, Australia, and the Molecular Mycology Research Laboratory at Westmead Hospital, Westmead, Australia. Clinical isolates were obtained from the Mycology Laboratory at Westmead Hospital. Isolates were identified using standard colonial and microscopic char- acteristics (for molds) (20, 37) and the VITEK I (bioMerieux Vitek, Hazelwood, MO) and/or ID 32C (bioMerieux, Marcy-l’Etoile, France) commercial systems (for yeasts). Canavanine-glycine bromothymol blue (CGB) agar was used to differentiate between Cryptococcus neoformans (Cryptococcus neoformans var. neoformans and Cryptococcus neoformans var. grubii) and Cryptococcus gattii (19). A total of 159 (32 reference and 127 clinical) isolates belonging to 22 fungal species were studied; all species were represented by species-specific probes in the RLB assay (Table 1). Isolates comprised 16 Candida species (101 strains; Candida albicans, Candida dubliniensis, Candida glabrata, Candida guilliermondii, Candida haemulonii, Candida kefyr, Candida krusei, Candida lusitaniae, Candida norvegica, Candida norvegensis, Candida parapsilosis, Candida pelliculosa, Can- dida tropicalis, Candida utilis, Candida viswanathii, Candida zeylanoides), C. neo- formans complex (five strains of C. neoformans var. grubii, four strains of C. neoformans var. neoformans, and eight strains of C. gattii), and five Aspergillus species (40 strains; Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Aspergillus terreus, and Aspergillus nidulans).
observed in the current study were by the median age of the pregnant women with clinical Candida vaginitis was 28 with interquartile range of 22–32 years. Physiological and tissue changes, due to reproductive hormones, which happen in young women especially during pregnancy, increase their susceptibility to Candida infection, in addi- tion to adverse factors such as risky sexual behaviors. Previous study conducted in North America suggested that 70–75% of women can get at least one episode of Candida vaginitis in life time .
It should be noted that conventional identification was not discriminatory enough to characterize the most recently de- scribed species, such as C. orthopsilosis, C. metapsilosis, C. bracarensis, and Lodderomyces elongisporus. Notably, most Candida haemulonii isolates (4 out 5 isolates) and most Issatch- enkia terricola isolates (2 out of 3 included) were not classified by conventional methods. MIC distributions are shown in Ta- ble 3. Most strains were considered susceptible in vitro to AMB, as defined by a MIC of ⬍ 1 mg/liter. It is worth noting that 4 out 5 of the C. haemulonii strains showed high MICs to AMB (1 to 4 mg/liter).
Vaginal Candidiasis is the most common disease in pregnant women all over the world. The risk of carriage to develop disease in pregnant women is two times higher than non-pregnant women particularly during the third trimester due to changes in the levels of reproductive hormones and deposition of glycogen in the vagina (23,24) . Studies show that Candidiasis is the most common cause of vaginitis; in United States it is the second leading cause of viginitis next to bacterial vaginosis and it is also common in Europe (23) . Study from Spain reported 28% prevalence of Candida species among pregnant women. Of these, Candida albicans was the major isolate 90.4% followed by Candida glabrata 6.3%; Candida parapsilosis 1.1% and Candida kefyr 1.1 % (7) . Other studies around the world also reveled high prevalence of the disease; Sarajevo 46.8% (17) , Cuba 42.3% (23) and Kenya 42.7% (2) . Most studies reported Candida albican as the most predominant cause of vaginal Candidiasis in pregnant women which accounts 80- 95% of Candida infection (2, 5, 11, 17,18) . From non albican group, Candida glabratais the leading cause of vaginal Candidiasis in pregnant women (17) . In Ethiopia however, epidemiological studies of Candidiasis are limited to HIV patients and diabetic patients. Hence, literatures about the epidemiology of Candidiasis among pregnant women are lacking. Table No. 01: Prevalence of Candidiasis in pregnant women among different countries around the world
eral days by conventional methods, physicians often choose broad-spectrum antifungal therapy as initial treatment for all yeast-positive blood cultures, since delayed therapeutic inter- vention has been demonstrated to be associated with a poorer outcome (8). The C. albicans peptide nucleic acid fluorescent in situ hybridization (PNA FISH) assay was first described in 2002 (19) and later was cleared by the FDA as an in vitro diagnostic kit for identification of yeast directly from positive blood cultures. The assay uses PNA probes targeting C. albi- cans-specific rRNA in a standardized FISH assay format. PNA is a nucleic acid mimic whose hybridization properties make it particularly well-suited for FISH assays (20). C. albicans PNA FISH has been shown to have very high sensitivity and speci- ficity (12, 19, 22); the cost-benefit advantage of the assay has been established, with demonstrated savings accrued through avoidance of unnecessary echinocandin therapy (3, 7). C. albi- cans/C. glabrata PNA FISH (AdvanDx, Inc., Woburn, MA), described here, is a novel second-generation in vitro diagnostic test for identifying C. albicans and C. glabrata from blood culture bottles that have signaled positive and demonstrate yeast on Gram staining. The test differentiates the two Candida species by indicating green fluorescence for C. albicans and red fluorescence for C. glabrata. This study evaluated the use of the PNA FISH assay for accurate real-time identification of C. albicans and C. glabrata from blood cultures newly positive for yeast.
In the present study MALDI-TOF MS proved to be a rapid and reliable procedure for the accurate identification of patho- genic Candida strains and required minimal hands-on time or time for the interpretation of the results. The test procedure was generally completed within 10 min per isolate and within 3 h for 96 samples, starting from single yeast colonies on the agar plate. In contrast, the identification of germ tube-negative Candida species by phenotypic methods requires incubation periods of up to 72 h. Overall, MALDI-TOF MS did not produce any misidentifications, provided that spectra for the appropriate reference strains were present in the database (Table 2). Incorrect species assignments as best hits, due to the absence of reference strains, were clearly indicated by log(score) values that were too low, according to the thresholds of the MALDI Biotyper software.