Persistent Candida conglobata bloodstream infection in a preterm neonate successfully treated by combination therapy with amphotericin B and caspofungin.

Fungemia due to uncommon/rare Candida species is an emerging problem of global clinical significance. Here, we describe a case of Candida conglobata bloodstream infection in a preterm neonate. The diagnosis was established by repeated isolation of C. conglobata in blood cultures and by detection of rDNA of the fungus in serum samples. The identity of the isolate as C. conglobata was confirmed by sequencing of ITS region and D1/D2 domains of rDNA. Despite initial treatment with a liposomal amphotericin B (AmBisome) for 7 days, the blood culture remained positive. The neonate was successfully treated by combination therapy with caspofungin for 25 days. To the best of our knowledge, this is the first proven report unequivocally proving the etiologic role of C. conglobata in bloodstream infection.

Candida kefyr as a cause of bloodstream infection and adjunctive role of biomarkers in its diagnosis.

A rare case of bloodstream infection caused by Candida kefyr is described. The diagnosis was established by repeatedly isolating the yeast in blood cultures and by detecting C. kefyr-specific DNA in serum samples. Demonstration of elevated serum levels of ß-D-glucan and Candida mannan also provided additional diagnostic evidence. The identity of the isolates was confirmed by PCR sequencing of the ITS region of rDNA. This is the first report of C. kefyr candidemia from Kuwait and the Middle East. The report highlights emerging clinical significance of rare Candida spp. in etiology of candidemia and reinforces the adjunctive role of biomarkers in diagnosis.

Comparative in vitro susceptibility of clinical isolates of Candida paparsilosis complex and other Candida species to caspofungin and anidulafungin by Etest.

In vitro susceptibility of 141 clinical isolates of Candida species to caspofungin and anidulafungin is reported. the Etest was performed according to recommended procedure and minimum inhibitory concentrations (MICs) were read after 24 h of incubation at 35 °C. Applying a breakpoint of <2 mg/ml, all Candida spp. isolates, except those belonging to C. parapsilosis complex, were susceptible. The geometric mean for caspofungin and anidulafungin for different Candida spp. were as follows: Candida parapsilosis, 0.438 and 3.355 µg/ml; Candida orthopsilosis, 0.210 and 1.456 µg/ml; Candida albicans, 0.049 and 0.007 µg/ml; Candida dubliniensis, 0.077 and 0.009 µg/ml; Candida tropicalis, 0.061 and 0.027 µg/ml; Candida glabrata, 0.120 and 0.032 µg/ml; and Candida krusei, 0.288 and 0.052 µg/ml, respectively. Anidulafungin was significantly more active than caspofungin (p <0.001) except for C. parapsilosis complex spp. isolates. In conclusion, our Etest MICs compared well with epidemiological cutoff values derived from a large number of Candida spp. isolates tested by CLSI method in previous studies. However, considering the differences in MICs of the two echinocandins for C. parapsilosis complex isolates, the Etest needs further evaluation for its suitability.

Sunflower seed husk agar: a new medium for the differentiation of Candida dubliniensis from Candida albicans.

A sunflower (Helianthus annuus) seed husk agar medium has been developed and evaluated for differentiation of Candida dubliniensis from Candida albicans on the basis of colony morphology and chlamydospore production. All C. dubliniensis isolates (n=40) produced rough colonies with hyphal fringes and abundant chlamydospores whereas 101 of 105 (96.2%) C. albicans isolates produced smooth colonies with no evidence of chlamydospore production. Since this medium is free from oil droplets, chlamydospores can be examined with greater clarity by Dalmau plate technique. This medium provides a simple and cost-effective tool for the presumptive differentiation of C. dubliniensis from C. albicans and is particularly suited for clinical microbiology laboratories where biochemical or molecular methods for the differentiation of these two species are not available.

Simplified sunflower (Helianthus annuus) seed agar for differentiation of Candida dubliniensis from Candida albicans.

This study evaluated sunflower (Helianthus annuus) seed agar (SSA) for differentiation of Candida dubliniensis from Candida albicans on the basis of colony characteristics and chlamydospore production. Simplified SSA without creatinine and KH(2)PO(4) was also used. On both media, C. dubliniensis isolates (n = 25) developed rough colonies and formed abundant chlamydospores after incubation for 24-48 h at 28 degrees C, while C. albicans isolates (n = 53) showed smooth colonies with no evidence of chlamydospore formation. Cryptococcus neoformans isolates (n = 10) formed brown colonies on both media. Simplified SSA offers a simple and inexpensive tool for presumptive differentiation of C. dubliniensis from C. albicans in clinical microbiology laboratories.

Candida albicans strain carriage in patients and nursing staff of an intensive care unit: a study of morphotypes and resistotypes.

Candida albicans carriage of patients and nursing staff of an intensive care unit (ICU) was studied over an 8-month period. Swabs were taken at weekly intervals from multiple sites from patients. None of the patients had clinical Candida infection at the time of the first sampling. The hands and mouth of the nursing staff were sampled at fortnightly intervals. Of the 68 patients investigated for varying periods, 37 (54%) yielded C. albicans from one or more body sites, resulting in the isolation of 269 strains. Nosocomial acquisition of C. albicans was recorded in seven (19%) patients. The frequency of C. albicans isolation increased with extended stay in ICU. Sixteen of the 180 samples taken from hands and mouth of nursing staff, yielded C. albicans, 12 of which came from the mouth. Morphotyping of 88 randomly selected strains of C. albicans originating from 31 patients yielded 34 morphotypes. There appeared to be no preference for any morphotype to colonize a particular anatomic site. Based on the susceptibility results, nine resistotypes were recognized. No correlation was apparent between any specific morphotype and resistotype patterns. The differences in morphotype and resistogram patterns of C. albicans isolates originating from same patients over a period of time suggest that some of the patients were colonized with more than one strain. Similarities in the morphotype and resistotype patterns of C. albicans strains isolated from patients and nursing staff tend to suggest possibility of exogenous acquisition.

Disseminated cryptococcosis in an AIDS patient caused by a canavanine-resistant strain of Cryptococcus neoformans var. grubii.

A case of disseminated cryptococcosis caused by Cryptococcus neoformans var. grubii is presented in a male diabetic who had AIDS. The diagnosis was based upon the isolation and identification of the aetiological agent from a lymph-node biopsy, cerebrospinal fluid and sputum. The isolate formed spherical, encapsulated yeast cells, produced cherry-brown colonies on niger-seed agar, grew on canavanine-glycine-bromothymol blue (CGB) medium, changing its colour from greenish yellow to blue, and hydrolysed urea weakly in the presence of 100 microM EDTA. The strain was unable to assimilate D-proline and, serologically, it was untypable. The identity of the isolate as C. neoformans var. grubii, serotype A, possessing a mating-type allele A alpha, was confirmed by crossing with standard laboratory test strains and by performing PCR with the mating-type alpha allele-specific primer of the STE12 gene and with serotype (A and D)- and mating type (a and alpha)-specific primers of the STE20 gene. To the best of our knowledge, this is the first report of disseminated cryptococcosis in an AIDS patient caused by a canavanine-resistant strain of C. neoformans var. grubii, serotype A, possessing mating type allele A alpha; the strain is probably a hybrid. The report suggests that, in the absence of a clear-cut serotyping result, a positive CGB reaction alone is not sufficient for intervarietal discrimination and additional confirmatory evidence is required.

Teratoma of the placenta.

Primary non-trophoblastic tumours of the placenta reported to date are chorioangioma and teratoma, both of which are extremely rare. A case of teratoma in a term placenta is reported.

Basidiobolus ranarum as an etiologic agent of gastrointestinal zygomycosis.

Basidiobolus ranarum is a known cause of subcutaneous zygomycosis. Recently, its etiologic role in gastrointestinal infections has been increasingly recognized. While the clinical presentation of the subcutaneous disease is quite characteristic and the disease is easy to diagnose, gastrointestinal basidiobolomycosis poses diagnostic difficulties; its clinical presentation is nonspecific, there are no identifiable risk factors, and all age groups are susceptible. The case of gastrointestinal basidiobolomycosis described in the present report occurred in a 41-year-old Indian male who had a history of repair of a left inguinal hernia 2 years earlier and who is native to the southern part of India, where the subcutaneous form of the disease is indigenous. Diagnosis is based on the isolation of B. ranarum from cultures of urine and demonstration of broad, sparsely septate hyphal elements in histopathologic sections of the colon, with characteristic eosinophilic infiltration and the Splendore-Hoeppli phenomenon. The titers of both immunoglobulin G (IgG) and IgM antibodies to locally produced antigen of the fungus were elevated. The patient failed to respond to 8 weeks of amphotericin B therapy, and the isolate was later found to be resistant to amphotericin B, itraconazole, fluconazole, and flucytosine but susceptible to ketoconazole and miconazole. One other noteworthy feature of the fungus was that the patient's serum showed raised levels of Th2-type cytokines (interleukins 4 and 10) and tumor necrosis factor alpha. The present report underscores the need to consider gastrointestinal basidiobolomycosis in the differential diagnosis of inflammatory bowel diseases and suggests that, perhaps, more time should be invested in developing standardized serologic reagents that can be used as part of a less invasive means of diagnosis of the disease.

Bilateral pulmonary aspergilloma caused by an atypical isolate of Aspergillus terreus.

A case of bilateral pulmonary aspergilloma caused by an atypical isolate of Aspergillus terreus is described. The diagnosis was established by the presence of septate, dichotomously branched fungal elements in freshly collected bronchoalveolar lavage and sputum specimens and by repeated isolation of the fungus in culture. Specific precipitating antibodies against the A. terreus isolate were demonstrated in the patient's serum. The isolate was atypical as it failed to produce fruiting structures on routine mycological media, but it did so on extended incubation on potato flake agar and produced globose, relatively heavy-walled, hyaline accessory conidia (formerly termed aleurioconidia) on both vegetative and aerial mycelia. Also, it produced an intense yellow diffusing pigment in the medium. The report underscores the increasing importance of A. terreus in the etiology of pulmonary aspergillosis. It is suggested that A. terreus antigen be included in the battery of serodiagnostic reagents to facilitate the early diagnosis of infections caused by this species.

Antimicrobial susceptibility profile of soil isolates of Nocardia asteroides from Kuwait.

OBJECTIVES: To find the antimicrobial susceptibility profile of 42 soil isolates of Nocardia asteroides against 14 antimicrobial agents representing beta-lactams, aminoglycosides, ciprofloxacin, minocycline, erythromycin and third generation cephalosporins. METHODS: The antimicrobial susceptibility was determined by the disk diffusion method using Mueller-Hinton agar medium. A homogeneous suspension giving an inoculum of 106-108 CFU/mL was used to streak the plates. The zone of inhibition was read after 36-48 h of incubation at 37 degrees C. RESULTS: All the soil isolates of N. asteroides were susceptible to amikacin, imipenem and tobramycin. Susceptibility to cephalosporins was quite variable; 86% of the isolates were susceptible to cefotaxime, 57% to ceftriaxone and 40% to cefamandole. Fifty-seven per cent of the isolates showed intermediate susceptibility to cefamandole, 33% to ceftriaxone and 5% to cefotaxime. Ninety-three per cent of the isolates were resistant to sulfamethoxazole alone or in combination with trimethoprim. CONCLUSIONS: The study reports a wide variation in the antimicrobial susceptibility profile of soil isolates of N. asteroides originating from a single geographical area. Of interest is the finding that over 90% of N. asteroides isolates were resistant to sulfamethoxazole without any previous exposure to this drug. This may have serious therapeutic implications as sulphonamides or the combination of trimethoprim-sulfamethoxazole is the therapy of choice for nocardiosis. Demonstration of resistance to beta-lactam antibiotics may be attributed to the presence of beta-lactamases which was detectable in > 90% of the soil strains of N. asteroides. The study underscores the importance of antimicrobial susceptibility testing for clinical isolates of Nocardia since individual strains show considerable differences in their susceptibility patterns necessitating therapeutic adjustments.

Aspergillus and other moulds in the air of Kuwait.

A one-year survey was carried out to study the aerial prevalence of Aspergillus species and other moulds in the outdoor and indoor environments of Kuwait. Petri plates containing rose-Bengal medium were exposed for 20 minutes twice a month using a six-stage Andersen air sampler at the pre-determined sites. The exposed plates were incubated at 28 degrees C +/- 1 degree C up to 5 days and colonies were enumerated and identified by colonial and microscopic morphology. The data revealed that Aspergillus species were the predominant component (27.7%) of the outdoor aerospora of Kuwait and A. fumigatus alone accounted for 21.3% of the total aspergilli. In contrast, Cladosporium species formed the major component of the indoor aerospora (22.8%), followed by Aspergillus species (20.9%), Penicillium species (14.6%), and Bipolaris species (10.6%). A comparison of the fungi recorded in the outdoor and in the indoor air revealed that Aspergillus, Alternaria and Fusarium were significantly higher in the outdoor environment, whereas Cladosporium, Penicillium, and Bipolaris were significantly higher in the indoor environment. The relative prevalence of Aspergillus species and other moulds in the outdoor and indoor air of Kuwait was as follows: A. fumigatus 5.9 and 9.8%, A. flavus 4.9 and 3.9%, other aspergilli 16.8 and 7.0%, Alternaria species 19.8 and 7.9%, Cladosporium species 13.7 and 22.8%, Penicillium species 7.6 and 14.6%, and other moulds 31.2 and 34.1%, respectively. During the study, 25 different genera were identified, indicating a wide diversity in the spectrum of local fungal aerospora. The study provides useful information on the prevalence of allergenic fungi in the outdoor and indoor environments of Kuwait.

A study of the enzymatic profile of soil isolates of Nocardia asteroides.

In this study, using the API-ZYM system, we have reported the enzyme profile of 42 soil strains and 2 clinical strains of Nocardia asteroides isolated locally. Of the 19 enzymes tested, only 7 were demonstrable in over 90% of the soil isolates. These included alkaline phosphatase, esterase lipase, leucine arylamidase, acid phosphatase, phosphohydrolase, alpha-glucosidase and beta-glucosidase. In addition, beta-galactosidase activity was demonstrated in all the strains by the O-nitrophenyl-beta-D-galactopyranoside (ONPG) test. The enzymes which were not demonstrable in > 95% of the strains included valine arylamidase, cystine arylamidase, trypsin, chymotrypsin, alpha-galactosidase, beta-glucoronidase, N-acetyl-beta-glucosaminidase, alpha-mannosidase and alpha-fucosidase. With the exception of valine arylamidase, which was lacking in all but one isolate, the enzyme profiles of the soil isolates were comparable with the clinical isolates of N. asteroides reported in previous studies. The reasons for this difference in the two sets of isolates is not clear. The study reinforces the view that specific differences in the enzymatic profiles of Nocardia species could be used for their rapid identification. However, more extensive studies are needed to establish the reproducibility of this method. To the best of our knowledge, this is the first study of the enzymatic profile of soil isolates of N. asteroides originating from a single geographic region.

Nocardia asteroides in the soil of Kuwait.

A pilot study was undertaken to determine the occurrence and distribution of pathogenic nocardiae in Kuwaiti soil. A total of 102 soil samples collected from two localities were investigated by the paraffin bait technique. Nocardia asteroides was the only species isolated from 42 (41%) soil samples. None of the isolates fulfilled the criteria required for identification of N. farcinica or N. nova. Thirty one (73.8%) isolates showed equivalent growth at 45 degrees C and 35 degrees C, 17 (40.4%) isolates utilized acetamide for carbon and nitrogen requirements and 3 (7.1%) isolates showed delayed arylsulphatase activity. Only a solitary isolate was resistant to cefamandole. Soil samples originating from the Kuwait University Campus, Shuwaikh, which were rich in humus/organic matter, were more productive for N. asteroides (67%) than the samples which were devoid of it but were mixed with crude oil (39%). Sand samples that lacked organic matter and crude oil samples were least productive of N. asteroides. These preliminary findings do not suggest that massive oil contamination of soil in the Ahmadi oil field area during the Gulf war promoted the natural occurrence of N. asteroides. However, isolation of N. asteroides in as many as 41% of the soil sample is a significant observation warranting further epidemiologic studies including its possible role in the operation desert storm sickness syndrome. This is the first report on the natural occurrence of N. asteroides in Kuwait.

Cell-constituent polyamines in Candida species and new biotyping of Candida albicans, Candida tropicalis and Candida parapsilosis strains.

Intracellular amines from three Candida species were extracted and chemically derivatized by a modification of the Seiler procedure. The results of qualitative determinations of 13 amines in 20 strains each of Candida albicans, Candida tropicalis and Candida parapsilosis are presented. This report is the first to describe the detection of amines other than the classical spermine, spermidine, putrescine and cadaverine in yeasts and fungi. Characteristic profiles due to the presence or absence of particular amines in the Candida species studied are demonstrated. Although these could not be used as strict differential markers at the species level, biotyping schemes based on amines are proposed to differentiate strains of C. albicans, C. tropicalis and C. parapsilosis.

Growth of fresh human isolates of some Candida species at 45 degrees C.

A total of 452 fresh isolates of six Candida species from human sources were tested for their ability to grow on Sabouraud's glucose agar at 45 degrees C. The percentage of each species which grew at this temperature was: C. albicans (99%), C. tropicalis (90%), C. glabrata (93%), C. krusei (100%), C. parapsilosis (19%) and C. lusitaniae (80%). Some variation in the amount of growth obtained after 48 h incubation was also observed. Comparison of the results with data from the literature revealed a possibility of a difference in the ability to grow at 45 degrees C between fresh human isolates and laboratory stock strains of Candida species.

Blood groups and types, hemoglobin variants, and G-6-PD deficiency among Abu Dhabians in the United Arab Emirates.

Some erythrocyte genetic factors were studied in the indigenous population of Abu Dhabi, the capital of the United Arab Emirates, on the southeastern coast of the Arabian peninsula. Determinations carried out included blood groups and types ABO, MNS, Rh0, KkJsa, FyaFyb, P1, Lea, Vela, hemoglobin variants, and screening for G-6-PD deficiency. Prevalence of most blood groups and types harmonized with that among neighboring Arabs and some Arabs elsewhere. The MS and NS gene complexes were noticeably high. African admixture was expressed by the presence of Jsa and HbS and large numbers of Fy. G-6-PD deficiency was rather high.