Sabouraud Dextrose Agar Research Articles

Yeast species in the respiratory samples of COVID-19 patients; molecular tracking of Candida auris.

Although the existence of Candida species in the respiratory tract is often considered commensal, it is crucial to recognize the significance of Candida colonization in immunocompromised or COVID-19 patients. The emergence of Candida auris as an emerging pathogen further emphasizes the importance of monitoring yeast infection/colonization, particularly in COVID-19 patients. In this study, respiratory samples mainly from COVID-19 patients, primarily those suspected of having a fungal infection, were cultured on Sabouraud dextrose agar plates and the yeast colonies were identified using a two-step multiplex PCR method. The samples suspected of C. auris underwent specific nested PCR followed by sequence analysis. A total of 199 respiratory samples were collected from 73 women and 126 men, ranging in age from 1.6 to 88 years. Among the patients, 141 had COVID-19, 32 had cancer, 5 were hospitalized in ICU, 2 had chronic obstructive pulmonary disease)COPD(, and others were patients with combination diseases. From these samples, a total of 334 yeast strains were identified. C. albicans (n=132, 39.52%) was the most common species, followed by C. tropicalis (n=67, 20%), C. glabrata (n=56, 16.76%), C. krusei (n=18, 5.4%), C. parapsilosis (n=17, 5.08%), Saccharomyces cerevisiae (n=10, 3%), C. kefyr (n=9, 2.6%), C. dubliniensis (n=7, 2.1%), C. lusitaniae (n=5, 1.5%), C. auris (n=3, 0.9%), C. guilliermondii (n=2, 0.6%), C. rugosa (n=1, 0.3%), C. intermedia (n=1, 0.3%), and Trichosporon spp. (n=1, 0.3%). C. auris was detected in a patient in ICU and two COVID-19 patients. While its presence was confirmed through sequence analysis, our extensive efforts to isolate C. auris were unsuccessful. While C. albicans colonization remains prevalent, our study found no evidence of Candida lung infection. Since the role of Candida colonization in airway secretions remains ambiguous due to limited research, further studies are imperative to shed light on this matter.

Distribution and antifungal susceptibility profile of oropharyngeal Candida species isolated from people living with HIV in the era of universal test and treat policy in Uganda.

Despite the increased frequency of oropharyngeal candidiasis among people living with human immunodeficiency virus (HIV), its management is no longer effective due to empirical treatment and emergence of antifungal resistance (AFR). This study sought to investigate the prevalence of oropharyngeal candidiasis and assess the antifungal susceptibility profile of oropharyngeal Candida species isolated from people living with human immunodeficiency virus. Additionally, we evaluated the correlation between oropharyngeal candidiasis and CD4 T cell as well as viral load counts. A descriptive cross-sectional study was carried out from April to October 2023 in which 384 people living with HIV underwent clinical examination for oral lesions. Oropharyngeal swabs were collected and cultured on Sabouraud Dextrose agar to isolate Candida species which were identified using the matrix assisted laser desorption ionization time of flight mass spectrometry. Additionally, the antifungal susceptibility profile of Candida isolates to six antifungal drugs was determined using VITEK® (Marcy-l'Étoile, France) compact system. Data on viral load were retrieved from records, and CD4 T cell count test was performed using Becton Dickinson Biosciences fluorescent antibody cell sorter presto. The prevalence of oropharyngeal candidiasis was 7.6%. Oropharyngeal candidiasis was significantly associated with low CD4 T cell count and high viral load. A total of 35 isolates were obtained out of which Candida albicans comprised of 20 (57.1%) while C. tropicalis and C. glabrata comprised 4 (11.4%) each. C. parapsilosis, C. dubliniensis and C. krusei accounted for 2 (5.7%) each. Additionally, 7 (20%) isolates were resistant to fluconazole, 1 (2.9%) to flucytocine and 0.2 (5.7%) isolates were intermediate to caspofungin. However, specific specie isolates like C. albicans showed 20% (4/20), C. glabrata 50% (2/4) and C. krusei 50% (1/2) resistance to fluconazole. Additionally, C. krusei showed 50% resistance to flucytosine. The prevalence of oropharyngeal candidiasis (OPC) among people living with HIV was low, and there was a significant association between OPC and CD4 T cell count as well as viral load. C. albicans was the most frequently isolated oropharyngeal Candida species. C. glabrata and C. krusei exhibited the highest AFR among the non-albicans Candida species. The highest resistance was demonstrated to fluconazole.

Antifungal Properties of Grape Seed Extract on Various Species of Candida Organisms: An In vitro Study

Purpose: Oral candidiasis is one of the most common fungal opportunistic infections in the mouth caused by a fungus called Candida, and it presents as commensals in all humans. Over the last few decades, the increasing incidence of Candida infections parallels the growing numbers of immunocompromised patients. This trend is aggravated by the fact that there is a reckless use of antifungals. To combat this trend, the use of natural antifungals which have fewer side effects is recommended. Black grape seed extract (GSE) has shown a promise in this regard. Materials and Methods: Candida species such as Candida krusei, Candida glabrata, Candida tropicalis, and Candida albicans were used in the study. Fungal strains were passaged in the agar Sabouraud dextrose environment from 24 h to have live and fresh strains for the test. Black GSE was obtained. Minimum inhibitory concentration (MIC), of GSE, amphotericin B and fluconazole drugs on the included fungi was examined using disc diffusion method. Then, the plates were incubated for 24 h at 37°C. Finally, growth inhibitory zone was examined. Results: The analysis of the results revealed that the growth of C. albicans and C. krusei had been stopped at the concentration of 6.25 μg/mL, but the growth of C. glabrata and C. tropicalis was inhibited at a concentration of 12.25 μg/mL and 0.8 μg/mL respectively. Conclusion: The results obtained from the present study showed that both disc diffusion method and the maximum inhibitory concentration of MIC (Macro dilution) confirms the antimicrobial susceptibility of four species of candida to GSE. We conclude the possibility of GSE as an effective alternative measure for commercial antifungal agents, especially among immunocompromised patients, However, more laboratory and clinical studies need to be conducted to validate our findings.

Characterization and Isolation of Fungi from Domestic Pigeon Droppings in the Governorate of Erbil and its Suburban Area

A total of 200 samples of domestic bird droppings were collected from 54 regions in Erbil Governorate and its suburbs, and 23 of the isolates were fungi obtained by using the Niger bird seed medium and Sabouraud dextrose agar medium. The fungal genera were identified through macroscopic, microscopic, and phenotypic properties on agar plates and slides by staining with Lactophenol cotton blue stain. Findings belong to nine genera (Aspergillus, Alternaria, Chrysosporium, Cunninghamella, Helminthosporium, Mucor, Penicillium, Rhizomucor, and Rhizopus). Among the isolates, six species belonging to Aspergillus were identified which also had the highest frequency (44.4 %) in total genera, while the lowest frequency was (0.1 %) for Alternaria sp., Chrysosporium sp., Cunninghamella sp. Results reveal that 17 sites' samples of bird droppings produced negative results when tested on Niger bird seed agar, while only two locations produced no results when tested on Sabouraud dextrose agar. Thus, Sabouraud dextrose agar (SDA) is thought to be better than Niger bird seed agar (NBSA) for isolating fungi since it is considered a generally rich media for them.

Potencial of Rice Bran Extract as a Basic Ingridient in Candida albicans Culture Media

Rice bran is a natural material that contains nutrients that serves as a fungal growth factor, can be used as an alternative media base for fungal growth. The aim of this research is to determine the potential of rice bran as an alternative medium for the growth of Candida albicans. The research design employed an experiment that compared the growth of Candida albicans on rice bran extract media and Sabouraud Dextrose Agar (SDA) media. The sample is rice bran powder obtained from rice mills and rice bran extract was made using a multi-stage pretreatment method. The rice bran extract was used to create Rice bran Agar (RBA) and Rice bran Liquid (RBC) media, while SDA dan (SDB) media were also prepared. The media were inoculated with Candida albicans, incubated, and observed for 5 days. Data analysis included the measurement of fungal colony diameter using One Way Anova and Candida albicans biomass using an independent t-test. The research findings indicate that the average diameter of fungal colonies on RBA extract media was 10.4 mm, whereas it was 12.3 mm on SDA media. The Anova and Post Hoc Tukey HSD results demonstrated significant daily differences in colony diameter on RBA media, indicating notable Candida albicans growth. Conversely, there was no significant difference in colony diameter on SDA media. The biomass average of Candida albicans on RBC media was 0.82 g, compared to 0.45 g on SDB media. The Mann-Whitney test (p < 0.05) indicated a significant contrast in the average dry weight of Candida albicans biomass, with RBC media being heavier than SDB media. The conclusion is RBA and RBC media have the ability to grow Candida albicans which can be used as an alternative to SDA and SDB media.

Oral Candida Colonization Among Adults in Morocco: Prevalence and Risk Factors

Candida is a commensal microorganism of the oral cavity and part of the normal microbial flora. Oral carriage of Candida can be influenced by several factors and may lead to the development of oral candidiasis. Oral swabs were collected from 46 onsultants at the stomatology department and then sent to the mycology laboratory for identification of isolates. The specimens were cultured on CandiSelect medium plates as well as on Sabouraud dextrose agar plates containing chloramphenicol. 38% of cultures were positive. Candida albicans was the most encountered specie (87.5% of positive cultures). Oral Candida colonization was more frequent in the following categories of patients: immunocompromised subjects, patients with malignancy, those undergoing chemotherapy, type 2 diabetics patients, smokers, and patients brushing their teeth irregularly (less than twice a day or occasionally). All of these associations were statistically significant. (p <0,05). The diagnosis of oral Candida carriage prior to the presentation of clinical symptoms could prevent patients at risk of serious complications. The promotion of oral health and hygiene education could help stop the spread of yeast colonization.

Molecular identification of Candida spp. isolated from women withvulvovaginal candidiasis in Koya District, Iraq

Vulvovaginal candidiasis (VVC) is the second most common vaginal infection among women of reproductive age. It is an opportunistic mucosal infection. Candida species causes VVC, which affects millions of women annually. The objective of this study was to isolate and identify the species of Candida from married women who had VVC attending Shahid Doctor Khalid Hospital & Haji Qader Health Centre in Koya, Iraq using traditional and molecular methods. This research was carried out, from December 2021 to May 2022; 150 vaginal swabs from married patients with symptoms of VVC were collected. Swabs were microscopically examined, grown on Sabouraud Dextrose Agar (SDA), and HiCrome™ Candida Differential agar and Germ Tube Test were used to identify Candida isolates. Next, the results were validated using the molecular method called polymerase chain reaction (PCR) to amplify and detect the ITS region in this study. The chitin synthase gene (CHS1) confirmed all C. albicans isolates. Out of the150 vaginal swabs gathered, 84 (56%) cases were positive for vaginal candidiasis. Ages26-30 were found to have the most significant proportion of positive patients, and Women who were pregnant had a more positive culture than women who were not. Identifying six different species of Candida was accomplished through the application of both traditional and molecular techniques in the study; they are C. albicans (88.1%), C. glabrata (4.8%), C. kefyr (2.4%), C. krusei (2.4%), C. parapsilosis (1.2%), and C. tropicalis (1.2%). Finally, we concluded that VVC was shown to be prevalent in Koya city, with the predominant species as C. albicans among all other species.

Microbiological evaluation of conjunctival anopthalmic flora after using digital 3D-printed ocular prosthesis compared to conventional one: a randomized clinical trial

BackgroundThis study aims to assess the influence of using 3D-printed acrylic resin versus conventional Poly-methyl methacrylate (PMMA) for fabricating ocular prostheses on the biofilm and microbial flora of anophthalmic socket.MethodsA randomized controlled trial was designed as a parallel group study. Participants were allocated randomly into two groups: the control group, which received conventionally fabricated ocular prostheses (CG, n = 11), and the test group, which received digitally 3D-printed ocular prostheses (DG, n = 11). Microbiological analysis was conducted before prosthesis insertion and three months after using the ocular prosthesis. Swab samples were inoculated on blood agar, MacConkey’s agar, and Sabouraud’s dextrose agar (SDA) for isolating Gram-positive, Gram-negative, and fungal organisms, respectively. Subsequently, the plates were incubated at 37 degrees Celsius for 48 h. Additionally, a validated questionnaire was used for subjective clinical evaluation, including parameters such as comfort level, socket discharge, lacrimation, and frequency of lubrication for each ocular prosthesis patient in both groups.ResultsTest group (DG, n = 11) exhibited a positive, though statistically insignificant, difference (p > 0.001) in microbial growth when compared to the control group (CG, n = 11). A statistically significant difference was observed in comfort levels between the two groups, with more comfort level within group II (test group) patients. While parameters such as discharge amount, discharge location, lacrimation and lubrication frequency displayed statistically insignificant differences between the two groups, all parameters showed improved results after three months of prosthesis use.ConclusionsThe choice of ocular prosthesis fabrication technique did not yield a statistically significant difference in anophthalmic flora. However, the 3D-printed acrylic resin, as an artificial eye material, displayed potential advantages in reducing the colonization of opportunistic pathogens. All subjective clinical evaluation parameters exhibited enhanced outcomes after three months of prosthesis use, emphasizing the need for an adaptation period during which patients complains are alleviated. In comparison with PMMA, 3D-printed acrylic resin showcased a certain degree of anti-colonization ability against pathogenic bacteria, along with a significant level of patient comfort, suggesting its potential as a promising material for ocular prostheses.Trial registrationThis parallel double-blinded RCT has been registered at ClinicalTrials.gov with identification number: NCT05584865, 18/10/2022.

Clinico Mycological Profile of Dermatophyte Infections at A Tertiary Care Hospital from South India

The prevalence of Dermatophytosis is similar in several continents of the Asian country. The varied state of weather is favourable for mycosis that may lead to various clinical symptoms and infection spreads speedily if left untreated with time. The aim of the present study is to determine the clinical and mycological pattern of dermatophyte infections in patients arriving at our dermatology outpatient department and also to correlate the formal clinical diagnosis with KOH positivity and culture positivity. This descriptive observational study was conducted among 300 patients presenting with dermatophyte infections who came to the dermatology outpatient Department of SRM Medical College and research centre, Kattankulathur, Chengalpattu. The clinical specimens were put through direct microscopy by Potassium Hydroxide (KOH) mount and with the culture on Sabouraud’s Dextrose Agar (SDA). The study showed males were affected mostly and most of the participants were students. The common symptoms observed were Itching, Scaling and Discolouration. Commonly, patients had a primary diagnosis of Tinea corporis infection followed by T. cruris. Trichophyton mentagrophytes were the commonly isolated organism followed by Trichophyton rubrum and Trichophyton violaceum. This study indicates the need for personal hygiene and the disadvantage of as only few participants had zoophilic infections of Tinea species. These methods of diagnosing and identification will further aid in better patients management.

Phenotypic characterization and antifungal susceptibility profile of Candida isolates from women with vulvovaginal candidiasis at a community health center linked to a teaching institution in the Sub-Himalayan region of North India.

Vulvovaginal candidiasis (VVC) is considered a common gynecological problem among females of reproductive age group. 70-75% of women report having had candidal vulvovaginitis at some point in their lifetimes and 40-50% suffer recurrent candidal vulvovaginitis. This study aims to identify the Candida species involved in VVC and to determine their antifungal susceptibility pattern. The present study was a cross-sectional study conducted on 257 females (18-55 yr) with complaints of abnormal vaginal discharge. For detection of Candida, the swab samples were subjected to Gram stain, 10% KOH mount, and culture on Sabouraud dextrose agar (SDA). Candida species identification was done by subculturing Candida isolates onto CHROMagar, corn meal agar (Himedia), and further confirmation was done by MALDI-TOF MS. Antifungal testing was done using the disk diffusion method. A total of 257 females with complaints of abnormal discharge were enrolled in this study. Out of 257, C. albicans 37 (58.7%) and 26 (41.3%) isolates were identified as non-albicans Candida. Out of 63 positive cases, a maximum number of study subject belongs to the age group 26-35 years (50.8%). Along with vaginal discharge, itching (65.37%) is the most common complaint. VVC was found to be most predominant in patients with prolonged antibiotic therapy (38.1%), and in pregnant females (15.9%). Understanding the emerging fungal pathogens and their drug susceptibility patterns is essential for the effective management of infections. Drug resistance can lead to treatment failure and highlights the need for alternative treatment options or strategies.

Biological Characteristics of Beauveria majiangensis Strain MJ1015 and Optimization of Solid Medium Technology for Sporulation.

The entomopathogenic fungus Beauveria majiangensis strain MJ1015, recently isolated from white grubs on a blueberry farm in Guizhou, China, could be used as a biocontrol agent. As a first step toward determining the effect of different solid culture media, temperature, and pH on colony growth rate and sporulation, we evaluated the optimum solid medium for mycelial growth and conidia production on a commercial scale. Subsequently, we also used single-factor analysis and response surface optimization to optimize the composition of the solid culture medium. On potato dextrose agar (PDA) medium, MJ1015 grew fastest and produced the highest spore yield at 29°C and pH 5. The best solid medium for the growth and sporulation of strain MJ1015 comprised 64.70 g/l of rice, 13.00 g/l of wheat, 0.30 g/l of NaNO3, 0.36 g/l of K2HPO4 · 3H2O, and 1.00 g/l of CaCO3. Rice, NaNO3, and K2HPO4 · 3H2O were the main influencing factors. The predicted value of cultured spores using the optimal medium was 4.56 x 1010 conidia/l. The validation test results showed that the average growth rate of strain MJ1015 on the optimal medium was 85% and 96% faster than that on Sabouraud dextrose agar with yeast extracts medium (SDAY) and PDA, respectively. Sporulation was 43.90 times and 9.65 times of that produced on SDAY and PDA, respectively. Our findings provide a theoretical basis for the commercial production of B. majiangensis to control white grubs.

806. Candida Gut Colonization in Critically Ill Patients

Abstract Background Candida spp. colonization is a risk factor for candidemia in intensive care unit (ICU) patients. Recent data suggest that the epidemiology of Candida infections is shifting away from C. albicans, but prospective data on the epidemiology of Candida spp. gut colonization are limited. Methods We conducted a prospective cohort study of adult patients admitted to an ICU at a tertiary care hospital. Patients had stool samples collected twice weekly for up to four weeks or until ICU discharge. A convenience sample of stool samples was plated on CHROMagar Candida Plus for 48 hours. Colonies of different morphologies were isolated on Sabouraud dextrose agar and identified with MALDI-ToF. Results A total of 102 patients were included in the cohort. We performed screening for stool Candida colonization in 12 patients: 8 (67%) men, median age 65 (IQR 54.5-68.5). Three (25%) were in shock and eight (67%) required mechanical ventilation on ICU admission. In-hospital mortality was 25%. Seven patients (58%) were colonized with Candida at ≥1 time point (Figure 1). Of 30 total time points tested (median per patient=2, range 1-5), 15 (50%) were positive for Candida spp. Most time points (9/15, 60%) were positive for C. glabrata; 4/15 (27%) were positive for both C. glabrata and C. albicans. A minority (1/15; 7%) were positive for C. albicans and C. parapsilosis alone, respectively. None had C. auris colonization. Most colonization was persistent: Of 6 colonized patients with ≥1 time point tested, 5 (83%) were colonized at multiple timepoints. Eleven patients had data collected on anti-fungal administration during the study period, and five of these received an antifungal (Figure 1). Of these five, three (60%) had no Candida colonization. One lost C. albicans colonization but had persistent C. glabrata colonization despite azole therapy, and another developed C. parapsilosis colonization despite azole and amphotericin administration. Figure 1. Results of serial stool samples tested for Candida colonization per individual patient. Conclusion Most ICU patients in this study had Candida spp. gut colonization, commonly with intrinsically azole-resistant C. glabrata. This study is limited by small sample size, and further data are needed to determine the clinical impact of antifungal-resistant Candida in ICU patients at high risk of candidemia. Disclosures All Authors: No reported disclosures

2485. Candida auris Hospital Cluster: Were Our Unwelcome Guests Driven Away by the COVID-19 Delta Surge?

Abstract Background Candida auris (CA) is an MDR pathogen that persists in the environment, causing outbreaks. NUH is an academic medical centre in Singapore. We describe a cluster of CA cases from April 2019 – October 2021. Our last case was during our Delta surge, just before we changed our routine inpatient cleaning to bleach based from quarternary-ammonium. Methods Any CA from the micro lab is escalated to the Infection Prevention Team to initiate isolation, cleaning, and contact tracing (CT) up to 30 days prior to positive cultures. In September 2020, we started enhanced surveillance via NAG (Nasal, axilla and groin) swabs in all new CRE (Carbapenem-Resistant Enterobacterales) patients. Surveillance cultures used sabouraud dulcitol agar with 10% saline incubated at 40°C. Identification and susceptibility testing done using Bruker MALDI Biotyper and Etest (Biomerieux). Hospital onset (HO) is defined as positive cultures sent after day 3 of admission. CA isolates were sent for whole genome sequencing (WGS). Results The 10 cases included 7 from clinical isolates, 2 from enhanced surveillance and 1 from CT. 8 were HO. 7 deaths were during the CA admission, but only 1 due to CA, and 1 occurred > 60 days after positive culture [Table 1]. Of 231 contacts swabbed all were negative, except for Case 8, in whom one contact (Case 9) was positive. Positive yield from enhanced surveillance of CRE patients, was 0.84% (2/239). WGS results indicated all except Case 9 belonged to Clade I with Y132F mutation in ERG111 gene [Table 2]. Cases 2, 3, 5, 6 were closely related with 2-7 SNPs difference, but spaced in years. Cases 7 and 8 differed by 3 SNPs hence, closely related. There were 2 related clones being transmitted, and 4 unrelated sub-types of CA. Case 8 and 9 were not related, though 9 was a contact of 8. Possible links were shared equipment or admission to a common room however, admissions were either non-overlapping or non-successive [Table 3]. Case 5 was admitted to the same room after a long stay by Case 3, with one patient in between. No links found between Cases 7 and 8. Conclusion WGS results suggest that various clones of CA remain in the environment as reservoirs, and related cases are not often spatiotemporally related. There have been no CA cases since Oct 2021, possibly due to the change to bleach-based cleaner during our Delta surge. Disclosures All Authors: No reported disclosures

Phenotypic identification of Candida species and relative expression of candida drug resistance (CDR1) gene in fluconazole resistant and sensitive Candida albicans

ABSTRACT:
 Background and objective:
 Candida albicans belongs to the genus Candida. It is a commensal yeast in healthy individuals and can cause wide variety of infections in immune-compromised patients. The aim of the present study was to observe the frequency of Candida species, resistant profile of C. albicans against fluconazole and over-expression of CDR1 gene in both fluconazole resistant and sensitive C. albicans.
 Methodology:
 A total number of 206 Candida species were collected on Sabouraud Dextrose agar from Jinnah hospital Lahore. The isolates were re-confirmed by using various microbiological techniques like wet preparation, Gram stain and Germ tube test. Identification upto the species level was done by APIC 20AUX. Antifungal susceptibility profile of C. albicans against fluconazole was determined by broth microdilutiom method. Real Time PCR was performed on both the sensitive and resistant C. albicans to observe the role of CDR1 gene against fluconazole resistance. Results:
 
 tropicals was the most common species with the frequency of 66 (32.0%) followed by C. albicans 41(19.9), C. glabrata 24 (11.7%), C. lusitaniae 14 (6.8%) and other less common species. The majority of infected patients were females (59.2%) than males (41%). Antifungal susceptibility testing showed that 80% C. albicans were sensitive to fluconazole. Results of gene expression showed that CDR1 gene play a major role towards fluconazole resistance in C.albicans.Conclusion: Fluconazole was found to be effective in treating infections caused by C. albicans. 
 
 Keywords: Candida albicans, Candida Drug Resistant Gene 1, Polymerase Chain Reaction, Candida Tropicalis, Candida glabrata, and Candida lusitaniae

Antifungal susceptibility of Aspergillusgenus determined by the Etest® method: eleven years of experience at the Instituto Médico La Floresta. Caracas, Venezuela.

Abstract. This research aimed to determine the susceptibility of Aspergil-lus spp. to four antifungal agents using the Etest® method in several clinical samples (respiratory samples, soft tissue, otic tissue, and ocular tissue, among others) from a private health center in Venezuela. Thirty-three strains were evaluated: 11 Aspergillus section Flavi, eight Aspergillus section Fumigati, six Aspergillus section Nigri, four Aspergillus section Terrei, and four Aspergillus spp. A 0.5 McFarland standard suspension of a 5-day culture of each Aspergillusstrain was prepared on Potato Dextrose agar and then inoculated on Sabouraud agar plates with 2% glucose. Voriconazole (VCZ), amphotericin B (AMB), caspo-fungin (CAS), and posaconazole (PCZ) were tested. Minimal inhibitory concen-trations (MIC) in μg/mL were determined after 24 and 48 hours of incubation at 35 °C and th range (R), geometric mean (GM), MIC50, and MIC90 were calcu-lated. The results for the 33 Aspergillus spp. tested after 24 h were the follow-ing: VCZ (R = 0.031- 16; GM = 0.145; MIC50 = 0.125 and MIC90 = 0.5), AMB (R = 0.031-16; GM = 0.644; MIC50 = 0.5 and MIC90 = 8), CAS (R = 0.031-16; GM = 0.1076; MIC50 = 0.063 and MIC90 = 1), PCZ (R =0.031 - 0.5; GM = 0.0755; MIC50 = 0.063 and MIC90 = 0.25). This investigation allowed assessing the an-tifungal susceptibility profiles of Aspergillus spp. isolated from clinical samples by the Etest® method, which is practical, reproducible and easy to perform in microbiology laboratories.

Mucormycosis Associated with Global Coronavirus Disease-19 Pandemic: A Study of Fungal Profile from a Tertiary Health-care Center

Background and Aim: In this era of the coronavirus disease (COVID) pandemic, new health issues appear quickly and cause an outbreak in India. Here, we focus on invasive fungal infections (IFA), namely mucormycosis, aspergillosis, and candidiasis. Hence, in this article, we focus on conventional techniques for early diagnosis of fungus to help clinicians start empirical treatment for the patient and take necessary steps thereby. Materials and Methods: This observational study was conducted for 1 month, from May 2021 to June 2021, on 200 patients admitted to our mucormycosis ward. Samples were first examined microscopically using potassium hydroxide (KOH) wet mounts followed by culture on sabouraud dextrose agar and the gross appearance of the growth was observed. Further identification of species was achieved through the use of lactophenol cotton blue mounts and slide culture technique. Results: Out of 200 patients, 65.5% were male and 34.5% female. We divided these patients into three age groups, i.e., 20–30 years (15%), 40–60 years (54%), and 60–80 years (31%). Seventy-four percent of patients presented with a history of diabetes mellitus, 50% had a history of steroids taken in their past, and 48% had a history of oxygen therapy taken at the time of COVID-19 positivity (52%) samples positive for mucormycosis. Forty-one percent were KOH-positive for Mucorales and 35.5% were culture positive. The most commonly isolated fungus was Rhizopus, followed by Mucor, Lichthemia, Aspergillus spp., and Candida spp. Conclusion: To improve test sensitivity and enable early diagnosis, other serum biomarkers such as galactomannan, beta-D-glucan, cryptococcal antigenemia, and blood quantitative polymerase chain reaction for Aspergillus or Mucorales should be implemented.

Dermatophytosis is uncommon in asymptomatic wild eastern cottontail rabbits in Illinois, Midwestern United States.

To investigate the presence of dermatophytes on the haircoat of wild eastern cottontail rabbits (ECR) (Sylvilagus floridanus) with and without skin lesions. 2-week-old or older ECR admitted to a Wildlife Medical Clinic (WMC) in central Illinois, Midwest United States, from September 2021 to August 2022. ECR were surveyed over a 1-year period to assess the prevalence and seasonality of dermatophytosis. A Wood's lamp exam was performed over the haircoat. Hairs were sampled with a sterile toothbrush and plated on Sabouraud dextrose agar. The plates were photographed twice weekly for 3 weeks, and colonies were identified as contaminants or dermatophytes. 523 ECR were admitted to WMC, 141 ECR met the age inclusion criteria, and 121 samples were plated. ECR presented as a litter were sampled together. None of the sampled ECR presented skin lesions other than acute traumatic wounds. No fluorescence was observed on any ECR during the Wood's lamp examination. Based on culture colony morphology, 115/121 of the samples were identified as contaminants and no growth was observed in 6/121 of plates. Dermatophytes' colonies were not identified on any of the culture plates. Dermatophytes are zoonotic fungi and can potentially be carried by wild animals. The fungal infection poses a health concern to humans and domestic pets through direct interaction. Our current results suggest that dermatophytosis may not be prevalent in asymptomatic wild rabbits in the studied areas of the Midwestern United States.

Kali carbonicum 6cH controls Malassezia pachydermatis growth in vitro

External otitis caused by Malassezia pachydermatis is a common disease in the veterinary practice routine, whose treatment has been challenging due to the risk of antimicrobial resistance. Thus, Homeopathy is seen as a possible alternative to control the infection. To screen potential medicines to be used in a non-individualized clinical homeopathy approach, a series of in vitro assays were performed, in which a suspension of M. pachydermatis colonies in 0.5 McFarland scale was diluted 1:1000 in sterile saline solution, and 10 µL of pre-selected homeopathic medicines was added. Sulphur 6cH, Dolichos pruriens 6cH, and Kali carbonicum 6cH were chosen from a pilot study, and the last potency was prepared in pure, sterile water. The controls were pure, sterile water and succussed pure, sterile water. An amount of 50 µL of each suspension has seemed in plates containing Sabouraud dextrose agar medium, with or without 1% Tween 80, for counting of colony-forming units (CFU) and cytomorphological analysis of M. pachydermatis samples taken from the same colonies. The analysis was made using Image J 1.53 by manually counting quiescent and germinative forms. Kali carbonicum 6cH treatment decreased the germinative/quiescent ratio in relation to the controls, independent of Tween 80 presence in the medium. The UFC counting, however, did not show statistically significant differences among groups, probably by the lack of cytotoxicity against the yeasts. In conclusion, Kali carbonicum 6cH was considered a promising medicine to be tested in clinical conditions, probably due to its capacity to control yeast growth without creating a selective pressure able to induce resistance mechanisms.

Pulmonary Aspergillosis in Naïve Non-neutropenic Lung Cancer Patients

Abstract: Pulmonary aspergillosis is the most common mould infection of the lungs. Patients with chronic lung disease or mild immunodeficiency are at risk. Lung cancer represents an oncological challenge. Limited data are available about aspergillosis in newly diagnosed non-neutropenic lung cancer patients. Objectives:: The aim of this work was to evaluate naïve lung cancer patients for the presence of as-pergillosis, identify fungal isolates, and determine antifungal susceptibility. The inhibitory effect of secondary metabolites of fungal isolates on the human fibroblast cell line was also assessed. Results:: Aspergillus isolates were detected in 33 BAL samples out of the 69 patients (47.8%). The commonest isolate was A. niger. There was no significant correlation between total fungal count and age of patients. The antifungal Micafungin had a broad-spectrum and potent fungistatic activity against all fungal species, with A. niger being the most sensitive. MTT assay was performed to evaluate the cytotoxic effect of both A. niger and A. fumigatus on normal lung cells (WI-38) and re-vealed that toxicity was concentration-dependent and A. niger had significantly lower IC 50, indi-cating more toxicity. Conclusion:: In the context of the diagnosis of lung cancer, maintaining vigilance is crucial to diag-nose fungal infection as aspergillosis. Initiation of proper management can help improve patient outcomes.

Occurrence and Antifungal Susceptibility of Candida Species Isolated from Pregnant Women in a Tertiary Hospital

Candida is the leading cause of infection among pregnant women. This study was aimed to determine the occurrence and antifungal susceptibility of Candida species isolated from apparently healthy pregnant women receiving antenatal care at Bingham University Teaching Hospital. High vaginal swab was inoculated onto Sabouraud Dextrose Agar and Gram staining was carried out. Candida isolates were sub- cultured on Himedia CHROM Candida differential agar and identified according to their color, morphology, and appearance. Disc antifungal susceptibility test was carried out using standard protocols. The occurrence rate of vulvovaginal candidiasis (VVC) in this study was 42.5%. In this study, Candia albicans was the most predominant Candida species (51.2%) isolated. Pregnant women within the age range of 20-24 years had the highest infection occurrence of 100% while the lowest occurrence of the infection (25%) was observed among women within the age range of 40-44 years. There was a significant relationship between age and VVC infection (P>0.05). In relation to trimester, pregnant women in the second trimester had a high infection rate of 87.5% while women in the third trimester had the least occurrence rate of 10.9%. There was a significant relationship between vulvovaginal candidiasis infection and trimester (P>0.05). All Candida species were resistant to the antifungal drugs tested except for Candida kefyr which was susceptible to fluconazole and voriconazole. Therefore, early diagnosis and appropriate treatment among pregnant women receiving antenatal care can reduce the infection and prevent complications during childbirth