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Background and Purpose: Onychomycosis due to dematiaceous fungi is rarely reported and the identification is also quite tricky due to poor sporulation.The recent emergence of dematiaceous fungi as a cause major of onychomycosis is a matter of concern in the field of mycology. Therefore, this study aimed to understand the dematiaceous fungi as a possible cause of onychomycosis, especially among agricultural workers. In addition, the evaluation of the antifungal susceptibility patterns gave an idea of the accurate drug that will help in the treatment challenge and prevent antifungal resistance. Materials and Methods: The standard procedure was followed for direct microscopic examination and isolating fungi. Furthermore, antifungal susceptibility testing (AFST) was conducted in accordance with the Clinical and Laboratory Standards Institute (CLSI) M-38-A2 protocol.Results: Both KOH and fungal positivity were recovered in 275 workers out of 356 suspected cases, of which 52% were NDMs, 4.3% were yeast, 28.7% were dermatophytes and 14.9% were sterile hyphae. Among NDMs (52%, n=143), 45.5% (n=65) were hyaline hyphomycetes and 54.5% (n=78) were dematiaceous hyphomycetes. Among dematiaceous fungi, Pestalotiopsis spp., Arthrinium spp., etc. were commonly isolated. Additionally, azoles, amphotericin-B and anidulafungin showed excellent antifungal activity against tested isolates. Conclusion: Dematiaceous fungi are now becoming a potential cause of onychomycosis. A more detailed study is needed on the identification of these emerging isolates and the mode of action of antifungal drugs for a better treatment strategy.

Fungi infections are becoming more prevalent and burdensome on a global scale leading to an important concern for immunocompromised patients. Hospitals often become infected with serious, invasive Candida infections. Higher frequency of Non-albicans Candida (NAC) species are found in the hospital setting, and some of these fungi can become opportunistic. Pathogens after a change in the host environment trigger them to move from a commensal to a pathogenic phase. Various clinical symptoms of Candida species, which are common human commensals, range from mucocutaneous overgrowth to bloodstream infections. In many hospitals, phenotypic methods are still considered the gold standard method for identification. Among the 112 isolates, Candida albicans (n=47; 52.64%) was noted as a significant etiology isolated from clinical samples. Further, C. albicans accounted the principal etiology in urine (n=28; 31.36%), and vaginal swab (n=13; 14.56%), followed by C. tropicalis (urine: n=15; 16.8% and vaginal swab: n=5; 5.6%). In blood C. pelliculosa (n=14; 15.68%) was found to be predominant followed by C. tropicalis (n=11; 12.32%). Antifungal susceptibility pattern was performed for (n=51) samples by VITEK AST and 100% susceptibility (voriconazole, and micafungin) was recorded in C. tropicalis and C. albicans. Whereas, fluconazole resistance was observed in C. tropicalis (n=3; 15%), and C. pelliculosa (n=1; 11.11%) and amphotericin B resistance in C. tropicalis (n=1; 5%) and C. albicans (n=1; 9.1%).

IntroductionThe emergence of the Omicron SARS-CoV-2 variant from various states of India in early 2022 has caused fear of its rapid spread. The lack of such reports from Chhattisgarh (CG), a central state in India, has prompted us to identify the Omicron circulating lineages and their mutational dynamics.Materials and methodsWhole-genome sequencing (WGS) of SARS-CoV-2 was performed in 108 SARS-CoV-2 positive combined samples of nasopharyngeal and oropharyngeal swabs obtained from an equal number of patients.ResultsAll 108 SARS-CoV-2 sequences belonged to Omicron of clade 21L (84%), 22B (11%), and 22D (5%). BA.2 and its sub-lineages were predominantly found in 93.5% of patients, BA.5.2 and its sub-lineage BA.5.2.1 in 4.6% of patients, and B.1.1.529 in 2% of patients. Various BA.2 sub-lineages identified were BA.2 (38%), BA.2.38 (32%), BA.2.75 (9.25%), BA.2.56, BA.2.76, and BA.5.2.1 (5% each), BA.2.74 (4.6%), BA.5.2.1 (3.7%), BA.2.43 and B.1.1.529 (1.8% each), and BA.5.2 (0.9%). Maximum mutations were noticed in the spike (46), followed by the nucleocapsid (5), membrane (3), and envelope (2) genes. Mutations detected in the spike gene of different Omicron variants were BA.1.1.529 (32), BA.2 (44), BA.2.38 (37), BA.2.43 (38), BA.2.56 (30), BA.2.74 (31), BA.2.75 (37), BA.2.76 (32), BA.5.2, and BA.5.2.1 (38 similar mutations). The spike gene showed the signature mutations of T19I and V213G in the N-terminal domain (NTD), S373P, S375F, T376A, and D405N in receptor-binding domain (RBD), D614G, H655Y, N679K, and P681H at the furin cleavage site, N764K and D796K in fusion peptide, and Q954H and N969K in heptapeptide repeat sequence (HR)1. Notably, BA.2.43 exhibited a novel mutation of E1202Q in the C terminal. Other sites included ORF1a harboring 13 mutations followed by ORF1b (6), ORF3a (2), and ORF6 and ORF8 (1 mutation each).ConclusionBA.2 followed by BA.2.38 was the predominant Omicron lineage circulating in Chhattisgarh. BA.2.75 could supersede other Omicron due to its mutational consortium advantage. The periodical genomic monitoring of Omicron variants is thus required for real-time assessment of circulating strains and their mutational-induced severity.

Taxonomic delineation of etiologic agents responsible for recalcitrant dermatophytosis causing an epidemic in India is still debated. The organism responsible for this epidemic is designated as T. indotineae, a clonal offshoot of T. mentagrophytes. To evaluate the real identity of the agent causing this epidemic, we performed a multigene sequence analysis of Trichophyton species isolated from human and animal origin. We included Trichophyton species isolated from 213 human and six animal hosts. Internal transcribed spacer (ITS) (n = 219), translational elongation factors (TEF 1-α) (n = 40), ß-tubulin (BT) (n = 40), large ribosomal subunit (LSU) (n = 34), calmodulin (CAL) (n = 29), high mobility group (HMG) transcription factor gene (n = 17) and α-box gene (n = 17) were sequenced. Our sequences were compared with Trichophyton mentagrophytes species complex sequences in the NCBI database. Except for one isolate (ITS genotype III) from animal origin, all the tested genes grouped our isolates and belonged to the “Indian ITS genotype”, currently labeled as T. indotineae. ITS and TEF 1-α were more congruent compared to other genes. In this study, for the first time, we isolated the T mentagrophytes ITS Type VIII from animal origin, suggesting the role of zoonotic transmission in the ongoing epidemic. Isolation of T. mentagrophytes type III only from animal indicates its niche among animals. Outdated/inaccurate naming for these dermatophytes in the public database has created confusion in using appropriate species designation.

This study was designed to understand the molecular mechanisms of azole resistance in Candida tropicalis using genetic and bioinformatics approaches. Thirty-two azole-resistant and 10 azole-susceptible (S) clinical isolates of C. tropicalis were subjected to mutation analysis of the azole target genes including ERG11. Inducible expression analysis of 17 other genes potentially associated with azole resistance was also evaluated. Homology modeling and molecular docking analysis were performed to study the effect of amino acid alterations in mediating azole resistance. Of the 32 resistant isolates, 12 (37.5%) showed A395T and C461T mutations in the ERG11 gene. The mean overexpression of CDR1, CDR3, TAC1, ERG1, ERG2, ERG3, ERG11, UPC2, and MKC1 in resistant isolates without mutation (R-WTM) was significantly higher (p

Increasing reports of azole resistance in Candida tropicalis, highlight the development of rapid resistance detection techniques. Nonsynonymous mutations in the lanosterol C14 alpha-demethylase (ERG11) gene is one of the predominant mechanisms of azole resistance in C. tropicalis. We evaluated the tetra primer-amplification refractory mutation system-PCR (T-ARMS-PCR), restriction site mutation (RSM), and high-resolution melt (HRM) analysis methods for rapid resistance detection based on ERG11 polymorphism in C. tropicalis. Twelve azole-resistant and 19 susceptible isolates of C. tropicalis were included. DNA sequencing of the isolates was performed to check the ERG11 polymorphism status among resistant and susceptible isolates. Three approaches T-ARMS-PCR, RSM, and HRM were evaluated and validated for the rapid detection of ERG11 mutation. The fluconazole MICs for the 12 resistant and 19 susceptible isolates were 32-256 mg/L and 0.5-1 mg/L, respectively. The resistant isolates showed A339T and C461T mutations in the ERG11 gene. The T-ARMS-PCR and RSM approaches discriminated all the resistant and susceptible isolates, whereas HRM analysis differentiated all except one susceptible isolate. The sensitivity, specificity, analytical sensitivity, time, and cost of analysis suggests that these three methods can be utilized for the rapid detection of ERG11 mutations in C. tropicalis. Additionally, an excellent concordance with DNA sequencing was noted for all three methods. The rapid, sensitive, and inexpensive T-ARMS-PCR, RSM, and HRM approaches are suitable for the detection of azole resistance based on ERG11 polymorphism in C. tropicalis and can be implemented in clinical setups for batter patient management.

Dermatophytosis, caused by dermatophytes is becoming difficult to treat due to various reasons. Accurate diagnosis is essential for the accurate management of this infection and prevention of relapse or recurrence. Although this condition is easy to diagnose clinically, due to overlapping signs and symptoms of few dermatological conditions it may be misdiagnosed necessitating laboratory confirmation. Isolation, identification of the dermatophytes and the antifungal susceptible profile may further help to initiate appropriate antifungal agent. The classical conventional techniques such as direct microscopic examination and isolation of fungi from the clinical specimens are still considered as an important modality of diagnosis. With the rise of the molecular era, molecular techniques are increasingly being applied to diagnose dermatophytosis and identify the dermatophytes. The present review provides an overview and update on the laboratory diagnosis of dermatophytosis.

We report invasive candidiasis caused by Candida viswanathii over 2 time periods during 2013–2015 in a tertiary care hospital in Chandigarh, India. Molecular typing revealed multiple clusters of the isolates. We detected high MICs for fluconazole in the second time period.

Introduction: Haemophilus influenzae is responsible for wide range of localized and invasive lower respiratory tract infections (LRTI) with the highest burden of disease in low and middle income countries. Aim: The aim of the present study was to characterize the H.influenzae isolates from suspected LRTI. Materials and Methods: A prospective study was conducted over a period of one and half years (December 2012 to May 2014) including patients with LRTI. H.influenzae was isolated from lower respiratory specimens following standard procedures. Complete characterization of the isolates was performed by bio typing, capsular serotyping, molecular genotyping and antibiotic susceptibility testing. The predisposing factors and clinical presentation were studied in the infected patients. Results: A total of 8995 samples were received during the study period, out of which growth was significantly observed in 2848 (31.7%) samples. Among the various respiratory pathogens, H.influenzae was isolated from 175 (6.14%) patients. Majority (78.9%) of the patients presented with acute exacerbations of chronic obstructive pulmonary disease. The isolates most frequently were of Biotype II (35.42%). Only four of the 50 isolates subjected to capsular serotyping were typeable and were of type b, e and f. All the 50 isolates tested were found to be non-typeable by PCR for capsular genotyping. Maximum resistance was found against ampicillin (9.71%). Conclusion: H.influenzae was found to be a significant cause of LRTI. Majority of the isolates were found to be non typeable strains. Non typeable H. influenzae isolates should not be neglected as they can colonize the respiratory tract in COPD patients and can lead to biofilm formation and treatment failure.