sporothrix brasiliensis atcc  (ATCC)

Journal: Animals : an Open Access Journal from MDPI

Article Title: Clinical and Molecular Characterization of Feline Sporotrichosis in the Brazilian Amazon: PCR-Based Identification of Sporothrix brasiliensis

doi: 10.3390/ani15152318

Figure Lengend Snippet: Clinical lesion, fungal culture, and microscopic morphology of Sporothrix brasiliensis isolated from a feline case. ( A ) Feline with a lesion on the lateral facial region showing cutaneous involvement with dissemination to the ocular area, ear, and nasal planum swelling; ( B ) colony of S. brasiliensis grown on Sabouraud agar after incubation, obtained from a sample collected from the ocular and ear regions; ( C ) microscopic morphology of S. brasiliensis displaying the characteristic “daisy-like” arrangement, observed at 400× magnification.

Article Snippet: Reference strains used in this study included Sporothrix schenckii ATCC CFP-00746, Sporothrix brasiliensis ATCC MYA-4606, Candida albicans ATCC 10231, and Aspergillus niger ATCC 16404, obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Isolation, Incubation

Journal: Animals : an Open Access Journal from MDPI

Article Title: Clinical and Molecular Characterization of Feline Sporotrichosis in the Brazilian Amazon: PCR-Based Identification of Sporothrix brasiliensis

doi: 10.3390/ani15152318

Figure Lengend Snippet: Comparative evaluation of DNA extraction methods applied to fungal isolates of Sporothrix schenckii ATCC CFP-00746, Sporothrix brasiliensis ATCC MYA-4606, a regional Sporothrix isolate (Sample 2), Candida albicans ATCC 10231, and Aspergillus niger ATCC 16404. ( A ) Mean DNA concentration (ng/µL) obtained using the phenol–chloroform, Silica Column, and Acetate Precipitation methods. ( B ) Mean DNA purity, expressed as the 260/280 nm absorbance ratio. Error bars represent the standard deviation from measurements performed in triplicate.

Article Snippet: Reference strains used in this study included Sporothrix schenckii ATCC CFP-00746, Sporothrix brasiliensis ATCC MYA-4606, Candida albicans ATCC 10231, and Aspergillus niger ATCC 16404, obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: DNA Extraction, Concentration Assay, Standard Deviation

Journal: Animals : an Open Access Journal from MDPI

Article Title: Clinical and Molecular Characterization of Feline Sporotrichosis in the Brazilian Amazon: PCR-Based Identification of Sporothrix brasiliensis

doi: 10.3390/ani15152318

Figure Lengend Snippet: Phylogenetic analysis based on the ITS region using the neighbor-joining method with bootstrap support, performed applying the MEGA (Molecular Evolutionary Genetics Analysis) X software package. The tree was constructed using the internal transcribed spacer ( ITS ) molecular marker, based on the alignment of Sporothrix species sequences available in the NCBI database. Regional isolates from the Amazon region included in this study are shown in green. Aspergillus niger and Candida albicans were used as outgroup controls and are shown in red. All sequence data are available in the NCBI database.

Article Snippet: Reference strains used in this study included Sporothrix schenckii ATCC CFP-00746, Sporothrix brasiliensis ATCC MYA-4606, Candida albicans ATCC 10231, and Aspergillus niger ATCC 16404, obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Software, Construct, Marker, Sequencing

Journal: The Cell Surface

Article Title: Sporothrix brasiliensis Gp70 is a cell wall protein required for adhesion, proper interaction with innate immune cells, and virulence

doi: 10.1016/j.tcsw.2024.100139

Figure Lengend Snippet: Sporothrix schenckii strains used in this study.

Article Snippet: HSB6 , 5110 ATCC MYA 4823 transformed with pBGgHg-GP70 , 9.2 ± 1.1 , 3.1 ± 0.3 , This work.

Techniques: Expressing, Transformation Assay

Journal: The Cell Surface

Article Title: Sporothrix brasiliensis Gp70 is a cell wall protein required for adhesion, proper interaction with innate immune cells, and virulence

doi: 10.1016/j.tcsw.2024.100139

Figure Lengend Snippet: Sporothrix brasiliensis yeast-like cell morphology . Representative light microscopy images of S. brasiliensis wild-type (WT) strain, and three mutant strains with high levels of GP70 silencing (HSB6, HSB7, and HSB8). WT, strain 5110 ATCC MYA 4823.Scale bars = 5.0 μm.

Article Snippet: HSB6 , 5110 ATCC MYA 4823 transformed with pBGgHg-GP70 , 9.2 ± 1.1 , 3.1 ± 0.3 , This work.

Techniques: Light Microscopy, Mutagenesis

Journal: The Cell Surface

Article Title: Sporothrix brasiliensis Gp70 is a cell wall protein required for adhesion, proper interaction with innate immune cells, and virulence

doi: 10.1016/j.tcsw.2024.100139

Figure Lengend Snippet: Analysis of 3-carboxy-cis,cis-muconate cyclase activity in Sporothrix brasiliensis wild-type, control, and GP70 -silenced strains. Cell homogenates prepared from yeast-like cells (cell-bound enzyme activity) were used to measure enzyme activity using 3-carboxy-cis-cis-muconate as substrate. Alternatively, the dialyzed and concentrated media where yeast-like cells were grown (cell-free enzyme activity) was used in enzyme assays. Data are means ± SD of three biological replicates performed in duplicates. The Dunnett's test and then unpaired t -test were used for data analysis. * P < 0.05 when compared to WT or control cells. ** P < 0.05 when compared to strains HSB3-HSB5. † P < 0.05 when compared to cell-bound enzyme activity. WT, strain 5110 ATCC MYA 4823. Strains HSB1 and HSB2 were transformed with pBGgHg; while HSB3-HSB8 with pBGgHg-Gp70.

Article Snippet: HSB6 , 5110 ATCC MYA 4823 transformed with pBGgHg-GP70 , 9.2 ± 1.1 , 3.1 ± 0.3 , This work.

Techniques: Activity Assay, Control, Transformation Assay

Journal: The Cell Surface

Article Title: Sporothrix brasiliensis Gp70 is a cell wall protein required for adhesion, proper interaction with innate immune cells, and virulence

doi: 10.1016/j.tcsw.2024.100139

Figure Lengend Snippet: Adhesion to extracellular matrix components of Sporothrix brasiliensis wild-type, control, and GP70 -silenced strains. In A , ELISA-based analysis of cell adhesion. The solid phase contained the extracellular matrix protein in 96-well plates, then yeast-like cells were added, and adherent cells were labeled with rabbit anti-rHsp60 antibodies. The presence of antibodies was detected by a colorimetric assay as described in the Material and methods section. WT, strain 5110 ATCC MYA 4823. In B , yeast-like cells were preincubated with preimmune sera or an anti-Gp70 antibody before their inclusion in the ELISA to detect adhesion to laminin. In C , the same legend as in panel B , but adhesion to fibronectin was analyzed. Absolute values are similar to those reported in panel A . Results are means ± SD of three biological replicates performed by duplicate. The Dunnett's test and then the unpaired t-test were used for data analysis. In A , * P < 0.05 when compared to WT or strains HSB1 and HSB2. In B and C , * P < 0.05 when compared to the non-treated (NT) condition of the same strain.

Article Snippet: HSB6 , 5110 ATCC MYA 4823 transformed with pBGgHg-GP70 , 9.2 ± 1.1 , 3.1 ± 0.3 , This work.

Techniques: Control, Enzyme-linked Immunosorbent Assay, Labeling, Colorimetric Assay

Journal: The Cell Surface

Article Title: Sporothrix brasiliensis Gp70 is a cell wall protein required for adhesion, proper interaction with innate immune cells, and virulence

doi: 10.1016/j.tcsw.2024.100139

Figure Lengend Snippet: Polysaccharide exposure at the cell wall surface and analysis of N -linked and O -linked glycans in Sporothrix brasiliensis wild-type, control, and GP70 -silenced strains. In ( A ), yeast-like cells were labeled with fluorescein isothiocyanate conjugated-wheat germ agglutinin or IgG Fc-Dectin-1 chimera for labeling of chitin and β-1,3-glucan, respectively. The fluorescence associated with 300 cells was acquired and normalized to that obtained with heat-killed cells, which represent 100 % of labeling. In ( B ), cells were treated either with endoglycosidase H or β-eliminated to trim N -linked glycans or O -linked glycans, respectively. Both were quantified by high-performance anion-exchange chromatography coupled with pulsed amperometric detection and sugar content normalized to that obtained with 1 × 10 9 yeast-like cells. In both panels, data are means ± SD of three biological replicates performed in duplicates. The Dunnett's test and then the unpaired t-test were used for data analysis. * P < 0.05 when compared to the WT strain or control strains HSB1 or HSB2. WT, strain 5110 ATCC MYA 4823.

Article Snippet: HSB6 , 5110 ATCC MYA 4823 transformed with pBGgHg-GP70 , 9.2 ± 1.1 , 3.1 ± 0.3 , This work.

Techniques: Control, Labeling, Fluorescence, Chromatography

Journal: The Cell Surface

Article Title: Sporothrix brasiliensis Gp70 is a cell wall protein required for adhesion, proper interaction with innate immune cells, and virulence

doi: 10.1016/j.tcsw.2024.100139

Figure Lengend Snippet: Cytokine stimulation by human peripheral blood mononuclear cells. Interactions between Sporothrix brasiliensis yeast-like cells and human peripheral blood mononuclear cells were incubated for 24 h and the secreted cytokines were recovered from supernatants and quantified by ELISA. Data are means ± SD obtained with samples from eight donors, each assayed in duplicate wells. Results were analyzed with the Dunnett's test and then Mann-Whitney U test. * P < 0.05 when compared to WT, HSB1, or HSB2 cells. † P < 0.05 when compared to the “no treatment” group from the same strain. No treatment, human cells preincubated with 5 μg mL −1 polymyxin B; + anti-MR Ab, human cells preincubated with 5 μg mL −1 polymyxin B and 10 μg mL −1 anti-mannose receptor (MR) antibodies; + anti-TLR4 Ab, human cells preincubated with 5 μg mL −1 polymyxin B and 10 μg mL −1 anti-TLR4; + laminarin, human cells preincubated with 5 μg mL −1 polymyxin B and 200 μg mL −1 laminarin. WT, strain 5110 ATCC MYA 4823.

Article Snippet: HSB6 , 5110 ATCC MYA 4823 transformed with pBGgHg-GP70 , 9.2 ± 1.1 , 3.1 ± 0.3 , This work.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: The Cell Surface

Article Title: Sporothrix brasiliensis Gp70 is a cell wall protein required for adhesion, proper interaction with innate immune cells, and virulence

doi: 10.1016/j.tcsw.2024.100139

Figure Lengend Snippet: Phagocytosis of GP70 -silenced S. brasiliensis by human monocyte-derived macrophages. In A, Acridine Orange-labeled yeast-like cells and human monocyte-derived macrophages were incubated for 2 h at 37 °C and 5 % ( v /v) CO 2 , and macrophages were analyzed by flow cytometry. Fifty thousand cells were counted per sample, and only those macrophages interacting with at least one fluorescent yeast cell were included in the analysis. Depending on the positivity in different fluorescent channels, the interactions were classified as in the early, intermediate, and late stages of the uptake process. Control, mock interactions where only macrophages were included. * P < 0.05 when compared to WT, HSB1 or HSB2 strains. In B , the same legend as in panel A but yeast-like cells were either live (No treatment) or heat inactivated (HK) before the interaction with immune cells. Alternatively, the monocyte-derived macrophages were preincubated with 200 μg mL −1 laminarin and then challenged with live yeast-like cells (+ Laminarin). Cell numbers are from the late stage of phagocytosis. Control, macrophages interacting with no yeast-like cells. Results from both panels were analyzed with the Dunnett's test and then the Mann-Whitney U test. * P < 0.05 when compared to the “no treatment” condition of the same strain. In both panels, results are means ± SD from eight donors assayed by duplicate. WT, strain 5110 ATCC MYA 4823.

Article Snippet: HSB6 , 5110 ATCC MYA 4823 transformed with pBGgHg-GP70 , 9.2 ± 1.1 , 3.1 ± 0.3 , This work.

Techniques: Derivative Assay, Labeling, Incubation, Flow Cytometry, Control, MANN-WHITNEY

Journal: The Cell Surface

Article Title: Sporothrix brasiliensis Gp70 is a cell wall protein required for adhesion, proper interaction with innate immune cells, and virulence

doi: 10.1016/j.tcsw.2024.100139

Figure Lengend Snippet: Analysis of virulence in Galleria mellonella larvae. In A, Groups containing 30 larvae were inoculated with 1 × 10 5 yeast-like cells and survival followed for two weeks. Data are shown in Kaplan–Meier plots. The Log-rank test showed no differences among the WT and HSB1-HSB5 strains ( P = 0.87). Mortality curves generated with strains HSB6-HSB8 were significantly different from those generated with WT cells ( P < 0.05). In B . hemolymph was collected and used to measure cell-free lactate dehydrogenase. The 100 % value corresponds to that obtained from lysed hemocytes. This hemolymph was also used to quantify hemocytes ( C ) or phenoloxidase activity ( D ). PBS refers to an animal group inoculated with phosphate saline buffer. WT, strain 5110 ATCC MYA 4823. In A , each group contained 30 larvae, In B , C , and D , data are shown as means ± SD from three biological replicates. Each animal group contained 10 larvae, and results were analyzed with the Dunnett's test and then the Mann-Whitney U test. * P < 0.05 when compared to the animal group inoculated with the WT, HSB1, or HSB2 control strains.

Article Snippet: HSB6 , 5110 ATCC MYA 4823 transformed with pBGgHg-GP70 , 9.2 ± 1.1 , 3.1 ± 0.3 , This work.

Techniques: Generated, Activity Assay, Saline, MANN-WHITNEY, Control