Evaluation of Various Media and Antimicrobial Agents for Isolation of Soil Fungi

Papavizas, G. C.; Davey, C. B.

doi:10.1097/00010694-195988020-00010

Cited by 123 publications

(70 citation statements)

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“…Plates were shuffled in the field to ensure media types were not inoculated consecutively. In 2012, dextrose-peptone-yeast extract agar (DPYA; Papavizas and Davey, 1959) and Sabouraud-dextrose agar (SAB) were used, with five plates of each media type for Dorchester Mine and Glebe Mine and six of each for Berryton Cave. In 2015, DPYA, SAB, malt extract (MEA), and modified DPYA were used, with five plates of each media type for each of the four hibernacula.…”

Section: Methodsmentioning

confidence: 99%

Detecting viable Pseudogymnoascus destructans (Ascomycota: Pseudeurotiaceae) from walls

Vanderwolf¹,

Malloch²,

McAlpine³

2016

JCKS

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Pseudogymnoascus destructans (Pd) causes the fungal disease white-nose syndrome (WNS), which has led to high mortality in some hibernating bat species in eastern North America. The ability to detect viable Pd in hibernacula is important for understanding the role the environment plays as a reservoir for infectious Pd. Previous studies have generally used the high-sugar medium Sabouraud-dextrose (SAB) and have had low yields of viable Pd from environmental samples of Pd-positive hibernacula. While cultureindependent methods (i.e., molecular genetics) have previously shown much better success in detecting Pd, these methods cannot determine viability. In 2012 and 2015, we swabbed walls in four hibernacula with WNS-positive bats in New Brunswick, Canada, and cultured the samples using dextrose-peptone-yeast extract agar (DPYA), SAB, and Malt extract (MEA) media. Samples cultured on DPYA produced viable Pd 43.7 to 50.0 % more frequently than SAB, with a maximum overall return for DPYA among sites of 62.5 % Pd-positive samples over both years. During the initial outbreak of WNS in our study region, Pd-positive swabs were produced from 40.0 to 83.3 % of samples on DPYA, whereas SAB produced a maximum of 40.0 %. At one site we detected Pd from 83.3% of swabs cultured on DPYA and 0 % on SAB. MEA produced no viable Pd. Our figures for Pd detection are as high as or higher than previously published culture-independent methods, while also confirming the viability of the Pd present. We found that the yield of viable Pd from hibernacula walls decreased from 2012 to 2015 as the hibernating bat population decreased due to WNS mortality, but patterns varied amongst hibernacula, and overall, were not statistically different. It is possible that environmental growth of Pd contributes to its persistence within hibernacula. We suggest that future studies on the environmental persistence of viable Pd discontinue the use of high-sugar media that lack inhibitory fungal growth ingredients, such as SAB and MEA, as they favor fast-growing fungal species that overgrow and mask slowergrowing fungi such as Pd.

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Section: Methodsmentioning

confidence: 99%

Detecting viable Pseudogymnoascus destructans (Ascomycota: Pseudeurotiaceae) from walls

Vanderwolf¹,

Malloch²,

McAlpine³

2016

JCKS

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“…The remaining spikes were dried, threshed, and the seeds were surface-sterilized in70%ethanolfor30sand in 1% sodium hypochlorite for 2 min, rinsed several times with stérile water, allowed to air dry aseptically for a few minutes, then placed on two sélective agar média in Pétri dishes. Mannitol agar (Bôhm-Shraml et al 1993) was used to detect bacteria, fungi and F. graminearum, while pentachloro-nitrobenzene (PCNB) modified agar (Papavizas 1967) was used to detect Fusarium spp. The concentration of agar in both média was reduced (7 g L 1 ) to facilitate the embedding of the seeds.…”

Section: Methodsmentioning

confidence: 99%

Occurrence of the orange wheat blossom midge [Diptera :Cecidomyiidae] in Quebec and its incidence on wheat grain microflora

Mongrain¹,

Couture²,

Dubuc

et al. 2005

phyto

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Samples of wheat spikes (Triticum aestivum) were collected in the summer of 1995 from different crop districts in Quebec and the occurrence of orange wheat blossom midge (Sitodiplosis mosellana) and seed microflora were determined. Estimated yield loss caused by wheat midge larvae averaged 6.3%. The percentage of infested spikes was significantly correlated with total seed contamination by fungi and bacteria (r = 0.79). The specific occurrence of Fusarium graminearum in grains was also significantly correlated with number of larvae per spike (r = 0.67) or per spikelet (r = 0.67). Consequently, the wheat midge might play a role in dissemination of F. graminearum.

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“…Diluting streaks were completed in the cave within 1 h of the initial streak, after which plates were sealed in situ with Parafilm (Pechiney Plastic Packaging, Chicago, IL). Dextrose-peptone-yeast extract agar (DPYA) was used (Papavizas and Davey, 1959), infused with the antibiotics chlortetracycline (30 mg/L) and streptomycin (30 mg/L). We have previously documented this as a superior medium for isolating P. destructans from the environment (Vanderwolf et al, 2016b).…”

Section: Fungal Samplingmentioning

confidence: 99%

Lack of cave-associated mammals influences the fungal assemblages of insular solution caves in eastern Canada

Vanderwolf¹,

Malloch²,

Ivanova³

et al. 2016

JCKS

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The biogeography of cave fungi and factors that influence community composition are poorly known. The movement of animals into caves from the outside environment is thought to be one factor that affects cave mycota. Islands often have different faunal assemblages from the mainland, and this may affect the fungal diversity of island caves. In 2014 we swabbed walls in three natural solution caves on Anticosti Island, Quebec, to determine the composition of cave fungal assemblages present relative to well-studied nearby mainland sites. At least one of these caves, Grotteà la Patate in Anticosti National Park, appears to support overwintering bats. Culture-dependent methods were used to establish pure cultures, and fungi were identified by a combination of morphology and genetic sequencing. A total of 54 fungal taxa were identified, with a mean of 7.4 6 3.9 taxa per swab. The most common taxa isolated were Penicillium spp., Pseudogymnoascus pannorum sensu lato, P. roseus, Trichoderma sp., Cladosporium spp., Thysanophora spp., Mucor sp., and Trichosporon dulcitum. Pseudogymnoascus destructans (Pd), the causative agent of the fungal disease white-nose syndrome in bats, was not detected, and we conclude that Pd was not present in the three sampled caves as of summer 2014. Two of the caves did not appear to be suitable bat hibernacula based on microclimate, although diverse fungal assemblages were detected on the walls. Several other fungal taxa common to bat hibernacula on the mainland, in addition to Pd, were lacking from Anticosti Island caves. We suggest that fungal assemblages on Anticosti Island are influenced by the absence of non-volant cave-visiting mammals on the island, particularly porcupines (Erethizon dorsatum) and raccoons (Procyon lotor), both frequent cave associates elsewhere in Maritime Canada.

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Evaluation of Various Media and Antimicrobial Agents for Isolation of Soil Fungi

Cited by 123 publications

References 0 publications

Detecting viable Pseudogymnoascus destructans (Ascomycota: Pseudeurotiaceae) from walls

Detecting viable Pseudogymnoascus destructans (Ascomycota: Pseudeurotiaceae) from walls

Occurrence of the orange wheat blossom midge [Diptera :Cecidomyiidae] in Quebec and its incidence on wheat grain microflora

Lack of cave-associated mammals influences the fungal assemblages of insular solution caves in eastern Canada

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