Clinical spectrum, phenotypic and molecular characterization, and antifungal susceptibility of an emerging human pathogen, Acrophialophora, from India.

Acrophialophora is implicated in superficial and invasive infections, especially in immunosuppressed individuals. The present study was undertaken to provide clinical, microbiological, phylogenetic, and antifungal susceptibility testing (AFST) profile of Acrophialophora isolated from India. All the isolates identified as Acrophialophora species at the National Culture Collection for Pathogenic Fungi, Chandigarh, India were revived. Phenotypic and molecular characterization was performed, followed by temperature studies, scanning electron microscopy (SEM), and AFST. We also performed systematic review of all the cases of Acrophialophora species reported till date. A total of nine isolates identified as Acrophialophora species were identified by molecular method as A. fusispora (n = 8) and A. levis (n = 1), from brain abscess (n = 4), respiratory tract (n = 3), and corneal scraping (n = 2). All patients but two had predisposing factors/co-morbidities. Acrophialophora was identified as mere colonizer in one. Temperature studies and SEM divulged variation between both species. Sequencing of the internal transcribed spacer ribosomal DNA and beta-tubulin loci could distinguish species, while the LSU ribosomal DNA locus could not. AFST showed the lowest minimum inhibitory concentrations (MICs) for triazoles and the highest for echinocandins. Systematic literature review revealed 16 cases (11 studies), with ocular infections, pulmonary and central nervous system infections, and A. fusispora was common species. All the patients except three responded well. High MICs were noted for fluconazole, micafungin, and caspofungin. This is the first study delineating clinical, phenotypic, and genotypic characteristics of Acrophialophora species from India. The study highlights microscopic differences between both species and emphasizes the role of molecular methods in precise identification. Triazoles appear to be the most effective antifungals for managing patients.

We describe clinical, phenotypic, and genotypic characteristics of Acrophialophora species. This species causes mild infection to fatal infection in immunosuppressed individuals. Triazoles are effective in treating such infections.

Antifungal Susceptibility Profile of Clinical and Environmental Isolates of Aspergillus Species From a Tertiary Care Center in North India.

INTRODUCTION: Aspergillus species are ubiquitously found in the environment worldwide and are important causative agents for infection. Drug resistance among Aspergillus species is emerging, hence the present study was undertaken to look for antifungal susceptibility profiles of clinical and environmental isolates of Aspergillus species. MATERIALS AND METHODS: During the period from January 2018 to June 2019, a total of 102 Aspergillus isolates (40 clinical, 40 hospital, and 22 community environment) were tested for antifungal susceptibility testing for determination of minimum inhibitory concentration (MIC)/minimum effective concentration (MEC) as per Clinical and Laboratory Standards Institute (CLSI) M38-A3 method for itraconazole, voriconazole, amphotericin B, and caspofungin. RESULTS: Out of these 102 Aspergillus isolates, A. flavus was the most common species present. Aspergillus species were found to have low MIC values to azoles such as itraconazole and voriconazole except for one clinical isolate, which showed a MIC value of 2 µg/ml to voriconazole. Two isolates were non-wild-type for amphotericin B, but all isolates were wild-type for caspofungin. CONCLUSION: Antifungal susceptibility testing among clinical Aspergillus isolates and environmental surveillance studies in view of emerging drug resistance should be undertaken at a larger scale.

From Spores to Solutions: A Comprehensive Narrative Review on Mucormycosis.

Mucormycosis is an infrequent but fatal illness that mainly affects patients with uncontrolled diabetes mellitus, diabetic ketoacidosis, solid and hematologic neoplasms, organ transplantation, chronic steroid intake, prolonged neutropenia, iron overload states, neonatal prematurity, severe malnutrition, and HIV. Many cases were reported across the world recently following the COVID-19 pandemic. Recent research has led to a better understanding of the pathogenesis of the disease, and global guidelines are now available for managing this serious infection. Herein, we comprehensively review the etiological agents, pathogenesis, clinical presentations, diagnosis, and management of mucormycosis.

Artificial Intelligence: Exploring utility in detection and typing of fungus with futuristic application in fungal cytology.

Artificial Intelligence (AI) is an emerging, transforming and revolutionary technology that has captured attention worldwide. It is translating research into precision oncology treatments. AI can analyse large or big data sets requiring high-speed specialized computing solutions. The data are big in terms of volume and multimodal with the amalgamation of images, text and structure. Machine learning has identified antifungal drug targets, and taxonomic and phylogenetic classification of fungi based on sequence analysis is now available. Real-time identification tools and user-friendly mobile applications for identifying fungi have been discovered. Akin to histopathology, AI can be applied to fungal cytology. AI has been fruitful in cytopathology of the thyroid gland, breast, urine and uterine cervical lesions. AI has a huge scope in fungal cytology and would certainly bear fruit with its accuracy, reproducibility and capacity for handling big data. The purpose of this systematic review was to highlight the AI's utility in detecting fungus and its typing with a special focus on future application in fungal cytology. We also touch upon the basics of AI in brief.

Open-labeled Randomized Controlled Trial on Efficacy of Azithromycin Versus Doxycycline in Pediatric Scrub Typhus.

BACKGROUND: Doxycycline (DX) is the first-line therapeutic agent for scrub typhus. Macrolides, especially azithromycin (AZ), have been found to be equally efficacious as DX for treating scrub typhus in adults. We conducted this study to compare the efficacy of AZ versus DX in pediatric scrub typhus. STUDY DESIGN: Open-label randomized controlled trial. METHODS: Children 1-14 years of age suffering from acute febrile illness of ≥5 days and with a positive scrub IgM serology were randomized to receive either DX (2.2 mg/kg/dose twice daily in <40 kg; 100 mg BD in >40 kg for 7 days) or AZ (10 mg/kg/day for 5 days). The primary outcome was defervescence within 7 days of DX or 5 days of AZ. RESULTS: We had 75 children randomized and analyzed using an intention-to-treat approach and 60 children analyzed via per-protocol analysis. The proportion of children achieving defervescence was comparable in 2 groups [per-protocol analysis: 90.9% in the DX group vs. 96.3% in the AZ group, RR: 0.94 (95% CI: 0.83-1.08)]. On Kaplan-Meier curves, AZ showed a significantly higher probability of defervescence at any time point during treatment as compared with DX (log-rank test P value 0.035). CONCLUSION: AZ and DX had comparable rates of defervescence among children with scrub typhus. Trial registration No.: CTRI/2020/01/022991.

Primary Cutaneous Mucormycosis: A Necrotising Soft Tissue Infection with Poor Prognosis.

BACKGROUND: Cutaneous mucormycosis is an unusual fungal infection that continues to occur. It needs aggressive surgical debridement and timely administration of antifungals due to its high fatality rate. High clinical suspicion on the part of a surgeon is required to prevent the same. CASE PRESENTATION: We present two cases of cutaneous mucormycosis in which the patients succumbed to death, highlighting the seriousness of the condition. One patient had a lower leg ulcer and was diabetic, and the other patient had a gluteal abscess following an intramuscular injection. Tissue samples grew Rhizopus arrhizus and Apophysomyces sp., respectively. Both patients were treated with amphotericin B, and extensive debridement was performed. DISCUSSION: Cutaneous mucormycosis can be reported in immunocompetent people, and there is a need for early recognition of the entity as a differential diagnosis of any nonhealing necrotic ulcer. CONCLUSION: Proper training and education of technical and clinical staff should be done at peripheral primary and secondary care centres so as not to miss out on cases of mucormycosis and for better prognosis in a cutaneous variety of mucormycosis in surgical patients.

Phaeohyphomycosis: A study from tertiary health care center in North India.

Objectives: Phaeohyphomycosis refers to infections caused by phaeoid/dematiaceous or darkly pigmented fungi. This study was undertaken to further increase our knowledge about the incidence of phaeohyphomycosis and its causative agents. Materials and Methods: The present study was conducted over a period of one and a half years (January 2018-June 2019) on specimens received from patients with varied clinical manifestations ranging from superficial infections, subcutaneous cysts, pneumonia, brain abscess to a disseminated infection. These specimens were processed in the Department of Microbiology for potassium hydroxide (KOH) examination and culture and in Pathology for cytology/histopathological examination (HPE). All specimens positive on direct examination for dark grey, brown or black fungi were included in the study. Results: A total of 20 specimens were confirmed as phaeohyphomycosis. Most of the patients belonged to the age group of 41 to 50 years. Male: Female ratio was 2.3:1. Trauma was the most common risk factor. Spectra of the isolated fungal pathogens comprised of Bipolaris species, Exophiala species, Curvularia geniculata, Phialemonium species, Daldinia eschscholtzii, Hypoxylon anthochroum, Phaeoacremonium species, Leptosphaerulina australis, Medicopsis romeroi, Lasiodiplodia theobromae, Eutypella species, Chaetomium globosum, Alternaria species, Cladophialophora bantiana and 2 unidentified dematiaceous fungi. Recovery from phaeohyphomycosis was seen in 12 patients, 7 were lost to follow up and one patient succumbed to the illness. Conclusion: Infections caused by phaeoid fungi can no longer be viewed as rare. In fact, phaeohyphomycosis can have myriad of presentations spanning from mild cutaneous infections to fatal brain disease. Therefore, a high index of clinical suspicion is needed to diagnose such infections. The primary treatment modality remains surgical removal of the lesion in cutaneous or subcutaneous infections however disseminated disease with a guarded prognosis requires aggressive management.

Acrophialophora fusispora as an Agent of Mycotic Keratitis: A Case Report and Review of Literature.

BACKGROUND: Acrophialophora species is an infrequent human opportunistic pathogen. It is widely distributed in temperate as well as tropical regions. Here, we present a rare case of fungal keratitis caused by A. fusispora. CASE PRESENTATION: A 26-year male driver presented with pain, watering, redness, whitish discoloration, and blurring of vision in the left eye for the last 3-4 days. Upon examination, he had a dry-looking corneal ulcer with infiltration and satellite lesions. Corneal scrapings were positive for septate fungal hyphae by Gram staining and KOH mount. After five days, the growth observed was presumptively identified as genus Acrophialophora and finally identified as Acrophialophora fusispora by genetic sequencing. The patient failed to respond medically and was planned for therapeutic keratoplasty. DISCUSSION: To date, four cases of ocular involvement due to Acrophialophora have been described. Amongst which one case was associated with an immunocompromised state. Three of the cases were resolved medically, while one required therapeutic keratoplasty, indicating possible strong pathogenicity to the eye. CONCLUSION: As Acrophialophora seems to have a predilection for eye infections, an early diagnosis with timely appropriate treatment is the best way to restore the normal vision of a patient.

Species Distribution and Antifungal Susceptibility Profile of Dermatophytes from a Tertiary Care Centre in North India.

Objective Dermatophytoses, one of the most ancient diseases, is becoming a menace in recent times. This has made the knowledge of antifungal susceptibility a priority in today's times. Material and Methods This is a prospective study conducted over 18 months including all dermatophytes isolated during the period. Dermatophytes were identified by routine phenotypic methods. Antifungal susceptibility testing was performed for griseofulvin, terbinafine, and itraconazole as per the Clinical Laboratory Standard Institute M38 A2, and minimum inhibitory concentrations (MICs) were read after 5 days. Results Patient details and associated risk factors were recorded. Fixed dose combinations with steroids were associated with 79.3% (46 out of 58) of patients with dermatophytosis of skin. Among the 72 dermatophytes isolated during the study period, 58 (80.5%) were isolated from skin scrapings and 14 (19.4%) from nail samples. Tinea corporis with cruris was the most common presentation. The most common dermatophyte isolated from skin scrapings was Trichophyton mentagrophytes complex (70.6%, 41 out of 58), while from nail samples it was Trichophyton rubrum complex (78.57%, 11 out of 14). Based on the MIC 50 and MIC 90 results, itraconazole showed the lowest MICs, followed by terbinafine and then griseofulvin. Conclusion With the changing epidemiology of species distribution and antifungal resistance, there is a need for continuous surveillance of these parameters of dermatophytes.

COVID-19 and Plethora of Fungal Infections.

Purpose of Review: Severe-acute respiratory coronavirus 2 (SARS-CoV-2) causing corona virus disease 2019 (COVID-19) has been the single most important pathogen driving health care delivery system for the last one and half years. Now, as the time is passing, many issues related to co-infections/secondary infections/superinfections in COVID-19 patients are emerging. The literature is getting enriched everyday by addition of reports from all over the world for the same. The purpose of this review is to decipher the plethora of fungal infections in COVID-19. Recent Findings: COVID-19 infection along with it brought many risk factors namely lung injury, immunosuppression, need for oxygen therapy, monoclonal antibodies, steroid therapy, etc. which are known predisposing factors for fungal infections. Rather the extent and severity of fungal pathogens has been so much that it has led to new terminologies like CAC (COVID-19-associated Candida), CAPA (COVID-19-associated pulmonary aspergillosis) and CAM (COVID-19-associated mucormycosis). There is increase in invasiveness of Candida, prevalence of aspergillosis in COVID-19 damaged lung and outbreak of mucormycosis in COVID-19 patients resulting in "double trouble," keeping laboratory personnel, clinicians, and intensivists on their toes in managing these patients. Summary: Awareness and understanding regarding these possible complications is necessary to decrease the morbidity and mortality among patients. The COVID-19 and fungal coinfections may bring more insight into ways of pathogenesis of fungal infections, need for better antifungal agents, quick diagnostic modalities, and better management policies in the near future.

Seroprevalence of hepatitis A and hepatitis E in patients at a teaching hospital of northern India over a period of 8 years.

Background: Considering the diverse socio-economic and demographic factors in a vast country like India, it is important to study the long-term trends of hepatitis A (HAV) and hepatitis E (HEV) viruses. This study describes their seroprevalence and long-term trends in a tertiary care center of North India. Methods: The present retrospective observational study was conducted over a period of 8 years (January 2011-December 2018). Serological testing was done for detecting IgM antibodies against HAV and HEV using enzyme-linked immunosorbent assay. Results: A total of 5319 samples were received during the study period, of which 903 (16.9%) were reactive for anti-HAV IgM antibodies and 795 (14.9%) were reactive for anti-HEV IgM antibodies. Majority of the cases occurred from June to October while HEV cases had a constant presence during the later years. Among HAV group, 534 (59.1%) were children, 336 (37.2%) were adults, and 33 (3.7%) were pregnant females. In HEV group, 632 (79.5%) were adults, 114 (14.3%) were pregnant females, whereas only 49 (6.2%) were children. Among those who were co-infected (n = 87), 48 (55.2%) were adults, 22 (25.3%) were pregnant females, and the rest 17 (19.5%) were children. Conclusions: The shift in seroprevalence toward adults, along with an increasing trend of the number of cases reporting to the hospital, warrants active surveillance of HAV. Similarly, screening protocols for HEV should be set up as part of the antenatal management for early detection of the cases among pregnant females.

Bacterial Colonization of Home Nebulizers Used by Children With Recurrent Wheeze.

OBJECTIVE: To look for bacterial colonization of parts of home nebulizers used for children with recurrent wheeze and asthma. METHODS: Children aged 1 mo-12 y, using home nebulizers for recurrent cough and wheeze were enrolled from May to October, 2019. Caregivers were administered a structured questionnaire by a single researcher, during their hospital visit, to elicit information on their nebulizer cleaning practices. Samples were taken from nebulizer medicine chamber and tubing for bacterial culture and sensitivity. RESULT: Bacterial growth was observed in 17 culture samples obtained from medicine chamber and/or tubing of nebulizers used by 12 (20.3%) out of the 59 enrolled children. The bacteria isolated were Klebsiella pneumoniae, Pseudomonas aeruginosa and Staphylococcus aureus (Methicillin resistant S. aureus and Coagulase negative staphylococci) and these were resistant to many of the commonly used antimicrobials. Almost 20% parents had never cleaned the nebulizers. Diluent re-use was significantly associated with bacterial colonization of nebulizer parts [AOR (95% CI) 20.6 (2.26-188.5); P=0.007]. CONCLUSION: Home nebulizers, if not cleaned properly as per set protocols, may get colonized with potentially harmful bacteria. There is a need to increase awareness about their proper use amongst parents of children with recurrent wheeze.

Acute disseminated encephalomyelitis due to a zoonotic co-infection.

Acute disseminated encephalomyelitis (ADEM) is a post-infectious, autoimmune, demyelinating neurological illness, usually attributed to infection with viruses. We describe a case of ADEM occurring in a child with Leptospira-Brucella co-infection. The 12-year-old girl developed a biphasic febrile illness with encephalopathy. On evaluation, she was found to have serological evidence of Brucella and Leptospira infections. Persistence of neurological symptoms after initiating treatment for the co-infection led us to do a magnetic resonance imaging scan of the brain which showed typical findings suggestive of ADEM. Patient responded appropriately to treatment of ADEM with glucocorticoids. The high prevalence of these zoonotic infections in developing countries, and the risk that these may lead to ADEM highlights the importance of detailed evaluation of such cases for proper treatment and better outcomes.

ADEM is a serious neurological disease which occurs as an uncommon complication of certain infections that lead to formation of antibodies which attack the cells of the nervous system. It usually occurs after viral infections, but we came across a 12-year-old girl with ADEM who tested positive for simultaneous infection with two different micro-organisms, both not viruses. These microbes, called Leptospira and Brucella, are common in developing countries and usually lead to infection in individuals in close contact with animals, or with consumption of infected, unpasteurized animal products. Neurological symptoms are uncommon in both infections. However, our case highlights that both infections can occur together and lead to serious neurological illness which needs proper evaluation and a different kind of treatment so that patient has better recovery.

A Comprehensive Review of Trichosporon spp.: An Invasive and Emerging Fungus.

Trichosporon species are basidiomycetous yeast-like organisms found ubiquitous in nature. They are increasingly been recognized as opportunistic pathogens capable of causing life-threatening invasive diseases (trichosporonosis), especially in immuno-suppressed patients and rarely in immuno-competent patients too. Earlier multiple members of the genus Trichosporon were clubbed together as T. beigelli but after the advent of molecular techniques, more than 50 different subspecies and around 16 different strains causing human diseases are reported. It is known to cause a wide range of diseases, from superficial to probable and proven invasive diseases to summer hypersensitivity. The ability of Trichosporon strains to form biofilms on implanted devices, glucuronoxylomannan in their cell walls, and production of proteases and lipases lead to the virulence of this genus. This ubiquitous fungus exhibits intrinsic resistance to echinocandins, variable minimum inhibitory concentrations (MIC) for amphotericin B, and moderate susceptibility to fluconazole and Itraconazole, which are the commonly used anti-fungal agents for any invasive fungal infections which lead to the re-emergence of this notorious yet neglected pathogen and hence the reports of breakthrough infections among patients receiving these antifungals. This review is to understand the epidemiological, clinical details, and antifungal susceptibility pattern of various Trichosporon infections and it highlights the importance of early detection and treatment for this emerging yeast and also will add to the ongoing surveillance for the anti-fungal susceptibility pattern for this fungus.

Epidemiological profile and antifungal susceptibility pattern of Trichosporon species in a tertiary care hospital in Chandigarh, India.

BACKGROUND AND PURPOSE: Trichosporon species are ubiquitous in nature which are associated with fatal opportunistic invasive infections, especially in immunocompromised patients. The present study aimed to evaluate the epidemiological and clinical details, as well as the antifungal susceptibility pattern of the patients with Trichosporon infections. MATERIALS AND METHODS: In total, 50 clinical isolates of Trichosporon species from various samples were included in this study. The samples were isolated over a period of 18 months from patients in a tertiary hospital in North India. The isolates were characterised phenotypically with Vitek MS (bioMérieux, France). Trichosporon spp. were isolated from urine (30%), nail (30%), tissue (16%), pleural fluid (14%), and sputum (5%). In total, majority of the isolates were of Trichosporon asahii (92%), followed by Trichosporon mucoides (6%), and Trichosporon ovoides (2%). It is noteworthy that most of the reported cases were from intensive care unit (34%). RESULTS: Intravenous catheters, antibiotics, and antifungal uptake were significantly associated risk factors with Trichosporon infection. All invasive isolates were observed to be resistant in vitro to caspofungin and exhibited high minimum inhibitory concentration (MIC) values against amphotericin B, fluconazole, and 5-flucytosine. The MICs for voriconazole and posaconazole were low. CONCLUSION: Trichosporonosis is being increasingly reported all around the world, including India. The results of this study highlighted the importance of early detection and treatment for this emerging yeast and also added to the ongoing surveillance for the antifungal susuceptibility pattern for this fungus.

Spectrum of Multiorgan Dysfunction in Scrub Typhus Infection.

OBJECTIVES: We planned this study to determine the clinical spectrum and compare incidence of multiorgan dysfunction in children hospitalized with scrub typhus with other etiologies of tropical fever. DESIGN: Prospective cohort study. SETTING: Pediatric emergency and PICU services of a university teaching hospital situated in the sub-Himalayan region. PATIENT: Children aged 2 months to 14 years with acute undifferentiated fever of more than 5 days. INTERVENTIONS: Detailed fever workup was performed in all children. We compared scrub typhus IgM positive children (cases) with remaining febrile children who were negative for scrub IgM assay (controls) for mortality and morbidity. MAIN RESULTS: We enrolled 224 febrile children; 76 children (34%) were positive for scrub typhus IgM ELISA. Scrub typhus group had a significantly higher incidence of multiorgan dysfunction [OR 3.5 (95% CI 2.0-6.3); p < 0.001] as compared to non-scrub typhus group requiring supportive care. The incidence of altered sensorium [OR 8.8 (95% CI 3.1-24.9)], seizures [OR 3.0 (95% CI 1.1-8.3)], acute respiratory distress syndrome [OR 17.1 (95% CI 2.1-140.1)], acute renal failure (5% vs. 0%), meningitis [OR 6.2 (95% CI 1.2-31.6)], thrombocytopenia [OR 2.8 (95% CI 1.5-5.1)], transaminitis [OR 2.7 (95% CI 1.6-4.8)], requirement of oxygen [OR 17.8 (95% CI 4.0-80.3)], positive pressure support [OR 3.7 (95% CI 1.2-10.5)] and shock requiring inotropes [OR 3.0 (95% CI 1.3-6.7)] was significantly higher in scrub typhus group as compared to the non-scrub typhus group (Table 1). CONCLUSIONS: Pediatric scrub typhus who were hospitalized had severe systemic manifestations when compared to other causes of fever.