Sabouraud Dextrose Agar Medium Research Articles

Strategies to Reduce the Use of Antibiotics in Fresh and Chilled Equine Semen.

The study assessed the impact of four equine semen processing techniques on sperm quality and microbial load immediately post-processing and after 48 h of refrigeration. The aim was to explore the potential reduction of prophylactic antibiotic usage in semen extenders. Semen from ten adult stallions was collected and processed under a strict hygiene protocol and divided into four aliquots: Simple Centrifugation with antibiotics (SC+), Simple Centrifugation (SC-), Single-Layer Colloidal Centrifugation (CC-), and Filtration (with SpermFilter®) (F-), all in extenders without antibiotics. Sperm motility, viability, and microbial load on three culture media were assessed. No significant differences were observed in the main in the sperm quality parameters among the four protocols post-processing and at 48 h (p < 0.05 or p < 0.1). Microbial loads in Columbia 5% Sheep Blood Agar and Schaedler vitamin K1 5% Sheep Blood Agar mediums were significantly higher (p < 0.10) for raw semen than for CS+, CC-, and F- post-processing. For Sabouraud Dextrose Agar medium, the microbial load was significantly higher (p < 0.10) in raw semen compared to CS+ and F-. No significant differences (p < 0.10) were found in 48 h chilled samples. Regardless of antibiotic presence, the evaluated processing methods, when combined with rigorous hygiene measures, maintained semen quality and reduced microbial load to the same extent as a traditional protocol using antibiotics.

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 &lt; 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.

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.

The effect of ethanol extract of &lt;i&gt;Cosmos caudatus&lt;/i&gt; leaves on the percentage of the cell cycle in &lt;i&gt;Candida albicans&lt;/i&gt; culture

Candidiasis is a common fungal gynecological disease among humans. The use of antifungal agents, such as Fluconazole, has been reported to increase resistance to candidiasis by 7%. This study aimed to investigate the effect of antifungal flavonoids from Cosmos caudatus leaf extract on the cell cycle percentage in C. albicans culture. This research employed a true experimental post-test only with a control group design. The C. albicans isolate was obtained from the Microbiology Laboratory under the code C. albicans SV-1148. The isolates were cultured in Sabouraud Dextrose Agar (SDA) medium and Sabouraud Dextrose Broth (SDB). The sample group was divided into a negative control group, a positive control group with fluconazole (60 µg/ml), and a treatment group with various concentrations of ethanol extract from Kenikir leaves (C. caudatus Kunth.) - 5%, 10%, 20%, 40%, and combinations (20% ethanol extract from Kenikirleaves + 30 µg/ml Fluconazole). This study revealed a reduction in the percentage of cell cycles in the S phase (DNA synthesis) in the treatment group receiving ethanol extract from Kenikir leaves (C. caudatus Kunth.) and the combination treatment group compared to the negative control group. The study suggests that this decrease in the percentage of cell cycles results from DNA damage caused by the presence of flavonoids in Kenikir leaves (C. caudatus Kunth.). Kenikirleaves (C. caudatus Kunth.) have the potential to decrease the percentage of S-phase cell cycles (DNA synthesis) in the culture of C. albicans. This research demonstrates that Kenikir leaves (C. caudatus Kunth.) contain natural flavonoids with antifungal properties and have the potential to be used as an alternative medicine for candidiasis in humans. It is hoped that incorporating Kenikir as a food ingredient can serve as an alternative prevention and treatment approach for candidiasis.

ANTIFUNGAL ACTIVITY OF RAMANIA LEAF EXTRACT (Bouea Macrophylla Griff) AGAINST Candida albicans

Background: Root canal infection is a polymicrobial infection that causes pulpitis or localized pulp inflammation. Root canal infection occurs due to the presence of pathogenic microorganisms in the root canal. One of the most common pathogenic microorganisms found in root canals is the fungus Candida albicans. Treatment that can be done to treat pulp tissue infection is endodontic treatment. Chlorhexidine gluconate 2% is one of the irrigation materials that can be used in root canal treatment. Ramania leaf extract contains compounds such as triterpenoids, phenols, flavonoids, steroids, saponins, alkaloids, and tannins that can function as antifungals. Purpose: Measuring and analyzing the antifungal activity of ramania leaf extract (Bouea Macrophylla Griff) on the growth of Candida albicans. Method: This study is a true experimental study with a post-test-only design with a control group design consisting of 7 treatment groups and repeated 4 times. The antifungal activity was assessed from the inhibition zone formed on Sabouraud Dextrose Agar media by the diffusion method. Results: Based on the Mann-Whitney test showed that there was only 1 pair of groups that did not have a significant difference, namely 100% concentration of ramania leaf extract with 2% chlorhexidine gluconate while the other groups had significant differences from each other. Conclusion: Ramania leaf extract concentrations of 6.25%, 12.5%, 25%, 50%, and 100% had antifungal activity in inhibiting the growth of the fungus Candida albicans. Keywords: Antifungal Activity, Candida albicans, Chlorhexidine gluconate 2%, Ramania Leaf Extract.

Isolation and identification of Candida spp. isolated from patients with oral candidiasis in Kerbala, Iraq.

83 oral swabs were collected from patients clinically diagnosed with oral candidiasis and Who attended the Children’s Teaching Hospital in Kerbala City. Samples were cultured directly on Sabouraud Dextrose Agar medium(SDA) supplemented with amoxicillin, and it was found that there was growth in 63 samples (76%) classified according to the type of pathogen 58 samples (92%) yeast and 5 samples (8%) bacteria. Most isolated yeasts belonged to the genus Candida, which amounted to 93.1% (54 samples), while the non-Candida yeasts recorded 6.9% (4 samples). Candida albicans was recorded as the dominant among the Candida species at 53.7% (29 isolates), followed by C. dubliniesis (18.5%) (10 isolates), C.lusitaniae 12.9% (7 isolates), C.tropicalis 7.4% (4 isolates), C.parapsilosis 5.6%, and C.kefyr 1.9% (1 isolate). The results of the physiological examinations showed the ability of all C.albicans, and C.dubliniesis isolates to produce germ tubes and chlamydo spores on cornmeal agar medium compared to the other species. All species showed the ability to produce pseudohyphae, but C.albicans isolates are the only ones that can grow under a temperature of 45 m. The growth results on HiCrome™ Candida Differential Agar (M1297A) confirmed the physiological tests, where all C. albicans isolates appeared in light green and C. difficiensis in dark green. In contrast, C.lusitaniae isolates appeared in purple, C.tropicalis in metallic blue, C.parapsilosis in white to pale pink, and C.kefyr in pink to purple. The diagnosis of the isolated species was confirmed using the Vitek device by the Vitek 2 YST card, and the results were interpreted by comparing the results of biochemical reactions and information available in the system database. The Vitek system was also used to examine the sensitivity of the isolated species to six antifungals, which are Fluconazole, Flucytosine, Capsufungin, Amphotericin B, Micafungin, and Voriconazole, using the Vitek 2AST-YS08 card. All species isolates are sensitive to Capsufungin, except one isolate of C.tropicalis and one of C.parapsilosis showed moderate sensitivity to this antibiotic. As for Amphotericin B, two isolates of C. albicans showed resistance (6.9%), and one isolate of C. lusitaniae showed resistance to Amphotericin B by 14% compared to the other species that showed sensitivity to this antibiotic by 100%. In comparison, all species were sensitive to Flucytosine at 100%, except 3 isolates of C. albicans were resistant to this antibiotic (10.3%) and 1 C. dubliniesis (10%). Keywords: Candida sp., Oral candidiasis, Fungal pathogens

Antifungal Activity of Endophytic Bacteria Isolated from Miana Plants (Coleus scutellarioides (L.) Benth.) against Candida albicans

Highlights: 1.Due to the adverse effects associated with current antifungal drugs, research on traditional medicine is necessary to explore other options for candidiasis treatment.2.Following the findings of this study, it is recommended to conduct further research by incorporating Coleus scuttellarioides filtrate into the growth medium, since this may improve bacterial growth by producing optimal secondary metabolites. Abstract Candida albicans is the most common organism responsible for both mucosal and systemic infections, accounting for approximately 70% of fungal infections worldwide. Miana, scientifically known as Coleus scuttellarioides (L.) Benth., is recognized for its use in traditional medicinal practices. Miana plants contain endophytic bacteria that possess the ability to produce secondary metabolites with potential antifungal agents. The objective of this study was to assess the antifungal activity of nine endophytic bacteria isolates derived from Coleus scuttellarioides against Candida albicans. This study was laboratory-based qualitative experimental research that applied the Kirby-Bauer diffusion method and several modifications. The Candida albicans specimens were spread throughout the entire potato dextrose agar medium. Afterwards, paper discs that had been soaked in a liquid culture of endophytic bacterial isolates were carefully placed on the surface of the medium. The complete setup was then incubated for 1–2 days. The potential antifungal activity of endophytic bacteria was assessed by observing the emergence of a clear zone surrounding their growth, which would indicate inhibition. An additional observation was performed in the follow-up test, involving the use of Sabouraud dextrose agar medium to confirm the initial test result. The results from the inhibitory test revealed that none of the bacterial isolates exhibited any inhibition zone. Conversely, ketoconazole as the positive control showed an inhibition zone with an average diameter of 28.5 mm. In conclusion, endophytic bacterial isolates obtained from Coleus scuttellarioides have no discernible antifungal properties against Candida albicans. This study implies that ketoconazole remains effective in treating infections caused by Candida albicans.

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 &#x0D; isolated from dry dates in Yemen Republic. Thirty four date samples &#x0D; collected from different shops and markets in Sana’a city were &#x0D; analyzed mycologically for the presence of fungi cultivated on three &#x0D; types of media. Eight species belonging to 7 genera were isolated &#x0D; from analyzed date samples on the three cultural media. Aspergillus&#x0D; genera was the most dominant fungi and grew well on the three type &#x0D; of media of which A. niger is the most common species, A. flavus was &#x0D; isolated in rare, low and rare frequency on 1% and 20% sucrose &#x0D; Czapek’s and Sabouraud dextrose agar media. Rhizopus stolonifer &#x0D; was isolated in rare, low and low frequency on 1, and 20% sucrose &#x0D; Czapek's and Sabouraud dextrose agar media. The date samples were &#x0D; analyzed for the presence of total aflatoxin using ELISA technique &#x0D; which revealed that 4 out of 5 date samples were contaminated with &#x0D; aflatoxin ranged from 3059.89-7585.96 ppt (ng Kg-1). &#x0D;