Utility of galactomannan diagnostic assay in rhino-orbito-cerebral mycosis in COVID-19 patients.

An increasing number of fungal infections were reported post COVID-19 and many of them were caused by fungi of mixed aetiologies. This study was carried out to assess the utility of serum galactomannan (GM) assay in establishing the etiology of acute rhino-orbito-cerebral mycosis caused by Aspergillus spp. Two serum samples were obtained from 41 suspected post COVID-19 rhino-orbito-cerebral mycosis patients to perform GM assay. Serum GM assay was positive in 68.7% of the cases of proven aspergillosis at cut off OD = 1.0. Serum GM assay can be used as a supplementary test in the diagnosis of rhino-orbito-cerebral mycosis caused by Aspergillus spp.

Influence of Hepatitis C virus genotype and other factors on the viral load.

Anti-HCV reactive subjects were selected and relevant data was collected. Viral load and genotype were determined for all patients and were divided into low (<800,000 IU/mL) and high viral load (>800,000 IU/mL). Correlation of viral load with parameters like age, gender, risk factors and genotype etc. was determined by binomial regression. Higher viral load was noted with genotype 4, males and high risk groups like People Who Inject Drugs (PWIDs), blood transfusion before routine testing or frequent transfusion, Intravenous drug therapy and MTP by unregistered medical practitioners (P ≤ 0.5). Prevention and treatment strategies for HCV should be tailored around these areas.

Post-COVID-19 rhino-orbito-cerebral mucormycosis-A clinico-mycological study from North India.

The second wave of coronavirus disease 2019 (COVID-19), during the early 2021, lead to a devastating outbreak of mucormycosis in India. This study aimed to determine the aetiology, clinical features, comorbidities, and risk factors of rhino-orbito-cerebral mucormycosis (ROCM) and antifungal susceptibility pattern for the isolates. The study included all suspected cases of ROCM in post-COVID-19 patients attending the hospital from May to December 2021. A total of 70 patients were diagnosed with mucormycosis during the study period. The commonest presentations were rhino-orbital and rhino-orbito-cerebral in 35.7% of cases each. Diabetes mellitus was the commonest associated risk factor in 95.7% of all patients, while 78.5% of the patients were treated with corticosteroids in the recent past, and 25.7% presented with active COVID-19 pneumonia. The commonest isolate was Rhizopus arrhizus n = 14, followed by Aspergillus flavus n = 16, A. fumigatus n = 4, A. niger n = 3, Fusarium oxysporumn = 1, and Apophysomyces variabilisn = 1. Fungal species identification was done by phenotypic methods for all the isolates and DNA sequence analysis of 18 isolates, and antifungal susceptibility testing of 30 isolates was performed by commercially prepared HiMIC plate (HiMedia, Mumbai, India) using broth microdilution for amphotericin B, isavuconazole, itraconazole, voriconazole, and posaconazole. The MIC50 and MIC90 of amphotericin B for R. arrhizus strains were 0.25 and 4 µg/ml, respectively; and the MIC50 and MIC90 results for itraconazole, posaconazole, and isavuconazole were 8 and 8, 2 and 2, and 2 and 8 µg/ml, respectively. In vitro data showed that amphotericin B was the most effective antifungal against most species. The commercially available ready-to-use minimum inhibitory concentration plates are user-friendly for performing antifungal susceptibility, which may be useful in choosing appropriate regimens and monitoring emerging resistance.

Agricultural diversification and intra-household dietary diversity: Panel data analysis of farm households in Bangladesh.

This paper investigates the associations between agricultural diversification and dietary diversity among men, women and children of farm households in Bangladesh. Using three waves of nationally representative Bangladesh Integrated Household Survey (BIHS- 2011-12, 2015 and 2018) data and a panel data model, the study found that agricultural production diversification can emphasize the dietary diversification across the life cycle of farm household members. The balanced panel data reveals that agricultural production diversification has a statistically significant and positive impact on dietary diversity of individual members (men, women and children) of farm households. Moreover, other important factors that impact on agricultural diversification and improve dietary diversity like women's education and employment, commercialization of farm households, access to non-farm income sources, and access to information facilities also have a strong association in improving dietary status, food and nutritional security as well. Although there is currently limited diversity in the diets of adult men, women and children of rural farm households, the situation can be improved over time with proper and consistent support. The findings suggest policy interventions should target not only agricultural diversification but also improve women's education and on and off farm employment opportunities and facilitate better access to information for rural households. These recommendations can support improved dietary diversity for all household members under different settings.

A study on the clinical spectrum of Hepatitis C virus infection from a tertiary care center in Sub-Himalayan India-spontaneous clearance, a neglected phenomenon.

Hepatitis C virus (HCV) infection has varied clinical manifestations. Noninvasive tools can be useful to assess the severity of liver disease and the rate of spontaneous clearance. HCV infection was determined by antibody or RNA-based tests over a period of 18 months in 8030 samples from the Gastroenterology department. Noninvasive indicators (AST-to-platelet ratio index and fibrosis-4 index) were computed. HCV RNA load was compared with Child-Turcotte-Pugh score. Rate of spontaneous clearance was estimated. About 3.2% of patients were found to have HCV. Fatigue, anorexia, and nausea were the primary complaints followed by ascites and encephalopathy. Extrahepatic features such as autoimmune hepatitis and non-Hodgkin's lymphoma were rare. There was an absence of advanced liver cirrhosis (κ = 0.96) in the majority of cases. Spontaneous HCV resolution was seen in 10.37%.

Rhizopus homothallicus, an emerging pathogen causing cavitary lung lesions.

Introduction: Rhizopus homothallicus is an emerging pathogen that causes pulmonary mucormycosis. Case Presentation: We report a case of pneumonia caused by R. homothallicus in a 54-year-old type 2 diabetic patient. The organism was isolated from bronchoalveolar lavage fluid and preliminarily identified by fungal morphology and finally by sequencing of the internal transcribed spacer region. Conclusion: Mucormycosis may be associated with cavitary lung lesions against a backdrop of poorly controlled diabetes or other immunosuppressed states. Pulmonary mucormycosis may have variable clinical and radiological presentations. Therefore, strong clinical suspicion and prompt management can address the high fatality associated with the disease.

Antifungal activity of silver nanoparticles on fungal isolates from patients of suspected mucormycosis / Actividad antifúngica de nanopartículas de plata en aislamientos fúngicos de pacientes con sospecha de mucormicosis

Introduction: The present study aimed to investigate the antifungal activity of silver nanoparticles (SNPs) against agents of suspected rhino orbital mucormycosis. Methods: Thirty-two strains were isolated from endoscopy-guided nasal swab and/or tissue biopsy after debridement/surgery on Sabouraud dextrose agar without cyclohexamide. Antifungal activity was conducted according to Clinical and Laboratory Standards Institute’s (CLSI) guidelines, M38-A2. The average size of silver nanoparticle was less than 10 nm. Results: Minimum inhibitory concentration (MIC) of nanoparticles of all strains was in the concentration range of 1–64 μg/ml and minimal fungicidal concentration (MFC) at 16–512 μg/ml. Conclusion: The SNPs revealed significant antifungal activity against agents of invasive mycosis.(AU)

Antifungal activity of silver nanoparticles on fungal isolates from patients of suspected mucormycosis.

INTRODUCTION: The present study aimed to investigate the antifungal activity of silver nanoparticles (SNPs) against agents of suspected rhino orbital mucormycosis. METHODS: Thirty-two strains were isolated from endoscopy-guided nasal swab and/or tissue biopsy after debridement/surgery on Sabouraud dextrose agar without cyclohexamide. Antifungal activity was conducted according to Clinical and Laboratory Standards Institute's (CLSI) guidelines, M38-A2. The average size of silver nanoparticle was less than 10 nm. RESULTS: Minimum inhibitory concentration (MIC) of nanoparticles of all strains was in the concentration range of 1-64 µg/ml and minimal fungicidal concentration (MFC) at 16-512 µg/ml. CONCLUSION: The SNPs revealed significant antifungal activity against agents of invasive mycosis.

Interference of Antinuclear Antibody (ANA) in Indirect Immunofluorescence Assay (IIFA)-Based Perinuclear Antineutrophil Cytoplasmic Antibody (pANCA) Interpretation.

Background: Indirect immunofluorescence assay (IIFA) based on antineutrophil cytoplasmic antibody (ANCA) testing is a commonly employed test for diagnosing autoimmune vasculitis. Antinuclear antibody (ANA) can give rise to a false interpretation of perinuclear-ANCA (pANCA) in ethanol-fixed granulocyte substrates. Analytical interference could frequently occur in setups where ethanol-fixed substrates are used alone. Here, we intend to investigate this ANA interference in pANCA interpretation. Methods: In this retrospective study, we studied anti-MPO-negative but ANA-positive and pANCA (IIFA based) samples. We also correlated immunoblot results (where data were available) and checked the association between grades of blot positivity (an indicator of the concentration of ANA) and frequency of pANCA interpretation. Data were analyzed by appropriate statistical techniques (Chi-square and kappa statistics). Results: About 19.2% of ANA blot (ENA-blot) positive samples displayed a pANCA positive pattern in the ethanol-fixed substrate, while this positivity in ENA-blot negatives was 6.5%. In positive ANA-IIFA samples, about 14.7% yielded pANCA patterns (on ethanol fixed substrates). Out of this, nuclear homogenous pattern yielding samples gave the highest frequency pANCA, that is, in 31.5% followed by speckled (11.1%), DFS (10.3%), and centromere (6.7%).The association of the nuclear homogenous pattern was statistically significant. Conclusions: ANA-positive results may interfere with the interpretation of pANCA as observed in ANA-IIFA and ENA-blot positive samples. ANA-IIFA patterns like nuclear homogenous may strongly associate this pANCA interpretation. This can help laboratories perform ANCA testing more effectively, ruling out ANA interference in ANCA screening.

Invasive Mucormycosis and Aspergillosis Coinfection Associated with Post-COVID-19 Pneumonia in a Tertiary Care Hospital.

Acute invasive fungal rhinosinusitis is a rare infection primarily affecting patients with co-morbidities like immunosuppression and poorly controlled diabetes. Mucormycosis is increasingly being reported in patients with SARS-CoV-2 (COVID-19). However, reports of coinfection of aspergillosis and mucormycosis involving nose, paranasal sinuses, orbit, and brain are rare in literature. We aimed to evaluate the patient demographics, clinical presentation, and management of cases presenting with mixed infection. We carried out retrospective analysis of 12 patients with confirmed diagnosis of mixed invasive fungal infections post-COVID-19 disease out of 70 cases of COVID-19-associated mucormycosis (CAM) presenting to a tertiary-level hospital in North India from May to June 2021. All patients had diabetes mellitus; the mean age was 48 years. The common presenting features were headache, nasal congestion, palatal ulcer, and vision loss accompanied by facial pain and swelling. Two patients developed cerebral abscess during the course of treatment; three patients had concurrent COVID-19 pneumonia. All patients received systemic liposomal amphotericin B and serial surgical debridements. The overall mortality rate was 16.7%. Our study demonstrates that mucormycosis and aspergillosis are angioinvasive mycoses that are clinically and radiologically identical. KOH direct mount of clinical sample showing septate hyphae should be extensively searched for aseptate hyphae after digestion and clearing of the tissue. A high index of suspicion of mixed infection post-COVID-19 and early initiation of liposomal amphotericin B followed by prompt surgical intervention can reduce the overall morbidity and mortality among patients with this condition.

Hepatitis C virus subtyping in Uttarakhand, India: a comparative study.

The objective of this study was to compare Reverse Hybridisation Assay with conventional sequencing for determination of Hepatitis C Virus Genotype and Subtypes. Anti-HCV antibody was determined followed by HCV RNA extraction which was used for (1) viral load determination (2) qualitative real-time PCR RHA for genotyping and (3) conventional sequencing. Compared to conventional sequencing, accuracy of RHA results was 96.55% for determination of genotype (κ = 0.93) and 89.66% for subtype (κ = 0.85). Sensitivity, specificity, negative predictive value (NPV) and positive predictive value (PPV) of the qualitative PCR were 82.29%, 100%, 44.44% and 100% respectively with an accuracy of 86.84%. RHA is a less time consuming and cheaper method for determination of HCV genotype and subtype yet results must be interpreted with caution and quality control monitoring should be strictly followed to ensure validity. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13337-021-00729-9.

Genotypic distribution of hepatitis C virus in Uttarakhand.

PURPOSE: Hepatitis C virus is a major cause of chronic hepatitis with seven known genotypes. Uttarakhand is a north Indian state in the Sub-Himalayan region where the genotypic distribution of HCV is largely unknown. This study was undertaken in order to assess the pattern of genotype and subtype and understand the risk factors leading to transmission of Hepatitis C virus in this understudied region. METHODS: Anti-HCV reactive cases were selected for determination of the circulating genotypes. Viral RNA was confirmed by real-time PCR. Strains were amplified and sequenced using Sanger's methods. Phylogenetic tree was constructed to determine the genotype. RESULTS: Genotype 3 was found to be the predominant genotype majority being subtype 3a and 3b followed by genotype 1. Subtypes 3g and genotype 4a were also observed. Major risk factor found was parenteral injection therapy from unregistered medical practitioners for minor ailments. CONCLUSIONS: Findings of our study will help in tailoring management and prevention protocols for HCV for the people of this region.

Increased Isolation of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli From Community-Onset Urinary Tract Infection Cases in Uttarakhand, India.

BACKGROUND: Management of community-acquired urinary tract infection (CA-UTI) relies heavily on empirical antibiotic therapy. Knowledge of the proportion of drug-resistant isolates especially extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli (E. coli), and various risk factors for acquisition are essential. METHOD:  Outpatient-treated CA-UTI cases were enrolled (continuously for three months), and microbiological analysis of urine sample was performed for significant bacterial growth followed by identification of conventional and matrix-assisted laser desorption/ionization-time of flight (MALDI-ToF) spectrometry method. Subsequent drug resistance and phenotypic ESBL detection were as per guidelines of the Clinical Laboratory Standard Institute (CLSI, USA). Univariate and multivariate analyses (logistic regression) of known and relevant risk factors of ESBL E. coli were performed as per standard statistical technique, using the SPSS computer package (IBM Corp., Armonk, NY).  Results: Two hundred and forty-one samples (of 694 samples) yielded significant growth. Sixty-one of 131 (46.6%) E. coli isolates were found to be ESBL producers. Non-beta-lactam antibiotic resistance in ESBL producers was high compared to non-ESBL producers (e.g., 88.5% vs 42.3% for quinolone resistance, 80.3% vs 34.3% for gentamicin resistance, etc.). Multivariate analysis (after univariate analysis detected probable factors of a likely ESBL model) indicated significant associations of ESBL-producing E. coli with advancing age (>55 years), prior hospitalization in last one year, use of antibiotics in previous six months, and presence of comorbid illness such as diabetes mellitus and chronic lung disease. CONCLUSION: High proportion of our community-acquired uropathogens are ESBL-producing E. coli and likely resistant to important antimicrobial agents such as quinolones, gentamicin, etc. Factors like advancing age, prior hospitalization, and antibiotic use, as well as comorbidities such as diabetes and chronic lung disease, may be strongly associated with ESBL E. coli and should be remembered while administering or preparing guidelines for empiric management of CA-UTI subjects.

Cryptococcal meningitis: An under-reported disease from the hills of Uttarakhand: A hospital-based cross-sectional study.

BACKGROUND: Cryptococcal meningitis is a fatal opportunistic neuroinfection and an AIDS defining illness. It can also occur in non-HIV patients who are immunodefecient due to chronic glucocorticoid use, organ transplantation, malignancy and sarcodiosis. MATERIALS AND METHODS: A cross-sectional study was conducted in a tertiary care hospital from July to December 2018. CSF samples of 364 patients were received by Microbiology laboratory during this period for the purpose of aerobic bacterial, fungal and TB culture, respectively. All samples were subjected to examination by direct wet mount, Gram stain and India ink preparation. Ziehl Neelsen staining, solid culture for Mycobacterium tuberculosis on Lowenstein Jensen medium and Gene Xpert was also performed on all CSF samples. These samples were further subjected to fungal culture on Sabouraud's dextrose agar. Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS) was used for identifying all bacterial (except M. tuberculosis) and fungal isolates. RESULTS: Out of 364 CSF samples received, 288 were sterile after 48 hours of aerobic incubation. Bacterial isolates, M. tuberculosis and Cryptococcus spp. were obtained in culture from 51, 21 and 4 samples, respectively. The prevalence of cryptococcal meningitis in our study was 1.09% (4/364). Cryptococcus neoformans var grubii was the most common isolate (2/4; 50%) followed by Cryptococcus neoformans var neoformans (1/4; 25%) and Cryptococcus neoformans var gattii (1/4; 25%), respectively. CONCLUSION: Cryptococcal meningitis is a rapidly fatal condition which requires a high index of suspicion and calls for a collective effort from family physicians and diagnosticians alike. This disease is under-reported from Uttarakhand and therefore calls for further research from this region.