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.

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.

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.

COVID-19-associated mucormycosis - "The Black fungus" with associated invasive aspergillosis and actinomycosis infection: Series of 5 cases in the Indian scenario.

COVID-19 caused by SARS-CoV-2 has been studied and reported widely since November 2019, after its first case was detected in Wuhan, China. It has proven to be a fatal disease worldwide. COVID-19 causes severe disease in patients with immunosuppression and has always been associated with high mortality in such patients. Immunocompromised patients are always at a higher risk of getting co-infections too, and the same is true with COVID-19. It predisposes to the development of many fungal infections of which, mucormycosis is one of the most common ones, especially in the Indian population where a large group of the population is predisposed to diabetes mellitus. India has the world's largest population of diabetic patients; therefore, the prevalence of COVID-19-associated mucormycosis (CAM) is also the highest in India. Also, the use of corticosteroids over a long duration of time predisposes the patients to the development of mucormycosis owing to immunosuppression. Hypoxia, low total leukocyte count, and high ferritin are the other predisposing factors that lead to the growth of mucormycosis associated with COVID-19. Here, we present five cases within a span of 2 months, of mucormycosis-associated COVID-19 with mixed infections of aspergillosis and actinomycetes. Four cases had mucormycosis with aspergillosis and one with mucormycosis with Actinomyces. Three patients recovered fully after being treated with intravenous amphotericin B; however, unfortunately, two of our patients could not be saved.

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.

A critical appraisal of mucormycosis in COVID- 19 patients in a tertiary care centre in India.

Background and Purpose: Morbidity and mortality of opportunistic fungal infections in COVID-19 patients are less studied and defined. The patients receiving immunosuppressive therapy, broad-spectrum antibiotics, corticosteroids, and invasive and non-invasive ventilation are the high-risk groups. Materials and Methods: The demographic profile as well as clinical and radiological findings of all the patients with COVID-19 suspected of Mucormycosis (MM) were recorded. The tissue samples from all the patients were sent for microbiological (KOH mount and culture) and histopathological analysis for confirmation of MM. Results: In total, 45 COVID-19 patients suspected of MM were included in the study and MM was confirmed in 42 patients. The mean age of the patients was 50.30±14.17 years with a female: male ratio of 1.1:1. The most common symptom was headache (52.38%) followed by purulent nasal discharge (38.09%) and facial pain in 33.33% of the cases. The ocular symptoms included a diminution of vision (33.33%) and redness of the eye (2.38%). The most common site of involvement was rhino-orbital (42.85%) followed by sinonasal (23.80%) and rhino cerebral (19.04%). Majority (38.09%) of the patients were diagnosed with stage II of Rhino-orbital-cerebral Mucormycosis (ROCM) based on radiology. A history of diabetes mellitus and steroids was present in 97.61% and 85.71% of the cases, respectively. Moreover, KOH was positive for MM in 97.61% of the cases while the culture was positive in only 35.71% of the cases. In addition, on histopathology, MM was confirmed in 64.28 % of the cases. Mixed growth with Aspergillus species and Rhizopus species was observed in 14.28% of the cases in culture and 11.90% of the cases in histopathology test. Furthermore, angioinvasion was found in 23.80% of the cases according to the histopathology test. Conclusion: Based on the results, the most common conditions associated with MM in COVID-19 patients were diabetes mellitus and steroid therapy. A high level of clinical suspicion aided with diagnostic tests, including KOH mount, culture, histopathology, and radiology which helped the early detection of opportunistic fungal infection in COVID-19 patients to ensure timely treatment.

Patient satisfaction survey of microbiological tests done in G.B. Pant Hospital.

PURPOSE: Measuring patient satisfaction plays an increasingly important role in the growing push toward healthcare provider accountability. This study seeks to evaluate G.B. Pant Hospital (a North Indian tertiary care centre) patient satisfaction with clinical laboratory services. DESIGN/METHODOLOGY/APPROACH: A total of 100 out- and in-patients were randomly selected and interviewed about microbiological services using a standard format, a method which can be easily used to compare patient satisfaction with laboratory services elsewhere. FINDINGS: Patients represented all age groups: females and males were balanced. Few were from poor socio-economic backgrounds. Patients do not have problems getting tests done, but the laboratory's inconvenient location caused dissatisfaction. Patients do not have problems communicating with staff, but medical terms are not understood by patients. Hospital cleanliness needs improving, especially toilets, which causes the most patient dissatisfaction. Hospital staff were deemed highly competent and judged to give excellent technical help to patients. The questionnaire's financial subscale shows 100 per cent satisfaction because all tests in the microbiology department are free. The overall satisfaction with services stood at 83 per cent. Satisfaction scores for G.B. Pant Hospital appear to be satisfactory. RESEARCH LIMITATIONS/IMPLICATIONS: This study does not compare patient satisfaction in two or more hospitals and findings may not be generalisable. PRACTICAL IMPLICATIONS: Patient satisfaction surveys are the best way to identify deficiencies and improve hospital services. Repeating studies at six monthly intervals is a useful managerial intervention aimed at delivering and maintaining quality healthcare. ORIGINALITY/VALUE: This laboratory satisfaction survey is the first of its kind for government hospitals in India. The survey revealed a positive feedback and helped to identify the areas of concern along with estimating the patient satisfaction scores. This is the best way to identify the areas of deficiencies and improving the services provided by the hospital. The authors feel that repeating such studies at a regular interval of six months would be a useful guide for the managerial interventions.

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.

Inactivated whole-virion vaccine BBV152/Covaxin elicits robust cellular immune memory to SARS-CoV-2 and variants of concern.

BBV152 is a whole-virion inactivated vaccine based on the Asp614Gly variant. BBV152 is the first alum-imidazoquinolin-adjuvanted vaccine authorized for use in large populations. Here we characterized the magnitude, quality and persistence of cellular and humoral memory responses up to 6 months post vaccination. We report that the magnitude of vaccine-induced spike and nucleoprotein antibodies was comparable with that produced after infection. Receptor binding domain-specific antibodies declined against variants in the order of Alpha (B.1.1.7; 3-fold), Delta (B.1.617.2; 7-fold) and Beta (B.1.351; 10-fold). However, pseudovirus neutralizing antibodies declined up to 2-fold against the Delta followed by the Beta variant (1.7-fold). Vaccine-induced memory B cells were also affected by the Delta and Beta variants. The SARS-CoV-2-specific multicytokine-expressing CD4+ T cells were found in ~85% of vaccinated individuals. Only a ~1.3-fold reduction in efficacy was observed in CD4+ T cells against the Beta variant. We found that antigen-specific CD4+ T cells were present in the central memory compartment and persisted for at least up to 6 months post vaccination. Vaccine-induced CD8+ T cells were detected in ~50% of individuals. Importantly, the vaccine was capable of inducing follicular T helper cells that exhibited B-cell help potential. These findings show that inactivated vaccine BBV152 induces robust immune memory to SARS-CoV-2 and variants of concern that persists for at least 6 months after vaccination.

Effectiveness of ChAdOx1 nCoV-19 vaccine against SARS-CoV-2 infection during the delta (B.1.617.2) variant surge in India: a test-negative, case-control study and a mechanistic study of post-vaccination immune responses.

BACKGROUND: SARS-CoV-2 variants of concern (VOCs) have threatened COVID-19 vaccine effectiveness. We aimed to assess the effectiveness of the ChAdOx1 nCoV-19 vaccine, predominantly against the delta (B.1.617.2) variant, in addition to the cellular immune response to vaccination. METHODS: We did a test-negative, case-control study at two medical research centres in Faridabad, India. All individuals who had a positive RT-PCR test for SARS-CoV-2 infection between April 1, 2021, and May 31, 2021, were included as cases and individuals who had a negative RT-PCR test were included as controls after matching with cases on calendar week of RT-PCR test. The primary outcome was effectiveness of complete vaccination with the ChAdOx1 nCoV-19 vaccine against laboratory-confirmed SARS-CoV-2 infection. The secondary outcomes were effectiveness of a single dose against SARS-CoV-2 infection and effectiveness of a single dose and complete vaccination against moderate-to-severe disease among infected individuals. Additionally, we tested in-vitro live-virus neutralisation and T-cell immune responses to the spike protein of the wild-type SARS-CoV-2 and VOCs among healthy (anti-nucleocapsid antibody negative) recipients of the ChAdOx1 nCoV-19 vaccine. FINDINGS: Of 2379 cases of confirmed SARS-CoV-2 infection, 85 (3·6%) were fully vaccinated compared with 168 (8·5%) of 1981 controls (adjusted OR [aOR] 0·37 [95% CI 0·28-0·48]), giving a vaccine effectiveness against SARS-CoV-2 infection of 63·1% (95% CI 51·5-72·1). 157 (6·4%) of 2451 of cases and 181 (9·1%) of 1994) controls had received a single dose of the ChAdOx1 nCoV-19 vaccine (aOR 0·54 [95% CI 0·42-0·68]), thus vaccine effectiveness of a single dose against SARS-CoV-2 infection was 46·2% (95% CI 31·6-57·7). One of 84 cases with moderate-to-severe COVID-19 was fully vaccinated compared with 84 of 2295 cases with mild COVID-19 (aOR 0·19 [95% CI 0·01-0·90]), giving a vaccine effectiveness of complete vaccination against moderate-to-severe disease of 81·5% (95% CI 9·9-99·0). The effectiveness of a single dose against moderate-to-severe disease was 79·2% (95% CI 46·1-94·0); four of 87 individuals with moderate-to-severe COVID-19 had received a single dose compared with 153 of 2364 participants with mild disease (aOR 0·20 [95% CI 0·06-0·54]). Among 49 healthy, fully vaccinated individuals, neutralising antibody responses were lower against the alpha (B.1.1.7; geometric mean titre 244·7 [95% CI 151·8-394·4]), beta (B.1.351; 97·6 [61·2-155·8]), kappa (B.1.617.1; 112·8 [72·7-175·0]), and delta (88·4 [61·2-127·8]) variants than against wild-type SARS-CoV-2 (599·4 [376·9-953·2]). However, the antigen-specific CD4 and CD8 T-cell responses were conserved against both the delta variant and wild-type SARS-CoV-2. INTERPRETATION: The ChAdOx1 nCoV-19 vaccine remained effective against moderate-to-severe COVID-19, even during a surge that was dominated by the highly transmissible delta variant of SARS-CoV-2. Spike-specific T-cell responses were maintained against the delta variant. Such cellular immune protection might compensate for waning humoral immunity. FUNDING: Department of Biotechnology India, Council of Scientific and Industrial Research India, and Fondation Botnar.

Longitudinal Serology of SARS-CoV-2-Infected Individuals in India: A Prospective Cohort Study.

Clinical and epidemiological characteristics of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are now widely available, but there are few data regarding longitudinal serology in large cohorts, particularly those from low-income and middle-income countries. We established an ongoing prospective cohort of 3,840 SARS-CoV-2-positive individuals according to RT-PCR in the Delhi-National Capital Region of India to document clinical and immunological characteristics during illness and convalescence. The immunoglobulin G (IgG) responses to the receptor binding domain (RBD) and nucleocapsid were assessed at 0 to 7 days, 10 to 28 days, and 6 to 10 weeks after infection. The clinical predictors of seroconversion were identified by multivariable regression analysis. The seroconversion rates during the postinfection windows of 0 to 7 days, 10 to 28 days, and 6 to 10 weeks were 46%, 84.7%, and 85.3%, respectively (N = 743). The proportion with a serological response increased with the severity of coronavirus disease 2019 (COVID-19). All participants with severe disease, 89.6% with mild to moderate infection, and 77.3% of asymptomatic participants had IgG antibodies to the RBD antigen. The threshold values for the nasopharyngeal viral RNA RT-PCR of a subset of asymptomatic and symptomatic seroconverters were comparable (P = 0.48) to those of nonseroconverters (P = 0.16) (N = 169). This is the first report of longitudinal humoral immune responses to SARS-CoV-2 over a period of 10 weeks in South Asia. The low seropositivity of asymptomatic participants and differences between assays highlight the importance of contextualizing the understanding of population serosurveys.

Acute lower respiratory tract infections in children.

Acute lower respiratory tract infection (ALRTI) is a common illness, but there have been relatively few studies of the bacterial etiology in developing countries. Nasopharyngeal aspirates of 70 children under 10 years of age with ALRTI were cultured for aerobic bacterial pathogens. Klebsiella pneumoniae was the commonest organism (32.2%) isolated followed by S. pneumoniae (10%), E. coli (10%), P. aeruginosa (5.7%), S. aureus (2.8%) and H. influenzae (1.4%). There were significantly more bacterial pathogens isolated in children <1 year of age (73.7%) than in those > 1 year of age (56.2%) (P=0.03). A shift in spectrum from Gram-positive cocci to Gram negative bacilli in ALRTI was observed in our study.

Cryptococcal granulomas in an immunocompromised HIV-negative patient.

Disseminated cryptococcosis usually occurs in immunocompromised individuals with defective cell-mediated immunity, most commonly seen with HIV infection. We present a case of disseminated cryptococcosis in an HIV-negative male patient who presented with headache, fever, altered sensorium of short duration and multiple cutaneous lesions. An emergency CT scan of the head showed multiple intracranial and intraventricular granulomas. Routine laboratory investigations were within the normal range. A CSF examination revealed capsulated yeasts on India ink and a culture yielded cryptococcus neoformans. A cryptococcal antigen test by latex agglutination kit was positive. A biopsy revealed multiple capsulated yeasts cells in the cutaneous lesions, which were consistent with cryptococcus neoformans. The patient was successfully treated with Amphotericin B and Fluconazole with regression of cranial and cutaneous lesions.

Methicillin and Vancomycin Resistant S. aureus in Hospitalized Patients.

S. aureus is the major bacterial cause of skin, soft tissue and bone infections, and one of the commonest causes of healthcare-associated bacteremia. Hospital-associated methicillin-resistant S. aureus (MRSA) carriage is associated with an increased risk of infection, morbidity and mortality. Screening of high-risk patients at the time of hospital admission and decolonization has proved to be an important factor in an effort to reduce nosocomial transmission. The electronic database Pub Med was searched for all the articles on "Establishment of MRSA and the emergence of vancomycin-resistant S. aureus (VRSA)." The search included case reports, case series and reviews. All the articles were cross-referenced to search for any more available articles. A total of 88 references were obtained. The studies showed a steady increase in the number of vancomycin-intermediate and vancomycin-resistant S. aureus. Extensive use of vancomycin creates a selective pressure that favors the outgrowth of rare, vancomycin-resistant clones leading to heterogenous vancomycin intermediate S. aureus hVISA clones, and eventually, with continued exposure, to a uniform population of vancomycin-intermediate S. aureus (VISA) clones. However, the criteria for identifying hVISA strains have not been standardized, complicating any determination of their clinical significance and role in treatment failures. The spread of MRSA from the hospital to the community, coupled with the emergence of VISA and VRSA, has become major concern among healthcare providers. Infection-control measures, reliable laboratory screening for resistance, appropriate antibiotic prescribing practices and avoidance of blanket treatment can prevent long-term emergence of resistance.

Nosocomial blood-stream infections from extended-spectrum-beta-lactamase-producing Escherichia coli and Klebsiella pneumonia from GB Pant Hospital, New Delhi.

BACKGROUND: Nosocomial septicemia due to extended spectrum beta-(Beta)-lactamase (ESBL) producing Klebsiella pneumoniae and Escherichia coli are a therapeutic challenge due to resistance. Knowledge of disease burden and resistance patterns is required for proper and timely management. We report the prevalence and antimicrobial susceptibility of ESBL producing E. coli and K .pneumoniae from septicemia at a tertiary care hospital. METHODOLOGY: A total of 2,870 blood samples of suspected cases of septicemia were studied between January and December 2009. Antimicrobial susceptibility was determined by Kirby Bauer's disc diffusion method and MICs for imipenem, meropenem, and ertapenem were determined using the E-test. All isolates of E. coli and K. pneumoniae were tested for ESBL production by E-test method. RESULTS: Forty-one (70.7%) K. pneumoniae isolates and ten (41.7%) E. coli isolates were ESBL producers. Two (5%) of ESBL producing K. pneumoniae isolates, but no E. coli isolates, were resistant to carbapenems. In vitro, all ESBL producers were sensitive to tigecycline. CONCLUSION: Our data indicated that the prevalence of ESBL-producing E. coli and K. pneumonia strains isolated from blood cultures from hospitalized patients is high. ESBL-producing organisms were found to be more susceptible to meropenem than to imipenem and ertapenem. Tigecycline is active against all the ESBL or multidrug resistant (MDR) E. coli and Klebsiella spp. isolates.

Increasing trends of leptospirosis in northern India: a clinico-epidemiological study.

BACKGROUND: Leptospirosis, a zoonosis associated with potentially fatal consequences, has long been a grossly underreported disease in India. There is no accurate estimate of the problem of leptospirosis in non-endemic areas such as north India. METHODS/PRINCIPAL FINDINGS: In order to understand the clinical spectrum and risk factors associated with leptospirosis, we carried out a retrospective study in patients with acute febrile illness in north India over the last 5 years (January 2004 to December 2008). There was increased incidence of leptospirosis (11.7% in 2004 to 20.5% in 2008) as diagnosed by IgM ELISA and microscopic agglutination titer in paired acute and convalescent sera. The disease showed a peak during the rainy season (August and September). We followed up 86 cases of leptospirosis regarding their epidemiological pattern, clinical features, laboratory parameters, complications, therapy, and outcome. Mean age of patients was 32.6 years (2.5 years to 78 years) and males (57%) outnumbered females (43%). Infestation of dwellings with rats (53.7%), working in farm lands (44.2%), and contact with animals (62.1%) were commonly observed epidemiological risk factors. Outdoor workers including farmers (32.6%), labourers (11.6%), para-military personnel (2.3%), and sweepers (1.2%) were commonly affected. Modified Faine's criteria could diagnose 76 cases (88.3%). Renal failure (60.5%), respiratory failure (20.9%), the neuroleptospirosis (11.6%), and disseminated intravascular coagulation (DIC) (11.6%) were the commonest complications. Five patients died, giving a case fatality rate of 5.9%. CONCLUSIONS/SIGNIFICANCE: There has been a rapid rise in the incidence of leptospirosis in north India. Severe complications such as renal failure, respiratory failure, neuroleptospirosis, and DIC are being seen with increasing frequency. Increased awareness among physicians, and early diagnosis and treatment, may reduce mortality due to leptospirosis.

Evaluation of knowledge and practice amongst nursing staff toward infection control measures in a tertiary care hospital in India.

PURPOSE: A study to assess the level of knowledge and practice of 100 staff nurses on infection control measures was carried out in a tertiary care center. METHOD: A structured questionnaire was used to collect the data. RESULTS: The mean knowledge of staff nurses regarding infection control measures was 75.5% and the mean reported infection control practice was 57.5%. After conducting exhaustive lectures on infection control related topics, a significant decline in the hospital-acquired infection (HAIs) rates was seen in the high-risk areas. CONCLUSION: Training of nursing staff is needed to improve knowledge and practice in infection control.