Antifungal Sensitivity Testing Research Articles

Understanding chromoblastomycosis in Sri Lanka; insights into fungal aetiology and antifungal susceptibility patterns

Introduction: Chromoblastomycosis is a chronic disfiguring fungal infection of the subcutaneous tissue found mainly in the socio-economically marginalized population in the tropical and sub-tropical regions, characterized by the presence of sclerotic cells. This study was done with the aim of identifying the antifungal sensitivity patterns of the causative agents of chromoblastomycosis in Sri Lanka from 1992 to 2019.Methodology: Study was done on the stored isolates received from patients with chromoblastomycosis since 1992 to 2019, to the Mycology Reference Laboratory of Medical Research Institute, Colombo. Isolates were revived by subculture on Potato dextrose agar. Identification was done phenotypically. Antifungal sensitivity testing was performed by E test method. Available demographic and clinical data were analysed.Results: Of the 74 isolates, 71 (96%) were caused by Fonsecaea pedrosoi. Mean MIC for the antifungals tested were lowest for posaconazole 0.015 μg/ml followed by itraconazole 0.225 μg/ml, and voriconazole 0.768 μg/ml while amphotericin B demonstrated complete resistance with mean MIC of 30.591 μg/ml. No statistically significant differences were noted on the antifungal sensitivity of the isolates from 1992 to 2004 and 2005 to 2019.Conclusions: Fonsecaea pedrosoi was the commonest causative agent causing chromoblastomycosis in the studied population. Posaconazole demonstrated the lowest MIC values for the tested isolates followed by itraconazole and voriconazole. Amphotericin B demonstrated very high MIC values which were likely to result in clinical failure upon treatment. MIC values demonstrated minimal fluctuations throughout the tested time.

Molecular Detection of Candida aaseri in Oral Cavity of Immunocompromised Patients

The impaired human immune system, resulting from diabetes mellitus, can lead to the transition of Candida from a commensal to a pathogenic status, causing oral fungal infections. Candida aaseri is a dimorphic yeast with lipolytic activity which has not been previously reported in clinical infections. This study reports an extremely rare oral fungal infection associated with C. aaseri in a patient with type II diabetes mellitus. The patient, a 42-year-old female with diabetes mellitus type II from Basrah, Iraq, was and identified with a fungal oral infection. Swab samples were collected from the patient's oral cavity for microbial investigation and cultured on Sabouraud Dextrose Agar (SDA) at 37°C for 48 hours. The results revealed pure yeast growth. Blue colonies were observed on CHROMagar Candida. The genomic DNA of the isolated yeast was extracted for molecular-level diagnosis, utilizing the mRNA gene's internal transcribed spacer (ITS) region. C. aaseri was the sole yeast species identified in this clinical case. Anti-fungal sensitivity tests indicated that C. aaseri was sensitive to Nystatin, Posaconazole, and Fluconazole while exhibiting significant resistance to Colistin Sulfate. This study represents the first global report of isolating C. aaseri from an oral infection in an immunocompromised patient.

Clinicomycological Study of Dermatophyte Infections in Patients Attending Dermatology Outpatient Department in a Tertiary Care Hospital

Context: Dermatophytosis is a fungal disease characterized by the infection of skin, hair, and nails. There is an upsurge of complicated superficial dermatophytosis and an increase in the incidence of chronic, relapsing, recurrent cases in India that are also often unresponsive to conventional doses of drugs. Aims: The aim of this study was to study the prevalence of dermatophyte species in the particular study area and it is susceptibility to itraconazole, terbinafine, and fluconazole. Settings and Design: All dermatophytes isolates obtained were subjected to 10% KOH mount, fungal culture, and antifungal sensitivity testing for fluconazole, itraconazole, and terbinafine. Materials and Methods: Newly diagnosed cases of dermatophytosis without treatment for the past 3 months were included in the study. KOH preparation was observed and subjected to fungal culture. Then, the susceptibility of the dermatophytes to oral antifungals (itraconazole, terbinafine, and fluconazole) was done. Results: Out of 28 patients with tinea, 14 males and 12 females were tested KOH positive. Out of them, 23 patients showed positive for dermatophyte growth. In them, 15 isolates were Trichophyton mentagrophytes and 8 isolates were Trichophyton rubrum. Antifungal drug susceptibility testing data indicated that, for the two species, MIC of itraconazole ranged between 0.03 and 16 μg/ml, terbinafine between 0.03 and 16 μg/ml, and fluconazole between 0.25 and 64 μg/ml. Conclusions: The study provided information regarding the prevalent species of dermatophyte in our study area and it is high sensitivity to itraconazole compared to terbinafine and the lowest for fluconazole. This gives us guidance as to which antifungal to use empirically in our clinical practice to treat tinea infection.

Pan-Indian Clinical Registry of Invasive Fungal Infections Among Patients in the Intensive Care Unit: Protocol for a Multicentric Prospective Study.

Fungal infections are now a great public health threat, especially in those with underlying risk factors such as neutropenia, diabetes, high-dose steroid treatment, cancer chemotherapy, prolonged intensive care unit stay, and so on, which can lead to mycoses with higher mortality rates. The rates of these infections have been steadily increasing over the past 2 decades due to the increasing population of patients who are immunocompromised. However, the data regarding the exact burden of such infection are still not available from India. Therefore, this registry was initiated to collate systematic data on invasive fungal infections (IFIs) across the country. The primary aim of this study is to create a multicenter digital clinical registry and monitor trends of IFIs and emerging fungal diseases, as well as early signals of any potential fungal outbreak in any region. The registry will also capture information on the antifungal resistance patterns and the contribution of fungal infections on overall morbidity and inpatient mortality across various conditions. This multicenter, prospective, noninterventional observational study will be conducted by the Indian Council of Medical Research through a web-based data collection method from 8 Advanced Mycology Diagnostic and Research Centers across the country. Data on age, gender, clinical signs and symptoms, date of admission, date of discharge or death, diagnostic tests performed, identified pathogen details, antifungal susceptibility testing, outcome, and so on will be obtained from hospital records. Descriptive and multivariate statistical methods will be applied to investigate clinical manifestations, risk variables, and treatment outcomes. These Advanced Mycology Diagnostic and Research Centers are expected to find the hidden cases of fungal infections in the intensive care unit setting. The study will facilitate the enhancement of the precision of fungal infection diagnosis and prompt treatment modalities in response to antifungal drug sensitivity tests. This registry will improve our understanding of IFIs, support evidence-based clinical decision-making ability, and encourage public health policies and actions. Fungal diseases are a neglected public health problem. Fewer diagnostic facilities, scanty published data, and increased vulnerable patient groups make the situation worse. This is the first systematic clinical registry of IFIs in India. Data generated from this registry will increase our understanding related to the diagnosis, treatment, and prevention of fungal diseases in India by addressing pertinent gaps in mycology. This initiative will ensure a visible impact on public health in the country. DERR1-10.2196/54672.

Observation of a Bone Invasion Model of Aspergillus fumigatus In Vitro and Analysis of the Antifungal Susceptibility.

Recently, the prevalence of invasive fungal infections has been on the rise, and one of the prevalent symptoms frequently observed is bone deterioration and bone loss. Using an in vitro model we studied how Aspergillus fumigatus invades the bone. Pathological analysis was then employed to observe the structure and distinctive features of the invading fungal elements within the bone invasion model. Meanwhile, the antifungal effects of itraconazole, voriconazole, posaconazole, and amphotericin B were evaluated. The pathological findings showed that in the experimental group, fungal spores and hyphae invaded the bone tissue or were observed growing in the vicinity of the bone edge tissues, as indicated by both HE and PAS staining. In contrast, no fungal elements were observed in the control group, indicating that the in vitro bone invasion model of A. fumigatus was successfully constructed. Furthermore, the findings from the antifungal sensitivity test demonstrated that the lowest effective concentrations of antifungal drugs against the bone invasion model were as follows: 4μg/ml for itraconazole, 0.5μg/ml for voriconazole, 2μg/ml for posaconazole, and 2μg/ml for amphotericin B. The successful construction of the bone invasion model of A. fumigatus has provided a solid basis for future investigations into the mechanisms underlying A. fumigatus bone invasion and the study of its virulence factors. Utilizing bone models is of utmost importance in advancing the development of novel antifungal treatment approaches, as well as in effectively preventing and treating fungal bone invasion and osteolytic diseases.

Etiology and Antifungal Sensitivity Test in Otomycosis Caused by Candida Sp

Otomycosis is a common fungal infection of the external auditory meatus frequently diagnosed in otolaryngology outpatient clinics. Resistance to antifungals is currently a significant concern, with intrinsic and acquired resistance increasing among isolates that cause fungal infections. The purpose of this research was to identify Candida species causing otomycosis and determine the pattern of antifungal susceptibility among these Candida species. A prospective study was conducted in the Margono Soekarjo General Hospital and Department of Microbiology, Faculty of Medicine, Universitas Jenderal Soedirman Purwokerto, Indonesia, from April – September 2022. Forty-seven (47) clinical samples of otomycosis were collected from 41 patients and then isolated bedside on fungal culture media and was prepared on an object glass for direct microscopic examination of the specimens. Fungal identification was performed using 10% potassium hydroxide (KOH) to observe fungal elements. Samples were cultured on Saboraud dextrose agar (SDA) media with chloramphenicol and Czapek dox agar. The Germ Tube Test was used to identify Candida while yeast-specific identification and antifungal susceptibility assay using a rapid commercial kit was applied for specific identification of the fungus. Antifungal susceptibility patterns were obtained using the Integral System Yeast Plus (ISYP) media pack. Candida parapsilosis was the most prevalent Candida species discovered in this study, accounting for approximately 41.66%, which was followed by Candida tropicalis (25%) and Candida krusei (12.5%), whereas Candida albicans only accounted for 4.1% of the specimens. All Candida species were sensitive to flucitosine and ketoconazole, whereas the voriconazole sensitivity rate reached 96%. This study concludes that Candida parapsilosis is the most prevalent species of Candida in otomycosis, and that all Candida species are sensitive to Flucitosine, ketoconazole, and voriconazole.

Phytochemical Screening and Antifungal Evaluation of Leaf Extracts of Nephrolepis undulata Afzel. Ex Sw. (Nephrolepidaceae) against selected Fungi

Nephrolepis undulata (sword fern) is a medicinal plant used traditionally for treatment of various ailments including stomach ache, infections and cough. Therefore, the objective of this study is to undertake a phytochemical screening and antifungal activity evaluation of leaf extracts of Nephrolepis undulata Afzel. Ex Sw. (Nephrolepidaceae) against selected fungi (Aspergilus fumigatus, Rhodotorula mucilagenosa, Trichophyton rubrum, Mucor indicus, Candida parapsilosis and Cryptococcus neoformans) using appropriate methods. Phytochemical screening was done using standard methods of analyses while antifungal evaluation of the extracts was done using the agar diffusion method. Phytochemical screening of the extracts revealed the presence of alkaloids, anthracene derivative, carbohydrate, flavonoid and saponin in the methanol, chloroform and petroleum ether extracts. Tannins was present in the methanol extract but absent in chloroform and petroleum ether extracts. The antifungal sensitivity test revealed that the extracts had concentration dependent inhibitory activities against the clinical isolates used in the study; with the methanol extract having the best and broadest spectrum of activity. It had the lowest Minimum Inhibitory Concentration (MIC) of 0.3 mg/mL against A. fumigatus. However, its activity was only next to Ketoconazole, the reference drug. The extracts contained active phytochemicals which may be responsible for the varied antifungal actions of the extracts. These positive finding validates the ethnomedicinal application of the plant in the management of various ailments and further demonstrates the applicability of the plant as a possible raw material, in the pharmaceutical industry.

Prevalence of Dermatophytic Infections and Their Antifungal Susceptibility Pattern in the Rajshahi Region

Background: Dermatophytes are the most significant fungi because of their widespread involvement in the population and their prevalence worldwide. Bangladesh's hot and humid climate, overcrowded population, poverty, malnutrition, and ignorance make dermatophytosis a common cutaneous infection. These infections occur in both healthy and immunocompromised patients. Dermatophytes are responsible for most cutaneous fungal infections, and the estimated lifetime risk of acquiring a dermatophyte infection is 10-20%.
 Objective: To isolate and identify different dermatophytes and their antifungal susceptibility pattern in the Rajshahi region. Materials and Methods: Different clinical samples (e.g., skin scrapings, nail clipping, and hair plucking) were collected under aseptic precautions. The isolation and identification of dermatophytes were performed through a microscopic examination using 10% KOH mount, mycological culture, and species identification by lactophenol cotton blue mount from positive culture. In addition, all dermatophytes isolates were subjected to antifungal susceptibility testing using the agar-based disk diffusion method in Mueller Hinton agar media.
 Results: Out of 171 samples, Trichophyton rubrum was the predominant dermatophyte species with 76(71.7%), followed by T.mentagrophyte were 15(14.2%), E. floccosum were 12(11.3%), and M. canis were 03(2.8%). voriconazole, clotrimazole, and itraconazole were more effective drugs. Griseofulvin was the least effective drug, followed by fluconazole.
 Conclusion: This study indicates dermatophytosis is a common skin disease in northern Bangladesh. Due to the increasing trend of antifungal drug resistance among dermatophytes, treatment should be based on antifungal sensitivity testing.
 TAJ 2022; 36: No-1: 119-125

Clinico-Mycological Profile of Recurrent Dermatophytosis with Drug Sensitivity in a Tertiary Care Center in Southern Odisha

Background: Recurrent dermatophytosis is a pressing problem worldwide due to inadequate treatment, drug resistance, and indiscriminate use of topical steroids. This scenario increases the need for the study of risk factors and antifungal susceptibility testing. There is a paucity of information on this subject. Objective: We aimed to find the clinico-mycological pattern of recurrent dermatophytic infections, the factors responsible for recurrence and determine the sensitivity pattern of isolates against four antifungals (fluconazole, itraconazole, terbinafine, and amphotericin B). Materials and Methods: Patients with recurrent dermatophytosis attending the outpatient Department of Our Tertiary Care Institute were enrolled in this study. A detailed history was taken; clinical examination was done; samples were collected for mycological examinations, and in-vitro antifungal sensitivity testing was done by broth microdilution method as per Clinical and Laboratory Standard Institute M38-A guidelines. Results: One hundred and twenty-eight patients were included in the study with a male:female ratio of 1.38:1. The most commonly affected age group was 31–40 years. Tinea corporis with tinea cruris was the most common type of clinical presentation. Potassium hydroxide mount was positive in 53 samples and culture was positive in 59 samples. Trichophyton rubrum was the predominant species isolated, followed by Trichophyton mentagrophytes. On in-vitro antifungal susceptibility testing, itraconazole and amphotericin B had the lowest minimum inhibitory concentrations (MIC), followed by terbinafine. Fluconazole had the highest MIC among the drugs tested. Conclusion: Topical corticosteroid abuse, lack of personal hygiene, and habit of sharing clothes were seen in many patients with recalcitrant dermatophytosis. T. rubrum was the most common isolated species. Antifungal susceptibility testing revealed itraconazole and amphotericin B to have the lowest MIC value and fluconazole to have the highest MIC value.

Prevalence and Antifungal Susceptibility of Sporothrix species in Guangzhou, Southern China.

Sporotrichosis is a subcutaneous and chronic infection caused by traumatic inoculation of pathogenic sporothrix species, usually infecting the skins and subcutaneous tissues of humans and animals. However, the lack of epidemiological data required further molecular identification to describe the distribution of this fungus in our region. In this study, forty-eight clinical sporothrix isolated from Sun Yat-Sen Memorial Hospital were classified, and the susceptibility of each strain to seven antifungal agents was determined. Forty strains of S. globosa and eight strains of S. shenkshii were identified via colony morphology and PCR sequencing of calmodulin gene. Antifungal susceptibility tests of the mycelial phase in vitro showed terbinafine (TRB) and luliconazole (LULI) were the most effective, followed by itraconazole (ITZ) and amphotericin B (AMB). By contrast, voriconazole (VCZ), 5-flucytosine (5FC) and fluconazole (FCZ) have low efficacy with high MIC. Our results showed a predominantly S. globosa infection trend in southern China. Simultaneously, sporothrix is sensitive to TRB, LULI, ITZ and AMB whereas resistant to FCZ. This study firstly reports antifungal sensitivity test in vitro and epidemiological correlation analysis of sporothrix in southern China, and also the first time to find that sporothrix is sensitive to LULI.

Succinylation modification provides new insights for the treatment of immunocompromised individuals with drug-resistant Aspergillus fumigatus infection.

Invasive Aspergillus fumigatus infection poses a serious threat to global human health, especially to immunocompromised individuals. Currently, triazole drugs are the most commonly used antifungals for aspergillosis. However, owing to the emergence of drug-resistant strains, the effect of triazole drugs is greatly restricted, resulting in a mortality rate as high as 80%. Succinylation, a novel post-translational modification, is attracting increasing interest, although its biological function in triazole resistance remains unclear. In this study, we initiated the screening of lysine succinylation in A. fumigatus. We discovered that some of the succinylation sites differed significantly among strains with unequal itraconazole (ITR) resistance. Bioinformatics analysis showed that the succinylated proteins are involved in a broad range of cellular functions with diverse subcellular localizations, the most notable of which is cell metabolism. Further antifungal sensitivity tests confirmed the synergistic fungicidal effects of dessuccinylase inhibitor nicotinamide (NAM) on ITR-resistant A. fumigatus. In vivo experiments revealed that treatment with NAM alone or in combination with ITR significantly increased the survival of neutropenic mice infected with A. fumigatus. In vitro experiments showed that NAM enhanced the killing effect of THP-1 macrophages on A. fumigatus conidia. Our results suggest that lysine succinylation plays an indispensable role in ITR resistance of A. fumigatus. Dessuccinylase inhibitor NAM alone or in combination with ITR exerted good effects against A. fumigatus infection in terms of synergistic fungicidal effect and enhancing macrophage killing effect. These results provide mechanistic insights that will aid in the treatment of ITR-resistant fungal infections.

Mitochondrial nanomotion measured by optical microscopy.

Nanometric scale size oscillations seem to be a fundamental feature of all living organisms on Earth. Their detection usually requires complex and very sensitive devices. However, some recent studies demonstrated that very simple optical microscopes and dedicated image processing software can also fulfill this task. This novel technique, termed as optical nanomotion detection (ONMD), was recently successfully used on yeast cells to conduct rapid antifungal sensitivity tests. In this study, we demonstrate that the ONMD method can monitor motile sub-cellular organelles, such as mitochondria. Here, mitochondrial isolates (from HEK 293 T and Jurkat cells) undergo predictable motility when viewed by ONMD and triggered by mitochondrial toxins, citric acid intermediates, and dietary and bacterial fermentation products (short-chain fatty acids) at various doses and durations. The technique has superior advantages compared to classical methods since it is rapid, possesses a single organelle sensitivity, and is label- and attachment-free.

PHENOTYPIC CHARACTERIZATION OF SOME PATHOGENIC FUNGI ISOLATED FROM POULTRY AND THEIR SURROUNDINGS IN EL–GHARBIA GOVERNORATE, EGYPT

Poultry production is affected by several fungal diseases. Such fungal infection occurs in poultry farms via using a moldy litter, or ingestion of contaminated drinking water or moldy ration. In this study, a total of 210 birds with a history of respiratory distress of different breeds were collected randomly from sporadic different private farms and hatcheries in El–Gharbia Governorate, Egypt. The birds were sacrificed, then a total of 1050 tissue specimens from lung, air sacs, liver, crop and trachea were collected. In addition, 40 samples of poultry ration, 14 bedding materials, 4 air samples and 29 water samples were also collected. The collected samples were cultured on Sabouraud’s agar plates. Macromorphological and micromorphological fungal examinations were performed for phenotypic characterization. Histopathological examinations were also performed using with hematoxylin and eosin stains. Antifungal sensitivity testing was screened using Mueller’s Hinton Agar for studying the susceptibility of the recovered fungal isolates to the most commonly used antifungal drugs in Egypt, namely amphotericin B, clotrimazole, fluconazole, itraconazole, ketoconazole, and nystatin. The obtained results demonstrated that mold isolation was the highest in the collected samples from birds at 36.84%, followed by drinking water (31.57%). The highest incidence of mold isolation was recorded at the lungs of broilers and baladi birds followed by the air sacs. While in saso birds, the highest incidence was at the air sac. Collectively, 97 mold strains were identified from the lung, 74 from the air sacs, 30 from the liver, 61 from the trachea, and 44 from the crop. In addition, 19 mold isolates were recovered from the bird surroundings. Aspergillus niger as well as Penicillium chrysogenum were recovered and showed resistance to ketoconazole, while Cladosporium perangustum was resistant to fluconazole. All of the isolated molds were sensitive to itraconazole and nystatin except A. flavus that was resistant to nystatin. All Aspergillus spp. were resistant to fluconazole except A. niger. In conclusion, Aspergillus spp. was the most associated mold with poultry species and their surroundings in Egypt farms. Itraconazole and nystatin could be applied as proper antifungal drugs the control of for Aspergillus infection in birds.

Case report: candidemia in a child with cystic fibrosis with fatal course

In the present study we report a case of candidemia in an infant with cystic fibrosis admitted to a Pediatric Intensive Care Unit (PICU) with a fatal course. Six-month-old male patient with a history of productive cough and persistent fever. He was admitted to the PICU of a public hospital in the city of Recife, PE, Brazil, with a diagnosis of dyspnea and cystic fibrosis, with a respiratory infection. Antibiotics were administered, however without success, peripheral blood and transcatheter blood cultures were requested, the isolated agents were Staphylococcus epidermidis and Pseudomonas aeruginosa, respectively. After seven days, the patient presented lesions on the genitalia and persistent fever. Blood and genital lesions were collected, the samples were identified as Candida albicans through the classical taxonomy and the VITEK 120 automated system. The antifungal sensitivity test followed the protocol by the broth microdilution method (CLSI- Clinical and Laboratory Standard Institute, 2008b). The isolates from blood samples were sensitive to amphotericin B with a Minimal Inhibitory Concentration (MIC) of 0.03 µg/mL, 0.12 µg/mL for anidulafungin and 0.25 µg/mL for voriconazole, exhibiting resistance to fluconazole with MIC of 64 µg/mL. C. albicans isolated from the genitalia lesion was sensitive to all the drugs used. The patient was administered nystatin 4 times a day and amphotericin B. The patient improved from the genital lesion and fever. However, after three days, the patient presented cardiac and respiratory deficit that led to cardiac arrest, leading to death.

A Case of Tinea pseudoimbricata Caused by Microsporum Audouinii

Tinea pseudoimbricata has morphological similarity to Tinea imbricata which is characterized by multiple concentric rings, but it is caused by dermatophytes other than Trichophyton Concentricum. We report a case of tinea pseudoimbricata caused by Microsporum Audounii that presented with multiple concentric scaly annular erythema. The most common clinical presentation of pseudoimbricata was tinea corporis followed by Tinea cruris. However, tinea imbricate-like lesions are very rare. Tinea pseudoimbricata is considered a unique sub-type of Tinea incognito which caused by inappropriate use of topical steroid. Both Fungal culture and microscopic findings confirmed Microsporum Audouinii infection in this case. The patient was treated with oral terbinafine according to culture and antifungal sensitivity testing at a dose of 250 mg/day for 8 weeks.

Correlation Between Drug Resistance and Virulence of Candida Isolates from Patients with Candidiasis.

This article aims to provide a theoretical basis for new or adjuvant strategies to facilitate the early diagnosis and treatment of candidiasis and to determine if drug-resistant Candida would affect virulence. Our strains were collected from patients diagnosed with candidiasis in our hospital. The strains were identified by MALDI-TOF system and ITS sequencing. Antifungal sensitivity testing in vitro was performed to evaluate susceptibility of these isolates to current widely used antifungal drugs. The Galleria mellonella larvae model infected by Candida spp. was used to compare the virulence of drug-resistant and susceptible Candida spp. A total of 206 Candida strains were collected from clinical specimens. Candida albicans was the most common species among them, and was predominantly isolated from male patients aged over 40 years in ICU environments suffering from pulmonary and/or cerebral conditions. The accuracy rate of MALDI TOF-MS identification was 92.72% when compared with ITS sequencing as the standard method. Most Candida species, except for C. tropicalis which showed high resistance to micafungin, showed high susceptibilities to voriconazole, itraconazole, amphotericin B and micafungin but were highly resistant to terbinafine. For each specific Candida species, the G. mellonella larvae model revealed that the virulence of drug-resistant Candida isolates did not markedly differ from that of the drug-susceptible isolates, however, the virulence was dose-dependent on inoculated fungal cells in this model. The possibility of Candida infection should not be neglected in patients at critical care hospital settings and C. albicans is the most common causative agent. MALDI-TOF MS has the advantages of rapidity and high accuracy, and should be a preferred method for identification of Candida spp. in a clinical laboratory. Voriconazole, itraconazole, amphotericin B and micafungin can still be recommended as the first line antifungals to treat candidiasis.

Evaluation of antifungal susceptibility and clinical characteristics in fungal keratitis in a tertiary care center in North India.

To study the antifungal susceptibility of common corneal pathogenic fungi to antifungal agents in the North Indian population. Prospective study of the antifungal sensitivity testing (natamycin, amphotericin B, voriconazole, itraconazole, fluconazole, posaconazole, caspofungin, micafungin) of fungal isolates from 50 cases of culture positive fungal keratitis by using E test method. Details noted included demographic data, visual acuity, clinical details, grade of keratitis, healing time, and success in medical management. Of 50 patients with fungal keratitis (mean age: 40.28 ± 16.77 years), 12 eyes healed within 3 weeks, 14 had a delayed healing response, and 24 had chronic keratitis. Among the 15 cases of Fusarium isolates, 93.3% were sensitive to natamycin, while 40% to amphotericin B; 66.6% to voriconazole, 13.4% to itraconazole and fluconazole each. 80% of Fusarium cases (n = 12) showed susceptibility to posaconazole. Among Aspergillus flavus isolates, 53.4% (n = 8) were sensitive to natamycin, with only 40% (n = 7) showing sensitivity to amphotericin B and good susceptibility to azoles. MIC against susceptible Fusarium spp. for natamycin was 3-16 μg/mL, amphotericin B: 1-8 μg/mL, voriconazole: 0.5-1.5 μg/mL, itraconazole: 0.5-12 μg/mL, posaconazole: 0.094-1.5 μg/mL. MIC against Aspergillus flavus was natamycin: 8-32 μg/mL, amphotericin B: 0.5-16 μg/mL, voriconazole: 0.025-4 μg/mL, itraconazole: 0.125-8 μg/mL, posaconazole: 0.047-0.25 μg/mL; against Aspergillus niger isolates, to natamycin was 6 μg/mL (n=1), amphotericin B 8-12 μg/mL (n = 3), voriconazole: 0.125-0.19 μg/mL (n = 3), itraconazole: 0.38-0.75 μg/mL, posaconazole: 0.064-0.19 μg/mL and against Aspergillus fumigatus (n = 1), was natamycin4 μg/mL, amphotericin B - 8 μg/mL, voriconazole 0.25 μg/mL, itraconazole 1 μg/mL, and posaconazole 0.19 μg/mL. MIC against susceptible Acremonium spp. for natamycin was 1.5-16 μg/mL, amphotericin B: 0.5-8 μg/mL, voriconazole: 0.19-3 μg/mL, itraconazole: 0.125 μg/mL, posaconazole: 0.125-0.5 μg/mL and against susceptible Curvularia was natamycin 0.75-4 μg/mL, amphotericin B 0.5-1 μg/mL, voriconazole 0.125-0.19 μg/mL, itraconazole 0.047-0.094 μg/mL, posaconazole 0.047-0.094 μg/mL. MIC against Mucor spp.+ Rhizopus spp. (n = 1) was natamycin: 8 μg/mL, amphotericin B: 0.75 μg/mL, posaconazole: 1.5 μg/mL. MIC against of Alternaria (n = 1) was voriconazole: 0.19 μg/mL, posaconazole: 0.094 μg/mL. MIC against Penicillium (n=1) was natamycin: 8 μg/mL, voriconazole: 0.25 μg/mL, itraconazole: 0.5 μg/mL, and Posaconazole: 0.125 μg/mL. Our observations highlight the variations in susceptibility to antifungal agents. Posaconazole seems to be effective with low MIC against common corneal pathogenic fungal isolates.

An Overview of Candidemia Patients in Tertiary Hospital, Surabaya, Indonesia

Candidaemia is a bloodstream infection (BSIs) caused by Candida spp. common in hospitalized patients. Candidemia has a high mortality and morbidity. This study aimed to give an idea about incidence, patient characteristics, species and patterns of candida susceptibility, and risk factors in candidaemia patients at Dr. Soetomo Hospital from January to December 2020. This is descriptive research using an observational research design. This study used a medical record instrument for candidemia patients in the inpatient ward of Dr. Soetomo and data in the form of records of blood culture results at the Clinical Microbiology Unit of RSUD Dr. Soetomo Surabaya for the period January-December 2020. The observed variables include incidence, sex, age, the origin of hospitalization, species distribution, frequency of antifungal sensitivity testing, and risk factors for candidemia. In 12 months, 53 candidemia patients were treated at Dr. Soetomo Hospital. The incidence of candidemia is 1.58 cases per 1000 person-years. Most of them are male (64.15%) and aged less than <1 year (28,30). Most of the medical units at the time of the diagnosis were intensive care units and pediatric wards (28.30%). Candida albicans was the most commonly found species (33.96%), then Candida tropicalis (26.41%) and Candida parapsilosis (24.53%). Two Candida glabrata isolates (40%) were susceptible to caspofungin, and all isolates (100%) of Candida albicans, Candida parapsilosis, Candida rugosa, Candida lusitaniae, Candida tropicalis, Candida dubliniensis, and were susceptible to the antifungals tested. Patients who use the Central Venous Catheter (CVC) have the highest level of risk factor, followed by patients with diabetes mellitus.

P004 Neglected risk for of invasive candidiasis: s tudy of distribution, species differentiation and antifungal susceptibility pattern of Candidemia among patients with liver disease

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM ObjectivesPatients with liver disease (LD) are more predisposed to candidemia due to the dysfunctional Kupffer cells which fails to capture the circulating yeasts, thereby causing fungal dissemination. Candidemia is ten times more common among end-stage liver disease patients compared with other LD patients. Compared to those with abdominal candidiasis, ICU admission and mortality are found to be higher in liver transplant recipients (LTRs) with candidemia. Though a majority of Candida infection among LD patients are due to C. albicans, there has been an increasing prevalence of non-albicans isolates. This study was mainly aimed at species differentiation and antifungal susceptibility. Our objective was to study the pattern of Candida isolates from blood among patients with different LD states admitted to tertiary care Liver Hospital. To determine the distribution of Candidemia in patients with different liver disease states. To determine the various species of Candida isolated from blood in patients with liver diseases. To determine the anti-fungal susceptibility pattern of Candida isolates in blood in patients with liver disease. To determine the year-wise changing trends in Candida spp. isolated in blood and its anti-fungal susceptibility pattern in patients with liver disease.Materials and MethodsThis is a retrospective observational study conducted in the department of Microbiology at Institute of Liver and Biliary Sciences (ILBS), New Delhi. Ethical approval for this study (IEC/2021/84/NA06) was provided by the Institutional ethics committee/Institutional review board. A total of 118 LD patients with candidemia who were admitted to ILBS between January 2017 to December 2020 were included in the study. Clinical details of these patients were collected from the hospital information system. Patients with LD were divided into 4 groups—acute liver failure (ALF), acute on chronic liver failure (ACLF), chronic liver disease (CLD), and post-liver transplantation (Post-LT). Candida speciation as identified by VITEK® 2 (Biomérieux) and also its antifungal susceptibility pattern by broth microdilution.ResultsThe mean age of the 118 LD patients was 48.6 ± 14.9 years [± standard deviation (SD)]. Among them 107 (90.7%) were males and 11 (9.3%) were females. Among the 118 LD patients, 6 (5.1%) were ALF patients, 32 (27.1%) were ACLF patients, 66 (55.9%) were CLD patients and 14 (11.9%) were post-LT patients. The most common Candida species isolated was C. tropicalis (22.9%), followed by C. glabrata (17.8%), C. albicans (16.9%), C. parapsilosis (12.7%), C. auris (8.5%), C. krusei (6.8%) and other Candida species (14.4%). The other Candida species isolated were C. haemulonii, C. kefyr, C. lusitaniae, C.pelliculosa, C. duobushaemulonii, C. lipolytica, and C. rugosa. The sensitivity to fluconazole (55.1%), voriconazole (86.4%), caspofungin (82.2%), micafungin (97.5%), flucytosine (80.5%), amphotericin B (86.4%). Mortality was 66%.ConclusionOur study shows that chronic liver disease, acute liver failure and liver transplantation are important risk factors for invasive candidiasis. There was a rise in the non-albicans Candida, especially C. tropicalis and C. glabrata. There is rising antifungal resistance to azoles and can lead to therapy failure. Hence antifungal sensitivity testing is essential in patients with these neglected risk factors to prevent mortality.

P141 Study of prevalence, antifungal profile and predictors of mortality in chronic pulmonary Aspergillosis in a cohort of liver disease patient

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM ObjectivesChronic pulmonary aspergillosis (CPA) is a neglected cause of respiratory disease even in high-TB burden countries like India. It is a term that includes simple aspergilloma, chronic cavitary pulmonary aspergillosis which is the most common variant that presents with formation of lung cavities with or without an aspergilloma or nodules, and chronic fibrosing pulmonary aspergillosis. CPA is a chronic progressive infection that destroys lung tissue in non-immunocompromised patients. So, we studied the prevalence of CPA in liver disease patients with respect to spectrum of infections, fungus isolated, and outcome.MethodsA cohort of 2250 liver disease patients with isolation of Aspergillus in respiratory samples, was analyzed retrospectively for the diagnosis of CPA, from June 2018 to June 2020. The demographic characteristics, clinical, radiological, and laboratory objective and subjective variables including age, sex, previous pulmonary conditions, dyspnea score, quality of life, serum albumin, ESR, etc were assessed using Kaplan-Meier curves, log-rank tests, and Cox proportional hazards modeling. In samples of patients, retrospective review of time from likely onset of CPA to referral and cause of death were also investigated.ResultsCPA was diagnosed in 16% of the patients. Survival was 72%, at 1 year. Increased mortality was associated with chronic obstructive pulmonary disease (hazard ratio 2.57; P = .029), nontuberculous mycobacterial infection (2.04; P <.001), increasing age (1.03, P <.001), lower albumin (0.92, per g/L), underlying chronic liver disease (2.07; P <.001) were significantly associated with CPA. The Aspergillus species isolated were A. flavus (66%), A. fumigatus (31%), Aspergillus nidulans (1%), A. terreus (1%), and other Aspergillus species (1%). All the isolates were sensitive to echinocandins and amphotericin B. Azole-resistance was detected in 3% isolates. Multiple pulmonary lesions were more common (73%) than single cavitary lesion. Mortality was seen in 64% of cases.ConclusionCPA pulmonary aspergillosis is a common complication and cause of mortality in post-cavitary pulmonary disease. Several factors impact on mortality of CPA, and can be evaluated as tools to assess CPA prognosis. Underlying liver disease is an important predictor of mortality. As there is a rise in azole-resistance, appropriate diagnosis, and antifungal sensitivity testing should be done in all suspected cases. Early diagnosis and treatment can prevent mortality in CPA patients.