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Diversity of fungi from the mound nests of Formica ulkei
and adjacent non-nest soils
Journal: Canadian Journal of Microbiology
Manuscript ID cjm-2015-0628.R2
Manuscript Type: Article
Date Submitted by the Author: 25-Feb-2016
Complete List of Authors: Duff, Lyndon B.; Brandon University, Biology Urichuk, Theresa M.; Brandon University, Biology Hodgins, Lisa N.; Brandon University, Biology Young, Jocelyn R.; Brandon University, Biology Untereiner, Wendy; Brandon University, Biology
Keyword: Aspergillus, fungal biodiversity, xerotolerant, mound-building ant
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Diversity of fungi from the mound nests of Formica ulkei and adjacent non-nest soils 1
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Lyndon B. Duff, Theresa M. Urichuk, Lisa N. Hodgins, Jocelyn R. Young, and Wendy A. 3
Untereiner1 4
Department of Biology, Brandon University, 270 18th Street, Brandon, Manitoba, R7A 6A9, 5
Canada 6
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L.B. Duff ([email protected]) 8
T.M. Urichuk ([email protected]) 9
L.N. Hodgins ([email protected]) 10
J.R. Young ([email protected]) 11
W.A. Untereiner ([email protected]) 12
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1 Corresponding author 22
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Abstract 23
Culture-based methods were employed to recover 3929 isolates of fungi from soils collected 24
in May and July 2014 from mound nests of Formica ulkei and adjacent non-nest sites. The 25
abundance, diversity, and richness of species from nest mounds exceeded those of non-26
mound soils, particularly in July. Communities of fungi from mounds were more similar to 27
those from mounds than non-mounds; this was also the case for non-mound soils with the 28
exception of one non-mound site in July. Species of Aspergillus, Paecilomyces and 29
Penicillium were dominant in nest soils and represented up to 81.8% of the taxa recovered. 30
Members of the genus Aspergillus accounted for the majority of Trichocomaceae from nests 31
and were represented almost exclusively by Aspergillus navahoensis and A. pseudodeflectus. 32
Dominant fungi from non-mound sites included Cladosporium cladosporioides, Geomyces 33
pannorum and species of Acremonium, Fusarium, Penicillium and Phoma. Although mound 34
nests were warmer than adjacent soils, the dominance of xerotolerant Aspergillus in soils 35
from mounds and the isolation of the majority of Trichocomaceae at 25˚C and 35˚C suggests 36
that both temperature and water availability may be determinants of fungal community 37
structure in nests of F. ulkei. 38
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Key words: Aspergillus, fungal biodiversity, mound-building ant, xerotolerant 41
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Introduction 48
The mound-building ant Formica ulkei Emery (Hymenoptera: Formicidae) ranges from Alberta 49
to Nova Scotia (Canada) and southward to Illinois, Indiana and Iowa (USA) (Holmquist 1928; 50
Sherba 1958; Glasier et al. 2013). This species builds conspicuous nests in meadows and 51
pastures along the margins of forests and sparsely wooded areas (Holmquist 1928; Dreyer 52
and Park 1932; Sherba 1958). Nests are composed of excavated soil and covered by a layer 53
of thatch (i.e., small pieces of grass and other plant material) (Sherba 1958, 1959, 1962). 54
The mound nests of F. ulkei are thermoregulatory in function and are constructed to 55
achieve and maintain higher temperatures than adjacent undisturbed soils during the months 56
when the ants are most active (Sherba 1962). Nests are built in exposed sites and are 57
oriented to maximize their exposure to solar radiation (Sherba 1958); they gain heat from 58
solar radiation in the early spring and maintain temperatures that are higher and more stable 59
than those of surrounding soils because of the insulating properties of thatch (Sherba 1962; 60
Frouz and Jilková 2008). This layer of organic material prevents the overheating of mounds 61
during the warmest parts of the year in other ant species that construct thatched nests 62
(Bollazzi and Rocces 2010; Kadochová and Frouz 2014) and it may serve the same function 63
in F. ulkei. 64
Although it is recognized that mound-building ants are capable of dramatically modifying 65
their environments and altering the chemical and physical properties of soils (Beattie and 66
Culver 1977; Frouz and Jilková 2008; Jilková et al. 2011), few studies have explored the 67
impact of microclimatic conditions on the composition of the communities of fungi in these 68
soils (Ba et al. 2000; Zettler et al. 2002; Rodrigues et al. 2014). Given the availability of a 69
large group of nests of F. ulkei in south-eastern Manitoba, we undertook a study to 1) confirm 70
the temperature characteristics of the mound nests of this species reported in previous 71
studies, and 2) test the hypothesis that the community of culturable fungi from soils from 72
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nests differs from adjacent, non-nest soils. We were also interested in comparing the species 73
richness and diversity of the communities of culturable fungi of separate mound nests of F. 74
ulkei. 75
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Materials and Methods 77
Collection of soils and temperature data 78
Thermocron iButton data loggers (DS1921G, Maxim Integrated Products, San Jose, USA) 79
that had been pre-set to measure temperature every two hours were coated in Performix 80
Plasti Dip (Plasti Dip International, Blaine, USA) to prevent moisture damage (Roznik and 81
Alford 2012). Data loggers were buried 5 cm deep in soil on the top, south side and north side 82
of three mound nests of Formica ulkei located on the un-forested edge of a cattle pasture that 83
had not been grazed in approximately 10 years, south of White Mud Falls, Manitoba (UTM 84
coordinates of mound 1 = 14U 0707355 5588945; mound 2 = 14U 0707363 5588913; mound 85
3 = 14U 0707367 5588908). One data logger was buried at a depth of 5 cm at one location 1 86
m south of each mound. Another data logger was also secured at a height of 2 m to the north 87
(i.e., the shaded) side of a tree located in the middle of the study area to collect air 88
temperatures. Data loggers recorded temperatures from 9 May to 18 September 2014. 89
Nests were sampled on 11 May and 14 July 2014 by collecting the uppermost 3 cm of 90
soil beneath the thatch from the top and south sides of each mound. Each site on all mounds 91
was sampled using a new plastic spoon. Soils to a depth of 3 cm were collected from 92
adjacent non-mound soil 1 m south of nests using a soil core sampler that was sterilized in 93
100% ethanol and rinsed in sterile distilled water between samples. Samples were placed into 94
separate, unused plastic freezer bags, sealed and transported in an ice cooler to the 95
laboratory. Each sample was emptied into a clean aluminum pan, air-dried at room 96
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temperature (18-21˚C), subjected to sieving using a 2 mm mesh to remove plant debris, and 97
stored in a new freezer bag. 98
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Isolation and identification of fungi 100
Individual soil samples were used within 3 days following collection to prepare ten-fold serial 101
dilutions in sterile distilled water ranging from 10-1 to 10-7. Each dilution was plated in triplicate 102
on Dextrose-Peptone-Yeast Extract agar (DPYA) (Papavizas and Davey 1959) lacking oxgall 103
and sodium propionate, and Dichloran Rose Bengal agar (DRBA) (King et al. 1979) 104
containing 25 mg Rose Bengal, 2 mg dichloran, and KH2PO4 rather than K2HPO4. Both media 105
were supplemented with 50 mg chlortetracycline hydrochloride and 50 mg streptomycin 106
sulphate. Duplicate sets of plates were incubated at 25˚C and 35˚C for 5 days. 107
All fungal colonies were transferred to Modified Leonian’s agar (MLA) (Malloch 1981), 108
incubated at room temperature and identified based on cultural and micro-morphological 109
characters. Isolates that could be discriminated as separate taxa within genera but not 110
identified to species were numbered. Sporulating fungi that could not be identified to the level 111
of genus were designated as “undetermined” whereas those taxa that did not sporulate on 112
MLA were labeled “sterile” (see supplemental Table S1). Non-filamentous fungi and 113
Zygomycota, which were isolated in very low numbers on both DRBA and DPYA, were 114
disregarded. Fungi recovered on DPYE were also excluded from analyses because of the 115
high levels of bacterial contamination, particularly in soils collected in July. 116
Dominant species of Aspergillus were characterized on Czapek Dox agar (CZ), Czapek 117
Yeast agar (CYA), Czapek Yeast agar with 20% sucrose (CY20S) and Malt Extract agar 118
following Klich (2002a) and on Creatine Sucrose agar (CREA) as described by Samson et al. 119
(2014). The thermotolerances of these taxa were determined by assessing their ability to 120
grow on CYA and MLA when incubated at 37˚C, 45˚C, and 50˚C. Cultures used for DNA 121
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extraction were grown as described previously (Untereiner et al. 2008) and total nucleic acids 122
were extracted from mycelia following the protocols of Lee and Taylor (1990). The nuclear 123
ribosomal internal transcribed spacer (nucITS) region and a portion of the gene encoding the 124
protein β-tubulin were amplified as described in Bogale et al. (2010) using the primers ITS4, 125
ITS5 (nucITS) (White et al. 1990) and Bt2a, and Bt2b (β-tubulin) (Glass and Donaldson 126
1995). PCR products were cleaned using a QIAquick PCR Purification Kit (Qiagen, 127
Mississauga, Canada). Sequencing reactions were performed using a Taq DyeDeoxy cycle 128
sequencing kit or a BigDye Terminator cycle sequencing kit (Applied Biosystems, Inc., Foster 129
City, USA) using the primers listed above. Confirmation of the identification of these taxa as 130
Aspergillus navahoensis (UAMH 11867; GenBank KU310972, KU310974) and A. 131
pseudodeflectus (UAMH 11868; GenBank KU310973, KU310975) was based on the 132
comparison of generated DNA sequences to the nucITS and β-tubulin barcodes provided by 133
Samson et al. (2014). 134
135
Statistical analyses 136
Daily temperature readings for Thermocron iButton data loggers placed in the south side of 137
each mound were averaged per day from May 6 to September 18, 2014. Data for the tops of 138
mounds were not included in averages because two iButtons from this location were 139
dislodged during the course of the study. Data from the north sides of mounds were also 140
excluded because these temperatures differed significantly from temperatures from the south 141
sides of mounds (data not shown). A one-way analysis of variance (ANOVA) of temperature 142
differences (mounds 1, 2, and 3, non-mounds 1, 2, and 3, and ambient temperature) was 143
conducted using PSPP v 0.8.4 (Pfaff 2015). The same software was used to perform a post-144
hoc Tukey HSD test. 145
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Numbers of isolates on DRBA were used to calculate colony-forming units (CFU) per g 146
of soil and the proportional abundance of each species or taxon within a group (i.e., “sterile” 147
and “undetermined”). Diversity indices (Shannon, Simpson and Simpson inverse) were 148
calculated using BiodiversityR (Kindt and Coe 2005). Rényi diversity profiles describing the 149
richness and evenness of sites were also generated using BiodiversityR. Between sites 150
comparisons of species-abundance data were measured using the Morisita-Horn index of 151
similarity in BiodiversityR. These data were converted into distance matrixes and employed to 152
generate dendrograms using hierarchical clustering R v 3.2.2 (R Core Team 2015). 153
154
Results 155
Maximum and mean average daily temperatures of soils from mounds exceeded those of 156
adjacent non-mound sites (Table 1) and results of an ANOVA (F(6, 924) = 61.90, p = 0.000) 157
(Supplemental Table S2) indicated significant differences in the mean average temperatures 158
between sites. Post-hoc Tukey HSD multiple comparisons revealed that the average daily 159
temperatures of mounds were higher than non-mound sites (Supplemental Table S3). The 160
temperatures of mound 2 and 3 did not differ significantly, nor were significant differences in 161
temperature seen among non-mound sites. All mound sites were warmer than ambient 162
temperature whereas non-mound sites 2 and 3 were cooler. Non-mound site 1 did not differ 163
significantly from ambient temperature. Differences in the average weekly temperatures of 164
soils from mound and non-mound sites are illustrated in Figure 1. 165
Excluding non-filamentous fungi and Zygomycota, a total of 3929 isolates representing 166
307 taxa were recovered at all dilutions from mound nest and adjacent non-mound sites on 167
DRBA (Table 2, Supplemental Table S1). Higher numbers of isolates and taxa were obtained 168
from DRBA incubated at 25 C. Soils collected in July contained a larger numbers of isolates 169
(Table 2) and had greater species richness (Table 3) than soils collected in May. 170
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The abundance (CFU g-1), diversity, and richness of species from soils of nest mounds 171
generally exceeded those of non-mounds, particularly in July (Table 3). Mound soils differed 172
in richness among sites in July, as did soils from non-mounds. Species richness in May did 173
not differ as dramatically between sites with the exception of mound 1 which was under-174
sampled because of an error in the preparation of soil dilutions. Rényi profiles did not 175
discriminate between mound and non-mound soils in May with respect to species diversity; 176
soils in July differed with the exception of non-mound 2 that intersected with mound 2 and 177
mound 3 (Figure 2). As illustrated in Figure 2, the evenness of species from soils from non-178
mound 2 was higher than at all other sites in May and July but the evenness of the remaining 179
sites could not be ranked. The evenness of soil from mound 1 in May likely reflects the 180
aforementioned under-sampling. Communities in soils from mounds were more similar to 181
species from mounds than non-mound sites in May and July (Figure 3). This was also the 182
case for taxa from non-mounds with the exception of the community from non-mound 1 in 183
July that more closely resembled the mycota from mounds. 184
The most abundant fungi in soil from mounds were Aspergillus navahoensis (ITS 99% 185
similarity to EF652424; β-tubulin 99% similarity to EF652248) and Aspergillus 186
pseudodeflectus (ITS 100% similarity to EF652507; β-tubulin 100% similarity to EF652331), 187
that represented 17.4 to 44.2% and 8.6 to 37.6% of the recovered taxa, respectively (Tables 188
4-5). Both species were recovered from all mounds in May and July. The proportional 189
abundances of these species were higher in May except that A. pseudodeflectus was more 190
abundant in mound 2 in July. Aspergillus pseudodeflectus was recovered from only a single 191
non-mound site in May but in very low abundance (0.3%) representing a single isolate 192
whereas A. navahoensis was never isolated from non-mound soils. Cultures of A. 193
navahoensis conformed to the description of this species provided by Christensen and States 194
(1982) and were distinctive in producing rapidly maturing ascomata, abundant Hülle cells, and 195
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crystal-encrusted hyphae. Aspergillus navahoensis grew at 37˚C and at 45˚C, but showed 196
better growth at 37˚C; it exhibited no growth at 50˚C. Aspergillus pseudodeflectus grew at 197
37˚C but exhibited no growth at 45˚C and 50˚C. 198
Additional taxa from mound soils with abundances higher than 5% included 199
Cladosporium cladosporioides, Geomyces pannorum, Myriothecium sp. and species of 200
Acremonium. However, these fungi were not the dominant members of the mycota of all 201
mounds nor were they equally abundant in the same mound in both May and July. 202
Undetermined species were dominant members of soils from mound 3 and were more 203
abundant in July. Sterile fungi comprised more than 5% of the isolates in soils from every 204
mound but only in July. 205
Species of Penicillium were dominant members of the mycota of soils from non-mound 206
sites but were less abundant in May than in July. Other taxa from non-mound sites with 207
abundances greater than 5% included Cladosporium cladosporioides, Geomyces pannorum, 208
undetermined and sterile fungi, and members of the genera Acremonium, Fusarium and 209
Phoma. However, only Geomyces pannorum, undetermined and sterile fungi, and species of 210
Penicillium represented more than 5% of the taxa recovered at more than one site at a given 211
sampling time. 212
213
Discussion 214
The results of the present study agree with Scherba (1962) who reported that the thatch-215
covered mound nests of Formica ulkei are warmer than surrounding undisturbed soils during 216
the months when these ants are most active. We also observed significant differences 217
between the temperatures of the north and south sides of mounds (data not included), a 218
phenomenon that can likely be attributed to variations in the dimensions of mounds, the 219
composition and density of thatch, and degree of shading (Scherba 1962; Frouz 2000; 220
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Kadochová and Frouz 2014). 221
Our investigation also demonstrates that the communities of fungi in soils from nest 222
mounds of Formica ulkei differ from non-mound soils with respect to the abundances of 223
species, species richness, and diversity. Soils of nest mounds of F. ulkei resemble those of 224
Solenopsis invicta (red imported fire ant) in containing greater numbers of fungal colonies 225
than adjacent, non-nest soils (Zettler et al. 2002) but they differ in having higher levels of 226
species richness. In July, two of the three mounds we sampled had higher levels of species 227
diversity than non-nest soils. In contrast, culture-dependent assessments revealed that below 228
ground nests of young colonies of Atta (leaf-cutting ants) contain lower to comparable 229
numbers of colonies of filamentous fungi as non-nest soils but have similar levels of species 230
diversity and richness (Rodrigues et al. 2014). 231
Members of the Trichocomaceae (species of Aspergillus, Paecilomyces and Penicillium) 232
were dominant in soils from mound nests of F. ulkei and represented 39.5% (mound 3) to 233
81.8% (mound 1) of the total numbers of taxa recovered. Trichocomaceae are among the 234
most common filamentous Ascomycota isolated from the nests of mound-building and leaf-235
cutting ants (Baird et al. 2007; Zettler et al. 2002; Sharma and Sumbali 2013; Rodrigues et al. 236
2014) but only a single study (Zettler et al. 2002) resembles ours in recovering different 237
representatives of this family from nests and non-nest soils. 238
Aspergillus accounted for more than 80% of Trichocomaceae isolated from mound nests 239
and were represented almost exclusively by Aspergillus navahoensis (section Nidulantes) and 240
A. pseudodeflectus (section Usti). Aspergillus navahoensis was described from soils from a 241
cool desert shrub community in northern Arizona (Christianson and States 1982) and belongs 242
to a section of the genus that occurs at greater than expected frequencies in desert soils 243
(Klich 2002b). This species was recovered originally in low numbers (Christianson and States 244
1982) and, apart from the present study, does not appear to have been collected since it was 245
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described. Aspergillus pseudodeflectus is an infrequently collected osmophilic species 246
described from desert soils in Egypt (Samson and Mouchacca 1975) that was reported to be 247
restricted to the tropics and subtropics (Christensen and Tuthill 1985). It is closely related to 248
A. calidoustus, a more commonly encountered species known from clinical and environmental 249
sources that is distinguished from A. pseudodeflectus based on its ecology and molecular 250
barcodes (Samson et al. 2011, 2014). 251
Trichocomaceae were also abundant in soils from non-mound sites but were 252
represented almost exclusively by species of Paecilomyces and Penicillium. These genera 253
were consistently more abundant in non-mound soils than in soils from mounds. Members of 254
the genus Aspergillus were absent from non-mound soils with the exception of a single colony 255
of A. pseudodeflectus that we suspect was a contaminant. 256
Differences in the fungal communities of the soils of nest mounds of Formica ulkei and 257
adjacent non-nest sites likely reflect environmental factors that are influenced by nest location 258
and architecture. For example, mound nests of F. ulkei in Illinois were shown to be restricted 259
to drier regions along forest margins and were constructed to maximize insolation (Dreyer and 260
Park 1932; Dreyer 1942). Nest construction also dramatically alters the physical 261
characteristics of soil that operate to regulate the moisture content and temperatures of 262
mounds relative to surrounding soils. Mound building can increase soil porosity and reduce 263
the bulk density of soils, both of which influence soil aeration and soil permeability (Frouz and 264
Jilková 2008). The moisture content in mounds of F. ulkei at 5 cm has been shown to be 265
lower than in adjacent soils throughout the year and lower than in mounds at 30 cm during the 266
warmer months when the ants were active (Sherba 1959). Although we did not determine the 267
moisture content of soils at our study site, we observed that the daily temperatures of nests of 268
F. ulkei peaked in the evening and decreased slowly during the night (data not shown) in 269
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agreement with the description of the drier and more exposed nests of Formica polyctena 270
(European red wood ant) (Frouz 2000). 271
The supposition that the nests of Formica ulkei at our study site were drier than adjacent 272
sites is also supported by the dominance of Aspergillus in nests as compared to soils located 273
1 m from each mound. Species of Aspergillus are common in soils from warmer regions of the 274
world (Domsch et al. 1993; Bills et al. 2004) and are among the most xerotolerant 275
Ascomycota (Dix and Webster 1995; Zak and Wildman 2004). Members of this genus are 276
particularly abundant in desert and grassland soils where they represent up to 20% of isolated 277
species (Christensen and Tuthill 1985). Although both A. navahoensis and A. 278
pseudodeflectus were capable of growth at the highest average daily temperatures recorded 279
for mound and non-mound soils, only the former species was determined to be thermotolerant 280
(i.e., it grows at temperatures below 20˚C and at 40˚C or higher). This finding, in conjunction 281
with our observation that all Aspergillus and Paecilomyces and nearly half of the species of 282
Penicillium were isolated at both 25˚C and 35˚C (Supplemental Table S1), suggests that 283
water availability is also be a determinant of fungal community structure in mound nests of F. 284
ulkei. 285
Factors such as nutrient availability, soil chemistry and the physical properties of soils 286
also likely influence the structure of fungal communities in the mound nests of F. ulkei and 287
adjacent non-nest soils. For example, soils in ant nests have higher levels of nutrients (Frouz 288
et al. 2005; Frouz and Jílková 2008; Ginzburg et al. 2008; Jílková et al. 2015) and differ from 289
surrounding soils in pH, porosity, and the content of organic matter (Frouz and Jílková 2008; 290
Jílková et al. 2011). Microbial activity is assumed to be higher in ant nests because of these 291
differences, but the mechanisms underlying the impacts of ants on soil processes and other 292
soil biota are not well understood (Frouz and Jílková 2008; Del Toro et al. 2012). 293
Nests of Formica ulkei are reservoirs of fungal diversity that should be explored further 294
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using the approaches presented here. Our understanding of these communities would be 295
improved with the more frequent sampling of nest mounds and adjacent non-nest soils, the 296
isolation of fungi over a longer period of time, the use of media designed to isolate 297
ecologically specialized taxa, and the determination of temperature differences from a larger 298
number of sites within nest mounds. And because the enumeration methods used in our 299
study are selective for fungi that produce abundant spores (Garrett 1981), it would be 300
valuable to examine the diversity of culturable fungi in these soils using alternative isolation 301
methods (described in Bills et al. 2004). The complementary use of sequence-based 302
approaches such as environmental metagenomics would also enhance our understanding of 303
these assemblages of fungi, particularly in recovering non-culturable species and taxa that 304
are under-sampled employing cultured-dependent methods (Bills et al. 2004; Karst et al. 305
2013; Rodrigues et al. 2014). Sequence based approaches would also facilitate the 306
identification of sterile fungi and many of the micro-morphologically simple or taxonomically 307
challenging species present in the mound nests of Formica ulkei. 308
309
Acknowledgments 310
We are indebted to Gary McNeely (Brandon University) and three anonymous reviewers for 311
their insightful editorial comments and suggestions for the improvement of this paper. We also 312
thank Dennis and Jacqueline Caya for their permission to access nest mounds located on 313
their property and David Caya for serving as a bear guard during soil sampling. Financial 314
support for this study was provided by a Natural Sciences and Engineering Research Council 315
(NSERC) of Canada Discovery Grant to W.A.U. Funding in the form of NSERC 316
Undergraduate Summer Research Awards to L.B.D. (2014, 2015), T.M.U. (2014) and J.R.Y. 317
(2015) is very gratefully acknowledged. 318
319
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References 320
Ba., A.S., Phillips, S.A., Jr., and Anderson, J.T. 2000. Yeasts in mound soil of the red 321
imported fire ant. Mycological Research 104: 969–973. 322
323
Baird, R., Woolfolk, S., and Watson, C.E. 2007. Survey of the bacterial and fungal associates 324
of black/hybrid imported fire ants from mounds in Mississippi. Southeastern Naturalist 6(4): 325
615–632. 326
327
Beattie, A.J., and Culver, D.C. 1977. Effects of the mound nests of the ant, Formica 328
obscuripes, on the surrounding vegetation. American Midland Naturalist 97(2): 390–399. 329
330
Bills, G., Christensen, M., Powell, M., and Thorn, G. 2004. Saprobic soil fungi. In Biodiversity 331
of fungi: Inventory and monitoring methods. Edited by G.M. Mueller, G.F. Bills and M.S. 332
Foster. Elsevier Academic Press, Burlington, MA. pp. 271–302. 333
334
Bogale, M., Orr, M.-J., O’Hara, M.J., and Untereiner, W.A. 2010. Systematics of Catenulifera 335
(anamorphic Hyaloscyphaceae) with an assessment of the phylogenetic position of 336
Phialophora hyaline. Fungal Biology 114: 396–409. 337
338
Bollazzi, M., and Roces, F. 2010. The thermoregulatory function of thatched nests in the 339
South American grass-cutting ant, Acromyrmex heyeri. Journal of Insect Science 10(137): 1–340
17. 341
342
Christensen, M., and States, J.S. 1982. Aspergillus nidulans Group: Aspergillus navahoensis, 343
and a revised synoptic key. Mycologia 74(2): 226–235. 344
Page 14 of 29
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Canadian Journal of Microbiology
Draft
Duff et al.; Fungi from nests of Formica ulkei 15
345
Christensen, M., and Tuthill, D.E. 1985. Aspergillus: an overview. In Advances in Penicillium 346
and Aspergillus systematics. Edited by R.A. Samson and J.I. Pitt. Plenum, New York, NY. pp. 347
195–209. 348
349
Del Toro, I., Ribbons, R.P., and Pelini, S.L. 2012. The little things that run the world revisited: 350
a review of ant-mediated ecosystem services and disservices (Hymenoptera: Formicidae). 351
Myremecological News 17: 133–146. 352
353
Dix, N.J., and Webster, J. 1995. Fungal ecology. Chapman and Hall, London. 354
355
Dreyer, W.A. 1942. Observations on the occurrence and size of ant mounds with reference to 356
their age. Ecology 23(4): 486–490. 357
358
Dreyer, W.A., and Park, T. 1932. Local distribution of Formica ulkei mound-nests with 359
reference to certain ecological factors. Psyche 39(4): 127–133. 360
361
Domsch, K.H., Gams, W., and Anderson, T.-H. 1993. Compendium of soil fungi. IHW-Verlag, 362
Regensburg. 363
364
Frouz, J. 2000. The effect of nest moisture on daily temperature regime in the nests of 365
Formica polyctena wood ants. Insects Sociaux 47: 229–235. 366
367
Frouz, J., and Jílková, V. 2008. The effect of ants on soil properties and processes 368
(Hymenoptera: Formicidae). Myrmecological News 11: 191–199. 369
Page 15 of 29
https://mc06.manuscriptcentral.com/cjm-pubs
Canadian Journal of Microbiology
Draft
Duff et al.; Fungi from nests of Formica ulkei 16
370
Frouz, F., Kalčík, J., and Cudlín, P. 2005. Accumulation of phosphorus in nests of red wood 371
ants Formica s. str. Annales Zoologici Fennici 42: 269–275. 372
373
Garrett, S.D. 1981. Soil fungi and soil fertility: an introduction to soil mycology, 2nd ed.. 374
Pergamon Press, Oxford, UK. 375
376
Ginzburg, O., Whitford, W.G., and Steinberger, Y., 2008. Effects of harvester ant (Messor 377
spp.) activity on soil properties and microbial communities in a Negev Desert ecosystem. 378
Biology and Fertility of Soils 45: 165–173. 379
380
Glasier, J.R.N., Acorn, J.H., Nielson, S.E., and Proctor, H. 2013. Ants (Hymenoptera: 381
Formicidae) of Alberta: a key to species. Canadian Journal of Arthropod Identification 22: 1–382
104. 383
384
Glass, N.L., and Donaldson, G.C. 1995. Development of primer sets designed for use with the 385
PCR to amplify conserved genes from filamentous ascomycetes. Applied and Environmental 386
Microbiology 61: 1323–1330. 387
388
Holmquist, A.M. 1928. Notes on the life history and habits of the mound-building ant, Formica 389
ulkei Emery. Ecology 9(1): 70–87. 390
391
Jílková, V., Matejícek, L., and Frouz, J. 2011. Changes in the pH and other soil chemical 392
parameters in soil surrounding wood ant (Formica polyctena) nests. European Journal of Soil 393
Biology 47(1): 72–76. 394
Page 16 of 29
https://mc06.manuscriptcentral.com/cjm-pubs
Canadian Journal of Microbiology
Draft
Duff et al.; Fungi from nests of Formica ulkei 17
395
Jílková, V., Frouz, J., Mudrák, O., and Vohník. M. 2015. Effects of nutrient-rich substrate and 396
ectomycorrhizal symbiosis on spruce seedling biomass in abandoned nests of the wood ant 397
(Formica polyctena): a laboratory experiment. Geoderma 259–260: 56–61. 398
399
Kadochová, S., and Frouz, J. 2014. Thermoregulation strategies in ants in comparison to 400
other social insects, with a focus on red wood ants (Formica rufa group) F1000 research 2: 401
280 (doi:10.12688/f1000research.2-280.v2). 402
403
Karst, J., Piculell, B., Brigham, C., Booth, M., and Hoeksema, J.D. 2013. Fungal communities 404
in soils along a vegetation ecotone. Mycologia 105(1): 61–70. 405
406
Kindt, R., and Coe, R. 2005. Tree diversity analysis. A manual and software for common 407
statistical methods for ecological and biodiversity studies. World Agroforestry Centre, Nairobi. 408
409
King, A.D., Jr., Hocking, A.D., and Pitt, J.I. 1979. Dichloran-rose bengal medium for 410
enumeration and isolation of molds from foods. Applied and Environmental Microbiology 411
37(5): 959–964. 412
413
Klich, M.A. 2002a. Identification of common Aspergillus species. Centraalbureau voor 414
Schimmelcultures, Utrecht, The Netherlands. 415
416
Klich, M.A. 2002b. Biogeography of Aspergillus species in soil and litter. Mycologia 94(1): 21–417
27. 418
Page 17 of 29
https://mc06.manuscriptcentral.com/cjm-pubs
Canadian Journal of Microbiology
Draft
Duff et al.; Fungi from nests of Formica ulkei 18
419
Lee, S.B., and Taylor, J.W. 1990. Isolation of DNA from fungal mycelium and single spores. In 420
PCR protocols: a guide to methods and applications. Edited by M.A. Innis, D.H. Gelfand, J.J. 421
Sninsky and T.J. White. Academic Press, San Diego, CA. pp. 282–287. 422
423
Malloch, D. 1981. Moulds: their isolation, cultivation and identification. University of Toronto 424
Press, Toronto, ON. 425
426
Papavizas, G.C., and Davey, C.B. 1959. Evaluation of various media and antimicrobial agents 427
for isolation of soil fungi. Soil Science 88(2): 112–117. 428
429
Pfaff, B. 2015. GNU PSPP Statistical Analysis Software [online]. Available at 430
http://www.gnu.org/software/pspp/ [accessed June 2015]. 431
432
R Core Team. 2015. R: a language and environment for statistical computing [online]. 433
Available at http://www.r-project.org/ [accessed August 2015]. 434
435
Rodrigues, A., Passarini, M.R.Z., Ferro, M., Nagamoto, N.S., Forti, L.C., Bacci, M., Jr., Setta, 436
L.D., and Pagnocca, F.C. 2014. Fungal communities in the garden chamber soils of leaf-437
cutting ants. Journal of Basic Microbiology 54: 1186–1196. 438
439
Roznik, E.A., and Alford, R.A. 2012. Does waterproofing Thermochron iButton dataloggers 440
influence temperature readings? Journal of Thermal Biology 37(4): 260–264. 441
442
Page 18 of 29
https://mc06.manuscriptcentral.com/cjm-pubs
Canadian Journal of Microbiology
Draft
Duff et al.; Fungi from nests of Formica ulkei 19
Samson, R.A., and Mouchacca, J. 1975. Additional notes on species of Aspergillus, Eurotium 443
and Emericella from Egyptian desert soil. Antonie van Leeuwenhoek 41: 343–351. 444
445
Samson, R.A., Varga, J., Meijer, M., and Frisvad, J.C. 2011. New taxa in Aspergillus section 446
Usti. Studies in Mycology 69: 81–97. 447
448
Samson, R.A., Visagie, C.M., Houbraken, J., Hong, S.B., Hubka, V., Klaassen, C.H.W., 449
Perrone, G., Seifert, K.A., Susca, A., Tanney, J.B., Varga, J., Kocsubé, S., Szigeti, G., 450
Yaguchi, T., and Frisvad, J.C. 2014. Phylogeny, identification and nomenclature of the genus 451
Aspergillus. Studies in Mycology 78: 141–173. 452
453
Sherba, G. 1958. Reproduction, nest orientation and population structure of an aggregation of 454
mound nests of Formica ulkei Emery (Formicidae). Insectes Sociaux 5(2): 201–213. 455
456
Sherba, G. 1959. Moisture regulation in mound nests of the ant, Formica ulkei Emery. The 457
American Midland Naturalist 61(2): 499–508. 458
459
Scherba, G. 1962. Mound temperatures of the ant Formica ulkei Emery. The American 460
Midland Naturalist 67(2): 373–385. 461
462
Sharma, V., and Sumbali, G. 2013. Occurrence and diversity indices assessment of 463
micromycetes associated with the ant-hills of parks and gardens. 2013. Current Research in 464
Microbiology and Biotechnology 1(4): 166–172. 465
466
Page 19 of 29
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Duff et al.; Fungi from nests of Formica ulkei 20
Untereiner, W.A., Angus, A., Réblová, M., and Orr, M.-J. 2008. Systematics of the 467
Phialophora verrucosa complex: new insights from analyses of β-tubulin and large subunit 468
nuclear rDNA and ITS sequences. Botany 86(7): 742–750. 469
470
White, T.J., Bruns, T., Lee, S., and Taylor, J. 1990. Amplification and direct sequencing of 471
fungal ribosomal RNA genes for phylogenetics. In PCR protocols: a guide to methods and 472
applications. Edited by M.A. Innis, D.H. Gelfand, J.J. Sninsky and T.J. White. Academic 473
Press, San Diego, CA. pp. 315–324. 474
475
Zak, J.C., and Wildman, H.C. 2004. Fungi in stressful environments. In Biodiversity of fungi: 476
Inventory and monitoring methods. Edited by G.M. Mueller, G.F. Bills and M.S. Foster. 477
Elsevier Academic Press, Burlington, MA. pp. 303–315. 478
479
Zettler, J.A., McInnis, T.A., Jr., Allen, C.R., and Spira, T.P. 2002. Biodiversity of fungi in red 480
imported fire ant (Hymenoptera: Formicidae) mounds. Annals of the Entomological Society of 481
America 95(4): 487–491. 482
483
484
485
486
487
488
489
490
491
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Figure captions 492
Figure 1. Weekly averages of ambient temperatures and temperatures of mound (M1, M2 and 493
M3) and non-mound (S1, S2 and S3) soils. Markers indicate the dates when soils were 494
collected. 495
496
Figure 2. Renyi profiles comparing the diversity of fungi found in mound and non-mound soils 497
from a) May 2014 and b) July 2014. Alpha = 0 is the species richness, alpha = 1 is the 498
Shannon-Weiner diversity index, and alpha = 2 is the log of the reciprocal of the Simpson 499
diversity index. 500
501
Figure 3. Dendrograms illustrating Morisita-Horn similarities between the communities of fungi 502
from mound (M1, M2 and M3) and non-mound (S1, S2 and S3) soils in a) May 2014 and b) 503
July 2014. 504
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Figure 1. Weekly averages of ambient temperatures and temperatures of mound (M1, M2 and M3) and non-mound (S1, S2 and S3) soils. Markers indicate the dates when soils were collected.
577x347mm (96 x 96 DPI)
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Figure 2. Renyi profiles comparing the diversity of fungi found in mound and non-mound soils from a) May 2014 and b) July 2014. Alpha = 0 is the species richness, alpha = 1 is the Shannon-Weiner diversity index,
and alpha = 2 is the log of the reciprocal of the Simpson diversity index.
397x725mm (96 x 96 DPI)
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Figure 3. Dendrograms illustrating Morisita-Horn similarities between the communities of fungi from mound (M1, M2 and M3) and non-mound (S1, S2 and S3) soils in a) May 2014 and b) July 2014.
133x236mm (300 x 300 DPI)
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Table 1. Descriptive statistics for average daily temperatures (°C) of mound (M) and non-mound (S) sites.
Site N Minimum Maximum Mean Std.
deviation Std. error
95% Confidence interval for mean
Lower bound Upper bound
M1 133 7.03 23.08 17.11 3.77 0.33 16.46 17.75
M2 133 8.52 29.08 22.26 5.13 0.45 21.38 23.14
M3 133 7.10 25.17 18.83 4.34 0.38 18.09 19.58
S1 133 7.46 19.92 14.96 3.03 0.26 14.44 15.48
S2 133 6.17 18.96 14.40 3.10 0.27 13.87 14.93
S3 133 6.25 18.58 13.94 2.82 0.24 13.46 14.43
All sites 798 6.17 29.08 16.92 4.70 0.17 16.58 17.25
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Table 2. Number of isolates of fungi recovered at 25 °C and 35 °C on DRBA from mound (M) and non-mound (S) sites in May and July 2014.
Sample time
Temperature of incubation (°C)
M1 S1 M2 S2 M3 S3 Total
May 25 134 135 191 80 391 237 1168 May 35 54 2 152 152 116 20 496 July 25 416 59 267 139 552 1 1434 July 35 328 59 152 35 240 17 831
Total − 932 255 762 406 1299 275 3929
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Table 3. Richness and diversity estimators of fungal communities of mound (M) and non-mound (S) sites calculated in BiodiversityR.
May 2014 Total CFU/g soil Species richness Shannon diversity Simpson diversity a Simpson inverse
M1 115.67 x 107 8 1.32 ± 0.14 0.668 ± 0.072 3.01 ± 0.65
1 10.83 x 107 44 2.75 ± 0.07 0.871 ± 0.014 7.72 ± 0.81
M2 29.55 x 107 36 2.32 ± 0.08 0.792 ± 0.029 4.81 ± 0.68
S2 36.66 x 107 33 2.81 ± 0.09 0.928 ± 0.003 13.86 ± 0.61
M3 57.75 x 107 30 2.27 ± 0.09 0.821 ± 0.022 5.59 ± 0.70
S3 11.4 x 107 37 2.21 ± 0.09 0.800 ± 0.022 5.00 ± 0.55
July 2014 Total CFU/g soil Species richness Shannon diversity Simpson diversity Simpson inverse
M1 38.7 x 107 49 2.11 ± 0.08 0.781 ± 0.021 4.57 ± 0.43
S1 5.77 x 107 36 2.00 ± 0.06 0.654 ± 0.056 2.89 ± 0.47
M2 42.45 x 107 66 3.09 ± 0.06 0.894 ± 0.008 9.46 ± 0.77
S2 6.15 x 107 50 3.40 ± 0.06 0.954 ± 0.002 21.7 ± 0.94
M3 82.88 x 107 85 3.33 ± 0.05 0.924 ± 0.005 13.2 ± 0.80
S3 3.57 x 107 19 0.955 ± 0.03 0.315 ± 0.157 1.46 ± 0.33 a Simpson diversity = (1-Simpson index)
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Table 4. Colony forming units per gram of soil (CFU/g) and the proportional abundancea of taxa recovered May 2014 from mound (M) and non-mound (S) sites.
Taxa (n = 126)
M1 S1 M2 S2 M3 S3
pi (%)
CFU/g pi
(%) CFU/g
pi (%)
CFU/g pi
(%) CFU/g
pi (%)
CFU/g pi
(%) CFU/g
Acremonium spp. (9)b − − 6.1 6.60 x 10
6 0.5 1.50 x 10
6 9.8 3.60 x 10
7 13.8 7.95 x 10
7 1.8 2.10 x 10
6
Aspergillus navahoensis 41.5 4.80 x 108 − − 44.2 1.31 x 10
8 − − 20.8 1.20 x 10
8 − −
Aspergillus pseudodeflectus 37.6 4.35 x 108 − − 8.6 2.55 x 10
7 − − 34.3 1.98 x 10
8 0.3 3.00 x 10
5
Aspergillus spp. 1.3 1.50 x 107 − − − − − − 0.3 1.50 x 10
6 − −
Aureobasidium sp. − − − − − − − − − − 0.3 3.00 x 105
Bipolaris spp. (2) − − − − 2.0 6.00 x 106 − − − − − −
Cladosporium cladosporioides 1.3 1.50 x 107
1.1 1.20 x 106 9.6 2.85 x 10
7 0.8 3.00 x 10
6 2.6 1.50 x 10
7 26.3 3.00 x 10
7
Cladosporium herbarum 2.6 3.00 x 107 1.7 1.80 x 10
6 2.0 6.00 x 10
6 − − 0.5 3.00 x 10
6 − −
Cladosporium macrocarpum − − − − − − − − − − 0.5 6.00 x 105
Curvularia brachyspora − − − − 0.5 1.50 x 106 − − − − 2.6 3.00 x 10
6
Curvularia geniculatus 1.3 1.50 x 107 − − − − − − − − − −
Curvularia spp. (2) − − − − 1.0 3.00 x 106 − − 1.6 9.00 x 10
6 2.6 3.00 x 10
6
Devresia sp. − − 0.3 3.00 x 105 − − − − − − − −
Doratomyces nanus − − 0.3 3.00 x 105 − − − − − − − −
Fusarium spp. (4) − − 5.5 6.00 x 106 5.1 1.50 x 10
7 13.1 4.80 x 10
7 1.3 7.50 x 10
6 − −
Geomyces sp. − − 2.7 3.00 x 106 − − − − − − − −
Geomyces pannorum − − 16.9 1.83 x 107 2.5 7.50 x 10
6 8.8 3.24 x 10
7 8.3 4.80 x 10
7 34.2 3.90 x 10
7
Humicola sp. − − − − 0.5 1.50 x 106 − − − − − −
Lecythophora sp. − − − − − − − − − − 0.5 6.00 x 105
Myrothecium sp. 13.0 1.50 x 108 0.3 3.00 x 10
5 0.5 1.50 x 10
6 − − − − − −
Paecilomyces spp. (2) − − 0.3 3.00 x 105 − − 0.8 3.00 x 10
6 − − − −
Paecilomyces marquandii − − 1.4 1.50 x 106 − − 8.2 3.00 x 10
7 − − 3.4 3.90 x 10
6
Penicillium spp. (20) − − 1.7 1.80 x 106 10.7 3.15 x 10
7 36.0 1.32 x 10
8 0.5 3.00 x 10
6 10.0 1.14 x 10
7
Phoma spp. (11) 1.4 1.67 x 107 33.8 3.66 x 10
7 2.5 7.50 x 10
6 − − 4.9 2.85 x 10
7 2.9 3.30 x 10
6
Sterile (20) − − 12.5 1.35 x 107 5.6 1.65 x 10
7 3.4 1.23 x 10
7 2.1 1.20 x 10
7 3.4 3.90 x 10
6
Tricellula sp. − − − − − − 0.8 3.00 x 106 − − − −
Trichocladium sp. − − − − − − − − 2.6 1.50 x 107 − −
Trichoderma spp. − − 2.7 3.00 x 106 0.5 1.50 x 10
6 − − − − 0.5 6.00 x 10
5
Undetermined (36) − − 12.7 1.38 x 107 3.6 1.05 x 10
7 18.2 6.69 x 10
7 6.5 3.75 x 10
7 10.5 1.20 x 10
7
Total Trichocomaceae 81.8 3.4 63.5 45 55.9 13.7 a pi = proportional abundance of the ith species;
b number of species within a genus or group.
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Table 5. Colony forming units per gram of soil (CFU/g) and the proportional abundancea of taxa recovered July 2014 from mound (M) and non-mound (S) sites.
Taxa (n = 197)
M1 S1 M2 S2 M3 S3
pi (%)
CFU/g pi
(%) CFU/g
pi (%)
CFU/g pi
(%) CFU/g
pi (%)
CFU/g pi
(%) CFU/g
Acremonium spp. (12)b 1.3 5.17 x 10
6 7.5 4.30 x 10
6 1.4 6.00 x 10
6 12.7 7.80 x 10
6 5.6 4.65 x 10
7 0.8 3.00 x 10
5
Acremonium-like spp. (5) − − 1.6 9.00 x 105 3.5 1.50 x 10
7 1.0 6.00 x 10
5 0.2 1.50 x 10
6 2.5 9.00 x 10
5
Alternaria alternata 0.0 1.67 x 105 − − 0.4 1.50 x 10
6 − − 1.8 1.50 x 10
7 − −
Aspergillus navahoensis 36.8 1.42 x 108 − − 25.4 1.08 x 10
8 − − 17.4 1.44 x 10
8 − −
Aspergillus niger 0.4 1.50 x 106 − − − − − − − − − −
Aspergillus pseudodeflectus 22.4 8.65 x 107 − − 17.0 7.20 x 10
7 − − 17.2 1.43 x 10
8 − −
Aspergillus spp. − − − − − − − − 1.8 1.50 x 107 − −
Aureobasidium pullulans 0.1 5.00 x 105 − − 0.7 3.00 x 10
6 − − 0.9 7.50 x 10
6 − −
Bipolaris sp. 0.8 3.00 x 106 − − − − − − − − − −
Chrysosporium spp. (2) − − 1.2 6.67 x 105 − − 1.0 6.00 x 10
5 − − − −
Cladosporium cladosporioides 0.5 2.00 x 106 − − 2.8 1.20 x 10
7 3.4 2.10 x 10
6 0.7 6.00 x 10
6 − −
Cladosporium herbarum 1.2 4.67 x 106 − − 0.4 1.50 x 10
6 0.5 3.00 x 10
5 0.9 7.50 x 10
6 − −
Cladosporium sphaerospermum 0.8 3.00 x 106 − − − − 0.5 3.00 x 10
5 − − − −
Clonostachys rosea 0.8 3.00 x 106 − − 3.5 1.50 x 10
7 1.5 9.00 x 10
5 − − − −
Clonostachys sp. − − − − 0.4 1.50 x 106 − − − − − −
Curvularia geniculatus 0.4 1.50 x 106 − − 0.7 3.00 x 10
6 − − 0.5 4.50 x 10
6 − −
Deverisea sp. − − − − − − 1.5 9.00 x 105 − − − −
Fusarium spp. (4) 0.0 1.67 x 105 0.6 3.33 x 10
5 0.4 1.50 x 10
6 1.5 9.00 x 10
5 0.4 3.30 x 10
6 0.8 3.00 x 10
5
Geomyces spp. (3) − − 0.6 3.33 x 105 0.4 1.50 x 10
6 − − 0.0 1.50 x 10
5 0.8 3.00 x 10
5
Geomyces pannorum 0.5 2.00 x 106 5.2 3.00 x 10
6 0.7 3.00 x 10
6 9.3 5.70 x 10
6 6.5 5.40 x 10
7 0.8 3.00 x 10
5
Humicola sp. − − − − − − 0.5 3.00 x 105 − − − −
Humicola-like sp. − − − − − − 0.5 3.00 x 105 − − − −
Idriella lunata − − − − 0.4 1.50 x 106 0.5 3.00 x 10
5 − − − −
Myrmecridium sp. − − − − − − − − 0.5 4.50 x 106 − −
Myrmecridium schulzeri − − 0.6 3.33 x 105 − − − − 0.2 1.50 x 10
6 − −
Myrothecium sp. 17.5 6.77 x 107 − − − − − − − − − −
Paecilomyces spp. (2) − − − − − − 2.4 1.50 x 106 − − − −
Paecilomyces marquandii − − 3.5 2.00 x 106 0.4 1.50 x 10
6 3.9 2.40 x 10
6 0.2 1.50 x 10
6 0.8 3.00 x 10
5
Penicillium sp. (26) 1.8 6.83 x 106 62.8 3.62 x 10
7 8.1 3.45 x 10
7 39.0 2.40 x 10
7 2.9 2.40 x 10
7 85.7 3.06 x 10
7
Pleurostomophora sp. − − − − 0.4 1.50 x 106 − − 0.5 4.50 x 10
6 − −
Pseudogymnoascus sp. − − − − − − 0.5 3.00 x 105 − − − −
Ramichloridium sp. − − − − − − − − 0.2 1.50 x 106 − −
Solosympodiella sp. − − − − − − − − 0.2 1.50 x 106 − −
Spicellum sp. − − − − − − − − − − 0.8 3.00 x 105
Stachybotrys eucylindriospora − − − − − − − − 0.2 1.50 x 106 − −
Sterile (72) 11.8 4.55 x 107 9.8 5.63 x 10
6 25.8 1.10 x 10
8 16.6 1.02 x 10
7 20.8 1.73 x 10
8 0.8 3.00 x 10
5
Trichoderma spp. 0.0 1.67 x 105 0.6 3.33 x 10
5 3.5 1.50 x 10
7 2.0 1.20 x 10
6 − − 0.8 3.00 x 10
5
Undetermined (42) 2.9 1.13 x 107 6.3 3.63 x 10
6 3.9 1.65 x 10
7 1.5 9.00 x 10
5 20.3 1.68 x 10
8 5.0 1.80 x 10
6
Total Trichocomaceae 61.4 66.3 50.9 45.3 39.5 86.5 a pi = proportional abundance of the ith species;
b number of species within a genus or group.
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