Sabouraud Dextrose Agar Research Articles

Efficiency of Persicaria salicifolia as a natural alternative antifungal against Aspergillus flavus infection in Oreochromis niloticus from Aswan Governorate, Egypt

Fungal diseases in fish cause economic losses all over the world, and knowledge about them is scarce and outdated in Aswan Governorate, Egypt, making interpretation, prevention, and treatment difficult. The necessity to find a fungicide that is natural, environmentally friendly, and does not emerge drug resistance is a must. Therefore, the current study aimed to isolate and diagnose fungal infection in farmed Oreochromis niloticus, causing mortalities, in Aswan Governorate. During 2021, 200 fresh O. niloticus samples were collected from the Sahary Fish Hatchery and Aswan General Authority for Fish Resources Development fish farm. Some fish showed hemorrhagic lesions all over the body, detachment of scales, and fin erosion. Collected tissue samples were cultured on potato dextrose agar and Sabouraud dextrose agar for phenotypic characterization. Macroscopic and microscopic analyses were used to identify the isolated fungi. A total of 18 fungal species and two varieties appertaining to ten fungal genera were recovered from 48 samples out of 200 examined O. niloticus (24%), with Aspergillus flavus being the most prevalent at a rate of 25.6%. The isolated A. flavus was proven to be pathogenic to farmed O. niloticus, as by experimental infection. The natural herb Persicaria salicifolia had an LC50 value of 41.68 mg/l in exposed O. niloticus and was used to treat A. flavus-infected O. niloticus. It can be concluded that A. flavus poses a major hazard to O. niloticus aquaculture and can be treated with 40 mg/kg in feed or 20 mg/l in water of P. salicifolia for 6 days.

Efficacy and Rapidity of Potassium Hydroxide Mount and Modified Chicago Sky Blue 6B Stain with Potassium Hydroxide in Fungal Keratitis Detection.

To compare the efficacy and rapidity of direct microscopic detection of fungal elements from corneal ulcers between 10% potassium hydroxide (KOH) and 1% Chicago Sky Blue 6B (CSB) in 10% KOH (CSB-KOH). Thirty patients with clinically suspected fungal keratitis were recruited. Participants with impending corneal perforation were excluded. Two slides were smeared with corneal ulcer scrapings from the ulcer's edge and base for comparison of fungal staining solutions. One slide was infused with KOH, and the other slide was filled with CSB-KOH. Additional scraping was collected for inoculation on Sabouraud dextrose agar for fungal culture. The sensitivity, specificity and rapidity of both stainings were analyzed. The sensitivity of fungal culture, KOH, and CSB-KOH were 43.75% (95% confidence interval [CI], 19.75%-70.12%), 62.50% (95% CI, 35.43%-84.80%), and 87.50% (95% CI, 61.65%-98.45%), respectively. The specificity were 100% (95% CI, 69.15%-100%) of both stainings and fungal culture which analyzed from 16 fungal keratitis cases by laboratory and clinical diagnosis. Mean CSB-KOH examination time was quicker than KOH with the mean time difference of 5.6 minutes (95% CI, 3.22-7.98 minutes) and p-value < 0.001. CSB-KOH was more effective and faster than KOH in detecting fungal elements from corneal ulcers. Therefore, CSB-KOH may be beneficial in diagnosing fungal keratitis and preventing blindness. Moreover, to the best of our knowledge, this is the first use of CSB stain in fungal keratitis detection.

Intracellular presence of Helicobacter pylori antigen and genes within gastric and vaginal Candida.

Helicobacter pylori infections are generally acquired during childhood and affect half of the global population, but its transmission route remains unclear. It is reported that H. pylori can be internalized into Candida, but more evidence is needed for the internalization of H. pylori in human gastrointestinal Candida and vaginal Candida. Candida was isolated from vaginal discharge and gastric mucosa biopsies. We PCR-amplified and sequenced H. pylori-specific genes from Candida genomic DNA. Using optical and immunofluorescence microscopy, we identified and observed bacteria-like bodies (BLBs) in Candida isolates and subcultures. Intracellular H. pylori antigen were detected by immunofluorescence using Fluorescein isothiocyanate (FITC)-labeled anti-H. pylori IgG antibodies. Urease activity in H. pylori internalized by Candida was detected by inoculating with urea-based Sabouraud dextrose agar, which changed the agar color from yellow to pink, indicating urease activity. A total of 59 vaginal Candida and two gastric Candida strains were isolated from vaginal discharge and gastric mucosa. Twenty-three isolates were positive for H. pylori 16S rDNA, 12 were positive for cagA and 21 were positive for ureA. The BLBs could be observed in Candida cells, which were positive for H. pylori 16S rDNA, and were viable determined by the LIVE/DEAD BacLight Bacterial Viability kit. Fluorescein isothiocyanate (FITC)-conjugated antibodies could be reacted specifically with H. pylori antigen inside Candida cells by immunofluorescence. Finally, H. pylori-positive Candida remained positive for H. pylori 16S rDNA even after ten subcultures. Urease activity of H. pylori internalized by Candida was positive. In the form of BLBs, H. pylori can internalize into gastric Candida and even vaginal Candida, which might have great significance in its transmission and pathogenicity.

Speciation of Candida species isolated from cutaneous skin lesions using CHROMagar and conventional methods

Background: Candida species are members of the normal flora of the skin, mucous membranes, and gastrointestinal tract. They are endogenous opportunists which cause secondary infection in individuals with some underlying immunocompromised conditions. Candidiasis is a common fungal disease found in humans affecting mucosa, skin, nails, and internal organs of the body. Candida albicans is generally considered the major pathogen among the Candida species. An increase in the prevalence of non-albicans species has been noted during the last decades. Aims and Objectives:The study was done to evaluate the performance of the conventional identification method and commercially available chromogenic Candida speciation media (CHROMagar) for the identification of medically important yeast and yeast-like organisms in a routine clinical microbiology laboratory. Materials and Methods: This study was conducted in the Department of Microbiology, Gandhi Medical College, Hamidia, and Associated Hospitals, Bhopal (M.P.), from January 2020 to October 2021. A total of 180 samples were collected from patients suspected of superficial mycoses which were then subjected to direct KOH examination and culture on plain Sabouraud’s dextrose agar (SDA), on SDA with antibiotics slants (cycloheximide and chloramphenicol). The growths were examined macroscopically as well as microscopically. Germ tube tests and inoculations on CHROMagar Candida medium were performed on all growths identified as yeasts and confirmed by morphology on CMA. Results: In our study, the majority of patients, belonged to the age group of 21–30 years (25%) with a male-to-female ratio of 2:1. Candida tropicalis (44.68%) was the most common Candida species, followed by C. albicans (38.30%), Candida krusei (12.77%), and Candida glabrata (4.25%). Conclusion: Along with C. albicans, non-albicans Candida spp such as C. tropicalis, C. krusei, and C. glabrata are increasingly being isolated from clinical specimens. Identification of Candida species by colony color on Hicrome correlated with the morphology on CMA. Thus, the incorporation of CMA in routine yeast identification is more judicious than Hicrome as it increases the accuracy in the identification of Candida species within the same time as that of Hicrome.

CHARACTERISTICS OF PH AND NUMBER OF MICROBES IN THE RETICULUM, OMASUM AND ABOMASUM OF BALI CATTLE

Bali cattle are native Indonesian cattle germplasm which must be preserved and cultivated. The research aims to determine the pH characteristics and number of microbes in the reticulum, omasum and abomasum of Bali cattle. The research sample was the stomach contents of the reticulum, omasum and abomasum of clinically healthy Bali cattle slaughtered at the slaughterhouse, Pesanggaran, Denpasar. Samples were taken aseptically from 30 cows. This research is observational with a cross-sectional study design. The number of bacterial and fungal colony populations was calculated using the pour plate method on nutrient agar plates and Sabouraud dextrose agar plates, respectively. The population of Enterobacteriaceae bacterial colonies was counted using the scatter method on Eosin Methylen Blue agar plates. pH measurements in cow stomach contents were carried out using pH indicator strips. The research data were analyzed descriptively. The results showed that the average pH of the reticulum, omasum and abomasum were 7,66; 6,50 and 5,26. The average total bacteria or ALTB in the reticulum, omasum and abomasum were 18,53 x 106 CFU/gr, 21,43 x 106 CFU/gr and 26,73 x 106 CFU/gr; the fungal population was 7,10 x 105 CFU/gr, 4,56 x 105 CFU/gr and 2,83 x 105 CFU/gr; Coliform counts were 60,03 x 103 CFU/gr, 57,70 x 103 CFU/gr and 61,60 x 103 CFU/gr; the number of non-Coliforms was 49,00 x 103 CFU/gr, 45,53 x 103 CFU/gr and 41,46 x 103 CFU/gr, the number of E.coli respectively 20,70 x 103 CFU/gr, 22, 83 x 103 CFU/gr and 22,86 x 103 CFU/gr.

Isolation and Identification of Yeasts from Slaughterhouses in Baghdad province

The aim of this research was isolation and identification of yeasts from slaughterhouse environment which including equipment specimens before slaughtered and after slaughtered as well as vital organs swabs in sheep carcasses were slaughtered inside of slaughterhouse , the specimens were collected in Baghdad province during winter season (2023);and two hundred (200) samples were collected from abattoirs divided in to (50) samples before slaughtered and (50) samples after slaughtered and finally another (100) samples were obtained from sheep carcasses which were distributed into twenty-five(25)swabs from each of lung,liver,heart and spleen; these samples were inoculated on Sabouraud Dextrose Agar (SDA),so the results revealed that the percentage of yeasts isolated before slaughtered was (52%) while after slaughtered it decreased to (42%),while percentage of isolation from vital organs of sheep carcasses (22%) in which most visible in abattoirs environment C. albicans was in percent of (10%) then Rhodotorula mucilaginosa was recorded (7.5%) , whereas after C. parapsilosis appeared in percent of (4.5%) while Cryptococcus albidus and C. guilliermondii appeared in percent of(3%) ; C. famata appeared in percent of (2.5%), both C. zeylanoides and C. sphaerica were recorded the same percent of (1%);finally each of C.rugose, C. Lusitaniae ,C.tropicalis, and C. krusei were appeared in percent of (0.5%).

In Vitro Assessment of Antifungal and Antibiofilm Efficacy of Commercial Mouthwashes against Candida albicans.

Candida albicans is the most critical fungus causing oral mycosis. Many mouthwashes contain antimicrobial substances, including antifungal agents. This study aimed to investigate the in vitro activity of 15 commercial mouthwashes against 12 strains of C. albicans. The minimal inhibitory concentrations (MICs), minimal fungicidal concentrations (MFCs), and anti-biofilm activity were studied. MICs were determined by the micro-dilution method using 96-well plates, and MFCs were determined by culturing MIC suspensions on Sabouraud dextrose agar. Anti-biofilm activity was evaluated using the crystal violet method. The mouthwashes containing octenidine dihydrochloride (OCT; mean MICs 0.09-0.1%), chlorhexidine digluconate (CHX; MIC 0.12%), and CHX with cetylpyridinium chloride (CPC; MIC 0.13%) exhibited the best activity against C. albicans. The active compound antifungal concentrations were 0.5-0.9 µg/mL for OCT products and 1.1-2.4 µg/mL for CHX rinses. For mouthwashes with CHX + CPC, concentrations were 1.56 µg/mL and 0.65 µg/mL, respectively. Products with polyaminopropyl biguanide (polyhexanide, PHMB; MIC 1.89%) or benzalkonium chloride (BAC; MIC 6.38%) also showed good anti-Candida action. In biofilm reduction studies, mouthwashes with OCT demonstrated the most substantial effect (47-51.1%). Products with CHX (32.1-41.7%), PHMB (38.6%), BAC (35.7%), Scutellaria extract (35.6%), and fluorides + essential oils (33.2%) exhibited moderate antibiofilm activity. The paper also provides an overview of the side effects of CHX, CPC, and OCT. Considering the in vitro activity against Candida albicans, it can be inferred that, clinically, mouthwashes containing OCT are likely to offer the highest effectiveness. Meanwhile, products containing CHX, PHMB, or BAC can be considered as promising alternatives.

Primary Screening for Fungi Isolates that Produce Mycotoxin from Sun-Dried Meat

This study aims to screen fungal isolates from sun dried meat samples to identify fungal presumed producing mycotoxins in the sun dried meat. A total of 8 fungal isolates from sun dried meat were used for the study. They are Aspergillus (2), Penicillium spp (3), Fusarium spp, Mucor spp and Rhizopus spp. The fungal isolates from eight sun dried-meat samples were sub-cultured on Potato Dextrose Agar and Sabouraud Dextrose agar in order to obtain pure colonies for further analysis. The detection of mycotoxins in the sun dry meat samples was carried out by the use of Coconut Agar Media (CAM), by which the fungal isolates were cultured and plates incubated for 3-7days at 28C. The results obtained show that some of the fungal isolates had the ability to produce mycotoxins. From the mycotoxin screening, only four (4) species (Aspergillus spps, and Penicillium spp) were positive (+) and Rhizopus spp, Fusarium spp and Mucor spp, were negative (-). Mycotoxin contamination level varied in different market locations in which Academy Market had the highest level of mycotoxin (71.4%) followed by Oje Market (14.3%) and Oremiji Market (14.3%). Mycotoxigenic producing fungi and mycotoxin levels in the sun dried meat samples are public health concern. Also these results calls for more sensitization for safety of sun dried meat.

Characterization of Poultry Gelatins Prepared by a Biotechnological Method for Targeted Changes at the Molecular Level.

Chicken collagen is a promising raw material source for the production gelatins and hydrolysates. These can be prepared biotechnologically using proteolytic enzymes. By choosing the appropriate process conditions, such changes can be achieved at the molecular level of collagen, making it possible to prepare gelatins with targeted properties for advanced cosmetic, pharmaceutical, medical, or food applications. The present research aims to investigate model samples of chicken gelatins, focusing on: (i) antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azinobis-3-etylbenzotiazolin-6-sulfonic acid (ABTS); (ii) the distribution of molecular weights via gel permeation chromatography with refractometric detection (GPC-RID); (iii) functional groups and the configuration of polypeptide chains related to molecular-level properties using Fourier transform infrared spectroscopy (FTIR); (iv) the microbiological populations on sabouraud dextrose agar (SDA), plate count agar (PCA), tryptic soy agar (TSA), and violet red bile lactose (VRBL) using the matrix-assisted laser desorption ionization (MALDI) method. Antioxidant activity towards ABTS radicals was more than 80%; activity towards DPPH radicals was more than 69%. The molecular weights of all gelatin samples showed typical α-, β-, and γ-chains. FTIR analysis confirmed that chicken gelatins all contain typical vibrational regions for collagen cleavage products, Amides A and B, and Amides I, II, and III, at characteristic wavenumbers. A microbiological analysis of the prepared samples showed no undesirable bacteria that would limit advanced applications of the prepared products. Chicken gelatins represent a promising alternative to products made from standard collagen tissues of terrestrial animals.

Effectiveness of Chitosan versus Natural Aloe Vera on Candida Adherence in Denture Soft Lining Material.

Soft denture lining materials act as a cushion between the denture base and tissues. Alongside having many advantages, its main problem is candida growth due to its rubbery and porous texture. Many interventions have been performed to halt the growth of candida within soft lining materials such as the use of antifungal therapy and strict oral and denture hygiene but there are consequences such as recurrence, drug resistance, and toxicity related to these interventions. Since natural agents such as aloe vera and chitosan have been proven to have antibacterial and antifungal properties with minimum adverse effects, this study aimed to study the effectiveness of chitosan and aloe vera powders incorporated within denture soft lining materials against candida adherence. Methodology. A total of 60 soft-lining material samples were prepared that were equally divided into three groups, viz., group 1 (chitosan incorporation), group 2 (aloe vera incorporation), and group 3 (control). Candida was obtained from the microbiology lab to form a candidal suspension, diluted in 0.9% NaCl to match the McFarland standard bacteriologic solution. Samples were incubated at 37°C for 24 hours in test tubes containing 100 mL of the candidal suspension and 9.9 mL of the previously prepared Sabouraud dextrose agar. Crystal violet stain was used to stain the adhering cells by fixing them with methanol 80%. For each sample, the adhering candida cells were counted on three standard fields by using an inverted light microscope, and the mean of those fields was recorded. The mean value for samples containing aloe vera was 41.15, while the mean values for samples containing chitosan and the control group were 16.05 and 79.1, respectively. Of all the three groups, aloe vera powder had a significant efficacy against candida growth as compared to the chitosan and control groups (P value = 0.001). Both herbal agents were effective against candida growth. In comparison, aloe vera was more effective against candida growth compared to chitosan.

Antifungal Activity of Morinda citrifolia Methanolic Extract against Candida albicans: An In Vitro Study.

Natural medicine used as an alternative and/or complementary treatment to counteract diseases is of great importance in public health. Therefore, the purpose of the present study was to assess the in vitro antifungal activity of Morinda citrifolia methanolic extract of peel, pulp, and seed against Candida albicans. The present study was experimental in vitro and cross-sectional. Eight replicates were prepared in Sabouraud dextrose agar with five wells each, where 0.12% chlorhexidine, distilled water, and methanolic extract of seed, peel, and pulp of Morinda citrifolia fruit were placed at concentrations of 10,690, 8,270, and 6,430 mg/mL, respectively, to evaluate sensitivity according to Duraffourd's scale. In addition, the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were determined by dilution and agar seeding method. Statistical analysis was performed by analysis of variance (ANOVA) and Tukey's post hoc test, considering a significance level of P < 0.05. The inhibition halos of Morinda citrifolia methanolic extract of seed, peel, and pulp against Candida albicans measured on average 15.94, 11.94, and 11.56 mm, respectively. The MIC of seed, peel, and pulp extract were 1366.25, 2067.5, and 1607.5 mg/mL respectively, whereas the MFC for seed, peel, and pulp extract were 2672.50, 2067.5, and 3215 mg/mL, respectively. Moreover, seed extract presented significantly higher antifungal activity than peel and pulp (P < 0.001). Morinda citrifolia methanolic extract of peel, pulp, and seed showed fungistatic and fungicidal effect against Candida albicans, being this very sensitive to seed extract with a MIC of 1366.25 mg/mL and a MFC of 2672.5 mg/mL, which allows recommending the development of effective pharmacological formulations for the control of candidiasis.

Isolation and investigation of external reproduction tract mycoflora in healthy male cats.

Various infectious agents can infect the male reproductive system. The aims of this study were to provide current data on fungal and yeast flora of the external organs of reproductive system of male short hair cats including penis and prepuce. In total 28 samples were taken from external genital system of male stray cats using sterilised cotton swabs. Samples were taken based on the absence of any reproductive complications using physical examinations. The samples were transferred to sterilised Stuart transport media and were then incubated in the Sabouraud dextrose agar with chloramphenicol at 32°C for 48h. The identification of fungi and yeasts was confirmed by germ tube formation test, CHROM agar, urease testing and Corn meal agar medium with Tween 80. Fungal agents were isolated from 7 (25%) out of 28 male stray cats. Fungal isolates were obtained from the penis and prepuce of 4 (36%) and 7 (100%) male cats, respectively. The most commonly recovered species samples were Candida krusei (3.75%) and Penicillum spp. (6.86%) from the penis and prepuce of the cats, respectively. The external organs of male reproduction of cats were infected with 2-4 different fungal agents. Only 5 (45%) cats were infected with one fungus; however, in 6 (55%) cats, mixed fungal infections were isolated. Cats 3-4 years old had the highest fungal contamination in the prepuce region (4/7), while the penis at this age had the least contamination (1/4). It is concluded that the external reproduction organs of male cats could be infected by different fungal agents.

Multilocus Sequence Analysis of Aspergillus Species from Clinical Samples and the Surge of Cryptic Aspergillosis in Southern India: A Descriptive Cross-sectional Study

Introduction: Aspergillus species cause a broad range of infections, from Allergic Bronchopulmonary Aspergillosis (ABPA) to invasive aspergillosis. Accurate species-level identification is crucial for diagnosis and treatment, especially since some rare Aspergillus species exhibit increased resistance to common antifungal agents. Aim: To identify Aspergillus species at the species complex level and determine the emergence of cryptic Aspergillus species using Multilocus Sequence Analysis (MLSA). Materials and Methods: This descriptive cross-sectional study was conducted over eight months, from January 2023 to August 2023, in three tertiary care centres in Chennai, India. Sixty Aspergillus strains were isolated from various clinical samples. The primary inclusion criteria were patients with suspected aspergillosis, categorised based on the European Organszation for Research and Treatment of Cancer Mycoses Study Group (EORTC-MSG) criteria. Isolates were grown on Oatmeal Agar or Sabouraud Dextrose Agar (SDA) and phenotypically identified using tease mount and slide culture techniques. Genomic fungal Deoxyribonucleic acid (DNA) was extracted and amplified using specific primers for Internal transcribed spacer (ITS), beta-tubulin, hydrophobin, and calmodulin genes, followed by nucleic acid sequencing. Results: Among the 60 Aspergillus isolates, the Aspergillus flavus complex comprised 26 (43.3%), the Aspergillus fumigatus complex 16 (26.7%), the Aspergillus niger complex 12 (20%), and the Aspergillus terreus complex 6 (10%). Initial speciation relied on microscopic and cultural characteristics. Isolates were recovered from various clinical specimens: bronchial wash 6 (10%), Bronchoalveolar lavage (BAL) fluid 10 (16.7%), sputum 3 (5%), ear swab 20 (33.3%), endotracheal aspirate 8 (13.3%), pus and tissue from the nasal cavity and paranasal sinus 11 (18.3%), and tissue from other sites 2 (3.3%). MLSA identified common species such as A. flavus, A. fumigatus, A. terreus, and A. niger, as well as rare species including A. awamori, A. carneus, A. lentulus, A. oryzae, A. tamarii, A. tubingensis, and A. welwitschiae. Sequence identity ranged from 99% to 100%, confirming species diversity. Phylogenetic analysis using ITS, beta-tubulin, hydrophobin, and calmodulin genes supported species identification with robust bootstrap support (1000 replications). Conclusion: Cryptic Aspergillus species are prevalent in clinical samples from the South Indian population, with a notable presence from the section Nigri and the first-time isolation of Aspergillus carneus in clinical samples from India. Molecular identification is essential for determining the invasive potential and guiding appropriate antifungal therapy.

In Vitro Effect of Low-level Laser Therapy on Candida albicans Colonies Isolated From Patients Undergoing Radiotherapy for Head and Neck Cancer.

Introduction: Studies on head and neck cancer (HNC) patients undergoing radiotherapy have revealed increased numbers of Candida spp., leading to oral candidiasis and oral mucositis. The effects of laser therapy on Candida spp. have been studied with varied results. This study aimed to investigate the effect of low-level laser therapy (LLLT) on C. albicans colonies isolated from HNC patients undergoing radiotherapy. Methods: This study included a treated group, wherein 11 C. albicans isolates were obtained from the saliva of HNC patients undergoing radiotherapy at a dose exceeding 30 Gy. The control groups consisted of an untreated negative control and a positive control treated with nystatin. The treatment groups were subjected to LLLT in continuous mode for 50 seconds at a wavelength of 976 nm, 0.1 W at a dose of 5 J/cm2 for Laser 1 (L1) group and 0.2 W at a dose of 10 J/cm2 for Laser 2 (L2) group. The tests were conducted using C. albicans inoculum equivalent to a 0.5-McFarland suspension on 96-well plates. All test group inoculums were then cultured on Sabouraud dextrose agar (SDA), and the colony count was assessed at 104 CFU/mL. Results: LLLT at 5 and 10 J/ cm2 reduced C. albicans colonies by 7.01% and 10.94%, respectively, but the reductions were not statistically significant (P>0.05). Nystatin eliminated all the colonies. Conclusion: LLLT shows potential for reducing C. albicans colonies in HNC patients undergoing radiotherapy. However, further studies with varied parameters and methods are necessary.

Anti-biofilm effectiveness of protocols for cleaning complete dentures in hospitalized patients: a randomized controlled trial.

Denture biofilm acts as a potential reservoir for respiratory pathogens, considerably increasing the risk of lung infections, specifically aspiration pneumonia, mainly 48h after hospital admission. The establishment of a straightforward, affordable, and applicable hygiene protocol in a hospital environment for the effective control of denture biofilm can be particularly useful to prevent respiratory infections or reduce the course of established lung disease. To evaluate the anti-biofilm effectiveness of denture cleaning protocols in hospitalized patients. The maxillary complete dentures (MCDs) of 340 hospitalized participants were randomly cleaned once using one of the following 17 protocols (n=20): brushing with distilled water, toothpaste, or neutral liquid soap (controls); immersion in chemical solutions (1% sodium hypochlorite, alkaline peroxide, 0.12% or 2% chlorhexidine digluconate), or microwave irradiation (650 W for 3 min) combined or not with brushing. Before and after the application of the protocols, the biofilm of the intaglio surface of the MCDs was evaluated using two methods: denture biofilm coverage area (%) and microbiological quantitative cultures on blood agar and Sabouraud Dextrose Agar (CFU/mL). Data were subjected to the Wilcoxon and Kruskal-Wallis tests (α=0.05). All 17 protocols significantly reduced the percentage area of denture biofilm and microbial and fungal load (P<0.05). The highest percentage reductions in the area of denture biofilm were observed for 1% hypochlorite solution with or without brushing and for 2% chlorhexidine solution and microwave irradiation only in association with brushing (P<0.05). The greatest reductions in microbial and fungal load were found for the groups that used solutions of 2% chlorhexidine and 1% hypochlorite and microwave irradiation, regardless of the association with brushing (P<0.05). A single immersion for 10 min in 1% sodium hypochlorite, even in the absence of brushing, proved to be a straightforward, rapid, low-cost, and effective protocol for cleaning the dentures of hospitalized patients.

Antimicrobial Efficacy of Calcium Hydroxide with Different Herbal Additives: An In-vitro Study

Introduction: Microorganisms such as Enterococcus faecalis, Enterobacteriaceae, and Candida albicans are associated with persisting infections and can cause endodontic failure. The addition of herbal additives such as Triphala, Clove, and Activated charcoal to Calcium hydroxide enhances the microbial efficacy within the root canal. Aim: To evaluate and compare the anti-microbial efficacy of calcium hydroxide with various herbal additives against E. faecalis and C. albicans using Agar Diffusion Test (ADT) and Direct Contact Test (DCT). Materials and Methods: The present in-vitro study was conducted in Bangalore, Karnataka, between July 2023 and September 2023. Ethical clearance was obtained from the institution. Sabouraud Dextrose Agar (SDA) and Mueller-Hinton Agar were prepared and used to culture C. albicans and E. faecalis, respectively. In each plate, four wells were filled with groups (n=10): Group-I: Nanocalcium hydroxide (NCH)+sterile distilled water, Group-II: NCH+Nanoparticle Clove, GroupIII: NCH+Nanoparticle Triphala, Group-IV: NCH+nanoparticle activated charcoal. The plates were incubated at 37°C, and zones of inhibition around the wells were measured on the first, third, and seventh day. DCT was evaluated in Colony Forming Units (CFUs)/ml by adding each bacterial suspension to four groups and inoculating on blood agar plates. One-way ANOVA test followed by Tukey’s post-hoc analysis was used to compare the mean Zone Of Inhibition (ZOI). Kruskal-Wallis Test followed by Mann-Whitney post-hoc test was used to compare the mean CFUs. Results: One-way ANOVA test showed that there was a significant difference in the mean ZOI between the four groups at p=0.005. The maximum mean ZOI against C. albicans was achieved by the combination of calcium hydroxide and clove with an average mean of 22.20±1.92 mm, 20±1.87 mm, and 17.20±2.28 mm on the 1st, 3rd, and 7th day, respectively. Calcium hydroxide with distilled water combination showed the highest mean ZOI against E. faecalis with an average mean of 18.20±0.84 mm on day 1, 17±1 mm on the 3rd and 7th day. The mean CFUs of E. faecalis and C. albicans showed a significant difference between the four groups at p&lt;0.001. Conclusion: The addition of herbals to Calcium hydroxide increased anti-microbial activity against Candida albicans and Enterococcus faecalis.

Emerging multidrug-resistant microbial bugs lurking in the ear canal – The hidden horror

Background: The ear discharge is a common presenting symptom in medical practice affecting all age groups but primarily children. It is a big menace in developing countries like India. Ear discharge can result from varied infective and clinical conditions of the ear. Aims and objectives: This study was done to determine the current infective etiological causes of ear discharge, their antibiograms, and associated risk factors. Materials and Methods: This was a retrospective study of laboratory records of all the ear swabs taken from patients with ear discharge in the Calcutta National Medical College and Hospital, for 15 months. A total number of 266 patients of both genders between 1 year and 80 years visiting the E.N.T department with ear discharge are taken as the study population. All pus swabs from infected ears were inoculated on blood agar, MacConkey agar, and Sabouraud Dextrose agar with chloramphenicol. Furthermore, smears for Gram staining and potassium hydroxide mount were made. Identification of the pathogens was done by standard biochemical tests. Sensitivity tests to antimicrobial agents were done on Mueller–Hinton Agar using the Kirby–Bauer disk diffusion method. Results: Out of 266 ear swabs, 231 (86.84%) had microorganisms isolated. Out of 266 patients, 140 (52.64%) were females and 126 (47.36%) were males. Pseudomonas species was the commonly isolated organism 98 (42.42%) followed by Staphylococcus aureus 92 (39.80%) and Klebsiella pneumoniae 17 (7.35%). Pseudomonas species were mostly sensitive to imipenem/cilastatin followed by meropenem and amikacin. Conclusion: Most cases of ear discharge were found in females and the age group of 11–18 years. The most common bacteriological cause of the ear discharge was Pseudomonas species followed by S. aureus. Carbapenems, cefepime, piperacillin-tazobactam, levofloxacin, linezolid, and voriconazole are effective antimicrobial agents for the empirical treatment of ear infections with discharge.

Characterization of the Beauveria bassiana ga fungal strain isolated from the muscardine infected silkworms (Bombyx mori L.)

In this study, the focus was on isolating and characterizing the fungal strain Beauveria bassiana from muscardine-infected silkworms (Bombyx mori L.). The fungal strain isolated from the muscardine-infected silkworm, designated as GA, closely resembled Beauveria bassiana isolate SBI-Bb04 based on morphological analysis and comparison of the 18S rRNA nucleotide sequence with sequences in the NCBI database. Beauveria bassiana GA demonstrates the ability to grow on Sabouraud Dextrose Agar (SDA) medium. Optimal conditions for its growth were determined to be a pH range of 6-7 and a temperature range of 20-25°C. The composition of SDA, containing glucose and peptone sources, was found to be ideal for both growth and sporulation of this fungal strain. Furthermore, Beauveria bassiana GA exhibits the capacity to synthesize various extracellular enzymes, including chitinase, protease, cellulase, and amylase. These enzymatic activities contribute to its pathogenicity and suggest its potential as a biocontrol agent against insect pests.

Molecular Identification of Trichophyton Rubrum Onychomycosis at Health Centres in Benue State, Nigeria

The molecular properties of Trichophyton rubrum associated with onychomycosis in adults attending selected tertiary and secondary health facilities in Benue State, Nigeria were investigated. The prevalence of onychomycosis, co-morbidities, onychomycosis related health histories, and infecting strains of Trichophyton rubrum were considered. Due ethical clearance was obtained, and informed consent granted by participants. The research employed purposive random sampling technique. Three hundred and eighty-four (384) adults aged 18 years and above, with visible symptoms of onychomycosis were recruited. Nail samples were collected, disinfected and examined. Sabouraud Dextrose Agar (SDA) was used for fungal cultures. Laboratory procedures included microscopy and morphological characterization of isolates. Toenails were more infected 210 (68.48%) than fingernails 8 (18.18%). The fungal species with the highest frequency of occurrence was T. rubrum 159 (70.36%), while the least was Candida spp. 19 (8. 49%). Allele-Specific Polymerase chain reaction (PCR) revealed new sub-repeat elements in the non-transcribed region of the rDNA spacer of T. rubrum. The study has documented onychomycosis prevalence rates and helped provide insight into the infecting species in Benue State. This will help develop appropriate treatment and intervention guidelines. Further and advanced molecular typing methods for enhanced understanding of infecting fungal strains is recommended.

A prospective observational study on species differentiation and antifungal susceptibility pattern in patients with genital candidiasis.

Candidial balanitis, balanoposthitis and vulvovaginitis can be diagnosed by direct microscopy, culture and treated with antifungals. Resistance to antifungals is emerging. Hence, we conducted a study to identify the causative species and antifungal susceptibility. To observe the species differentiation and antifungal susceptibility pattern in patients with genital candidiasis. A prospective observational study was carried out that included 54 patients of age group (18-60 years) diagnosed clinically and direct microscopically (KOH) for genital candidiasis. Culture was done using Sabouraud dextrose agar. Species identification and antifungal susceptibility were tested. Descriptive data were expressed in the form of frequency and percentage. Out of 54 patients, 41 had culture positive candidiasis. Among the isolated species, 68.3% were Candida albicans (28/41) and 31.7% were non- albicans Candida spp. Among non-albicans Candida species (13/41), Candida glabrata (19.5%), Candida tropicalis (7.3%), Candida guilliermondii (2.4%), Candida parapsilosis (2.4%) were identified. Antifungal susceptibility was tested for fluconazole (FLU), clotrimazole (CLTZ), itraconazole (ITZ), ketoconazole (KTZ), voriconazole (VOR), amphotericin-B (AMPH-B). Except C. glabrata and C.parapsilosis, all other species were sensitive to all tested antifungals. All isolated species were sensitive to KTZ, VOR, AMPH-B, and CLTZ. Nearly 22% of isolates were resistant to fluconazole. C. glabrata causes complicated, severe recurrent vulvovaginitis which is fluconazole resistant. Drug sensitivity prior prescribing antifungal agent identifies appropriate drug, decreases patient's disease morbidity and cross resistance.