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Introduction: In hospitalized patients, candidemia accounts for considerable morbidity and mortality. Knowledge of local species epidemiology and antifungal susceptibility is important when considering therapy. This study was performed to speciate Candida isolates from inpatients admitted with blood stream infections in our hospital and to assess their risk factors. Materials and Methods: This present study of 2 years' duration was performed from April 2022 to March 2024. BacT/Alert aerobic culture bottle (bioMérieux, France) was used for all routine blood cultures and incubated for 5 days at 37°C. Any blood culture growing yeast was identified by Gram stain and MALDI-TOF MS (Bruker Daltonics). Disk diffusion method was used for antifungal susceptibility testing and test was performed according to clinical laboratory standard institute (CLSI) guidelines. Results: About 132 Candida isolates from blood were isolated in 2 years. Predominance of males (59%) was noted with the median age of 52 years. Majority were nonalbicans Candida (81.06 %) with C. tropicalis (31.81%) and C. auris (13.63%). Susceptibility to fluconazole and voriconazole was 100% for both C. albicans and C. tropicalis isolates. Fluconazole resistance of 11.77% and 100% was observed in C. parapsilosis and C. auris isolates respectively. Conclusion: The predominance of non-albicans Candida with increasing fluconazole resistance and isolation of C. auris as the third most common non-albicans Candida species in this study is of concern. Accurate identification of non-albicans Candida specially C. auris and strict infection control policies should be in place to combat the spread of C. auris in the hospital. [ABSTRACT FROM AUTHOR]

Background: & objective: The existence of visually identical cryptic Aspergillus species that can be distinguished only by molecular techniques is becoming more widely acknowledged. For the majority of antifungal drugs, these are known to exhibit a greater minimal inhibitory concentration in vitro. For the purpose of receiving the proper care, it is crucial to identify these species at right time. Our aim in this work is to identify and describe the Aspergillus species that are cryptic from all of the clinical samples., Methods: Routine samples from inpatients and outpatients received in department of Microbiology, All India Institute of Medical Sciences, New Delhi, showing growth of Aspergillus species were included in this study. Phenotypic and Matrix Assisted Laser Desorption Ionisation - Time of Flight identified isolates were analysed for cryptic species, by PCR and ITS/ß - tubulin sequencing. In accordance with CLSI recommendations, antifungal susceptibility testing was conducted using micro broth dilution., Results: Of the 94 isolates, 54 A. fumigatus, 34 A. flavus, 3 A. nidulans, 2 A. terreus, and 1 A. niger were morphologically identified. MALDI-TOF misidentified 2 A. nidulans isolates and 1 A, stellatus isolate. The ß - tubulin sequence analysis revealed that 2 isolates (2.08 %) were cryptic, one was A. stellatus and another one was A. tubingensis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Indian Association of Medical Microbiologists. Published by Elsevier B.V. All rights reserved.)

A resistant and hypervirulent dermatophyte from India has been described as a taxonomic novelty, Trichophyton indotineae, a species of the Trichophyton mentagrophytes complex. Rapid detection and correct identification of closely similar dermatophytes with different predilections are essential for efficient clinical management. We evaluated the efficacy of rapid diagnostic methods clinical and environmental strains in the T. mentagrophytes complex. The methods included Real-time-PCR, DermaGenius, LAMP, and MALDI-ToF MS, using rDNA ITS sequences as taxonomic standard. The results show that only MALDI-ToF MS can distinguish 96.97% T. indotineae from other closely related species. The complex comprises numerous clones which may differ in anonymous markers but with similar evolutionary behavior. Therefore, we recommend to distinguish species only when they show an appreciable degree of adaptation and thus are clinically significant. The distinction of remaining clonal diversity is an epidemiological query and can be solved by haplotype numbering. [ABSTRACT FROM AUTHOR]

Cystic neutrophilic granulomatous mastitis (CNGM) is a histologically characterized variant of granulomatous lobular mastitis that is associated with lipophilic Corynebacterium species. It remains a largely underrecognized entity in India. Our aim was to study CNGM in the Asian Indian population and explore if 16s rRNA sequencing could be used on formalin-fixed paraffin-embedded (FFPE) tissue to identify the causative organism. We studied 24 cases with histological features of CNGM with hematoxylin and eosin, Gram, Ziehl-Neelsen, and Periodic acid–Schiff stains. Tuberculosis-polymerase chain reaction and 16s rRNA gene sequencing on DNA extracted from FFPE was attempted (N = 23). Gram-positive bacilli were seen in 20/24 cases. Routine culture with prolonged incubation yielded Corynebacterium species in 8 cases; 7 of these cases were evaluated by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for species identification. C matruchotti was identified in one case by BD Phoenix. MALDI-TOF MS identified the remaining 7 cases as C kroppenstedtii (N = 4) and C tuberculostearicum (N = 2), with no identification in one. Corynebacteria were identified by 16s rRNA sequencing on DNA extracted from FFPE in 12/23 cases using a primer targeting the V5-V6 region that was found to be more conserved in Corynebacterium species. All cases were negative for the diagnosis of tuberculosis. CNGM can be identified by routine stains. Culture using routine media with prolonged incubation is often adequate to isolate the organism. 16s rRNA sequencing on DNA extracted from FFPE tissue can help make an etiological diagnosis in some cases where only paraffin blocks are available. [ABSTRACT FROM AUTHOR]

• A Bacillus thuringiensis (Bt) strain, specific to tea looper was isolated from Danguajhar Tea Garden, West Bengal. • Large scale production of Bt was screened with four different carbon sources and barley was found to produce maximum spore. • Highly expressive sporulation associated toxins were identified as Immune inhibitor A of Bt by MALDI-TOF mass spectrometry. Tea, Camellia sinensis (L.) O'Kuntz is a cash crop, cultivated in tropical and subtropical climatic countries throughout the globe. The tea ecosystem hosts several species of arthropods, of them many species attack different parts of tea plants during their growth phases resulting in huge economical loses. Buzura suppressaria (Bs), Hyposidra talaca (HT), Hyposidra infixaria (HI) are members of order Lepidoptera, which are the most notorious tea pests, found in North-East India and make huge destruction of tea plantation due to their voracious eating habit and short life span. A potent indigenous strain of Bacillus thuringiensis (Bt) against the tea looper (TL) was isolated from Danguajhar Tea Garden, Goodricke Group Ltd. India, and characterized based on morphological, biochemical and genetic traits (16S rRNA gene NCBI GenBank Accession. No. JF966358). Virulence assay, to itemize the entomopathogenic nature of Bt against TL, was determined in laboratory and field conditions to enumerate its LC50 (1.11 × 107 spores/mL) and LT50 (54.64 h) values with a TL mortality rate of 99.43% in field. For large scale production, Bt was screened with four different mediums (based on barley, wheat, rice flour and nutrient broth) and was found to produce a maximum of 4.6 × 107 spores/mL, when barley was considered as a sole source of carbon. Highly expressive sporulation associated toxins was identified as an immune inhibitor A of Bt (NCBI Accession No.gi|9858110|AAG00998.1) by MALDI-TOF mass spectrometry which is an extracellular bacterial protein known for putrefaction of host tissue proteins with a wide-range of substrate specificity. The results were quite encouraging which can be adapted as a greener way of pest control. [ABSTRACT FROM AUTHOR]

Infective endocarditis (IE) is primarily a bacterial infection of the heart valves. The most common organisms implicated include Staphylococcus and Streptococcus species. However, with the advent of MALDI-TOF and molecular techniques, the reports of IE being caused by rare organisms are on a rise. Here we describe a case of IE due to Abiotrophia defectiva. This is the first report of simultaneous infection of both mitral and aortic valves by Abiotrophia defectiva from India. IE caused by Abiotrophia defectiva has been seen to be more severe, associated with higher failure rates and relapse. This emphasizes the accurate identification of nutritionally variant Streptococcus (NVS) species as the management of choice varies between Abiotrophia and Granulicatella. [ABSTRACT FROM AUTHOR]

Background & objectives: Tuberculosis is a major health problem in India, and the emergence of multidrug resistant (MDR) and extensively drug resistant (XDR) strains of Mycobacterium tuberculosis (Mtb) has further complicated the situation. Though several studies characterizing drug sensitive and drug resistant strains are available in literature, almost all studies are done on unrelated strains. Therefore, the objective of this study was to compare the proteomic data of four sequential isolates of Mtb from a single patient who developed MDR-TB during the course of anti-tuberculosis therapy (ATT). Methods: In this study, using two-dimensional (2D) gel electrophoresis and MALDI-TOF mass spectrometry, we compared and analyzed the cell lysate proteins of Mtb sequential clinical isolates from a patient undergoing anti-TB treatment. The mRNA expression levels of selected identified proteins were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Results: The genotypes of all four isolates remained homologous, indicating no re-infection. The initial isolate (before treatment) was sensitive to all first-line drugs, but the consecutive isolates were found to be resistant to isoniazid (INH) and rifampicin (RIF) and developed mutations in the katG, inhA and rpoB. the intensities of 27 protein spots were found to be consistently overexpressed in INH and RIF resistant isolates. The most prominent and overexpressed proteins found during the development of drug resistance were GarA (Rv1827), wag31 (Rv2145c), Rv1437 and Rv2970c. Interpretation & conclusions: This preliminary proteomic study provides an insight about the proteins that are upregulated during drug resistance development. These upregulated proteins, identified here, could prove useful as immunodiagnostic and possibly drug resistant markers in future. However, more studies are required to confirm these findings. [ABSTRACT FROM AUTHOR]

Background: Gastric Cancer (GC) remains a major global health problem due to a poor understanding of its progression at the molecular level and a lack of early detection or diagnosis. Early detection is highly crucial for improving prognosis. The incidence of GC is very high in countries, like India, due to the limitations among the established biomarkers for GC owing to poor sensitivity and specificity., Objective: This study aimed to identify the novel biomarkers from serum samples obtained from GC patients compared to healthy subjects., Methods: Serum samples from GC patients were analyzed by two-Dimensional Gel Electrophoresis (2DGE) coupled with tandem Mass Spectrometry (MS), including both Matrix-Assisted Laser Desorption/Ionization-Time of Flight (MALDI-ToF) and Liquid Chromatography-MS (LC-MS/MS) analysis. Identified proteins were further analyzed by gene ontology and protein interaction studies., Results: A total of 73 protein spots were detected in 2DGE image analysis. Among them, seven differentially-expressed proteins were identified using MS analyses, including serotransferrin/ transferrin, albumin, ceruloplasmin, C-reactive protein (CRP), fibrinogen γ-chain (FGG), and two unreported novel proteins, immunoglobulin kappa constant (IgκC) region and Homo sapiens zinc finger protein 28 (ZNF28) homolog. Among these proteins, serotransferrin, albumin, ceruloplasmin, FGG, and ZNF28 were down-regulated in GC samples (p<0.05), while IgκC region and CRP were up-regulated significantly., Conclusion: Most of the differentially expressed proteins were involved in angiogenesis, plasminogen-activating cascade, and blood coagulation pathways which are known to play a critical role in gastric tumorigenesis. Our current results provide a panel of candidate biomarkers for GC with novel biomarkers which have not been reported earlier., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Objectives: A molecular analysis of serologically RhD variant samples was conducted to find the incidence of various D variants in our blood donor population., Background: Determining a blood donor's RhD phenotype and genotype is important as transfusion of units with a weak D or partial D phenotype can result in immunisation of the recipients., Methods: Samples with discrepant D and weak D phenotypes identified on testing with at least five different monoclonal anti-D antisera were considered serological RhD variant and subjected to molecular testing (Massarray kit, Agena Bioscience, San Diego) for variant RHD gene., Results: A total of 39 samples, including 19 RhD discrepant samples and 20 weak D samples, were identified as serological RhD variant from a total of 4386 samples. Thirteen (13/39) samples carried variants leading to weak D phenotype, and eight samples had variants leading to partial D categories. Seven samples (7) could not be characterised, whereas 11 samples were identified as Rh negative (RHD*01N.01) after molecular testing. Overall incidence of D variants in the study population was 0.48%. RHD*weak D type 1(5, 0.1%) and RHD*DFR1 (5, 1%) were the most common variants identified., Conclusions: Few samples with weak reaction on serological testing were found to be partial D variant and vice versa. Donor centres should develop a protocol for genotyping of samples with aberrant results on serological testing for assessing the actual RhD status of an individual as results of serological testing may be misleading., (© 2020 British Blood Transfusion Society.)

Purpose. Acute bacterial meningitis continues to be a potentially life threatening condition. Hospital-acquired meningitis is rapidly increasing and adding an immense burden to the health system due to the emergence of multidrug resistance isolates. The purpose of this study is to find the antibiotic susceptibility pattern of the bacteria detected from hospital- and community-acquired meningitis. Methodology. A total of 400 Cerebrospinal fluid (CSF) samples from the suspected meningitis cases were collected and processed for cell count, biochemical examination, Gram staining, latex agglutination and culture. Bacteria grown on blood, chocolate and Mac-conkey agar were identified by matrix-assisted laser desorption/ionization-time of flight. Antibiotic susceptibility tests were performed as per Clinical and Laboratory Standard Institute guidelines. Results. Of the isolates, most prevalent Gram negative organisms in hospital-acquired bacterial meningitis were Escherichia coli 13 (27.08 %), Acinetobacter baumannii 12 (25 %), Klebsiella pneumoniae 5 (10.42 %), Pseudomonas aeruginosa 4 (8.33 %) and Gram positive organisms were Staphylococcus aureus 4 (8.33 %), Enterococcus faecium 3 (6.25 %) and CONS 2 (4.16 %). Streptococcus pneumoniae 3 (6.25 %) was the predominant organism in community-acquired bacterial meningitis. All the Gram negative isolates were multidrug resistance. Only colistin and imipenem were effective antibiotics against them. Likewise Gram positive organisms were susceptible to most of the antibiotics tested. However, E. faecium was only susceptible to Vanco+Teicoplanin. Conclusion. In hospital-acquired bacterial meningitis, multidrug resistance Gram negative bacteria are a huge challenge for the treatment of patients. Hence, antimicrobial stewardship should be followed to counteract with the emerging multidrug resistance isolates.