Development and validation of benomyl birdseed agar for the isolation of Cryptococcus neoformans and Cryptococcus gattii from environmental samples

Pham, Cau D.; Ahn, Stacey; Turner, Lance A.; Wohrle, Ron; Lockhart, Shawn R.

doi:10.1093/mmy/myt028

Cited by 14 publications

(13 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Cryptococcus species complex cannot produce melanin pigment without certain exogenous substrates (Casadevall et al, 2003;Eisenman et al, 2011;Garcia-Rivera et al, 2005). Based on this idea, several modified culture media were developed for the rapid identification of Cryptococcus species complex, such as mustard seed, chili pepper, Pinus halepensis seed, Eggplant, benomyl bird seed and blackberry agar (Menezes et al, 2011;Mseddi et al, 2011;Nandhakumar et al, 2006;Pedroso et al, 2007;Pham et al, 2014;Sengul et al, 2014). However, there are some limits.…”

Section: Discussionmentioning

confidence: 99%

Development of Modified Selective Media to Differentiate Cryptococcus Species Complex and its Serotypes using Natural Materials

Park

Kim

et al. 2017

BSL

View full text Add to dashboard Cite

The formation of brown colonies due to phenol oxidase activity on classic agar media containing natural material extracts of Helianthus annuus or on medium containing L-3,4-dihydroxyphenylalanine has been used to identify Cryptococcus species complex. In this study, various natural materials were used to develop a modified medium and to identify five major serotypes of Cryptococcus species complex. Serotypes A, D, and A/D were pigmented on medium using Perilla frutescens var. japonica Hara (PerJ agar) after a three-day incubation. Serotypes B and C were pigmented on PerJ agar after four-and five-day incubations, respectively. Growth time and pigmentation of the five serotypes occurred more rapidly on PerJ agar than on the other media. In addition, colony morphology, size, and pigmentation were specific by serotype. In conclusion, PerJ agar should be used in clinic settings to identify Cryptococcus species complex and its serotypes rapidly.

show abstract

Section: Discussionmentioning

confidence: 99%

Development of Modified Selective Media to Differentiate Cryptococcus Species Complex and its Serotypes using Natural Materials

Park

Kim

et al. 2017

BSL

View full text Add to dashboard Cite

show abstract

“…For isolation of Cryptococcus sp., two grams of each soil sample were suspended in sterile water, and the mixture was vortexed and allowed to settle for 1 minute. 20‐μL and 200‐μL aliquots of each supernatant were plated on benomyl birdseed agar plates (BBA) . Swabs were removed from the sample tube and gently rolled over the surface of multiple BBA plates.…”

Section: Methodsmentioning

confidence: 99%

“…20-μL and 200-μL aliquots of each supernatant were plated on benomyl birdseed agar plates (BBA). 13 Swabs were removed from the sample tube and gently rolled over the surface of multiple BBA plates. The plates were incubated at 25°C for up to 2 weeks.…”

Section: Introductionmentioning

confidence: 99%

Molecular typing of clinical and environmental isolates of Cryptococcus gattii species complex from southern California, United States

Hurst

Lysen

Cooksey

et al. 2019

Mycoses

View full text Add to dashboard Cite

Summary Background Two species complexes (SC) cause the majority of human Cryptococcus infections: Cryptococcus neoformans SC and Cryptococcus gattii SC. Infection is typically thought to be acquired following environmental exposure. In an urban setting, parks and other public spaces are a likely source of contact with C. gattii SC. Objectives The goals of this study were to describe the genetic diversity of C. gattii SC in the California environment, to determine the extent of environmental exposure in publicly accessed areas and to correlate the genotypes of environmental C. gattii SC isolates with those from patients in southern California. Methods Specimens from trees and soil from 13 parks and public areas of seven California counties were examined for C. gattii SC isolates. Isolates were sequence typed and compared to sequence types from human clinical isolates from the same area. Results Multilocus sequence typing identified C. gattii sensu stricto (VGI molecular type) as well as Cryptococcus bacillisporus (VGIII molecular type). Several C. bacillisporus but none of the C. gattii sensu stricto isolates shared sequence types with human clinical isolates from southern California. Conclusions C. gattii SC colonies exist in some California public parks. The presence of identical STs in environmental and human isolates of C. bacillisporus is suggestive of an arboreal origin of human infections. Two new tree species were documented as hosts for C. gattii SC in California, adding to the four species previously identified.

show abstract

“…Because of the nonselective properties of bird seed agar and caffeic acid agar, the yeast colonies are typically covered by rapid‐growing filamentous fungi during culture, directly affecting the rate of isolation. Some chemicals, such as benomyl, have been used to develop selective media that suppress the growth of Ascomycetes (Pham et al ., ). However, overgrowth of rapid‐growing filamentous fungi frequently occurs, particularly for Mucorales spp.…”

Section: Introductionmentioning

confidence: 97%

“…and Aspergillus spp. (Pham et al ., ). Modified dichloran Rose‐Bengal chloramphenicol (mDRBC) medium is used to isolate Cryptococcus , because it inhibits the growth of moulds, but considerable amounts of moulds are still observed on the surface of this agar (Castro e Silva Dde et al ., ).…”

Section: Introductionmentioning

confidence: 97%

Two‐step method for isolating Cryptococcus species complex from environmental material using a new selective medium

Lin

Chung

2019

Environ Microbiol Rep

View full text Add to dashboard Cite

Summary Cryptococcosis is an opportunistic infection caused by the Cryptococcus species complex. An outbreak of cryptococcosis caused by Cryptococcus gattii (AFLP6/VGII) in North America has indicated the need for studies of this organism and its environmental niche. Difficulties in isolating the Cryptococcus spp. because of the overgrowth of filamentous fungi onto culture media and its low fungal population size under natural conditions limit studies of these pathogenic yeasts. We designed a selective medium that inhibits the growth of environmental filamentous fungi but does not inhibit that of Cryptococcus cells. After enrichment in acidified YPD media and inoculation onto selective media, Cryptococcus cells in brown‐coloured colonies were isolated from environmental materials. This two‐step method is useful for isolating environmental members of the Cryptococcus species complex, which is essential for further studies involving diversity and the microbe–environment relationship of this yeast.

show abstract

Development and validation of benomyl birdseed agar for the isolation of Cryptococcus neoformans and Cryptococcus gattii from environmental samples

Cited by 14 publications

References 27 publications

Development of Modified Selective Media to Differentiate Cryptococcus Species Complex and its Serotypes using Natural Materials

Development of Modified Selective Media to Differentiate Cryptococcus Species Complex and its Serotypes using Natural Materials

Molecular typing of clinical and environmental isolates of Cryptococcus gattii species complex from southern California, United States

Two‐step method for isolating Cryptococcus species complex from environmental material using a new selective medium

Contact Info

Product

Resources

About