A six-year hospital-based surveillance study on burden of esophageal candidiasis in Gangtok, Sikkim.

Background and Objectives: Esophageal candidiasis once thought to be restricted amongst immunocompromised patients is being increasingly reported among non-immunocompromised individuals. It is debilitating and if not treated well may cause chronic long-lasting infections. The objective of this study was to identify the various species of Candida causing esophageal candidiasis and analyse their antifungal susceptibility pattern. Materials and Methods: This was an observational, prospective study. Total of 108 patients who attended the Gastroenterology Department of Sir Thutob Namgyal Memorial Hospital, Govt of Sikkim, Gangtok, India between July 2012 - May 2018 were included in the study. They had complaints of upper gastrointestinal disturbances and chronic dyspeptic symptoms that required an endoscopy. Esophageal biopsy and brushings were taken and were transported to Microbiology Department. They were subjected to microscopic observation, fungal culture on Sabourauds dextrose agar. Preliminary species identification was done by chlamydospore formation and growth characteristics on CHROMagar Candida. Species confirmation and antifungal susceptibility testing was done on VITEK 2 system at Microbiology Department, Kasturba Medical College and Hospital, MAHE, Manipal, Karnataka, India. Results: A total of 108 patients were screened among which 73 samples were positive for Candida species and species identification and antifungal susceptibility was performed. Forty fiveisolates were found to be C. albicans, 8 were C. glabrata, 4 were C. tropicalis, 3 were C. lusitaniae 2 were C. krusei, 2 were C. lipolyticaand 1 was C. parapsilosis. Eight isolates could not be identified and were recorded as Candida spp. C. albicans isolates were predominantly sensitive strain with susceptibility of 95% for both amphotericin B and fluconazole and 100% for caspofungin. C. glabrata showed high resistance to fluconazole with one isolate showing intermediate resistance to caspofungin. Conclusion: Upper gastrointestinal symptoms even in non-immunocompromised patients need to be screened by endoscopy to rule out esophageal candidiasis. With the emergence of drug resistant non albicans Candida species diagnostic testing laboratories should include Candida species identification and antifungal susceptibility testing facility to provide effective patient care.

Distal and lateral subungual onychomycosis of the finger nail in a neonate: a rare case.

BACKGROUND: Onychomycosis is extremely rare in neonates, infrequently reported in children and is considered to be exclusively a disease of adults. CASE PRESENTATION: We, herein report a case of fingernail onychomycosis in a 28-day-old, healthy, male neonate. The child presented with a history of yellowish discoloration of the fingernail of the left hand for one week. The etiological agent was demonstrated both by microscopic examination and culture of nail clippings. The isolate grown on culture was identified as Candida albicans by phenotypic characteristics and by matrix-assisted laser desorption ionization-time of flight mass spectrometry. Antifungal sensitivity testing was performed by broth dilution method as per the Clinical & Laboratory Standards Institute guidelines. An oral swab culture of the child also yielded C. albicans with the same antibiogram as the nail isolate. The case was diagnosed as distal and lateral subungual candida onychomycosis of severity index score 22 (severe) and was treated with syrup fluconazole 6 mg/kg body weight/week and 5% amorolfine nail lacquer once/week for three months. After three months of therapy, the patient completely recovered with the development of a healthy nail plate. CONCLUSIONS: The case is presented due to its rarity in neonates which, we suppose is the first case report of onychomycosis from Nepal in a 28-day-old neonate. Oral colonization with pathogenic yeasts and finger suckling could be risk factors for neonatal onychomycosis.

Corneal ulcer due to a rare coelomycetes fungus Chaetomium strumarium: Case report and global review of Chaetomium keratomycosis.

We present a rare case of corneal ulcer caused by a species of a coelomycetes fungus, Chaetomium strumarium. This fungal genus is a rare causative agent of keratomycosis, with only a handful of cases reported. The clinical presentation, investigative techniques, and preliminary management of our patient are reported. The cases reported in global literature are also summarized in a tabular form in the discussion.

Culture characterization of the skin microbiome in Type 2 diabetes mellitus: A focus on the role of innate immunity.

AIMS: The current study aimed at determining the differences between the cutaneous microbial flora of patients with Type 2 diabetes mellitus (T2DM) and those without, and thus evaluate for cutaneous microbiome dysbiosis in diabetes. METHODS: We employed a case-control study design with 41 participants in each group. The skin over the toe-web space was swabbed and cultured aerobically. Data was analyzed for differences in microbial prevalences and growths between the two groups. Predictors for heavy colonization by microbes were analysed using logistic regression. RESULTS: We found significantly higher prevalences of Staphylococcus epidermidis among patients with T2DM (77.5% vs. 53.7%, p=0.02). Further, when prevalent, these bacteria showed a significantly greater degree of skin colonization i.e. CFUs/cm2 among these patients, p=0.03. Highly pathogenic bacteria such as S. aureus were more prevalent among patients with T2DM. The regression model determined a significant association between T2DM status and heavy colonization by S. epidermidis (OR - 5.40, p=0.02). Also, agricultural workers were significantly more likely to have heavy colonization by S. epidermidis (OR - 3.75, p=0.02). Other predictor variables did not show significant association with heavy colonization by any of the isolated microbes. CONCLUSIONS: Our findings support the existence of cutaneous microbiome dysbiosis among patients with T2DM. Literature suggests that microbiome dybiosis in T2DM could stem from the same activated innate immune response that is thought to be central to the development of T2DM. This dysbiosis could increase the risk of developing skin infections.

Auxanographic Carbohydrate Assimilation Method for Large Scale Yeast Identification.

INTRODUCTION: The auxanographic carbohydrate assimilation had been an important method for differentiation of yeasts. Prevailing methods described in the literature for carbohydrate assimilation has limited scope for use in large scale yeast identification. AIM: To optimize the large scale auxanographic carbohydrate assimilation method for yeast identification. MATERIALS AND METHODS: A modified auxanographic carbohydrate assimilation method was developed and a total of 35 isolates of Candida species comprising of four ATCC (American Type Culture Collection) Candida strains (Candida albicans ATCC 90028, Candida tropicalis ATCC 90018, Candida parapsilosis ATCC 750, Candida krusei ATCC 6258) and 31 clinical isolates of Candida tropicalis (n=13), Candida krusei (n=7), Candida glabrata (n=3), Candida kefyr (n=3), Candida albicans (n=5) were validated. The carbohydrates tested were Glucose, Sucrose, Maltose, Lactose, Cellubiose, Raffinose, Trehalose, Xylose, Galactose and Dulcitol. RESULTS: A total of 35 Candida species were tested for their carbohydrate assimilative property and the results were consistent with the existing standard protocols. A well circumscribed opaque yeast growth indicated assimilation of the test carbohydrate and translucent to opalescent growth with the outline of initial inoculum alone indicated lack of assimilation. The control plate indicated no growth of the Candida species. CONCLUSION: The carbohydrate assimilation tests finds utility for yeast diversity studies exploring novel ecological niches. The technique described here facilitates testing of an extended range of carbohydrates and yeasts in a cost effective manner.

Diversity, in-vitro virulence traits and antifungal susceptibility pattern of gastrointestinal yeast flora of healthy poultry, Gallus gallus domesticus.

BACKGROUND: Poultry farming and consumption of poultry (Gallus gallus domesticus) meat and eggs are common gastronomical practices worldwide. Till now, a detailed understanding about the gut colonisation of Gallus gallus domesticus by yeasts and their virulence properties and drug resistance patterns in available literature remain sparse. This study was undertaken to explore this prevalent issue. RESULTS: A total of 103 specimens of fresh droppings of broiler chickens (commercial G domesticus) and domesticated chickens (domesticated G domesticus) were collected from the breeding sites. The isolates comprised of 29 (33%) Debaryozyma hansenii (Candida famata), 12 (13.6%) Sporothrix catenata (C. ciferrii), 10 (11.4%) C. albicans, 8 (9.1%) Diutnia catenulata (C. catenulate), 6 (6.8%) C. tropicalis, 3 (3.4%) Candida acidothermophilum (C. krusei), 2 (2.3%) C. pintolopesii, 1 (1.1%) C. parapsilosis, 9 (10.2%) Trichosporon spp. (T. moniliiforme, T. asahii), 4 (4.5%) Geotrichum candidum, 3 (3.4%) Cryptococcus macerans and 1 (1%) Cystobasidium minuta (Rhodotorula minuta). Virulence factors, measured among different yeast species, showed wide variability. Biofilm cells exhibited higher Minimum Inhibitory Concentration (MIC) values (µg/ml) than planktonic cells against all antifungal compounds tested: (fluconazole, 8-512 vs 0.031-16; amphotericin B, 0.5-64 vs 0.031-16; voriconazole 0.062-16 vs 0.062-8; caspofungin, 0.062-4 vs 0.031-1). CONCLUSIONS: The present work extends the current understanding of in vitro virulence factors and antifungal susceptibility pattern of gastrointestinal yeast flora of G domesticus. More studies with advanced techniques are needed to quantify the risk of spread of these potential pathogens to environment and human.

Online Databases for Taxonomy and Identification of Pathogenic Fungi and Proposal for a Cloud-Based Dynamic Data Network Platform.

The increase in public online databases dedicated to fungal identification is noteworthy. This can be attributed to improved access to molecular approaches to characterize fungi, as well as to delineate species within specific fungal groups in the last 2 decades, leading to an ever-increasing complexity of taxonomic assortments and nomenclatural reassignments. Thus, well-curated fungal databases with substantial accurate sequence data play a pivotal role for further research and diagnostics in the field of mycology. This minireview aims to provide an overview of currently available online databases for the taxonomy and identification of human and animal-pathogenic fungi and calls for the establishment of a cloud-based dynamic data network platform.

A modified micro chamber agar spot slide culture technique for microscopic examination of filamentous fungi.

The slide culture technique aids in the study of undisturbed microscopic morphological details of filamentous fungi. The existing methods for setting up of slide culture are quite cumbersome, time-consuming and require elaborate preparation. We describe a modified and easy to perform micro chamber agar spot slide culture technique.

A simplified touch tape preparation from tube cultures for microscopic examination of filamentous fungi.

Cellophane touch tape preparation provides reproducible results in minimal time when compared to tease mount and slide culture techniques for the identification of fungi from culture plates, but it is difficult to perform from tube cultures. Here, we describe an easy to perform touch tape preparation method that provided a better result from fungal tube culture.

In vitro scanning electron microscopic study on the effect of doxycycline and vancomycin on enterococcal induced biofilm.

INTRODUCTION: Enterococcus (E) faecalis bacteria adhere to dentine of teeth root canals to form the biofilm. E. faecalis has been shown to be resistant to antibiotics. This in vitro study aimed to investigate the efficacy of vancomycin and doxycycline in inhibiting E. faecalis biofilm formation. MATERIALS AND METHODS: A total of 60 extracted human teeth were incubated with E. faecalis (ATCC 35550 strain) for 45 days to allow biofilm formation. The teeth were equally divided into six groups (n=10): 1) positive control, 2) negative control, 3) doxycycline alone 4) doxycycline with filing, 5) vancomycin alone, 6) vancomycin with filing. The relevant canals were irrigated with 4µg/mL of either vancomycin or doxycycline antibiotic. Teeth were processed for scanning electron microscopy (SEM). Areas of biofilm remaining in the canals after antibiotic treatment were measured with Scion image analysis software using the SEM images. RESULTS: Vancomycin is more effective in reducing the overall biofilm area compared with doxycycline; moreover filing after antibiotic administration increased this effect. CONCLUSION: We can conclude that vancomycin had greater efficacy than doxycycline for inhibiting and reducing E. faecalis biofilms growth in root canals. However, it failed to completely eliminate biofilm formation.

Neuropsychiatric manifestation of confusional psychosis due to Cryptococcus neoformans var. grubii in an apparently immunocompetent host: a case report.

Cognitive disorders like dementia, delirium, depression, anxiety, psychosis and mania are the commonest neuropsychiatric manifestations. We discuss here a case of an adult women presenting with neuropsychiatric manifestations of confusional psychosis owing to Cryptococcosis. The principal cause was consequently established by culturing Cryptococcus neoformans from the cerebrospinal fluid confirmed as C. neoformans var. grubii (Serotype A) by genotypic methods. Antifungal therapy with IV Amphotericin B lead to sustained improvement and recovery of the patient from behavioural disorders. The case discussed here invokes the need for the vigilance it demands in delineating organic brain syndromes for a favourable treatment outcome.