{"title":"P415 High positive and rapid detection of clinical urine samples of fungal infection based on modified calcium fluorescence","authors":"Biao Chen, Dongmei Shi","doi":"10.1093/mmy/myac072.P415","DOIUrl":"https://doi.org/10.1093/mmy/myac072.P415","url":null,"abstract":"Abstract Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM Successful management of urinary fungal infection depends upon the detection positive and efficiency. The aim of this study was to evaluate the detection positive and efficiency of modified calcium fluorescent white (m-CFW) staining in direct detection of Candida spp. in urine samples of patients with suspected fungal infection. We collected 100 clinical urine samples from different departments and analyzed the detection positive rate of the methods of culture, KOH, sequence, and modified CFW. The results indicated that the positive rate of the methods was 12%, 8%, 14%, and 15%, respectively. The positive rate of modified CFW staining was significantly higher than that of ordinary microscopic examination and fungal culture (P <.05). Modified CFW in the detection of fungi in urine can significantly improve the positive rate of fungi in clinical urine samples and shorten the detection time. It has a certain reference value for clinical diagnosis and medication.","PeriodicalId":18325,"journal":{"name":"Medical mycology journal","volume":"1 1","pages":""},"PeriodicalIF":1.0,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91119827","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"P004 Neglected risk for of invasive candidiasis: s tudy of distribution, species differentiation and antifungal susceptibility pattern of Candidemia among patients with liver disease","authors":"Pradeep Kumar, Mohd Ayyub, P. Kale","doi":"10.1093/mmy/myac072.P004","DOIUrl":"https://doi.org/10.1093/mmy/myac072.P004","url":null,"abstract":"Abstract Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM Objectives Patients 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 Methods This 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. Results The 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%), ","PeriodicalId":18325,"journal":{"name":"Medical mycology journal","volume":"1 1","pages":""},"PeriodicalIF":1.0,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91139614","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Bram Spruijtenburg, Theun de Groot, A. Chowdhary, J. Meis
{"title":"P293 Development of a multiplex PCR short tandem repeat typing scheme for Candida tropicalis","authors":"Bram Spruijtenburg, Theun de Groot, A. Chowdhary, J. Meis","doi":"10.1093/mmy/myac072.P293","DOIUrl":"https://doi.org/10.1093/mmy/myac072.P293","url":null,"abstract":"Abstract Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM Candida tropicalis is a clinically relevant yeast that causes candidemia in humans with a high mortality rate. The yeast primarily infects immunocompromised patients and causes outbreaks in health care facilities. Antifungal-resistant isolates have been reported. Here, we report a short tandem repeat (STR) typing scheme, for C. tropicalis to enable fast, cost-effective, and high-resolution genotyping. This novel typing approach was applied to 117 clinical isolates. For the development of the typing scheme six novel STR markers were selected, combined into two multiplex PCRs, and used to type 117 C. tropicalis isolates, resulting in the identification of 104 different genotypes. The outcome of STR typing of 10 isolates was then compared to single nucleotide polymorphism (SNP) calling from whole genome sequencing (WGS). Isolates with >111 SNPs were also differentiated by the STR assay. Two isolates that were identical according to SNP analysis were separated by STR typing in one marker. To test specificity, STR typing was applied to 15 related yeast species and we found no amplification of these targets. For reproducibility testing, two isolates were typed independently 5-times, which showed identical results in each experiment. In summary, we developed a reliable, rapid, and high-resolution STR genotyping for C. tropicalis, which was found to correlate well to SNP calling by WGS.","PeriodicalId":18325,"journal":{"name":"Medical mycology journal","volume":"9 1","pages":""},"PeriodicalIF":1.0,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91362771","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"P037 Study of magnitude and risk factors in patients with candidemia at a tertiary care hospital with speciation and antifungal susceptibility of pathogenic Candida isolates.","authors":"B. Chauhan, C. Kumar, S. Baveja","doi":"10.1093/mmy/myac072.P037","DOIUrl":"https://doi.org/10.1093/mmy/myac072.P037","url":null,"abstract":"Abstract Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM Objectives Nosocomial candidiasis is associated with a mortality rate of over 60% while the attributable mortality rate is 49%. The present study was to determine the magnitude and risk factors in patients with candidemia at a tertiary care hospital with speciation and antifungal susceptibility of pathogenic Candida isolates. Methods The present study was a prospective, cross-sectional, observational study, conducted at a tertiary care hospital for a period of 1 year after approval from Institutional ethics committee. It included a total of 150 patients of all age groups, admitted to hospital for ˃48 h and diagnosed as proven Candidemia with isolation of Candida species from at least two blood culture samples or from a clinically significant single blood culture sample. A thorough history and clinical characteristics of each patient was noted. Blood was collected and processed as per standard protocol. Pathogenic Candida species were identified and their antifungal susceptibility testing was performed by disk diffusion method as per the standard method. The antifungal discs used were fluconazole (25 μg), itraconazole (10 μg), voriconazole (1 μg), and amphotericin B (100 units). Results were analyzed statistically using SPSS statistics 20. Results Candida species was isolated as the pathogen in 24/150 (16%) of clinically suspected cases of candidemia. Candida species isolated were non-albican Candida (NAC) species, mainly C. glabrata 11/24 (45.83%) followed by C. parapsilosis 8/24 (33.33%), and C. tropicalis 5/24 (20.83%). Candida species was isolated as the pathogen, predominantly in patients of age group 0-10 years [15/24 (62.5%)]. Majority of Candida species were isolated from patients who had prolonged ICU stays. Among 24 patients of proven candidemia, 2 (8.33%) patients were from NICU, 10 (41.6%) from PICU, and 3 (12.5%) from MICU. Other important risk factors observed in the present study were, recent major abdominal surgery, malignancy, and mechanical ventilation, each accounting for 2/24 (8.33%) cases. The resistance pattern of isolates of Candida species to antifungals showed that C. glabrata showed 100% resistance to fluconazole, 63.6% to itraconazole, and 45.4% to voriconazole. C. tropicalis showed 80% resistance to fluconazole, 60% to itraconazole, and 40% to voriconazole. Candida parapsilosis showed 87.5% resistance to fluconazole, 62.5% to itraconazole, and 37.5% to voriconazole. All three isolated pathogenic Candida spp. showed 100% susceptibility to amphotericin B. Mortality observed in present study was 7/24 (29.7%). A total of 5/7 patients were from ICU. Conclusion Non-albican Candida (NAC) species, mainly C. glabrata, C. tropicalis and C. parapsilosis were the causative agent of candidemia, seen to predominantly affect 0-10 year age group. Infections caused by Candida species remain a significant problem in ICU. An increase in resistance to azoles is a challenge","PeriodicalId":18325,"journal":{"name":"Medical mycology journal","volume":"30 1","pages":""},"PeriodicalIF":1.0,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89174091","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"S1.3c Diversity and hybridization in Malassezia furfur","authors":"B. Theelen","doi":"10.1093/mmy/myac072.S1.3c","DOIUrl":"https://doi.org/10.1093/mmy/myac072.S1.3c","url":null,"abstract":"Abstract S1.3 Malassezia: genetics, genomics, and biology, September 21, 2022, 11:00 AM - 12:30 PM The Basidiomycetous yeast Malassezia is the most abundant fungal genus on healthy human skin but may also cause various skin disorders such as seborrheic dermatitis, dandruff, and pityriasis versicolor. In recent years, Malassezia has increasingly been implicated in health and disease beyond the skin: as an underestimated cause of Malassezia bloodstream infections (BSIs) in immunocompromised patients and neonates, associated with Crohn's disease, promoting pancreatic oncogenesis, and exacerbating cystic fibrosis. Malassezia furfur is the number one Malassezia BSI cause and is also implicated in many skin disorders. With these new discoveries of Malassezia’s impact on human health, the need for a better understanding of its evolution and pathobiology also became more pressing. Hybridization has been suggested as a biological mechanism of adaptation to new hosts, and may lead to increased pathogenicity. Many examples of major hybrid yeast pathogens exist, such as Candida albicans, C. orthopsilosis, C. metapsilosis, and multiple examples in the Cryptococcus gattii/Cryptococcus neoformans species complex. Here the multiple hybridization events of the Malassezia furfur species complex will be discussed. Two distinct hybridization events occurred between the same parental lineages, and these parental strains were originally also hybrids. The identification of a pseudobipolar mating system and the analysis of the mating-type loci provide evidence that sexual liaisons of mating compatible cells from these parental lineages led to a diploid/aneuploid state in the hybrid lineages. Sequence similarity percentages suggest that both parental lineages in fact are two different species. The genetic diversity of ca 300 strains belonging to this species complex is evaluated in relationship to host background and phenotype.","PeriodicalId":18325,"journal":{"name":"Medical mycology journal","volume":"17 1","pages":""},"PeriodicalIF":1.0,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89243907","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sunita Gupta, H. Kaur, Anup K. Ghosh, S. Rudramurthy, A. Chakrabarti
{"title":"P206 Phenotypic and molecular characterization of emerging Basidiomycete Schizophyllum commune isolated from clinical samples from India","authors":"Sunita Gupta, H. Kaur, Anup K. Ghosh, S. Rudramurthy, A. Chakrabarti","doi":"10.1093/mmy/myac072.P206","DOIUrl":"https://doi.org/10.1093/mmy/myac072.P206","url":null,"abstract":"Abstract Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM Objective Schizophyllum commune is an environmental basidiomycete capable of causing human infections. Its identification is difficult as often it produces sterile, cottony white colonies without any conidia formation. S. commune is characterized by clamp connections and hyphal spicules. Though it produces basidiocarps with basidiospores it is difficult to induce in vitro. Molecular techniques are essential to confirm its identification. Here, we present the description of Schizophyllum commune isolates collected at our center over the last 5 years. Methods All the isolates used in this study was received from various part of India and accessioned at the National culture collection of pathogenic fungi (NCCPF), Chandigarh India. Isolates were grown on Sabouraud dextrose agar and malt-extract agar (MEA) at 25°C for 5-7 days. Lacto phenol cotton blue mounts were prepared and a microscopic examination was done. For observing basidiocarps, MEA plates were incubated for 6-8 weeks. DNA extraction was done using the phenol-chloroform method and ITS region was amplified using pan-fungal primers followed by Sanger sequencing. Results Among total of 22 Schizophyllum commune cultures, 19 were isolated from lower respiratory samples, two from corneal scrapings, and one from the brain. Upon microscopic examination, the major identification feature of this fungi, i.e., clamp connection and spicules were observed only in 8 (36%) isolates. The remaining 14 (64%) were identified up to species level using ITS sequencing. Basidiocarps could be induced only in two isolates. Conclusion Although rarely involved in human disease, Schizophyllum commune is being isolated from the clinical specimens. As microscopic identification is difficult and needs expertise, molecular identification is required for early diagnosis and treatment.","PeriodicalId":18325,"journal":{"name":"Medical mycology journal","volume":"28 1","pages":""},"PeriodicalIF":1.0,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87406532","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
T. Shrimali, Anuradha Sharma, Anup K. Ghosh, A. Morya, V. Tak, Vijayalakshmi Nag
{"title":"P444 Detection of causative agents of infectious keratitis in patients from western rajasthan","authors":"T. Shrimali, Anuradha Sharma, Anup K. Ghosh, A. Morya, V. Tak, Vijayalakshmi Nag","doi":"10.1093/mmy/myac072.P444","DOIUrl":"https://doi.org/10.1093/mmy/myac072.P444","url":null,"abstract":"Abstract Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM Objective To determine the spectrum of causative agents, the related risk factors, and their association in patients of infectious keratitis. Methodology It was a prospective study conducted over a period of 18 months from August 2018 to January 2020, which included 100 patients attending the Ophthalmology OPD with features of keratitis. Ophthalmological examination was followed by corneal scrapings’ collection, which were subjected to culture, microscopy, and molecular diagnostic tools. Bacterial isolates were identified by conventional methods and MicroScan Walkaway system while the isolated fungi were identified conventionally. Pan-fungal primers were used to detect fungal elements directly from the sample. Results Out of 100, 41 cases were positive by culture, of which 32 (78.04%) had fungal and nine (21.95%) had bacterial keratitis. Fusarium spp. accounted for 33.33% of fungal and Pseudomonas aeruginosa accounted for 55.55% of the bacterial isolates. Fungal material was detected in 41% using pan fungal primers. Cases were maximally recorded during July-October. Traumatic history was present in 78% patients caused by vegetative matter (49%). A male preponderance (67%) was also observed. Four patients underwent evisceration in spite-of rigorous management. Conclusion Poor prognosis emphasizes the need for faster diagnostics, which can detect the causative agents from the clinical specimen itself, reinforcing the concept of clinical metagenomics.","PeriodicalId":18325,"journal":{"name":"Medical mycology journal","volume":"11 1","pages":""},"PeriodicalIF":1.0,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88902621","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"S1.5a Epidemiology of myotic keratitis in developing countries","authors":"P. Thomas","doi":"10.1093/mmy/myac072.s1.5a","DOIUrl":"https://doi.org/10.1093/mmy/myac072.s1.5a","url":null,"abstract":"Abstract S1.5 Mycotic keratitis, September 21, 2022, 11:00 AM - 12:30 PM Mycotic keratitis (corneal infection due to a fungal etiology) is a well-recognized ophthalmological emergency warranting rapid initiation of specific antifungal therapy. However, the magnitude of the problem of mycotic keratitis in the community, especially in the Indian subcontinent and the developing world, is, perhaps, less apparent. A minimal annual incidence estimate of 1051, 787 cases (23.6/100 000 population [popln]) globally has recently been reported, with the highest rates being in Asia (33.9/100 000 popln, an absolute number of 939 895) and Africa (13.5/100 000; 75 196); if all culture-negative cases are assumed to be fungal, especially where the incidence of mycotic keratitis is known to be high, then the annual incidence would be about 1480 916 cases. A fungal etiology has been found to account for a very high proportion (> 45%) of microbial keratitis cases in countries in the Indian subcontinent. Countries where a fungal etiology accounts for >25% of microbial keratitis mostly tend to abut the equator. Interestingly, the proportion of microbial keratitis patients with a proven fungal etiology shows a significant negative correlation with the gross domestic product per capita. Although it is clear that the most common fungal species are Fusarium, Aspergillus, and Candida species, marked regional variations in fungal etiology have been noted. It is important to realize that sensitivity of the culture of ocular fungal pathogens can vary, depending on the pathogen, as well as the competence of the testing laboratory. For some countries, multiple reports over time have been noted, with there being some evidence of an increasing trend in the proportion of all microbial keratitis cases being diagnosed as mycotic keratitis. Even in a single geographical location, cases of mycotic keratitis may be higher than the yearly average at certain times of the year, such as during the harvest or windy seasons, or when there is increased relative humidity. A disturbing statistic to note is that, in 8%-11% of patients with mycotic keratitis, the affected eye needs to be removed, representing an irreversible annual loss of 84 143-115 697 eyes. It is recognized that many people suffering from mycotic keratitis in rural distant communities never present to health care workers due to financial and other constraints. Hence, the actual number of people afflicted by mycotic keratitis, man-days lost due to the disease and during therapy, and reduced quality of life due to persistent disability (corneal scarring) in the Indian subcontinent and developing countries requires further study.","PeriodicalId":18325,"journal":{"name":"Medical mycology journal","volume":"16 1","pages":""},"PeriodicalIF":1.0,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88927413","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sheetal Thakur, D. Shaw, T. Narang, S. Dogra, H. Kaur, Anup K. Ghosh, S. Rudramurthy
{"title":"P115 Reversed-phase high-performance liquid chromatography as rapid and simple method for quantifications of terbinafine drug in human serum from dermatophytosis cases","authors":"Sheetal Thakur, D. Shaw, T. Narang, S. Dogra, H. Kaur, Anup K. Ghosh, S. Rudramurthy","doi":"10.1093/mmy/myac072.P115","DOIUrl":"https://doi.org/10.1093/mmy/myac072.P115","url":null,"abstract":"Abstract Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM Objectives The alarming situation of treatment failure cases in dermatophytosis and resistance to the first-line drug (terbinafine) is a worrisome condition for the management of tinea cases. However, studies also reported non-responders to terbinafine treatment even when the isolates are susceptible to this drug in vitro. Thus, evaluating the pharmacokinetic profile of terbinafine might help better manage dermatophytosis. This study was conducted to standardize and validate a rapid and simple reversed-phase high-performance liquid chromatography (HPLC)-based protocol for terbinafine in serum/plasma of dermatophytosis cases. Methods HPLC analysis was standardized for terbinafine drug on an Agilent 1290 infinity system (Agilent Technologies InC., USA). Chromatographic parameters including mobile phase [acetonitrile (A), methanol (M), and water (W)], flow rate (0.7-1.5 ml/min), injection volume (20 μl), and various wavelengths ranging from 220 to 265 nm under isocratic conditions were assessed and optimized. The mobile phase consisted of a filtered and degassed mix of A: W and M: W with various ratios of 85:15, 60:40, 50:50, and M-100%, respectively. Quality control samples were prepared in drug-free serum by spiking with the terbinafine at 0.0312-100 μg/mL concentrations. An equal volume of serum and acetonitrile (A) were mixed. The mixture was vigorously vortexed for 30 s, followed by high-speed centrifugation at 13 000 rpm at 40°C for 10 min. The supernatant was transferred into the chromatographic vials and placed in the autosampler of HPLC for injection. The standardized method was tested in 6 dermatophytosis patients’ serum/plasma samples collected at 3-time points (first, second, and third week of start of antifungal). Results Linearity of calibration standard for terbinafine was optimized at 250C at a flow rate of 1.0 ml/min, injection volume 20 μl, 8 minutes run time with the standardized wavelength at 245 nm under isocratic conditions. The best suitable graph was determined by plotting the area under the curve (AUC) and peak height separately against the drug concentrations measured by reversed-phase- HPLC for terbinafine drugs (Fig. 1a and b). The standardized mobile phase consisted of filtered and degassed Methanol (100, v/v). The chromatographic separation was achieved on an Agilent C18 column, and 4.3 ± 1 time represents the peak for terbinafine drug. Based on the standardized protocol, six tinea cases were included for validation, and the therapeutic range achieved for terbinafine in clinical samples was 0.6 to 1.13 μg/ml. Conclusions The standardization of HPLC method was successfully applied to quantify terbinafine in spiked samples with terbinafine drug and showed no observable interferences at the standardized parameter. Further evaluation with larger number of samples is warranted.","PeriodicalId":18325,"journal":{"name":"Medical mycology journal","volume":"9 1","pages":""},"PeriodicalIF":1.0,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88936524","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Pradhan, Qinxi Ma, Emer Hickey, G. Avelar, D. Larcombe, J. Bain, Delma S. Childers, I. Dambuza, I. Leaves, L. J. de Assis, M. Netea, Gordon D. Brown, L. Erwig, N. Gow, A. J. Brown
{"title":"S9.4c Diverse environmental inputs mediate changes in β-glucan exposure at the Candida albicans cell surface thereby influencing tissue colonisation during systemic infection","authors":"A. Pradhan, Qinxi Ma, Emer Hickey, G. Avelar, D. Larcombe, J. Bain, Delma S. Childers, I. Dambuza, I. Leaves, L. J. de Assis, M. Netea, Gordon D. Brown, L. Erwig, N. Gow, A. J. Brown","doi":"10.1093/mmy/myac072.S9.4c","DOIUrl":"https://doi.org/10.1093/mmy/myac072.S9.4c","url":null,"abstract":"Abstract S9.4 Free oral presentations (late breaking), September 23, 2022, 4:45 PM - 6:15 PM Candida albicans adaptation to host niches affects the exposure of key pathogen-associated molecular patterns (PAMPs) on its cell surface and, consequently, the detection of C. albicans cells by the immune system. Focusing on β-(1,3)-glucan, we screened for host inputs that influence the exposure of this immune-stimulatory PAMP on the C. albicans cell surface. We used a combination of fluorescent microscopy, flow cytometry, and cytokine assays, and then analyzed certain conditions in more detail using transmission electron microscopy and time-lapse video microscopy of C. albicans-phagocyte interactions. We found that some nutrients, micronutrient limitation, stresses, and antifungal drugs trigger β-glucan masking, whereas other inputs, such as nitrogen sources and quorum sensing molecules, exert limited effects on β-glucan exposure. In particular, host- or bacterial-derived L-lactate, hypoxia, or iron limitation induce β-glucan masking, and this leads to attenuation of phagocytic responses [Nature Micro 2, 16 238; mBio 9, e01318-18; Nature Comms 10, 5315]. Lactate signals through Gpr1 to activate Crz1 in a calcineurin-independent manner, whereas hypoxia signals via mitochondrial ROS, and iron limitation signals through Ftr1 and Sef1. β-glucan masking also depends upon downstream signaling via the cAMP-PKA pathway. We conclude that C. albicans has evolved to exploit a range of specific host-derived signals to modulate the exposure of a major PAMP at its cell surface in an attempt to evade phagocytic uptake. Using barcode-sequencing in direct competition assays in vivo, we showed that preadaptation to specific β-glucan masking signals affects the ability of this fungus to colonize particular tissues during systemic infection in a murine model. This reinforces the view that β-glucan masking promotes C. albicans infection.","PeriodicalId":18325,"journal":{"name":"Medical mycology journal","volume":"224 1","pages":""},"PeriodicalIF":1.0,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88963624","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}