Project description:We investigated different escape and defense strategies of Candida parapsilosis against the natural fungivorous predator Protostelium aurantium using dual RNA-Seq. Trophozoites of the amoeba Protostelium aurantium were fed with C. parapsilosis and samples for RNA isolation were taken at time points 0 min (control), 30 min and 60 min. At indicated time points samples were shock frozen and used for RNA isolation. C. parapsilosis underwent rapid predation and intracellular killing of the yeast
Project description:We investigated different escape and defense strategies of Candida glabrata and Candida albicans against the natural fungivorous predator Protostelium aurantium using dual RNA-Seq. While C. albicans was found to be very succesfull in preventing from amoeba recognition, C. glabrata in turn, was rapidly taken up but remained undigested with a comparably low transcriptional activity. Trophozoites of the amoeba Protostelium aurantium were fed seperately with the two fungal species and samples for RNA isolation were taken at time points 0 min (control), 30 min and 60 min. At indicated time points samples were shock frozen and used for RNA isolation. Samples for RNAseq were taken at time points 0 min (control), 30 min and 60 min. RNA samples from identical timepoints for C. albicans and C. glabrata were pooled for RNA sequencing. We investigated different escape and defense strategies of Candida spp. against natural fungivorous predator. While C. parapsilosis underwent rapid predation and intracellular killing of the yeast, C. albicans was found to be very succesfull in preventing from amoeba recognition. C. glabrata in turn, was rapidly taken up but remained undigested with a comparably low transcriptional activity.
Project description:The incidence of fungal infections and, in particular, the incidence of fungal antibiotic resistance, which is associated with biofilm formation, have significantly increased, contributing to morbidity and mortality. Thus, new therapeutic strategies need to be developed. In this context, natural products have emerged as a major source of possible antifungal agents. Berberine is a protoberberine-type isoquinoline alkaloid isolated from the roots, rhizomes, and stem bark of natural herbs, such as Berberis aquifolium, Berberis vulgaris, Berberis aristata, and Hydrastis canadensis, and of Phellodendron amurense Berberine has been proven to have broad antibacterial and antifungal activity. In the present study, the potential antifungal effect of berberine against fluconazole-resistant Candida and Cryptococcus neoformans strains, as well as against the biofilm form of Candida spp., was assessed. The antifungal effect of berberine was determined by a broth microdilution method (the M27-A3 method of the Clinical and Laboratory Standards Institute) and flow cytometry techniques, in which the probable mechanism of action of the compound was also assessed. For biofilm assessment, a colorimetric 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was used to determine the susceptibility of sessile cells. The isolates used in the study belonged to the Laboratory of Bioprospection and Experiments in Yeast (LABEL) of the Federal University of Ceará. After 24 and 72 h, fluconazole-resistant Candida and Cryptococcus neoformans strains showed berberine MICs equal to 8 μg/ml and 16 μg/ml, respectively. Cytometric analysis showed that treatment with berberine caused alterations to the integrity of the plasma and mitochondrial membranes and DNA damage, which led to cell death, probably by apoptosis. Assessment of biofilm-forming isolates after treatment showed statistically significant reductions in biofilm cell activity (P < 0.001).
Project description:The Cse4p-containing centromere regions of Candida albicans have unique and different DNA sequences on each of the eight chromosomes. In a closely related yeast, C. dubliniensis, we have identified the centromeric histone, CdCse4p, and shown that it is localized at the kinetochore. We have identified putative centromeric regions, orthologous to the C. albicans centromeres, in each of the eight C. dubliniensis chromosomes by bioinformatic analysis. Chromatin immunoprecipitation followed by PCR using a specific set of primers confirmed that these regions bind CdCse4p in vivo. As in C. albicans, the CdCse4p-associated core centromeric regions are 3-5 kb in length and show no sequence similarity to one another. Comparative sequence analysis suggests that the Cse4p-rich centromere DNA sequences in these two species have diverged faster than other orthologous intergenic regions and even faster than our best estimated "neutral" mutation rate. However, the location of the centromere and the relative position of Cse4p-rich centromeric chromatin in the orthologous regions with respect to adjacent ORFs are conserved in both species, suggesting that centromere identity is not solely determined by DNA sequence. Unlike known point and regional centromeres of other organisms, centromeres in C. albicans and C. dubliniensis have no common centromere-specific sequence motifs or repeats except some of the chromosome-specific pericentric repeats that are found to be similar in these two species. We propose that centromeres of these two Candida species are of an intermediate type between point and regional centromeres.
Project description:The current study was conducted to survey the prevalence of pigeon candidiasis in diseased pigeons suspected to candidiasis by isolation, microscopic examination, and polymerase chain reaction (PCR) method and to characterize Candida spp. phylogenetically. For this purpose, samples were obtained from 100 suspected pigeons from September 2018 to February 2019 in Ahvaz, Iran. Cloacal and oropharyngeal swab samples were collected from each diseased pigeon with diarrhea resistant to the antibiotics, crop stasis, white diphtheritic membrane in the mouth, regurgitation, and vomiting. Sabouraud dextrose agar was used as a culture medium. Selected colonies were stained with lactophenol cotton blue stain. In the culture and direct microscopic observation, 19.00% of birds were suspected to candidiasis. Twenty-two isolates were identified. All 22 isolates were confirmed as Candida spp. By PCR method. The PCR test confirmed the presence of Candida spp. in 19.00% of pigeons. Based on the sequencing results of some PCR products, the isolates belonged to Candida albicans and Candida glabrata. The results revealed a 99.78% accordance when compared with other sequences of C. albicans which were formerly deposited in GenBank® from Colombia, Indonesia, China, and Sudan. The results revealed a 99.54% accordance when compared with other sequences of C. glabrata which were formerly deposited in GenBank® from the Netherlands and Spain. The symptoms such as diarrhea resistant to antibiotics, crop stasis, white diphtheritic membrane in the mouth, regurgitation, and vomiting were the most prevalent clinical symptoms in positive pigeons.