Sabouraud Dextrose Agar Medium Research Articles

Diversity of Fungal Colonization in Respiratory Tract of Naïve Lung Cancer and The Emergence of Voriconazole Resistant Aspergillus

Fungal spores in the air can be inhaled and enter the human respiratory tract. The entry of fungi into the respiratory tract can cause colonization or infection depending on the host immune response. Fungal colonization is the first step into debilitating fungal disease in humans, especially in immunocompromised groups. The increased rate of drug-resistant fungi has been reported in human disease and the environment. This study aims to examine the diversity of fungal colonization in humans and the rate of fungal resistance to voriconazole. This cross-sectional study was done in patients with naïve lung cancer who had not been previously treated with any cancer therapy nor given antifungal agent. Induced sputum from 70 subjects was collected and inoculated in the Sabouraud Dextrose Agar medium. Macroscopic and microscopic examinations were performed to identify fungal species. Voriconazole susceptibility tests were done using the disc diffusion method. This study found Candida albicans, Aspergillus niger, Aspergillus fumigatus, and Penicillium sp. among the most common lower respiratory tract colonies. This study also found the colonization of up to 5 species in a single subject. A high rate of voriconazole-resistant Aspergillus sp. was found (42.4%) among 59 isolates tested. Given that these subjects had never taken antifungal agents previously, the high rate of voriconazole resistance might be attributed to the environment, such as community and agriculture. Mitigation of antifungal use in the agricultural sector, fungal diversity in the environment, and clinical study of fungal colonization/ infection in other high-risk groups are needed.

A comparative study for optimization of MALDI-TOF MS identification of filamentous fungi.

To evaluate and compare the performance of three commercial culture media, two filamentous fungi libraries, and two different protein extraction procedures in MALDI-TOF MS fungal identification. A total of 21 quality control samples were cultured on Sabouraud dextrose agar (SDA), ID fungi plate medium (IDFP), and Sabouraud gentamicin chloramphenicol 2 agar (SGC2). For four consecutive days, fungal growths were inoculated on a MALDI target plate both by using a direct transfer technique (DT) and by using a formic acid-ethanol protein extraction procedure (EEP). The MALDI-TOF MS-generated spectra were identified by the MBT Bruker library and the MSI database. Selective culture media (IDFP and SGC2) significantly outperformed the non-selective SDA medium. IDFP was superior to the SGC2 medium for dermatophyte identification. The EEP only demonstrated a benefit over DT in the underperforming SDA medium. The MBT Bruker library outperformed the MSI database in Aspergillus identification while the MSI database outperformed the MBT library in dermatophyte identification. For non-Aspergillus fungi, the libraries performed comparably. The results of our study show the necessity of using selective culture media (IDFP and SGC2) for fungal identification with MALDI-TOF MS and demonstrate no significant benefit of the formic acid-ethanol protein extraction technique in these media. Given the relative strengths and weaknesses of the MBT library and the MSI database, it might currently be beneficial to consider these libraries as complementary and employ both databases to achieve optimal fungal identification.

Prevalence of Non-Dermatophytic Molds in Toenails among Diabetic Patients in Homs, Syria

Diabetes Mellitus (DM) is a risk factor for fungal infections, including onychomycosis. The study aims to determine the prevalence of toenail onychomycosis in diabetic patients and its correlation with associated factors, identify the causative agents and compare the fungal culture findings with the real-time polymerase chain reaction (RT-PCR) findings of diagnosing 6 fungal isolates. A total of 126 diabetic patients were included between November 2020 to June 2021. Nail samples were subjected to potassium hydroxide 20%. Culture was done on Sabouraud dextrose agar medium (SDA), both with and without cycloheximide. Identification of non-dermatophytic molds was based on colony characteristics, colony reverse, cello-tape flag method and slide culture technique. Molecular analysis was done to make sure of the identification of 6 fungal isolates. Out of 126 cases, 106 (84.1%) had toenails onychomycosis. Non-dermatophytic molds (NDM) were the most common isolates 75 (70.8%), followed by yeasts 8 (7.5%) and mixed infections 23 (21.7%). Distal lateral subungual onychomycosis (DLSO) was the most frequent clinical type 90 (85%). Onychomycosis was found to correlate significantly with advancing age, type 2 diabetes, its duration and clinically abnormal nails. However, there was no significant correlation with gender and glycosylated hemoglobin (HbA1C). When the results obtained by RT-PCR were compared with the results obtained by fungal culture, a 100% match of the isolates was found.

Reliable and rapid identification of terbinafine resistance in dermatophytic nail and skin infections.

Fungal infections are the most frequent dermatoses. The gold standard treatment for dermatophytosis is the squalene epoxidase (SQLE) inhibitor terbinafine. Pathogenic dermatophytes resistant to terbinafine are an emerging global threat. Here, we determine the proportion of resistant fungal skin infections, analyse the molecular mechanisms of terbinafine resistance, and validate a method for its reliable rapid identification. Between 2013 and 2021, we screened 5634 consecutively isolated Trichophyton for antifungal resistance determined by hyphal growth on Sabouraud dextrose agar medium containing 0.2 μg/mL terbinafine. All Trichophyton isolates with preserved growth capacity in the presence of terbinafine underwent SQLE sequencing. Minimum inhibitory concentrations (MICs) were determined by the broth microdilution method. Over an 8-year period, the proportion of fungal skin infections resistant to terbinafine increased from 0.63% in 2013 to 1.3% in 2021. Our routine phenotypic in vitro screening analysis identified 0.83% (n = 47/5634) of Trichophyton strains with in vitro terbinafine resistance. Molecular screening detected a mutation in the SQLE in all cases. Mutations L393F, L393S, F397L, F397I, F397V, Q408K, F415I, F415S, F415V, H440Y, or A398 A399 G400 deletion were detected in Trichophyton rubrum. Mutations L393F and F397L were the most frequent. In contrast, all mutations detected in T. mentagrophytes/T. interdigitale complex strains were F397L, except for one strain with L393S. All 47 strains featured significantly higher MICs than terbinafine-sensitive controls. The mutation-related range of MICs varied between 0.004 and 16.0 μg/mL, with MIC as low as 0.015 μg/mL conferring clinical resistance to standard terbinafine dosing. Based on our data, we propose MIC of 0.015 μg/mL as a minimum breakpoint for predicting clinically relevant terbinafine treatment failure to standard oral dosing for dermatophyte infections. We further propose growth on Sabouraud dextrose agar medium containing 0.2 μg/mL terbinafine and SQLE sequencing as fungal sporulation-independent methods for rapid and reliable detection of terbinafine resistance.

Isolation of Bacillus amyloliquefaciens M13-RW0 from Soil and Evaluation of its Antifungal Activity.

The frequency of observed invasive Aspergillosis has increased in recent years. Infection with other molds happens but does not lead to a large proportion of invasive infections. The present study aims to isolate Bacillus amyloliquefaciens M13-RW0 from soil and evaluate its antifungal effects against some saprophytic fungi, such as Aspergillus niger, Aspergillus flavus, and Mucor hiemalis. In this research, a total of 150 samples (from the soil, air, and surfaces) were prepared from different parts of Isfahan, Iran. Isolation and purification of growing bacteria were conducted using the nutrient agar medium. The inhibitory effects of 100 isolated bacteria were evaluated against the growth of A. niger, A. flavus, and M. hiemalis, 4 bacteria were isolated with inhibitory effects against the selected fungi, and consequently, one of the bacteria isolated from the soil samples was found to show the highest inhibition of fungal growth. Quantitative evaluation of the growth inhibitory effect was performed using linear culturing of fungal suspension (104 spore/ml) at distances of 5, 10, 15, 20, 25, and 30mm from bacterial isolate (0.5 McFarland) on Sabouraud Dextrose Agar (SDA) medium. Results were checked 24, 48, 72, and 96 hours later. The bacterial isolate with the most inhibitory effect was identified by phenotypic and molecular tests. According to the results, among the 4 inhibitory bacterial isolates, Bacillus amyloliquefaciens strain M13-RW01, isolated from the soil samples, was identified as the bacterium with the most significant potential antifungal activity. The strong inhibitory effect was revealed after 48 hours for all distances of 15mm and more between the fungi and the bacterium. The identified bacterium could not only be considered an inhibitor bacterium against saprophytic fungi but could also be put forward to help produce new antifungal drugs for controlling fungal diseases.

Evaluation of the Various Oral Candida Species among Patients on Hemodialysis: A Clinical Study.

Oral candidiasis develops as a result of an opportunistic infection. In patients on hemodialysis who may be immune-compromised, candida can change from a commensal to a pathogen. Identification and classification of Candida species are crucial for the treatment of these patients. To distinguish between distinct species of candida found in hemodialysis patients with chronic renal failure's oral cavity. A total of 100 people were investigated, including 50 patients with "Chronic Renal Failure (CRF)" on hemodialysis and 50 healthy controls. Using "Sabouraud's Dextrose Agar (SDA)" and "CHROM Agar Culture Media", salivary samples were incubated for 24, 48, or 72 hours at 37°C to develop candida species. Colony forming units/milliliter, a unit of measurement for colonies, were counted and calculated (CFU). On SDA and CHROM agar medium, positive candidal growth was seen in the CRF with HD and Control Groups. There was a significant variance in the growth and the subspecies distribution of the colonies among the groups. The most common species was c.albicans followed by c. cruzi and tropical. Other subspecies were negligible. The number of colonies was also greater than 400 CFU in the majority of the dialysis subjects, while it was only 200 CFU for the majority of the controls. Patients with chronic renal failure receiving hemodialysis showed significantly higher levels of candida isolation and differentiation than healthy individuals (P 0.05).

The Measurement of Candida albicans Tolerance under The Influence of Moringa oleifera

Natural ingredients such as M. oleifera have antibacterial and antifungal properties.The fungistatic and bacteriostatic properties are highly desirable for commensals.The tolerance properties of antifungals help create a balance among oral commensals such as Candida albicans (C. albicans). This study aimed to examine the tolerance properties of M. oleifera to C. albicans regarding biomass, pH stability, biostability, absorption, and opsonization properties. This study used Moringa leaves (M. oleifera) extracts and C. albicans as subjects. Tolerance properties were checked with analytical balance, pH meter, Sabouraud dextrose agar (SDA) medium, spectrophotometer, and 1% crystal violet. M. oleifera extracts at concentrations of 10% and 5% had excellent tolerance properties of index biomass properties, pH stability, biostability, absorption and residue, and opsonization. The M. oleifera have tolerance properties for C. albicans and significant differences between the concentrations tested. M. oleifera extract has good tolerance properties to C. albicans as an oral commensal.

Activity Test of Combination of Binahong Leaf Extract (Anredera Cordifolia) and Lemongrass Extract (Cymbopogon citratus) as Antifungal Againt Pityrosporum ovale

Pityrosporum ovale is a microorganism thought to be the main cause of dandruff. Binahong and lemongrass are herbal plants that can cure various diseases. Based on usage empirical leaf binahong and lemongrass kitchen nutritious for remove dandruff, hair loss, diabetes mellitus, anti- cancer and treatment on infection skin caused by bacteria nor mushrooms. Binahong leaf extract and extract lemongrass contain metabolites secondary wrong only flavonoids. Flavonoids are effective as antimicrobials. Destination from study this The purpose of this study was to determine the antifungal activity of the combination of binahong leaf extract (Anredera cordifolia) and lemongrass extract (Cymbopogon citratus) against Pityrosporum ovale . This study used 5 test groups, which consisted of a combination group of binahong leaf extract and lemongrass F1 = concentration 50%:50%, F2 = concentration 37,5%:12,5%, F3 = concentration 25%:25% and F4 = concentration 12,5%:37,5%, and the control group was positive (ketoconazole 2%) and negative ( aquadest), with 3 repetitions. The materials used in this study were binahong leaf simplicia and kitchen lemongrass simplicia macerated using 70% ethanol solvent, resulting in a thick extract which was then tested for antifungal activity against Pityrosporum ovale by the method diffusion paper disc on Sabouraud Dextrose Agar medium . The parameter observed was the diameter of the growth inhibition zone of the fungus Pityrosporum ovale . Analysis using ANOVA (Analysis of Variance). The results of this study were binahong leaf extract and kitchen lemongrass extract positive containing flavonoid compounds. The combination of binahong leaf extract and citronella extract has the ability to inhibit the growth of the fungus Pityrosporum ovale with visible inhibition zones formed. The most effective concentration to inhibit the growth of the fungus Pityrosporum ovale was at a concentration of F1 = 50%:50% of 17.40 mm with a strong category.

Comparison of different methods to determine the viability of dermatophytes in skin scrapings

One of the most prevalent infectious dermatoses seen in dermatology outpatient clinics is superficial fungal infections of the skin, hair, and nails. Filamentous fungi, dermatophytes, which invade and colonise the keratinised layers of the skin, are the main cause of skin fungal infection. Sometimes there is a discrepancy between the results of microscopy and culture for dermatophytes. If the microscopy of scrapings is positive due to presence of non-viable fungus, no growth will take place in culture. Culture is the method used to determine the presence of viable dermatophytes. However, the problem with culture is the long time, up to 4 weeks, that is required for the growth of fungus to take place. Aim of the present study is to develop a simple, rapid method(s) for identification of live dermatophytes in skin scrapings. The novel methods used for identification of live dermatophytes in skin scrapings were direct microscopic examination using periodic acid-Schiff (PAS) stain, trypan blue stain, neutral red stain, and methylene blue stain; and their results were compared with culture on Sabouraud Dextrose Agar (SDA) and Dermatophyte Test Medium (DTM). Out of a total 91 samples, fungi were isolated in 81 (89%) in SDA and 66 (72.5%) in DTM. Sixty-eight (74.7%) showed evidence of viable fungi on microscopic examination after staining with PAS and 77 (84.6%) with trypan blue. In 84 patients, the culture was positive either by SDA or DTM or both culture media, out of which 64 (76.2%) were detected viable on staining with PAS and 74 (88.1%) on staining with trypan blue. Three skin samples that failed to grow on both SDA and DTM demonstrated viable fungi on microscopic examination after staining with PAS. Four skin samples that failed to grow on both SDA and DTM demonstrated viable fungi on microscopic examination after staining with trypan blue. Taking culture positivity in SDA or DTM or both media, as the gold standard, the sensitivity of trypan blue stain was 88.1% and specificity was 57.1%. The sensitivity of PAS stain was 76.2% and specificity was 42.9%. Although culture remains the gold standard to identify viable dermatophytes; trypan blue and PAS stains, as described, may be used as simple and rapid alternatives to culture.

Effect of Pectin and Citric acid on Total Bacterial Load, Fungal Load, and Techno-economic Feasibility from processing to the storage of Guava Jelly

This study was conducted to investigate the Total bacterial load, fungal load, sensory evaluation, and techno-economic feasibility from processing to the storage of guava jelly. Fresh guava (Psidium guajava) jelly were prepared with 0.15%, 0.2%, and 0.25% pectin and with 1.1%, 1.15%, and 1.2% citric acid as a preservative. The bacterial count was determined through the Standard Plate Count (SPC) technique. The fungal analysis was done using a selective Sabouraud Dextrose Agar (SDA) medium. Lastly, the Inventory Theory’s model III was applied for the Techno-economic feasibility of guava jelly production. There was no yeast and mold growth detectable till day 90 of the storage period. However, the total bacterial load in jelly (acceptable limit ≤105cfu/ml) is satisfactory for consumption after 90 days of pectin and citric acid treatment. Furthermore, the production cost was approximately 65 Tk a bottle of 500g, which is considered economical compared to other local brands. Therefore, this formulation could be practiced for escalating the country’s development in the agriculture and food industries.

Comparison of in vitro activities of newer triazoles and classic antifungal agents against dermatophyte species isolated from Iranian University Hospitals: a multi-central study

BackgroundDermatophytes have the ability to invade the keratin layer of humans and cause infections. The aims of this study were the accurate identification of dermatophytes by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism method and sequencing and comparison between the in vitro activities of newer and established antifungal agents against them.MethodsClinical specimens of patients from five Iranian university laboratories were entered in this study. Samples were cultured on sabouraud dextrose agar medium. For molecular identification, extracted DNAs were amplified by the universal fungal primers ITS1 and ITS4, and digested with MvaI enzymes. The antifungal susceptibility test for each isolate to terbinafine, griseofulvin, caspofungin, fluconazole, itraconazole, luliconazole, and isavuconazole was performed, according to the microdilution CLSI M38-A2 and CLSI M61 standard methods.ResultsTwo hundred and seven fungi species similar to dermatophytes were isolated of which 198 (95.6%) were dermatophytes by molecular assay. The most commonly isolated were Trichophyton mentagrophytes (76/198), followed by Trichophyton interdigitale (57/198), Trichophyton rubrum (34/198), Trichophyton tonsurans (12/198), Microsporum canis (10/198), Trichophyton simii (3/198), Epidermophyton floccosum (3/198), Trichophyton violaceum (2/198), and Trichophyton benhamiae (1/198). The GM MIC and MIC90 values for all the isolates were as follows: terbinafine (0.091 and 1 μg/ml), griseofulvin (1.01 and 4 μg/ml), caspofungin (0.06 and 4 μg/ml), fluconazole (16.52 and 32 μg/ml), itraconazole (0.861 and 8 μg/ml), isavuconazole (0.074 and 2 μg/ml), and luliconazole (0.018 and 0.25 μg/ml).ConclusionTrichophyton mentagrophytes, Trichophyton interdigitale, and Trichophyton rubrum were the most common fungal species isolated from the patients. luliconazole, terbinafine, and isavuconazole in vitro were revealed to be the most effective antifungal agents against all dermatophyte isolates.

Mycoflora and Total Aflatoxin Isolated from Dry Dates in Sana'a, Yemen

This study is designed to record the mycoflora and total aflatoxin 
 isolated from dry dates in Yemen Republic. Thirty four date samples 
 collected from different shops and markets in Sana’a city were 
 analyzed mycologically for the presence of fungi cultivated on three 
 types of media. Eight species belonging to 7 genera were isolated 
 from analyzed date samples on the three cultural media. Aspergillus
 genera was the most dominant fungi and grew well on the three type 
 of media of which A. niger is the most common species, A. flavus was 
 isolated in rare, low and rare frequency on 1% and 20% sucrose 
 Czapek’s and Sabouraud dextrose agar media. Rhizopus stolonifer 
 was isolated in rare, low and low frequency on 1, and 20% sucrose 
 Czapek's and Sabouraud dextrose agar media. The date samples were 
 analyzed for the presence of total aflatoxin using ELISA technique 
 which revealed that 4 out of 5 date samples were contaminated with 
 aflatoxin ranged from 3059.89-7585.96 ppt (ng Kg-1). 

Isolation and Antifungal Susceptibility of Candida spp. from Pediatric Patients in Kurdistan of Iraq

Candida species are considered the most common opportunistic human fungal pathogen due to the presence of various virulence factors, including its ability to form a biofilm that aids in oral candidiasis. Pediatric patients are more susceptible to oral candidiasis than healthy pediatric because of the factors that encourage Candida oral carriage. This current study aims to isolate, evaluated the antifungal effect on Candida spp. and participated in oral candidiasis of pediatric patients in Sulaymaniyah and Kirkuk city. The study was performed from September 2021 to February 2022 on two groups of pediatrics which including pediatric patients (n=160) and healthy pediatric (n=50) as a control group. Oral swabs were obtained from 210 participants in the Kurdistan Region of Iraq at the Pediatric teaching hospital of the Sulaymaneyah and Kirkuk governorates. To culture the swabs, Sabouraud dextrose agar (SDA) medium was used. HiCrome™ Candida Differential agar used to identify Candida isolated, then depending on the internal transcribed spacer (ITS) region using confirmed polymerase chain reaction. Antifungal sensitivity was done for all C. albicans and non-albicans isolates using five common antifungal disks. Among of the 160 sample of pediatric patient 62 (38.8%) positive for Candida spp. which it is including 17 (27.4 %) and 26 (41.9%), 19 (30.6%) breast feeding, bottle feeding and mixing respectively. While within 50 samples of control group 13 (26%) were positive for Candida spp. which including 2 (15.4%), 9(69.2%), 2(15.4%) of same respectively feeding types. Regarding to the antifungal susceptibility the results showed that C. albicans had higher resistance rates against itraconazole, ketoconazole, and clotrimazole than the non-albicans Candida species However, highly resistant rate were detected in Itraconazole and Ketoconazole with 23.7% and 54.4% respectively, for all Candida species. The Current study concluded that oral candidiasis was more predominant in pediatric patients in compare to healthy pediatric also, and C. albicans is the most prevalent etiologic agent. However, higher rate of sensitivity were detected between Candida species for nystatin, which may suggest as the main treatment for oral Candida infections.

Microbiological quality assessment of fish origin food along the production chain in upper Blue Nile watershed, Ethiopia.

Pathogenic microorganisms can grow accidentally on fish origin human food and can be a cause of human food-borne illness. The purpose of this study was to estimate the occurrence and microbial load pattern of Escherichia coli, Salmonella, Staphylococcus aureus, and Shigella spp. along the fish origin food value chain. A total of 396 fish samples were collected by a systematic random sampling technique of cooked and raw in the three species of fish. Fish muscles were tested using selective media, followed by conventional biochemical tests. The bacterial load was assessed using a standard plate count method. Whereas the fungal load were measured by cultured in a Sabouraud's dextrose agar (SDA) medium. The overall prevalence was Escherichia coli 84 (21.21%), Salmonella 27 (6.82%), Staphylococcus aureus 19 (4.80%), and Shigella spp. 17 (4.29%). The average mean total coliform count was observed 1.2 × 102 cfu/g and 5.10 × 104 cfu/g in cooked and raw fish samples, respectively. Whereas total viable count mean of 8.05 × 104 cfu/g and 11.5 × 104 cfu/g in cooked and raw fish, respectively. The Fungal load counts under the range 5.6 × 101 cfu/g to 1.09 × 103cfu/g were observed. The study has revealed that fish food in the study area has the possibility of microbial public health risk. Hence, it could be wise to improve the knowledge of key actors from harvesting to consumption to enhance the meals protection practices and high-quality standards of fish foods.

DFT Investigation, Chemical Reactivity Identification and Molecular Docking of 2(E)˗1˗(3˗Bromothiophene˗2˗yl)˗3˗(Furan˗2˗yl)Prop˗2˗en˗1˗one

The Fourier transform infrared (FT-IR) and Raman spectra of 2(E)˗1˗(3˗bromothiophene˗2˗yl)˗3˗(furan˗2˗yl)prop˗2˗en˗1˗one (BTF) molecule have been recorded and analyzed. The equilibrium geometry, bonding features and harmonic vibrational frequencies have been investigated with the help of density functional theory (DFT) method under the B3LYP level of theory. The detailed assignments of the vibrational spectra have been carried out with the help of potential energy distribution (PED). The electronic properties were analyzed such as EHOMO energy (6.367 eV), ELUMO energy (2.705 eV), the energy difference (3.662 eV), softness (0.546 eV), and electrophilicity index (5.618 eV) were determined using UV˗Vis absorption spectrum by time dependent density functional theory (TD-DFT) approach, which helps to understand the reactivity, stability, and biological potency of the molecule. In addition, molecular electrostatic potential (MEP) was accounted in the identification of reactive sites. The intra-molecular interactions, charge delocalization and stability have been analyzed using natural bond orbital (NBO) analysis. Non-linear optical features were demonstrated from the first-order hyperpolarizability. Based on the electronic property the BTF compound was investigated for further biological investigation. The BTF compound was screened for their in vitro antifungal activity in Sabouraud Dextrose Agar Medium (SDA) and which shows moderate antifungal activity. In vitro anti˗inflammatory activity was evaluated using Bovine Serum Albumin (BSA) protein denaturation assay at different concentrations. Molecular docking was performed to investigate the interactions between BTF, co-crystal compound and the Mcl-1 protein, and their docking scores were 7.8, and 6.5, respectively, which confirms the molecule’s chemo-resistance properties.

Distribution and Fluconazole Susceptibility Pattern of Fungi Isolated from Saloon Equipment in Rivers State, Nigeria

The distribution and susceptibility pattern of fungal isolates from barbing equipment to Fluconazlole in barber shops distributed in three major cities; Bori, Omoku, and Port Harcourt in Rivers State, Nigeria were investigated. The fungal load including dermatophytes on the equipment (brush, clipper and comb) were determined using swab plate technique in which Sabouraud dextrose agar and dermatophyte test medium were used as the culture medium. Inoculated plates were incubated at 25°C. Antifungal Susceptibility testing was carried in which fluconazole was used as the antifungal agent at concentrations of 100mg/ml, 50mg/ml, 25mg/ml, and 12.5mg/ml concentration using two-fold dilution. The mean range of the fungal counts for clipper, brush and comb were: 34.34x102 to 49.0x102 CFU/cm2, 66.95 to 93.26 CFU/cm2 and 40.09 to 74.61 x102 CFU/cm2, respectively. Brush had the highest fungal counts followed by the comb while the clipper had the least fungal counts. The fungal counts recorded in brushes were significantly higher (p≥0.05) than those recorded for the clippers. Total of forty-four fungal isolates belonging to six genera and their percentage occurrence include; Aspergillus flavus (11.36%) Aspergillus terrus (11.36%), Fusarium solani (22.72%), Mucor indicus (15.91%), Rhizopus nigricans (15.91%), Trichophyton rubrum (4.54%) and Penicillium italicum (18.2%). The susceptibility pattern showed that except for Trichophyton rubrum, all fungal isolates were completely inhibited at 100mg/ml concentration of fluconazole. Also, except for Mucor indicus, the Minimal inhibitory Concentration of fluconazole was recorded at 12.5mg/ml. The occurrence of these organisms in barbing equipment calls for concern and the need for regulation of the barbing industry to prevent disease outbreaks is recommended.

PREVALENCE OF FUNGAL COINFECTIONS AMONG COVID 19 PATIENTS IN A TERTIARY CARE HOSPITAL

Coronavirus disease 2019 (COVID-19), caused by Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has been an emergency global public health events.COVID-19 patients, especially severely ill or immunocompromised, have higher probability of suffering from fungal coinfections. The study aims to investigate the prevalence of fungal coinfections among patients with conrmed severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).Aprospective study was undertaken in the Department of Microbiology, Government Medical College, and Amritsar for a period of 6 months from May 2021 to October 2021. Atotal of 216 conrmed cases of COVID 19 disease were enrolled for the study. Various clinical samples obtained were processed by direct microscopic examination in 10% KOH solution, Gram stain, Calcoour white staining and culture on Sabouraud dextrose agar media. Identication of fungi was done by lacto Phenol Cotton Blue, Germ tube test and Candida CHROM agar for species identication.Out of 216 cases, 138 were culture positive. Candida species were predominant isolates with Candida albicans (49%) being the major species followed by Aspergillus species (21.7%), Mucorales (19%) and mixed growth (1%). Predominantly males were affected in the age group of 46-55 years. Most common risk factor was hypertension (42%).An increasing prevalence (63.8%) of fungal coinfections has been reported among COVID 19 patients resulting in poor patient outcome and high mortality rates. Therefore, awareness of high risk of occurrence of fungal infections is crucial to downgrade any delay in diagnosis and treatment to support the prevention of complications related to these infections.

Susceptibility of airborne fungi to the mycocins produced by <em>Wickerhamomyces anomalus</em>

Atmospheric air is the most used and most successful dispersion medium for fungi. Airborne fungi can establish themselves and contaminate the air, reducing the quality of life of the organisms that circulate there. The broad spectrum of antimicrobial activity and great stability led to the use of Wickerhamomyces anomalus as a biocontrol agent, since it could be classified as a low-risk microorganism, rarely traced in human samples. In addition, the antimicrobial action of mycocins produced from this yeast in eukaryotic and prokaryotic microorganisms is already proven. The aim of the present study was to evaluate the inhibition of airborne fungi by mycocins. The passive sedimentation technique was used in Petri dishes exposed in an external environment. The exposed plates consisted of a control group, consisting only of Sabouraud Dextrose Agar medium, and another test, composed of medium and supernatant of mycocins from Wickerhamomyces anomalus. The growth of 6 genera of fungi was observed on the control plates: Cladosporium spp., Penicillium spp., Alternaria spp. Aspergillus spp., Chrysosporium spp. and Fusarium spp. While, on the test plates, there was no growth of microorganisms. Therefore, we concluded that the mycocins produced by Wickerhamomyces anomalus were able to inhibit the growth of airborne fungi.

S6.2d Dermatophytosis due to Trichophyton erinacei caused by contact with African hedgehogs as family pets

S6.2 Resurgence of dermatophytic infections, September 22, 2022, 4:45 PM - 6:15 PMObjectiveTo present a series of cases of various types of dermatophytosis caused by Trichophyton erinacei, caused by contact with African hedgehogs.MethodsPatients with clinical suspicion of ringworm caused by T. erinacei were included in the study. Each patient underwent a mycological study, with direct examinations (KOH), cultures in Sabouraud dextrose agar medium with and without antibiotics. With the cultures obtained, molecular identification was made using the PCR technique and Matrix Assisted Laser Desorption Ionization Time of Flight (MALDI-TOF) proteomics. Once confirmed, the patients were given treatment and follow-up. When the pets (hedgehogs) were accessible, they were cultured with a cytobrush and the quills removed.ResultsIn a period of 3.5 years, 16 verified cases were incorporated, 13 were females (72%) and 3 males (18%), the average age was 18.2 years, with the smallest case being 8 years old and the oldest 45 years old. The tineas manifested as follows: tinea manus 11 cases (68.7%); 2 respective cases of tinea corporis and tinea capitis (18.1 respectively) and one case of Majocchi's granuloma. The average evolution was 1.5 months with the case older than 3 months and the younger 15 days. All cases were given terbinafine at variable times and doses (according to weight and age) and clinical and mycological cure was obtained.DiscussionTineas can have epidemiological changes that are favored by a variety of factors, in general, pet ringworms are due to common domestic animals such as dogs and cats. Due to changing fashions, children and adolescents in recent years have changed pets such as hamsters, and recently hedgehogs, especially African variety (Atelerix albiventris) the latter may have T. erinacei as part of their skin microbiota, a similar dermatophyte or that was part of the T. mentagrophyes complex, in general of low virulence, however, its antigenic variants are little recognized by humans, which makes them present in a very inflammatory form, particularly in the hands, due to the handling of the animal, as well as the deeper inoculation (in the dermis) by the spikes left by the inoculum. So, they can generate deep tineas such as Majocchi's granuloma. In general, they respond well to systemic antifungals such as terbinafine and itraconazole, and sometimes the use of topical or systemic steroids is necessary.ConclusionsThis series of cases is a good example of how a change of habits when having pets can introduce new etiological agents that are poorly recognized antigenically, which give rise to more inflammatory and sometimes profound tineas.

P281 Healthy adult with arm swelling and new onset burning pain

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PMIntroductionCladosporium cladosporioides is a dematiaceous fungal agent that is known to produce a variety of clinical manifestations such as cutaneous and subcutaneous infection, corneal infection, pulmonary fungal infection, and rarely central nervous system infection. Several cases of subcutaneous mycosis have been reported from Assam, India. We are describing a similar case from Gujarat, India.Case HistoryA 36-year-old healthy, and muscular male farmer residing in central Gujarat presented with painless nodular swelling with normal overlying skin over the posterior aspect of the left arm for the last 3-4 years. He noticed a gradual increase in the size of this swelling and numbness for the last 4 months and burning sensations for the last 10 days before presentation to the surgeon. Local part ultrasound showed a subcutaneous soft tissue lesion. Excision biopsy reported invasive fungal infection and patient was referred for further evaluation to an infectious disease facility (Fig. 1a). He had no other symptoms and comorbidities. Patient didn't recall any trauma in the past. Direct microscopic examination with calcofluor white stain from surgical site scrapping revealed mycelial structure (Fig. 1b) and yielded pure growth of a dematiaceous fungus on Sabouraud dextrose agar medium, brown to blackish colony (Fig. 1c) with olive color on the backside (Fig. 1d) after 6 days. Lactophenol cotton blue stain preparation of culture isolate showed mycelial elements with morphology suggestive of C. cladosporioides (Fig. 1e), which was confirmed at the National Culture Collection for Pathogenic Fungi, PGI Chandigarh. Review of histopathology revealed a nodule with dense acute on chronic inflammation composed of lymphocytes, plasma cells with many neutrophils with abscess containing brightly eosinophilic structures with many PAS positive septate, thin and thick walled, branching, irregularly shaped, bullous hyphae surrounded by neutrophilic boarder (Splendor Hoeppli phenomenon) (Fig. 1f).Patient was treated with capsule itraconazole 200 mg three times a day for three days followed by 200 mg twice a day along with terbinafine and total surgical excision of the subcutaneous nodule. Patient achieved a therapeutic itraconazole level (1.4 mg/l) after 1 week. Patient is currently receiving treatment.DiscussionSubcutaneous mycosis is endemic in Assam. Several cases of subcutaneous chromoblastomycosis caused by C. cladosporidiosis mainly involving the lower limbs were reported from Assam. It mostly affects males engaged actively in outdoor activities during their productive age ranged from 20 to 50 years. It affects a relatively healthy, immunocompetent host and have a long history (in years) of asymptomatic nodule/swelling before diagnosis. Treatment comprises of prolonged antifungal treatment with itraconazole along with surgical excision.