Antifungal susceptibility of molecularly confirmed Aspergillus species from clinical samples.

Invasive aspergillosis is the second most common invasive human mycosis but susceptibility data of Aspergillus species is limited. Antifungal treatment of aspergillosis is often done empirically without knowing the true susceptibility. Therefore, we aimed to determine antifungal susceptibility of Aspergillus species isolated from various clinical specimens over a 1-year period. We identified 28 Aspergillus isolates by sequencing the internal transcribed spacer (ITS) and ß-tubulin genes and performed antifungal susceptibility testing on these isolates using Sensititre YeastOne. The isolates were identified as Aspergillus niger (60.7%), A. fumigatus (21.4%), A. flavus (10.7%), A. chevalieri (3.6%) and A. tubingensis (3.6%). Based on the European Committee on Antimicrobial Susceptibility Testing (EUCAST) Antifungal Clinical Breakpoint for Aspergillus spp., 16/17 (94.1%) A. niger isolates were susceptible to amphotericin B, all six isolates (100%) of A. fumigatus were susceptible to amphotericin B, itraconazole and voriconazole, but only 5/6 (83.3%) A. fumigatus were susceptible to posaconazole. Meanwhile, all three (100%) A. flavus isolates were susceptible to itraconazole. There are no other breakpoints established by the EUCAST for other antifungal-species combinations. In conclusions, Aspergillus niger remains the most commonly isolated species from clinical specimens and Aspergillus isolates at our centre are still largely susceptible to amphotericin B, echinocandins and most azoles. This information is valuable in guiding antifungal therapy in the treatment of aspergillosis.

Antifungal susceptibility profile and biofilm-producing capability of Candida tropicalis isolates in a tertiary medical centre.

BACKGROUND: Candida tropicalis is a globally distributed yeast that has been popping up in the medical literature lately, albeit for unenviable reasons. C. tropicalis is associated with substantial morbidity, mortality as well as drug resistance. The aims of this study were to ascertain the antifungal susceptibility profile and the biofilm-producing capability of this notorious yeast in our centre. METHODS: C. tropicalis isolates from sterile specimens were collected over a 12-month period. Conclusive identification was achieved biochemically with the ID 32 C kit. Susceptibility to nine antifungal agents was carried out using the colourimetric broth microdilution kit Sensititre YeastOne YO10. Biofilm-producing capability was evaluated by quantifying biomass formation spectrophotometrically following staining with crystal violet. RESULTS: Twenty-four non-repetitive isolates of C. tropicalis were collected. The resistance rates to the triazole agents were 29.2% for fluconazole, 16.7% for itraconazole, 20.8% for voriconazole and 8.3% for posaconazole-the pan-azole resistance rate was identical to that of posaconazole. No resistance was recorded for amphotericin B, flucysosine or any of the echinocandins tested. A total of 16/24 (66.7%) isolates were categorized as high biomass producers and 8/24 (33.3%) were moderate biomass producers. None of our isolates were low biomass producers. CONCLUSION: The C. tropicalis isolates from our centre were resistant only to triazole agents, with the highest resistance rate being recorded for fluconazole and the lowest for posaconazole. While this is not by itself alarming, the fact that our isolates were prolific biofilm producers means that even azole-susceptible isolates can be paradoxically refractory to antifungal therapy.

An unsuspected and unusual case of zoonotic sporotrichosis.

Sporotrichosis is an infection caused by a thermally dimorphic fungus, Sporothrix schenckii which is commonly reported in the tropics and subtropics. Infection usually occurs as a consequence of traumatic inoculation into the skin. A previously healthy 33-year-old Malay woman presented with a scratch wound over the lateral aspect of her left thigh for 2 months after being scratched by her pet cat. Inspection of the wound revealed a granulomatous plaque measuring 3cm x 2cm with hyperkeratotic scales, fissures and multiple blackish pigments. There was a tender, palpable left inguinal lymph node measuring 1cm x 1cm but the overlying skin looked normal. She was prescribed two courses of antibiotics empirically, without any clinical improvement seen. During follow up and further history taking, the patient informed that her pet cat was diagnosed with sporotrichosis a month before the scratching incident took place, but she claimed that her pet cat had been treated and was cured. She was then empirically started with oral itraconazole 200mg twice a day for two months. Histopathological examination of a skin biopsy showed acute on chronic suppurative granulomatous inflammation but was negative for any fungal elements. However, culture of the skin biopsy isolated Sporothrix schenckii. A final diagnosis of localised cutaneous sporotrichosis was made and the oral itraconazole was continued for another two months. The wound improved dramatically since itraconazole started.

Campylobacter jejuni bacteremia in a leukemic child: a nearly missed diagnosis.

Globally, Campylobacter spp. are responsible for most cases of bacterial gastrointestinal infections in humans and although rare, extraintestinal Campylobacter infections have been described. A 2-yearold neutropenic girl with underlying precursor B-cell acute lymphoblastic leukemia presented with a 3-day history of diarrhea. Her stool culture yielded no enteric bacterial pathogens. However, when her blood culture was flagged as positive for bacterial growth, no colonies could be observed on routine bacteriological isolation media. Nonetheless, gram-negative bacilli with seagull and spiral morphologies were seen when the surface of the isolation media used to subculture her blood was Gram-stained. Bacterial colonies were only visible when a subculture was attempted on a Campylobacter blood-free selective agar medium. The organism was identified as Campylobacter jejuni by matrix-assisted laser desorption ionization-time of flight mass spectrometry. Since the organism was erythromycin-resistant and the patient's age precluded the use of tetracycline and ciprofloxacin, an antibiotic regimen consisting of piperacillin-tazobactam and gentamicin was commenced. Her C. jejuni bacteremia resolved following eight days of antibiotic therapy.

Prevalence of toenail onychomycosis among diabetics at a primary care facility in Malaysia.

Onychomycosis increases the risk of developing secondary bacterial infection and cellulitis if left untreated. The aim of this study was to determine the prevalence of onychomycosis among diabetics and its associated factors. A cross sectional study using universal sampling of all type 1 and 2 diabetic patients attending a primary care facility of the Universiti Kebangsaan Malaysia (UKM) from January to March 2011 was conducted. Samples were taken from clinically abnormal nails and from the first right toenail in the absence of nail abnormalities and cultured for fungal elements. A total of 151 diabetics participated in the study. The mean patient age was 60.7 +/- 9.1 years. A total of 123 nail samples (81.5%) were culture positive for fungal elements. A positive correlation was found between onychomycosis and increasing age (p = 0.011) and clinically abnormal nails (p < 0.05). There were no significant correlations with gender, ethnic group, duration of diabetes, types of diabetes or glycemic control. The prevalence of onychomycosis among diabetics in our study was high.

Ochrobactrum anthropi catheter-related bloodstream infection: the first case report in malaysia.

A 60-year old Malay man presented with an intermittent lowgrade fever for 3 days and worsening of right ankle ulcers. He was known to have ischaemic heart disease, diabetes mellitus type 2, hypertension and end stage renal failure and was on regular haemodialysis. The organisms isolated from the peripheral and central blood specimens were identified as Ochrobactrum anthropi, a rarely encountered gramnegative bacillus with a unique antibiotic susceptibility profile. His condition improved after removal of the catheters and administration of antibiotic therapy. This case report highlights the importance of being wary of such rare, opportunistic pathogens and good communication between the microbiologists and the clinicians.

A case of mycotic aneurysm due to Burkholderia pseudomallei.

Burkholderia pseudomallei is an free-living gram-negative bacterium causing melioidosis and is endemic in Southeast Asia. A 56-year-old diabetic construction worker with a 1-month history of abdominal pain and 1-day history of high-grade fever was found to have a left non-dissecting infrarenal mycotic aortic aneurysm by abdominal computerized tomography scan. Bacteriological examination of his blood yielded Burkholderia pseudomallei. The patient was treated with right axillo-bifemoral bypass with excision of aneurysm and high-dose intravenous ceftazidime for two weeks, followed by oral trimethoprim/sulfamethoxazole and oral doxycycline for a minimum of five months.

Bullous cellulitis as an extraordinary manifestation of a Vibrio cholerae O1 Ogawa infection.

Vibrio cholerae is a gram-negative bacterium synonymous with its namesake disease, cholera. Thus, gastrointestinal symptoms are the norm and V. cholerae is very rarely associated with skin and soft tissue infections. We describe a case of a 63-year-old Chinese woman with multiple medical comorbidities on corticosteroid therapy who developed fever and a painful swelling on her left leg after being pricked by a branch while gardening. There was no abdominal pain, vomiting or diarrhea. A diagnosis of bullous cellulitis was made clinically, and blood was sent for bacteriological culture. A beta-hemolytic commashaped gram-negative bacillus was isolated from the blood. It was also oxidase-positive and produced an acid/alkaline (A/K) reaction on triple sugar iron agar. It was identified biochemically as Vibrio cholerae. After additional testing, it was found to be of the O1 serogroup and Ogawa serotype. The infection resolved following a 10-day course of high-dose co-trimoxazole therapy.

Antibacterial activities of the extracts, fractions and isolated compounds from Canarium patentinervium Miq. against bacterial clinical isolates.

BACKGROUND: Canarium patentinervium leaves are used by the local indigenous people of Malaysia for wound healing. The current study is undertaken to screen the comprehensive antibacterial activity of the leaves and barks extracts, fractions and isolated compounds from this plant. Bioassay guided fractionation was also undertaken to deeply evaluate the antibacterial activity of the water fraction of the leaves extract. This is to provide preliminary scientific evidence to the ethnopharmacology usage of this plant by investigating antibacterial properties of the plant and its isolated constituents. METHODS: Bio-assay guided fractionation and subsequent isolation of compounds using open column chromatography. The antibacterial activity against gram positive and gram negative ATCC strain and resistant clinical strains were evaluated using microtiter broth dilution method to determine minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and time-kill assay. The chemical structure of the isolated compounds from the water fraction of the ethanol extract of leaves was elucidated using Nuclear Magnetic Resonance (NMR). RESULTS: The ethanol extract of the leaves and barks showed antimicrobial activity against all four ATCC and eight clinical isolates. The ethanol extract of the leaves and the corresponding water fraction had good activity against MRSA S. aureus. (MIC: 250 µg/ml) and had bactericidal effect on eight of the clinical strains (MSSA,MRSA, oxacillin-resistant CONS, oxacillin-sensitive CONS, Enterococcus faecalis, Klebsiela species, Kleb pneumoniae ESBL and Candida parapsilosis). Further phytochemical investigation of the water fraction of the crude ethanol extract of leaves afforded compound 7 (hyperin) and compound 8 (cynaroside) that had bactericidal activity against tested bacterial species (MIC 50 µg/ml and 100 µg/ml). The two compounds were isolated from this genus for the first time. CONCLUSIONS: These results may provide a rational support for the traditional use of Canarium patentinervium Miq. in infections and wound healing, since the antimicrobial compounds isolated were also present in the leaves extract.

Candidaemia and antifungal susceptibility testing in a teaching hospital.

We reviewed cases of candidaemia at Universiti Kebangsaan Malaysia Medical Centre from 1st January 2005 to 30th June 2006. All blood cultures positive for Candida species or its teleomorphs within the study period were identified and antifungal susceptibility testing was performed. Out of 50 blood isolates, 20 (40%) were identified as Candida albicans, 16 (32%) C. tropicalis, five (10%) C. parapsilosis, three (6%) C. famata, two (4%) C. glabrata, two (4%) Pichia ohmeri, one (2%) C. krusei and one (2%) P. etchell/carsonii. Susceptibility to amphotericin B was 100%, fluconazole 90%, itraconazole 40%, ketoconazole 88%, 5-flucytosine 98% and voriconazole 98%.

Cryptococcal osteomyelitis of the femur: a case report and review of literature.

Fungal osteomyelitis is a rare opportunistic infection. It exhibits some clinical and radiological similarities to several other bone pathologies. A diagnostic delay may result in significant increase in morbidity. We report a case of a 37-year-old man with underlying hypogammaglobulinaemia presented with isolated cryptococcal osteomyelitis of the femur.