Fungal Culture Collections for the Biotechnology Industry

1988 ◽  
pp. 71-77
Author(s):  
Dennis Allsopp
Keyword(s):  
Biotechnology Industry ◽  
Fungal Culture ◽  
Culture Collections

In Vitro Establishment of Powdery Mildew (Microsphaera pulchra) on Microshoots of Flowering Dogwood

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 506a-506
Author(s):  
L.A. Klein ◽  
M.T. Windham ◽  
R.N. Trigiano
Keyword(s):  
Powdery Mildew ◽  
Woody Plant ◽  
Infected Individual ◽  
Callus Cultures ◽  
Fungal Culture ◽  
Culture Collections ◽  
Flowering Dogwood ◽  
Plant Parasite ◽  
In Vitro Establishment

Microshoot and callus cultures of Cornus florida (flowering dogwood), which were grown on woody plant medium amended with BA, were inoculated with Microsphaera pulchra (an obligate plant parasite) by gently shaking infected leaves bearing numerous conidia over the tissue. Culture dishes were sealed with parafilm and incubated at 24 °C with 25 mol·m–2·s–1 provided by cool fluorescent bulbs for 15 h. Cultures were examined with a dissecting scope every 24 h and cultures transferred when contaminating fungi were present. Specimens were prepared light microscopy and SEM. The fungus infected individual callus cells, but did not sporulate. In contrast, powdery mildew was well-established (both primary and secondary hyphae) in 70% of the microshoot cultures after 6 days and sporulated on 20% by 7 to 8 days. The cellular relationship between host and pathogen in vitro was similar to that found in greenhouse-grown plants. This technique has possible applications in maintaining fungal culture collections and studying host–pathogen relationships under more stringently controlled conditions.

scholarly journals ITEM - agro-food microbial culture collection: The importance of toxigenic fungi in the fight against mycotoxins

2014 ◽  
pp. 7-14
Author(s):  
Francesca Fanelli ◽  
Antonio Logrieco
Keyword(s):  
Biological Sciences ◽  
Fungal Pathogens ◽  
Industrial Applications ◽  
Culture Collection ◽  
Microbial Culture ◽  
Fungal Culture ◽  
Culture Collections ◽  
Fungal Isolation ◽  
Toxigenic Fungi ◽  
Fungal Biology

Fungal culture collections are important to biologists, microbiologists, epidemiologists and others involved in health and natural sciences. The improvement of techniques and methods for fungal isolation and preservation has contributed to maintain large microbial collections, which represent a rich source of biological sciences research, especially taxonomic, pathological and biodiversity studies as well as industrial applications. The collection centers are responsible for repository reference strains and for the maintenance of these microorganisms. The ITEM Microbial Culture Collection of ISPA (Institute of Sciences and of Food Production) includes more than 10,000 strains belonging to various agro-food microorganisms with phytopathological and toxicological significance. These microorganisms are mainly fungal pathogens belonging to toxigenic genera of Fusarium, Aspergillus, Alternaria, and Penicillium. This collection is a remarkable resource in the fight against mycotoxins: the increasing number of toxigenic fungi included in this collection ensures an original genetic source for biotechnological applications in several fields of research, contributing to knowledge improvement about fungal biology and strategies development for reducing mycotoxin contamination.

scholarly journals Molecular characterization of some lignicolous species from fungal culture collection

Genetika ◽  
2014 ◽  
Vol 46 (1) ◽  
pp. 235-242
Author(s):  
Nevena Stevic ◽  
Eleonora Capelja ◽  
Vladislava Galovic ◽  
Milana Novakovic ◽  
Maja Karaman
Keyword(s):  
Nuclear Dna ◽  
Culture Collection ◽  
Local Alignment ◽  
Fungal Culture ◽  
Culture Collections ◽  
Single Strain ◽  
Its1 Region ◽  
Industry And Education ◽  
Species Variability

Culture collections of microorganisms, including fungi, are strain deposits recognised as Biological Resource Centers (BRCs) with a great importance in science, industry and education. Their objective is to preserve the purity, viability and genomic integrity of every single strain as a member of such collection. Since improvement of molecular methods nowadays brought many novel approaches in manipulation with strains of microorganisms, they can also be useful for characterization of existing stored strains. ITS1 region in nuclear DNA is preferred barcoding marker for taxon identification, which can be explained by its great inter-species variability. This paper presents results from analysing ITS1 region sequences (17) obtained from fungal DNA of culture collection of autochthonous, lignicolous genera Piptoporus, Pleurotus, Ganoderma and Schizophyllum cultured on malt agar plates for 14 days at 25?C. BLAST (Basic Local Alignment Search Tool) was used for comparison with online databases, while alignment of sequences was made with MEGA 5.10 software. Morphological determination of species or genus was confirmed for 13 cultures, while the others were disproved. The resulting alignment indicated small intra-species variability of ITS1 region and pointed to it as an ideal marker for verification of fungal culture collections' authenticity.

scholarly journals 753. Performance Characteristics of Diagnostic Assays in Blastomycosis

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S424-S424
Author(s):  
Timothy O’Dowd ◽  
Jack McHugh ◽  
Nancy Wengenack ◽  
Elitza Theel ◽  
Paschalis Vergidis
Keyword(s):  
Granulomatous Inflammation ◽  
Cross Reactivity ◽  
Performance Characteristics ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fungal Culture ◽  
Noninvasive Test ◽  
Serum Antigen ◽  
Urine Antigen ◽  
Antigen Tests ◽  
Urine And Serum

Abstract Background Blastomycosis has historically been a difficult diagnosis to establish, often initially misdiagnosed as bacterial pneumonia. Serologic assays and polymerase chain reaction (PCR) tests are available, but their performance is not well defined. The objective of this study was to characterize their performance. Methods Subjects were identified via chart review of patients diagnosed with blastomycosis from 2005 to 2020. A definitive diagnosis was based on fungal culture, histopathology, or cytology. Performance characteristics of the Blastomyces antibody enzyme linked immunosorbent assay (ELISA), immunodiffusion (ID), complement fixation (CF), urine and serum antigen ELISAs, and PCR were evaluated in patients with confirmed blastomycosis. Data on patient demographics, location of disease, and mortality was also collected. Results We identified 193 patients with blastomycosis. The mean age was 51.8 years (range, 11-84) and 73.6% of patients were male. 42.5% resided in Minnesota, 18.1% in Wisconsin, and 12.9% in Iowa. Diagnosis was based on culture in 142 (73.2%) or histopathology/cytology in 67 (34.7%) patients. Granulomatous inflammation was present in 73.1% (38/52) while 21.2% (41/193) had evidence of extrapulmonary dissemination. The antibody, ID, and CF assays were positive in 43.5% (37/85), 35.1% (33/94) and 20.5% (8/39) of patients, respectively. Sensitivity of Blastomyces PCR was 40% (4/10) in sputum and 75% (21/28) in bronchoalveolar lavage (BAL) fluid. Blastomyces urine and serum antigen tests were positive in 68% (34/50) and 50% (9/18) of cases, respectively, while the urine antigen was positive in 63.6% (7/11) of disseminated cases. Patients had a positive Histoplasma urine antigen test in 54.1% (20/37) and Aspergillus galactomannan in BAL in 34.8% (8/23) of cases. Serum beta-D-glucan test was positive in 16.7% (2/12). 90-day mortality was 21/193 (10.9%) and median time from diagnosis to death was 18 days. Conclusion In this cohort, Blastomyces urine antigen was the most sensitive noninvasive test, with similar performance in pulmonary and disseminated disease. However, its utility is limited by poor specificity due to cross-reactivity. Blastomyces PCR from BAL fluid demonstrated the highest sensitivity. Blastomyces antibody, ID, and CF had poor sensitivity. Disclosures All Authors: No reported disclosures

Diagnostic Accuracy of a Direct Panfungal Polymerase Chain Reaction Assay Performed on Stained Cytology Slides

2021 ◽  
pp. 030098582199156
Author(s):  
Alexandra N. Myers ◽  
Unity Jeffery ◽  
Zachary G. Seyler ◽  
Sara D. Lawhon ◽  
Aline Rodrigues Hoffmann
Keyword(s):  
Polymerase Chain Reaction ◽  
Diagnostic Accuracy ◽  
Molecular Techniques ◽  
Fungal Culture ◽  
Chain Reaction ◽  
Identification Of Fungi ◽  
Fungal Dna ◽  
Polymerase Chain ◽  
Panfungal Pcr ◽  
Pcr Targeting

Molecular techniques are increasingly being applied to stained cytology slides for the diagnosis of neoplastic and infectious diseases. Such techniques for the identification of fungi from stained cytology slides have not yet been evaluated. This study aimed to assess the diagnostic accuracy of direct (without nucleic acid isolation) panfungal polymerase chain reaction (PCR) followed by sequencing for identification of fungi and oomycetes on stained cytology slides from dogs, cats, horses, and other species. Thirty-six cases were identified with cytologically identifiable fungi/oomycetes and concurrent identification via fungal culture or immunoassay. Twenty-nine controls were identified with no cytologically or histologically visible organisms and a concurrent negative fungal culture. Direct PCR targeting the internal transcribed spacer region followed by sequencing was performed on one cytology slide from each case and control, and the sensitivity and specificity of the assay were calculated. The sensitivity of the panfungal PCR assay performed on stained cytology slides was 67% overall, 73% excluding cases with oomycetes, and 86% when considering only slides with abundant fungi. The specificity was 62%, which was attributed to amplification of fungal DNA from control slides with no visible fungus and negative culture results. Direct panfungal PCR is capable of providing genus- or species-level identification of fungi from stained cytology slides. Given the potential of panfungal PCR to amplify contaminant fungal DNA, this assay should be performed on slides with visible fungi and interpreted in conjunction with morphologic assessment by a clinical pathologist.

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