Germ Tube Research Articles

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.

Prevalence and Antifungal Susceptibility of Candida Infections in Patients with Pulmonary Tuberculosis: A Cross-sectional Study from the Union Territory of Puducherry, India

Introduction: Pulmonary Tuberculosis (TB) is a bacterial infectious disease caused by Mycobacterium tuberculosis. Candida albicans is the most commonly isolated fungal pathogen in bronchopulmonary patients. Invasive candidiasis due to Candida species has been on the rise in recent years. However, drug sensitivity studies of C. albicans isolated from TB patients are not routinely conducted in India and other developing countries. Aim: To isolate and identify Candida albicans from patients with TB and to determine its antifungal susceptibility. Materials and Methods: This cross-sectional study was conducted on 129 patients suspected of having pulmonary TB, analysed from August 2016 to January 2018 at the Department of Chest Medicine, Aarupadai Veedu Medical College and Hospital, Puducherry, India. Sputum samples were collected and processed to identify Candida species through Gram staining, Ziehl-Neelsen staining, germ tube formation, and sugar fermentation. Antifungal resistance testing was performed using the disk diffusion method recommended by the Clinical and Laboratory Standards Institute (CLSI). The collected data were subsequently analysed using descriptive statistics and the Chi-square test to evaluate the results. Results: Candida infection was observed in 10 (7.75%) out of 129 patients with pulmonary TB. Among patients with co-infection, C. albicans was the most common, accounting for 4 (40%) cases, followed by C. krusei with 2 (20%) cases, C. tropicalis with 1 (10%) case, C. kefyr with 1 (10%) case, C. glabrata with 1 (10%) case, and C. parapsilosis with 1 (10%) case. Among the azole category, itraconazole and amphotericin B exhibited the highest sensitivity percentages against Candida. Conclusion: The current study revealed a high prevalence of coinfection between pulmonary TB and Candida spp., which may contribute to the symptoms of pulmonary lung disorders. The prevalence of Candida species could also be associated with an inadequate response to antitubercular drugs. Amphotericin B and itraconazole, two commonly used drugs in the community, are often employed as the preferred treatments for fungal infections. Early detection and treatment of this opportunistic candidiasis in patients with pulmonary TB will likely enhance outcomes for such individuals.

Candida albicans Infection and Associated Risk Factors among Pregnant Women Attending Antenatal Clinic at Federal Medical Centre, Birnin Kudu, Jigawa State

Candida albicans affects the quality of life and when present in pregnant women it causes complications especially in individuals that are immunocompromised and or poor attitude to hygiene. A total of 60 high vaginal swap (HVS) samples were taken from pregnant women and analyzed for the presence of C. albicans species. Swap sticks were used in the collection of white patches found around the posterior fornix of the vagina and inoculated in Sabouraud Dextrose Agar and incubated at 37oC for 24hours. Slides were prepared and viewed under reduced light microscope using ×10 and ×40 objective magnification. Germ tube test was carried out to distinguish C. albicans species from other Candida species, using test tubes containing 0.5ml human serum. The result shows that those in their second and third trimester have higher prevalence 12 (40.0%) of C. albicans and the secondi gravid 16 (53.3). Similarly, those in the age group of 15-20 years have prevalence 20 (66.7). Although, not significant (P> 0.05), third trimester, use of tights and age were important predisposing factors as revealed by the adjusted odd ratio of 3.3, 2.2 and 6.9 respectively. High prevalence of Candida albicans was observed and factors such as hygiene habit, age of the women, use of antibiotics, trimester and dressing among women played important role as risk factors. Destruction of epithelia cells of the vagina was significantly correlated with C. albicans infection.

Etiology and Antifungal Sensitivity Test in Otomycosis Caused by Candida Sp

Otomycosis is a common fungal infection of the external auditory meatus frequently diagnosed in otolaryngology outpatient clinics. Resistance to antifungals is currently a significant concern, with intrinsic and acquired resistance increasing among isolates that cause fungal infections. The purpose of this research was to identify Candida species causing otomycosis and determine the pattern of antifungal susceptibility among these Candida species. A prospective study was conducted in the Margono Soekarjo General Hospital and Department of Microbiology, Faculty of Medicine, Universitas Jenderal Soedirman Purwokerto, Indonesia, from April – September 2022. Forty-seven (47) clinical samples of otomycosis were collected from 41 patients and then isolated bedside on fungal culture media and was prepared on an object glass for direct microscopic examination of the specimens. Fungal identification was performed using 10% potassium hydroxide (KOH) to observe fungal elements. Samples were cultured on Saboraud dextrose agar (SDA) media with chloramphenicol and Czapek dox agar. The Germ Tube Test was used to identify Candida while yeast-specific identification and antifungal susceptibility assay using a rapid commercial kit was applied for specific identification of the fungus. Antifungal susceptibility patterns were obtained using the Integral System Yeast Plus (ISYP) media pack. Candida parapsilosis was the most prevalent Candida species discovered in this study, accounting for approximately 41.66%, which was followed by Candida tropicalis (25%) and Candida krusei (12.5%), whereas Candida albicans only accounted for 4.1% of the specimens. All Candida species were sensitive to flucitosine and ketoconazole, whereas the voriconazole sensitivity rate reached 96%. This study concludes that Candida parapsilosis is the most prevalent species of Candida in otomycosis, and that all Candida species are sensitive to Flucitosine, ketoconazole, and voriconazole.

Effectiveness of various nematode-trapping fungi for biocontrol of the Meloidogyne incognita in tomato (Lycopersicion esculentum Mill.)

The nine promising species of nematode-trapping fungi namely Dr. dactyloides, Dr. brochopaga, Da. gephyropaga, Da. phymatopaga, A. eudermata, A. oligospora, A. conoides, A. thaumasia and A. musiformis were isolated from the root-knots rhizosphere to compare their, in vitro nematode trapping ability against Meloidogyne incognita, ability to form conidial traps close to nematode infested soils and their effectiveness in reducing on M. incognita infection indices on tomato. The laboratory assays of nematophagous efficacy of nine nematode-trapping fungi against M. incognita revealed that Dr. dactyloides, Dr. brochopaga, D. gephyropaga and Da. phymatopaga seemed to be the most effective, with 73.2–99.8% trapping and killing of juveniles within 5 days of nematode and fungal interface. Conidial trap forming ability of different species of nematode-trapping fungi showed that Dr. dactyloides, Dr. brochopaga formed frequent formation of conidial traps without intermediate germ tubes whereas Da. phymatopaga and Da. gephyropaga formed adhesive traps on spore germ tubes in close vicinity of root-knot nematode infested soils and killed the nematode present on the agar discs. A. eudermata, A. oligospora, A. conoides, A. thaumasia and A. musiformis frequently germinated by germ tubes in soil and occasionally formed fewer traps. The pot experiment results showed that Dr. brochopaga and Dr. dactyloides, which dominantly formed conidial traps without an intermediate mycelial phase caused the highest decrease in the number of roots-knots (67.2–70.4%), females (80.5–85.8%) and eggs and juveniles (84.9–88.8%) of M. incognita followed by Da. gephyropaga and Da. phymatopaga which formed traps on conidial germ lings and mycelium. A. eudermata, A. oligospora, A. conoides, A. thaumasia and A. musiformis which formed poor conidial traps caused a moderate decrease in the number of root-knots (45.3–53.4%), females (55.5–61.1%), eggs and juveniles (52.9–60.1%) of M. incognita in comparison to control. This paper analyses the effective biocontrol of M. incognita by using different nematode-trapping fungi, especially conidial traps forming fungi and the fate of application of these species in the RKN-infested soil for management of M. ingocnita.

Study on the inhibitory effect of fermentation extract of Microporus vernicipes on Candida albicans.

Candida albicans is one of the most common species of Candida, which cause various mucosal and systemic infectious diseases. However, the resistance rate to existing clinical antifungal drugs gradually increases in C. albicans. Therefore, new antifungal drugs must be developed to solve the current problem. This study discovered that the solid fermented ethyl acetate crude extract of Microporus vernicipes had inhibitory activity on C. albicans. This study determined that the Mv5 components had significantly inhibited the activity of C. albicansusing column chromatography separation component screening. The components included 23 compounds of fatty acids and their derivatives, alkaloids, phenols, and other classes using ultra-high performance liquid chromatography tandem high-resolution mass spectrometry (UHPLC-HR-MS) analysis, with fatty acids constituting the primary components. The mechanism of action showed that the minimum inhibitory concentration (MIC) of Mv5 components against C. albicans was 15.63μg/mL, while minimum fungicidal concentration (MFC) was 31.25μg/mL. Mv5 components can inhibit the early biofilm formation and destroy the mature biofilm structure. It can inhibit the germ tube growth of C. albicans, thereby inhibiting the transformation of yeast morphology to hyphae. We detected 193 differentially expressed genes, including 156 upregulated and 37 downregulated genes in the Mv5 components of the MIC concentration group. We detected 391 differentially expressed genes, including 334 upregulated and 57 downregulated expression genes in the MFC concentration group. Among these differentially expressed genes, the genes related to mycelium and biofilm formation were significantly downregulated. GO enrichment analysis presented that single-organism process metabolic process, and cellular processes were the biological processes with the most gene enrichment. Kyoto Encyclopedia of Genes and Genomes (KEGG)of Mv5 components were mainly enriched in metabolic pathways, such as meiosis yeast and amino acid metabolism. Therefore, it is believed that the fermentation extract of M. vernicipes inhibits C. albicans, which can provide clues for developing effective antifungal drugs.

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.

Vulva Candidiasis among HIV Patients and Healthy Female Volunteers in Port Harcourt

Vulvo-rectal the most prevalent species of candida, Candida albicans, is the cause of candidiasis, a fungal infection of the vagina that affects the genital system as an adaptive and opportunistic pathogen.
 This study was aimed at examining the prevalence and risk factors of vulvo-vaginal candidiasis among HIV positive women and healthy volunteers in Port Harcourt. A total of 200 urine and HVS samples, 100 from each of the two groups, were collected. The samples were examined using culture, microscopy, and the germ tube identification method. The overall prevalence of Vulvo-vaginitis in HIV and healthy volunteers using urine and HVS was (32%), (34%), and (76%) and (54%) respectively. HIV recorded a higher of (76%) compared to that of healthy volunteers with (54%). There was no statistically significant difference in the two groups’ prevalence (p>0.05). The age group of 15–25 years old had the highest prevalence of (33%), while the age group of 40–45 had the highest prevalence of 80% for HIV. HIV-positive workers and retirees had higher rates of vulvo-vaginitis in urine samples (21% and 75%, respectively) than did healthy workers and retirees (15% and 25%, respectively). Individuals with tertiary education had the highest prevalence of (32%) and (14%) in HIV and healthy volunteers respectively. It was also reported that married individuals who are HIV positive had a higher incidence of (50%) while for healthy volunteers, single individuals recorded a higher frequency of (29%). Prevalence of vulvo-vaginal candidiasis was greater in HIV patients in Port Harcourt. To help combat fungal infections, it is proposed that antifungal medications be added to antiretroviral medications prescribed to HIV patients.

Novel Endophytic Pseudescherichia sp. GSE25 Strain Significantly Controls Fusarium graminearum and Reduces Deoxynivalenol in Wheat.

Fusarium heading blight (FHB) is a devastating disease in wheat, primarily caused by field invasion of Fusarium graminearum. Due to the scarcity of resistant wheat varieties, the agricultural sector resorts to chemical fungicides to control FHB incidence. On the other hand, biocontrol represents a promising, eco-friendly approach aligned with sustainable and green agriculture concepts. In the present study, a bacterial endophyte, Pseudescherichia sp. (GSE25), was isolated from wheat seeds and identified through complete genome sequencing and phylogenetic analysis. In vitro testing of this endophytic strain demonstrated strong antifungal activity against F. graminearum PH-1 by inhibiting spore germination, suppressing germ tube growth, and causing cell membrane damage. Under field conditions, the strain GSE25 significantly reduced the FHB incidence and the associated deoxynivalenol mycotoxin accumulation by over 60% and 80%, respectively. These findings highlight the potential of the isolated bacterial endophyte Pseudescherichia sp. GSE25 strain as a biocontrol agent in protecting wheat from FHB-caused F. graminearum. This is the first report showing a biocontrol effect of Pseudescherichia sp. a strain against phytopathogens.

Comparative physiological and transcriptome analysis provide insights into the inhibitory effect of osthole on Penicillium choerospondiatis

Blue mold induced by Penicillium choerospondiatis is a primary cause of growth and postharvest losses in the fruit of Phyllanthus emblica. There is an urgent need to explore novel and safe fungicides to control this disease. Here, we demonstrated osthole, a natural coumarin compound isolated from Cnidium monnieri, exhibited a strong inhibitory effect on mycelia growth, conidial germination rate and germ tube length of P. choerospondiatis, and effectively suppressed the blue mold development in postharvest fruit of P. emblica. The median effective concentration of osthole was 9.86 mg/L. Osthole treatment resulted in cellular structural disruption, reactive oxygen species (ROS) accumulation, and induced autophagic vacuoles containing cytoplasmic components in fungal cells. Transcriptome analysis revealed that osthole treatment led to the differentially expressed genes mainly enriched in the cell wall synthesis, TCA cycle, glycolysis/ gluconeogenesis, oxidative phosphorylation. Moreover, osthole treatment led to increase genes expression involved in peroxisome, autophagy and endocytosis. Particularly, the autophagy pathway related genes (PcATG1, PcATG3, PcATG15, PcATG27, PcYPT7 and PcSEC18) were prominently up-regulated by osthole. Summarily, these results revealed the potential antifungal mechanism of osthole against P. choerospondiatis. Osthole has potentials to develop as a natural antifungal agent for controlling blue mold disease in postharvest fruits.

Carvacrol-Induced Vacuole Dysfunction and Morphological Consequences in Nakaseomyces glabratus and Candida albicans.

With the prevalence of systemic fungal infections caused by Candida albicans and non-albicans species and their resistance to classical antifungals, there is an urgent need to explore alternatives. Herein, we evaluate the impact of the monoterpene carvacrol, a major component of oregano and thyme oils, on clinical and laboratory strains of C. albicans and Nakaseomyces glabratus. Carvacrol induces a wide range of antifungal effects, including the inhibition of growth and hyphal and biofilm formation. Using biochemical and microscopic approaches, we elucidate carvacrol-induced hyphal inhibition. The significantly reduced survival rates following exposure to carvacrol were accompanied by dose-dependent vacuolar acidification, disrupted membrane integrity, and aberrant morphology. Germ tube assays, used to elucidate the relationship between vacuolar dysfunction and hyphal inhibition, showed that carvacrol significantly reduced hyphal formation, which was accompanied by a defective C. albicans morphology. Thus, we show a link between vacuolar acidification/disrupted vacuole membrane integrity and compromised candidal morphology/morphogenesis, demonstrating that carvacrol exerts its anti-hyphal activity by altering vacuole integrity.

Prevalence and drug susceptibility of clinical Candida species in nasopharyngeal cancer patients in Vietnam

In the nature, Candida species are normal inhabitants and can be observed in a wide variety of vertebrates. In humans, especially for cancer patients who fall prey to opportunistic pathogens, this group of susceptible multi-drug resistant and biofilm-forming yeasts, are among the commonest ones. In this study, Candida species in 76 oral lesion samples from Vietnamese nasopharyngeal-cancer patients were isolated, morphologically identified using CHROMagar™, germ tube formation, and chlamydospore formation tests, and molecularly confirmed by PCR-RFLP. The drug susceptibility of these isolates was then tested, and the gene ERG11 was DNA sequenced to investigate the mechanism of resistance.The results showed that Candida albicans remained the most prevalent species (63.16% of the cases), followed by Candida glabrata, Candida tropicalis, and Candida krusei. The rates of resistance of non-albicans Candida for tested drugs were 85.71%, 53.57%, and 57.14% to fluconazole, clotrimazole, and miconazole, respectively. Although the drug-resistance rate of Candida albicans was lower than that of non-albicans Candida, it was higher than expected, suggesting an emerging drug-resistance phenomenon. Furthermore, ERG11 DNA sequencing revealed different mutations (especially K128T), implying the presence of multiple resistance mechanisms. Altogether, the results indicate an alarming drug-resistance situation in Candida species in Vietnamese cancer patients and emphasize the importance of species identification and their drug susceptibility prior to treatment.

The potentiality of Wickerhamomyces anomalus against postharvest black spot disease in cherry tomatoes and insights into the defense mechanisms involved

Our previous study revealed that Wickerhamomyces anomalus effectively inhibited Botrytis cinerea disease in cherry tomatoes. However, the potentiality of W. anomalus against black spot disease in cherry tomatoes as well as possible mechanisms related to this efficacy remain unclear. In our current study, we investigated the efficacy of W. anomalus against black spot disease in cherry tomatoes, and the molecular mechanisms related to disease-resistance response of W. anomalus treated cherry tomatoes were also investigated by transcriptome analysis. The results exhibited that W. anomalus successfully controlled postharvest black spot disease of cherry tomatoes caused by Alternaria alternata. Furthermore, W. anomalus vigorously controlled the spore germination and reduced the germ tube elongation of A. alternata. The physiological mechanisms by which W. anomalus inhibited the black spot decay in cherry tomatoes mainly through enhancing the ascorbate peroxidase (APX) and chitinase (CHI), and reducing the content of malondialdehyde (MDA). In addition, based on transcriptome technology, W. anomalus could up-regulate genes involved in several important KEGG pathways, including amino sugar and nucleotide sugar metabolism pathway, selenocompound metabolism pathway, linoleic acid metabolism pathway, and galactose metabolism pathway. These genes may help the fruit to enhance the enzyme activities and nutritional values of cherry tomatoes, synthesize the regulatory molecules in plant disease response as well as plant growth and development, increase the defense and oxidation resistance of the fruit during and after harvesting, and trigger ROS induced resistance in cherry tomatoes. Ultimately, W. anomalus has been proved as a biocontrol agent which could effectively control black spot disease in cherry tomatoes caused by A. alternata. This study can provide theoretical reference for the practical application of antagonistic yeasts in the biological control of postharvest diseases of cherry tomatoes.

First report of powdery mildew on Euonymus japonicus caused by Erysiphe euonymicola in Taiwan.

The Japanese spindle (Euonymus japonicus Thunb.) is commonly used as an ornamental hedge plant in Taiwan. In March 2020, a severe powdery mildew disease was observed on E. japonicus surrounding a city park spanning six hectares in Taichung city, Taiwan. Around 90% of the plants showed symptoms on the leaves and pedicels of young shoots. Similar symptoms were observed in other districts of Taichung city and Taipei city between March to June in subsequent years. Initial signs of infection manifest as circular chlorotic spots on the leaves, which are subsequently covered by white mycelia on either the upper or lower surfaces of the spots. In severe cases, both sides of the leaves become entirely covered by dense mycelia. Hyphal appressoria were solitary or in opposite paired, lobed to multilobed. Conidiophores grow erectly from the hyphae, consist of 2-3 cylindrical cells, 38.9 to 78.6 × 6.31 to 8.28 µm (n = 30). Foot cells are usually straight or slightly flexuous, 23.6 to 43.2 µm (n = 30), followed by 1 to 2 shorter cells. Ellipsoidal conidia are produced singly on the conidiophores, 24.1 to 36.3 × 10.6 to 14.97 µm (n = 30), without fibrosin bodies. Germ tubes are mostly subterminal, sometimes terminal, occasionally exhibiting a longitudinal pattern. Chasmothecia were not observed. These morphological characteristics correspond to the description of Erysiphe euonymicola U. Braun (Braun and Cook 2012), one of the Erysiphe species reported on E. japonicus. Genomic DNA was extracted from seven isolates obtained from different plants in the affected regions. The internal transcribed spacer (ITS) and 28S large subunit (LSU) of rDNA sequences (ITS accession nos.: OR073423-OR073429; LSU accession nos.: OR073448-OR073454) were amplified and sequenced using primer sets PMITS-1 / PMITS-2 (Cunnington et al. 2003) and NLP2 / PRM2 (Bradshaw and Tobin 2020), respectively. The resulting sequences exhibited identities ranging from 99.1 to 100% in ITS and 100% in LSU when compared to the corresponding sequences of E. euonymicola MUMH 133 (ITS: AB250228; LSU: AB250230) (Limkaisang et al. 2006). Phylogenetic analysis based on the concatenated sequences of ITS and LSU clustered the seven isolates within the same clade as three E. euonymicola isolates (MUMH 133, MUMH 6999 and MUMH 7012). Pathogenicity assays were conducted on one-meter tall E. japonicus plants by gently smearing infected leaves on all leaves of four healthy plants. Four uninoculated plants were used as control. All eight assayed plants were enclosed in plastic bags to maintain high humidity at 28 ± 2°C for 3 days. Chlorotic spots began to appear on leaves younger than one month old at 7 days post inoculation (dpi). By 28 dpi, all inoculated plants showed symptoms. Spots expanded or merged and formed a dense mycelial layer on leaves younger than three months, while mature dark green leaves were asymptomatic. No symptoms were observed on any leaves of the control plants. The morphological characteristics and sequences of ITS and LSU of the pathogen from the inoculated plants matched the above information. Based on these findings, E. euonymicola was identified as the causal agent of powdery mildew on E. japonicus, representing the first documented report of this disease in Taiwan. A voucher specimen TNM F0037001 (isolate EPM-1) was deposited in the National Museum of Natural Science, Taiwan. The pathogen has been frequently reported in recent years and significantly impacts the ornamental value of Euonymus spp. (Abbasi and Braun 2020; Lee et al. 2015; Li et al. 2011; Pei et al. 2022). This report also provides an evidence of an ongoing outbreak of the pathogen.

Antifungal Activity of Extracts From the Ferulago Pauciradiata in Vitro Against Botrytis Cinerea Pers

This is the first study to reveal the antifungal properties of Ferulago pauciradiata plant. In this context investigated the effects of methanol (FPM), ethanol (FPE), hexane (FPH) and water (FPW) extracts of the F. pauciradiata plant on the prevention of losses caused by gray mold (Botrytis cinerea Pers) in vitro. The effects of FPM, FPE, and FPH 10, 25, 50, 100, 300, 500, 1000, and 2000 µL doses were determined by mycelium growth, germ tube elongation, and spore germination in vitro. The water extract didn’t show antifungal activity against B. cinerea. Compared to the control, both FPM and FPH caused 100% inhibition at the dose of 2000 µL by suppressing mycelial growth due to dose increases, while FPE had a 97.3% effect on the same parameter at the dose of 2000 µL. While there was no elongation at the 2000 µL dose of FPM and FPH, there was an elongation of 8.4 µm at the same dose of FPM. In spore germination, 0% germination was observed in FPM and FPH 2000 µL dose, while 17.5% germination was observed in FPE. These results show that F. pauciradiata extracts, which are of biological origin and are not environmentally toxic, are a good alternative for use in the control of B. cinerea.

Effects of Temperature, Moisture, and Ultraviolet Light on Germination, Infection, and Survival of Gymnosporangium yamadae Teliospores and Basidiospores.

Apple rust caused by Gymnosporangium yamadae is a significant disease in China's main apple production areas. We evaluated the effects of temperature, moisture, and ultraviolet (UV) light on the germination, infection, and survival of teliospore horns and basidiospores under artificially controlled environmental conditions. The temperature required for the germination and infection of teliospores and basidiospores of G. yamadae ranged from 5 to 25°C, with an optimum temperature of approximately 17°C. The teliospore horns germinated after soaking in distilled water for 5 min and required at least 2.3 h of development to produce basidiospores under the most favorable conditions. The basidiospores germinated only in free water and produced germ tubes 0.8 h after being placed in the water. The half-life of the basidiospore was 72.5 h in the dark and only 9.5 h when exposed to intense UV light. The basidiospores inoculated on the host leaves required at least 2.3 h of water exposure to cause rust lesions. A revised Weibull model could describe the relationships between the germination and infection of teliospore horns and basidiospores with temperature and wetness duration. Collectively, these results can serve as a valuable guide for developing a model to predict future apple rust epidemics and establish a method for effective control strategies.

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

Effect of Propolis on Fungal Infection for Denture Wearers and Dry Socket

Propolis is consist of various amounts of resins and beeswaxes that produced by the honeybees from different natural plants, as leaf buds and flowers. It is a sticky and dark-brown color material that produced by honey bees from natural plants that the bees mixed it with wax and applied it in building, construction and adaptation of the nests specially in repair cracks and defects in the bees hive. They have been used in medical folk or medical traditional since times of ancient. It is now known to be natural medical treatment with, antifungal, antibacterial, antioxiditive, antitumor and mmunomodulatory properties. These medical properties of propolis are motivating isolation researches, identification of the possible relationship of these with its biological activity, and chemical compounds. It was used in dentistry for healing of surgical wound (or post-surgical mouth pain), tooth hypersensitivity, pulp capping and root canal treatment. Propolis has a promising goal in future medically. This research shows the application of propolis, clinically, as a natural dental medicine. Furthermore; systemic and clinical applications of these drugs may have some adverse effects like liver toxicity, drug interactions etc. Using propolis as antifungal by local application shown that it reduces the incidence of dry socket and denture stomatitis that associated with the Candida. In present study, the propolis has been used in the following manner. It is applied to the tissue surfaces of dentures, two times daily for two weeks. Propolis putty can also applied to dry socket cases after complete debridement and irrigation of it. The results showed after use propolis that the signs and symptoms including pain, redness and inflammation areas related stomatitis were gradually disappear, after 2 to 14 days the denture stomatitis is healing by using a propolis paste. Propolis putty has an effect on the fungal infection of dry socket that gradually reduced pain and inflammation until complete healing. Aim of the study, after the treatment of patients with denture stomatitis, they were subjected to examination and evaluation of the mucosa and socket, Candidal quantitative cultures and confirmation the diagnosis by three methods: Germ tube, Gram Stain, and ChromAgar media.

Antifungal Effect of Bacillus velezensis ZN-S10 against Plant Pathogen Colletotrichum changpingense and Its Inhibition Mechanism

In order to optimize crop production and mitigate the adverse impacts associated with the utilization of chemical agents, it is necessary to explore new biocontrol agents. Bacillus velezensis has been widely studied as a biocontrol agent because of its efficient and ecofriendly plant disease control mechanisms. This study shows that the strain ZN-S10 effectively reduces the area of leaf spots caused by the pathogen Colletotrichum changpingense ZAFU0163-1, which affects conidia production and germination, inhibits mycelium growth, and induces mycelium deformation. In antifungal experiments with crude extracts, we observed a delay in the cell cycle of conidia, which may be responsible for the inhibition of conidial germination. Among the bioactive metabolites detected through integrated LC-MS- and GC-MS-based untargeted metabolomics, 7-O-Succinyl macrolactin A, telocinobufagin, and surfactin A may be the main antifungal metabolites of strain ZN-S10. The presence of 7-O-Succinyl macrolactin A could explain the cell damage in germ tubes. This is the first report of telocinobufagin detected in B. velezensis. These results are significant for understanding the inhibitory mechanisms employed by B. velezensis and should serve as a reference in the production of biocontrol agents.

The impact of inter- and intra-species spore density on germination of the food spoilage fungus Aspergillus niger

Aspergilli can be used to produce food but can spoil it as well. Both food production and spoilage are initiated by germination of the conidia of these fungi that have been introduced by inoculation and contamination, respectively. Germination of these spores includes activation, swelling, establishment of cell polarity, and formation of a germ tube. So far, only quantitative single-species germination studies of fungal spores have been performed. Here, spore germination of the food spoilage fungus Aspergillus niger was studied quantitatively in mono-culture or when mixed with other food-relevant aspergilli (Aspergillus nidulans, Aspergillus terreus, Aspergillus clavatus, and Aspergillus oryzae). In the presence of the germination inducing amino acids proline or alanine, but not in the case of the lowly inducing amino acid arginine, the incidence of swelling and germ tube formation was reduced when 35,000 extra conidia of Aspergillus niger were added to wells containing 5000 of these spores. Adding 35,000 spores of one of the other aspergilli also did not have an effect on germination in the presence of arginine, but the germination inhibition was stronger when compared to the extra A. niger spores in the case of alanine. A similar effect was obtained with proline. Together, results show that the germination of A. niger conidia is impacted by the density of its own spores and that of other aspergilli under favorable nutritional conditions. These results increase our understanding of food spoilage by fungi and can be used to optimize food production with fungi.