Aerobiologia (2009) 25:99–109
DOI 10.1007/s10453-009-9115-9
ORIGINAL PAPER
The biodiversity of air spora in an Italian vineyard
Donát Magyar Æ Giuseppe Frenguelli Æ
Emma Bricchi Æ Emma Tedeschini Æ
Péter Csontos Æ De-Wei Li Æ János Bobvos
Received: 15 December 2008 / Accepted: 16 March 2009 / Published online: 7 April 2009
Ó Springer Science+Business Media B.V. 2009
Abstract Biodiversity indices are frequently used
to provide a numerical value of the diversity of
species within an ecological community. To study
fungal biodiversity in the air, a 7-day recording Hirsttype spore trap was used. The daily concentration of
124 taxa was recorded in an Italian vineyard. The
predominant fungi were: Cladosporium spp.,
unknown 3-septate fusiform spore, and Alternaria
spp. Shannon’s and Simpson’s biodiversity indices
and evenness were calculated first for air spora.
Meteorological circumstances affected the biodiversity; positive correlations were found between
Simpson’s biodiversity index and precipitation, but
no significant correlations were found with Shannon’s diversity index and evenness. To predict
D. Magyar (&) J. Bobvos
Institute of Environmental Health, Gyáli út 2-6, 1097
Budapest, Hungary
e-mail: magyar.donat@gmail.com
G. Frenguelli E. Bricchi E. Tedeschini
University of Perugia, via Borgo XX Giugno 74, 06120
Perugia, Italy
P. Csontos
Research Institute for Soil Science and Agricultural
Chemistry of the Hungarian Academy of Sciences,
Herman Ottó út 15, 1022 Budapest, Hungary
D.-W. Li
The Connecticut Agricultural Experiment Station, Valley
Laboratory, 153 Cook Hill Rd, P.O. Box 248, Windsor,
CT 06095, USA
Simpson’s index for airborne fungi, regression analysis was performed. It was shown that the best
estimator, sun hours, negatively affected the index.
The biodiversity of the dominant species was low on
dry days, since Cladosporium alone had much higher
abundance on such days than other species.
Keywords
Weather
Air spora Fungi Biodiversity
1 Introduction
The atmosphere is rich in propagula from different
fungal species. Many of them may cause a range of
allergic symptoms and plant diseases (Tilak and
Pande 2005; Gonianakis et al. 2006; Bousquet et al.
2007; Magyar 2007). Aerobiology, alongside its roles
in human and plant pathology, is becoming increasingly important in the study of biodiversity in indoor
environments as well as in natural and agricultural
habitats.
‘Biodiversity’ is a term frequently used in the
scientific press. In aerobiological works, biodiversity
descriptions are restricted to the list of spore types
including different species. Attempts to measure
airborne fungal biodiversity with biodiversity indices
have not yet been made. Such indices are frequently
used by ecologists to provide a numerical value of the
123
100
diversity of species within an ecological community
(Dighton 1998). A simple biodiversity index is
calculated as follows: the number of taxa in the area
(numerator) divided by the total number of individuals in the area (denominator). Therefore, this index
can give an acceptable result if as many taxa are
identified as possible.
Whereas the spores of many fungi are difficult to
identify, those of many others are highly characteristic and can be identified or distinguished even
without cultivation. Spore composition was extensively studied in streams and elsewhere (e.g., in
rainwater by Gönczöl and Révay 2004; in honeys by
Pérez-Atanes et al. 2001; in honeydew honeys
Magyar et al. 2005; on bark by Magyar 2008).
Several hundreds of spore types are identified also
from air samples; however, only a few works have
emphasized the richness of fungal species in the
atmosphere and aimed to identify rare airborne fungal
taxa (Shoemaker et al. 1974; Kendrick 1990; JáraiKomlódi and Tóth 1993; Lacey and West 2006;
Bustos Delgado et al. 2008). The present study was
carried out in a vineyard; from such habitat 13 fungal
taxa were reported by earlier air samplings (Corbaz
1972; Picco 1992; Diaz et al. 1998), 8 of them from
Italy (Picco 1992). These studies focused mostly on
common allergenic or plant pathogenic fungi (Alternaria spp., Botrytis cinerea, Cladosporium spp.,
Plasmopara viticola, Uncinula necator). In Italy,
the occurrence of 69 fungal species was documented
on vine plants by the USDA (Farr and Rossman
2008). Apparently, the species composition of the air
of vineyards has hardly been studied.
The aim of this paper is (1) to give a more detailed
insight of the spore composition of a vineyard, (2) to
apply biodiversity indices to the air spora, and (3) to
reveal the relationship between air spora biodiversity
and meteorological parameters.
2 Materials and methods
2.1 Fungal spore data
A 7-day recording air sampler (Hirst 1952; Lanzoni,
Bologna, Italy, catalogue code: VPPS 2000) was used
to record the daily concentration of airborne spores.
The sampler was located in the middle of a traditional
vineyard of Central Italy, near the city of Brufa which
123
Aerobiologia (2009) 25:99–109
is characterized by a mild Mediterranean climate. Air
sampling was conducted at the blooming period of the
vine plant for 3 years (from 27 May to 13 June 1994;
from 5 June to 3 July 1995; and from 13 to 24 June
1996). These months were selected because they
represented climatic extremes (as suggested by Troutt
and Levetin 2001), having high precipitation and
exceptionally windy conditions. The spore traps and
meteorological instruments were placed at 12 m height
on the top of a hill in Brufa (300 m a.s.l.), between the
valleys of Assisi and Torgiano. The surroundings of
Brufa are entirely dedicated to vine cultivation (Chardonnay, Pinot, Cabernet), its area, approximately
2,500,000 m2, is kept by the Luganotti Company.
Other plant species (Cupressaceae, Olea europaea L.,
Pinus spp., Quercus spp.) also occur sparsely.
The spore trap worked continuously, aspirating in
air at a rate of 10 l/min. The airborne fungal spores
impacted on a tape (MELINEXÒ strip) coated with a
thin adhesive layer (silicone oil). The greased tape was
mounted on a rotating drum within the trap, rotating
2 mm/h. The exposed tape was removed weekly and
cut into 48-mm segments, thus representing 24-h
periods. The segments were placed on microscope
slides and stained with basic fuchsine in mounting
medium (glycerine-jelly). To provide a full picture of
the air spora, 100% of the areas of the samples were
scanned. To count fungal content, two longitudinal
transverses along the length of the slide were scanned,
at 4009 magnification using a DIALUX 20 microscope. For each daily recording, fungal spores were
counted, classified into families, genera and sometimes into species, and expressed as number of spores
trapped per cubic metre of air. Because of their high
number, Cladosporium conidia were enumerated in
four random areas of a microscope field of sight (with
a diameter of 523 lm) in every 6 mm of air sample.
The Cladosporium counts were summarized and
multiplied by 16; the result of this calculation is the
daily concentration of spores per cubic metre of air.
We used the Leica QWin Image Analyser Programme to study spore morphology. Fungal spores
were identified on various levels. Uncertain taxa were
labeled according to Magyar (2007).
2.2 Weather data
Meteorological data were obtained from a weather
station (SIAP Bologna, Italy) located in the vicinity
Aerobiologia (2009) 25:99–109
of the sampling site. The daily records of temperature
(Tmin, Tmax, Tavg) (°C), dew point (°C) and relative
humidity were measured with a hygrothermometer.
Rainfall (mm) and the duration of precipitation (min)
were recorded with a rain gauge and rainfall intensity
(mm/min) was calculated. A 3-cup anemometer with
a standard wind vane and an electronic barometer
were used to measure wind speed (m/s) and wind
direction, and atmospheric pressure (Pa), respectively. Cloud cover (%), presence or absence of fog
and sun hours (min) were also recorded. Evaporation
(mm) was measured with the Wild evaporimeter.
Wind and temperature conditions were similar in
all the three sampling periods (avg 2.7 m/s, wind
gusts between 13.8 and 22.2 m/s, Tmax: 23.89°C;
Tmin: 14.70°C). The period in 1996 was slightly drier
(average precipitation 0.69 mm) than in the previous
year (average precipitation 2.19 mm).
2.3 Data analysis
Biodiversity was expressed by the Shannon’s index
(HS) and Simpson’s reciprocal index (D):
HS ¼
S
X
Pi ðlog Pi Þ
i¼1
D ¼
1
S
P
P2i
101
We constructed rank abundance curves to assess
how relative abundance and evenness depended on
weather. The species were sorted with respect to their
rank. The most abundant species were plotted first
then the next most common and so on until the array
is completed by the rarest of all. Plotting abundance
against rank yielded rank abundance curve. To show
the diversity dissimilarity among dry and wet (rainy)
days, right tail sum (RTS) biodiversity was used
(Patil and Taillie 1979)
RTSðiÞ ¼
S
X
pð jÞ; 1 i S:
j¼iþ1
3 Results
3.1 Air spora composition
A total of 189 different fungal taxa were observed in
our spore trap and 50 taxa appeared to be new for
air samples (Table 1). The majority, 73 taxa, were
classified as genus, and 67 of them were identified
to species level. More than 45% of the taxa were
mitosporic fungi. Ascomycetes represented also a
considerable proportion (24.6%) of the total air
spora. Percentages of airborne fungi belonging to
Basidiomycetes, Myxomycetes, Oomycetes, Urediniomycetes, and Ustilaginomycetes were lower
(1–5%).
i¼1
where S is the total number of genus in every air
sample (day) and pi is the proportion of individuals
that belong to the ith genus (Magurran 1988). Species
evenness was calculated by dividing the actual
diversity value with the potential maximum of
diversity. The Shapiro-Wilk W test was used in
testing for normality of variables (Shapiro et al.
1968). Since this statistic was significant (r 0.84,
P \ 0.00001), then the hypothesis that the respective
distribution is normal was rejected. Then, the variables were logarithmically transformed to normalise
data. To avoid zero values, we added 0.9 to the
concentrations according to Troutt and Levetin
(2001). To clarify the connection between biodiversity indices and weather variables, Pearson
correlation analysis was applied using the STATISTICA programme (StatSoft, 2003, version 6.1).
Linear regression was processed to predict the value
of the biodiversity index using the normalized data.
3.2 Air spora concentration
The daily concentrations of 124 different spore types
were determined (Table 1). The average of daily
spore concentration for all the studied period was
2,063 spores/m3 (max 3,919 spores/m3 on 14 June
1996). The most abundant fungal particles in our air
samples belonged to the genus Cladosporium (av
1,286.48 spores/m3, 62.33% of the total count).
Unknown 3-septate fusiform spore (104.16 spores/
m3, 4.65%), Alternaria spp. (92.03, 4.45%), unknown
hyalodidymae (65.94, 3.07%) and Ustilago tritici/
nuda (47.62, 2.08%) also made up a significant
portion of airborne fungi. Hyphal fragments (mostly
conidiophores belonging to different dematiaceous
hyphomycetes) reached a high concentration as well
(205.46). The common taxa (av above 1.00 spore/m3,
0.05%) comprised 88.52% of air spora. A wide range
of other identified fungal taxa was also recorded, but
123
102
Aerobiologia (2009) 25:99–109
Table 1 Spore types recorded in the air samples and numerical description of concentration data (spore/m3) in Brufa, Italy
Spore type
ID
no.
1994
Agrocybe spp.
2
1
4
1
18
7
28
6
182
1
4
2
12
Albugo spp.
3
2
10
3
28
2
15
3
43
3
6
2
29
Alternaria brassicae (Berk.) Sacc.a
1995
Avg Max
SD Sum
Avg
1996
Max
SD Sum
Avg
Max
SD Sum
5
0
0
0
0
0
4
1
8
1
4
1
7
Alternaria spp.
13
27
90
25
401
92
344
69
2,395
104
206
61
1142
Ampulliferina persimplex B. Suttona
15
2
17
5
34
1
9
2
13
3
13
4
32
Anthostomella/Herpotrichiella spp.
16
0
0
0
0
1
5
1
29
1
5
2
15
Arthrinium cuspidatum (Cooke and
Harkn.) Höhna
17
0
2
1
2
0
0
0
0
0
0
0
0
Arthrinium luzulae M.B. Ellisa
18
0
0
0
0
0
1
0
1
0
0
0
0
Arthrinium spp.
19
8
50
13
116
10
51
12
250
27
68
23
293
Aspergillus/Penicillium spp.
22
27
125
31
410
27
193
41
701
29
87
30
319
Badhamia sp.
Basidiomycetes
23
24
0
29
0
103
0
34
0
442
0
116
1
310
0
83
1
3,018
0
44
1
136
0
41
2
489
Belemnospora verruculosa P. M. Kirk
25
0
0
0
0
0
1
0
1
1
5
1
9
Beltrania rhombica Penz
26
0
1
0
5
0
1
0
3
0
2
1
5
Bipolaris spicifera (Bainier) Subram
27
0
2
1
3
0
2
1
10
2
6
2
17
Bipolaris spp.
28
0
1
0
3
0
2
1
9
1
3
1
9
Boletus spp.
29
1
3
1
9
2
7
2
61
1
2
1
7
Botrytis-type
30
6
36
10
96
15
88
19
382
13
30
9
141
Bovista spp.
33
0
1
0
2
0
2
0
5
0
1
0
1
Caloplaca sp.
34
0
0
0
0
1
9
3
30
1
5
2
9
Capnobotrys sp.a
35
0
1
0
4
0
1
0
1
0
1
0
1
Cercospora spp.
36
1
4
1
11
1
6
1
27
4
12
4
46
Chaetomium spp.
37
1
2
1
9
1
5
1
14
3
9
3
33
Chaetosphaerella sp.
38
0
0
0
0
0
2
1
12
1
2
1
6
Cheiromycella microscopica (P. Karst.) S. 39
Hughesa
0
0
0
0
0
1
0
1
0
0
0
0
Cladosporium aecidiicolaThüm ??
40
0
0
0
0
0
1
0
1
0
1
0
3
Cladosporium echinulatum (Berk.) G.A.
deVries-type
41
3
14
4
41
1
8
2
23
1
4
1
10
Cladosporium phlei (C. T. Greg.) G.A.
deVries-type
42
0
0
0
0
0
0
0
0
0
2
1
3
Cladosporium spp.
Claviceps sp.a
43
45
Clypeosphaeria notarisii Fuckela
46
0
1
0
1
0
1
0
3
0
3
1
3
Colletotrichum-type
47
1
9
3
19
12
72
17
319
4
31
9
40
Coprinus spp.
48
17
90
24
258
36
112
24
943
9
24
6
94
Cortinarius spp.
49
0
1
0
2
1
3
1
20
1
2
1
7
Coryneum spp.a
50
0
0
0
0
0
3
1
8
0
2
1
5
940 2,675 827 14,095 1,435 2,565 485 37,300 1,398 2,624 685 15,381
4
19
6
65
14
37
9
375
14
53 20
158
Cronartium/Melampsora spp.
51
1
9
3
20
0
2
1
4
0
2
1
5
Cucurbitaria spp.
52
0
1
0
4
0
3
1
8
1
3
1
7
Curvularia spp.
53
0
1
0
1
0
1
0
1
0
0
0
0
Delitschia sp.a
55
0
1
0
1
0
1
0
5
0
1
0
1
Dendrographium/Dendryphiella sp.
56
0
0
0
0
0
1
0
2
1
3
1
6
123
Aerobiologia (2009) 25:99–109
103
Table 1 continued
Spore type
ID
no.
1994
1995
1996
Avg Max SD Sum Avg Max SD Sum
Avg Max SD Sum
Dendryphion digitatum Subram.
57
0
0
0
0
0
1
0
1
0
1
0
1
Diapleella clivensis (Berk. and Broome) Munk.?
58
1
6
2
13
1
11
2
34
5
15
6
54
322 32
299
Diatrypaceae
59
3
15
5
50
12
87
19
Dictyoarthrinium sacchari (J.A. Stev.) Damon
60
1
8
2
15
1
21
4
Didymella spp.
61
1
5
2
21
43
411
Diplocladiella scalaroides G.Arnaud
62
0
0
0
0
0
1
0
1
0
0
0
Diplodia spp.
63
0
0
0
0
0
1
0
1
0
2
1
4
Drechslera biseptata (Sacc. and Roum.) M.J. Rich and
E.M. Fraser
64
0
0
0
0
0
1
0
4
0
1
0
3
Entomophthora-type
65
0
2
1
3
1
6
1
17
0
1
1
4
Epicoccum nigrum Link
66
5
14
4
80
15
41
11
391 22
52
Excipularia fusispora (Berk. and Broome) Sacca
Cercosporidium graminis (Fuckel) Deighton
68
69
0
0
0
1
0
0
0
4
0
0
1
2
0
0
Ganoderma spp.
70
5
14
4
81
8
26
6
212 10
38
Helicosporium sp.
71
3
6
2
39
9
50
10
229
5
30
9
55
Helminthosporium/Drechslera spp.
72
9
41
11 141
11
39
10
278
6
15
5
68
Inocybe sp.
77
0
0
0
0
0
1
0
3
0
0
0
0
1
a
30
89 356
7
30
12
83 1,123 10
51
18 111
3
4
0
1
2
4
79
0
13 240
1
1
2
7
10 110
Lasiosphaeria spp.
78
0
2
1
3
0
1
0
7
0
1
0
Leptosphaeria eustomoides Sacca
79
0
0
0
0
0
2
0
3
0
2
1
3
Leptosphaeria rubicunda Rehm
80
0
3
1
6
6
52
11
152
5
20
8
51
Leptosphaeria spp.
81
6
77
20
85
2
22
4
49
6
32
10
69
Lophiostoma vicinum Sacc.a
82
0
1
0
4
0
2
1
12
5
39
12
51
4
0
0
0
0
472 30
155
Massaria ? inquinans (Tode) Fr.
83
0
2
1
3
0
1
0
Massarina-type
84
2
6
2
27
18
140
29
Monilia sp.
86
1
7
2
18
0
4
1
5
0
1
0
1
Mucor-type
87
0
3
1
3
0
0
0
0
1
7
2
7
Mycosphaerella spp.
Myxomycetes
91
92
0
4
1
12
0
4
2
61
1
8
6
34
2
9
36 1
219 10
4
24
Neohendersonia kickxii(Westend.)B.Sutton and
Pollacka
93
0
1
0
2
0
1
0
1
0
2
1
4
Nigrospora oryzae (Berk. and Broome) Petch
94
0
1
0
1
0
0
0
0
0
0
0
0
Oidium spp.
97
35
102
8
31
8
218 10
23
Oncopodiella trigonella (Sacc.) Rifaia
98
0
0
0
0
0
0
0
0
0
1
0
2
Ophiobolus acuminatus (Sowerby) Dubya
99
2
26
7
32
2
19
5
63
8
36
11
83
34 525
50 326
1
9
7 115
7 114
Ovulariopsis sp.
100
0
0
0
0
0
2
0
2
0
1
0
2
Paraphaeosphaeria michotii (Westend.) O.E. Erikss.
101
1
5
1
8
1
5
1
30
6
22
7
69
Periconia macrospinosa Lefebvre and A.G. Johnsona
102
0
0
0
0
0
0
0
0
0
1
0
1
Periconia spp.
103
2
9
2
30
5
13
3
119
5
11
2
55
Plasmopara-type
Pestalotiopsis sp.
104
105
1
0
2
1
1
0
8
1
1
0
3
10
1
2
21
10
0
0
2
1
1
0
3
1
Phragmidium sp.
106
0
1
0
1
0
0
0
0
0
0
0
0
Pithomyces chartarum (Berk. and
M.A.Curtis)M.B.Ellis
107
0
2
1
6
1
3
1
13
0
1
0
2
123
104
Aerobiologia (2009) 25:99–109
Table 1 continued
Spore type
ID
no.
1994
1995
1996
Avg Max SD Sum Avg Max SD Sum Avg Max SD Sum
Pleospora herbarum (Pers.) Rabenh. ??
110
3
25
7
41
0
2
0
3
0
1
0
1
Pleospora oblongata Niessl.a
111
0
4
1
5
0
2
1
8
0
1
0
3
Pleospora rubelloides (Plowr. ex Cooke) J. Webster
112
0
2
1
2
0
3
1
8
1
3
1
9
Pleospora spp.
113
66
697
177 989
4
31
7
97
14
70
Polythrincium trifolii Speg
114
4
25
3
67
0
3
Puccinia spp.
117
47
178
18 492
60
124
54
3
9
51 708
7
19
67
24 157
1
5
38 663
Rebentischia unicaudata (Berk. and Broome) Sacc.a
118
0
1
0
2
0
1
0
6
0
1
0
3
Rosellinia sp.
119
0
2
1
2
0
1
0
1
0
0
0
0
17
Schroeteria delastrina (Tul. and C. Tul.) G. Wintera
121
1
2
1
10
1
7
1
13
2
6
2
Scopulariopsis sp.
122
0
0
0
0
1
15
3
17
0
0
0
0
Septonema sp.?
123
0
1
0
3
1
6
1
24
1
4
1
13
Sirosporium/Acrodictys sp.
124
0
3
1
6
0
1
0
4
1
5
2
15
Sordariaceae
126
0
0
0
0
0
1
0
2
0
1
0
1
0
Sphaeropsis sp.
127
0
0
0
0
0
0
Spilocaea spp.
Sporidesmium spp.
128
130
0
0
2
1
1
0
4
1
4
0
24
1
0
0
3
1
4
6 104
0
3
4
1
7
1
2
1
43
6
Sporormiella sp.
131
0
2
1
2
0
1
0
5
0
2
1
4
Stachybotrys sp.
132
2
11
3
23
0
3
1
6
1
2
1
7
5 174
Stemphylium spp.
133
5
13
5
70
7
24
10
27
Stigmina ? pallida (Ellis and Everh.) M.B. Ellis
134
0
2
1
4
0
1
0
2
0
1
1
4
Taeniolella breviuscula(Berk. and
M.A.Curtis)S.Hughes??
135
0
0
0
0
0
0
0
0
1
2
1
6
14 163
10 109
Taeniolella spp
136
11
53
8
21
6 202
11
17
Telephoraceae
137
2
12
3
28
10
42
11 249
1
3
1
14
Teloschistes/Xanthoria sp.
139
0
0
0
0
2
16
1
5
2
10
5
51
3 125
Tetraploa aristata Berk. and Broome
140
0
0
0
0
0
0
0
0
0
1
0
1
Tilletia sp.
Tilletiopsis-type
141
142
0
0
0
1
0
0
0
1
0
4
0
25
0
6
0
91
0
0
1
4
0
1
2
4
Torula spp.
143
13
31
10 194
8
41
9 220
13
22
5 138
Tranzscheliella hypodytes (Schltdl.)Vánky and
McKenziea
144
52
157
54 782
31
177
39 807
39
75
23 432
Trichothecium roseum (Pers.)Link
145
0
1
0
1
0
0
0
0
1
13
4
Trimmatostroma salicis Corda
146
0
3
1
6
0
1
0
1
0
1
0
13
3
Trimmatostroma scutellare (Berk. and Br.) M.B. Ellisa 147
0
3
1
5
0
2
1
6
2
10
3
18
Tripospermum spp.a
148
0
0
0
0
0
0
0
0
1
3
1
7
Triposporium elegans Corda
149
0
3
1
3
0
0
0
0
0
1
0
1
Urocystis spp.
159
0
2
1
3
0
3
1
4
0
2
1
4
Ustilago bromivora (Tul. and C. Tul.)
A.A.Fisch.Waldha
160
42
119
38 632
25
68
19 661
37
95
31 411
Ustilago tritici C. Bauhin/U. nuda (C N. Jensen) Rostr 161
54
248
71 807
25
131
29 661
70
156
56 770
Valsaria ?? insitiva (Tode) Ces. and De Not
163
0
0
0
0
0
1
0
2
1
4
1
8
Venturia spp.
164
1
4
1
15
2
35
7
63
2
6
2
20
123
Aerobiologia (2009) 25:99–109
105
Table 1 continued
Spore type
ID no. 1994
1995
1996
Avg Max SD Sum Avg Max SD
Sum
Avg Max SD
Sum
Xylariaceae
165
3
10
3
44
10
31
8
269
15
36
10
165
Zygophiala jamaicensis E.W. Mason
167
0
0
0
0
0
0
0
0
0
1
0
1
Unknown hyalodidymae
75
8
25
8
123
78
457
100 2,021 105
505
Unknown filiform spores
155
4
11
4
53
3
26
Unknown hyaline 3-septate fusiform spores 156
7
20
6
99
183
947
5
181 1,153
84
3
15
5
29
222 4,748
14
55
18
153
ID no. = identity number for spore types
Taxa found outside the counting area: Acrodictys sp., Aglaospora profusa (Fr.) De Not., Ampelomyces sp.a, Amphisphaerella
deceptiva M.E. Barra, Ascobolus sp., Asterosporium asterospermum (Pers.) S. Hughes, Bactrodesmium spilomeum (Berk. and
Broome) E.W. Mason and S. Hughesa, Chaetomium? murorum Cordaa, Coniosporium sp.a, Coremiella sp.a, Dendryphion nanum
(Nees) S. Hughesa, Drechslera bromi (Died.) Shoemakera, Drechslera tritici-repentis (Died.) Shoemakera, Endophragmiella fallacia
P.M. Kirka, Endophragmiella sp., Gilmaniella sp., Gliomastix sp., Helicoon sp., Heptameria uncinata (Niessl.) Rehm,
Hysterographium sp., Lepteutypa sp., Leptosphaeria dumetorum Feltgena, Leptosphaeria graminis (Fuckel) Sacc.a, Leptosphaeria
Niessliana Rabenh.a, Leptosphaeria thurgoviensis E. Müll.a, Melanographium citri (Gonz. Frag. and Cif.) M.B. Ellisa, Microbotryum
reticulatum (Liro) R. Bauer and Oberw.a, Monodictys castaneae (Wallr.) Hughesa, Navicella pileata (Tode) Fabre, Nodulosphaeria
ulnispora (Cooke) L. Holma, Podospora sp.?, Pollaccia sp.a, Pseudovalsa umbonata (Tul.) Sacc.a, Spegazzinia sp., Splanchnonema
quercicola M.E. Barra, Sporidesmium folliculatum (Corda) E.W. Mason and S. Hughesa, Sporidesmium leptosporum (Sacc. and
Roum.) S. Hughesa, Sporidesmium valdivianum (Speg.) M. B. Ellis, Taeniolella scripta (P. Karst.) S. Hughesa, Trematosphaeria
pertusa (Pers.) Fuckela, Ustilago hordei Bref., Ustilago striiformis (Westend.) Niessla, and 23 unknown taxa
a
First observation of the taxa in air
they made up fewer than 11% of the total fungal
spores.
3.3 Biodiversity
The daily average of the identified morphological
spore types calculated for the total studied period was
66; its highest number (112 taxa) was recorded in a
rainy day (22 June 1996). Simpson’s diversity index
of airborne fungal genera ranged between 1.08 and
8.56, its average was 2.75 (Table 2; Fig. 1).
Daily biodiversity value of the Simpson’s index
correlated positively with precipitation (Table 3). No
significant correlations were found with Shannon’s
diversity index and evenness.
The rank abundance curves for the wet days were
much shallower than those for dry ones, because the
relative abundances of genera were more evenly
distributed than on dry days. Rank abundance curves
of dry days were more steep than those of wet ones
(Fig. 1).
The regression analysis performed enabled the
development of a model to predict Simpson’s index
for airborne fungal genera showing the best estimator
of data variability to be sun hours (D = 4.405-
0.002 9 sun hours ? 0.49; 0.49 should be added
when wind speed [ 0.5 m/s). This model was statistically significant (F test, P \ 0.001); and accounted
for 21% of the variance (R 0.49, P \ 0.002).
Finally, RTS diversity ordering showed that wet
days had higher biodiversity concerning its dominant
taxa, than dry days (Fig. 2).
4 Discussion
The present study contributed to the knowledge of the
species composition of the air, adding several new
taxa, which appeared to be new to the air of
vineyards. From such a habitat only the following
fungi have been found by earlier air samplings:
Agrocybe, Alternaria, Botrytis, Cladosporium, Coelosporium senecionis, Epicoccum, Fusarium,
Helminthosporium-type, Leptosphaeria, Penicillium,
Plasmopara viticola, Polythrincium trifolii, Puccinia,
Rhizopus, Stemphylium, Torula, and Uncinula (Corbaz 1972; Picco 1992; Diaz et al. 1998). Most of
them were present on our slides, but Botrytis- and
Plasmopara-type spores (possibly belonging to the
fungal pathogens of grapevine) did not reached high
concentrations in the air.
123
106
Table 2 Numerical
description of air spora
composition and biodiversity
indices (Brufa, Italy)
Aerobiologia (2009) 25:99–109
Wet days
Shannon’s biodiversity index
Simpson’s biodiversity index
Species richness (number of species)
Relative abundance of genera of Ascomycetes
Relative abundance of genera of mitosporic fungi
Dry days
All days
Avg
1.80
1.65
Max
2.56
2.47
1.68
2.56
Min
0.27
1.03
0.27
SD
0.68
0.35
0.44
Avg
3.85
2.45
2.75
Max
8.56
5.13
8.56
Min
1.08
1.48
1.08
SD
Avg
2.08
61.36
0.80
66.90
1.30
65.73
Max
112
96
112
Min
20
23
20
SD
26.34
13.55
16.89
Avg
43.24
26.77
30.25
Max
60.23
53.90
60.23
Min
13.22
1.30
1.30
Avg
43.89
51.71
50.06
Max
54.43
76.47
76.47
Min
38.18
36.36
36.36
Fig. 1 The temporal patterns of rank abundance curves, the percentage of the number of genera of mitosporic fungi and
Ascomycetes, and Simpson’s biodiversity index (Brufa, Italy). Grey columns represent wet days
In our study, non-viable air sampling method was
applied, which ignored the morphologically indistinguishable species. Including all morphological spore
types (identified plus unknown) a total of 189 taxa
were recognized. Using a viable (cultural) air sampling method, a similar number of the taxa was
reported from the air of Turin by Airaudi and
Marchisio (1996), where 165 species of mesophilic
123
fungi and 26 thermotolerant species were isolated.
More than 30% of these airborne species belonged to
the genera of Aspergillus, Penicillium, and yeasts.
Our method is not suitable for the identification of
such taxa, because their spores cannot be distinguished and may be easily overlooked. It is preferred,
however, to identify many other taxa not growing on
agar media, e.g., obligate biotrophic fungi (Pyrri and
Aerobiologia (2009) 25:99–109
Table 3 Correlation
between meteorological
variables and biodiversity
indices of airborne fungi
(Brufa, Italy)
107
Meteorological factors
* Significant correlations
P \ 0.05
Simpson’s
biodiversity index
Evenness
Tmax
0.16
-0.12
-0.27
Tmin
0.09
-0.11
-0.17
Tavg
Sun hours
0.14
0.02
-0.12
-0.19
-0.24
-0.06
Evaporation
0.03
-0.12
-0.05
Cloud cover
-0.02
0.16
0.10
Relative humidity
0.02
0.20
0.04
Precipitation
0.06
0.34*
0.06
-0.04
0.29*
0.21
0.15
0.34*
-0.11
Rain hours
Tmax, temperature maximum;
Tmin, temperature minimum;
Tav, temperature average
Shannon’s
biodiversity index
Rain intensity
0.14
0.12
-0.15
Wind speed
Wind gust
-0.22
-0.21
0.18
Air pressure
-0.09
-0.14
0.06
Fig. 2 RTS-diversity
patterns comparing the air
spora diversity of dry and
wet days (Brufa, Italy)
Kapsanaki-Gotsi 2007). Therefore, it is not possible
to identify and enumerate the entire community with
a single method. To obtain the total number of fungal
taxa in an air sample, a combination of a non-viable
method and a wide variety of selective isolation
methods would be needed.
Statistical analysis showed that precipitation correlated strongly with the fungal diversity of the air.
Wet and dry periods had somewhat different spore
compositions. It was proved that Simpson’s biodiversity index was increased significantly by
precipitation. Rain caused species turnover, removing dry spores (mostly mitosporic fungi) and
aerosolizing wet ones (mostly Ascomycetes;
Fig. 1). The relative abundance of mitosporic fungi
in wet days was higher than that of Ascomycetes on
dry days (Table 2). On wet days, dry spores are also
present when the wash-out effect of precipitation is
incomplete, or their take-off occurs before and after
rains. On dry days, some Ascomycetes appear in the
night-time, when sufficient saturation provokes
spore discharge (Jones and Harrison 2004). However, lower species number and small relative
abundance was observed for ascospores on dry
days. In the analysis of rank abundance curves,
steep gradients of the dry days indicated low
123
108
evenness, as some high-ranking genera had much
higher abundance than the low ranking ones.
It should be mentioned that the correlation and
regression analyses of the biodiversity index did not
take into account temporal changes in species composition, such as species turnover. Hence, these
methods overlook the difference between the species
composition of wet and dry days. To resolve this
problem, we analyzed the RTS curves. It was
demonstrated that biodiversity of rainy days, concerning their common species, was higher than those
of dry days. The biodiversity of the dominant species
on dry days was reduced by the high concentration of
Cladosporium spp. On wet days, however, Cladosporium spp. were not so prevalent as on dry days,
possibly because rainfall washed out its spores, and
the dominance of this genus decreased. In consequence of this, rain correlated positively with
Simpson’s biodiversity index. According to the
regression analysis, sun hours became the most
important factor. Simpson’s biodiversity index was
decreased by this factor, possibly because it has a
positive effect on Cladosporium counts (P \ 0.001;
authors, unpublished results). The effect of wind is
positive on the biodiversity index, possibly by
aerosolization of spores of different taxa.
It is well established that inhalation of fungal
spores induces respiratory allergy symptoms in sensitized individuals. Among all patients suffering from
such diseases, 3–40% of them are sensitized to fungal
spores (Kurup et al. 2000; Bavbek et al. 2006;
Gonianakis et al. 2006). In Italy, it has been observed
that in patients with inhalation allergy the frequency
of positive skin test with mold extracts is about 10%.
Fungal spores involved in allergic diseases belong to
80 genera, the most significant of which belong
primarily to Deuteromycetes and Ascomycetes and,
secondly, to Basidiomycetes (Gioulekas et al. 2004).
Positive skin tests in asthmatic patients were found to
Alternaria, Botrytis, Helminthosporium, Epicoccum,
Aspergillus, Cladosporium, and Mucor (in descending
order of their frequency; Wilken-Jensen and Gravesen
1984). Some of them had high abundance in our air
samples, and had higher RTS rank on dry days than on
rainy ones. There were also several plant pathogenic
components in our air samples, e.g., Alternaria,
Puccinia, Stemphylium, Helmintosporium, and Ustilago species, which had also markedly higher rank in
the RTS curves of dry days.
123
Aerobiologia (2009) 25:99–109
5 Conclusions
Biodiversity indices could be applied for air spora.
Such indices show temporal dynamics, which are
strongly affected by meteorological factors, mostly
by sun hours. On dry days, the total biodiversity is
less than on wet ones, since the dominance of
Cladosporium reduces the biodiversity of the common species and the evenness of air spora.
Acknowledgments The authors are grateful to Maureen
Lacey (Rothamsted, UK), Sándor Tóth (Szt. Isván University,
Hungary) and Kálmán Vánky (Herbarium Ustilaginales Vánky,
Germany) for their precious help in the identification of spores.
The first author wish to acknowledge the Hungarian Scientific
Research Fund (OTKA, grant n. F67908) for financial support.
References
Airaudi, D., & Marchisio, V. F. (1996). Fungal biodiversity in
the air of Turin. Mycopathologia, 136, 95–102. doi:
10.1007/BF00437502.
Bavbek, S., Erkekol, F. O., Ceter, T., Mungan, D., Ozer, F.,
Pinar, M., et al. (2006). Senzitization to Alternaria and
Cladosporium in patients with respiratory allergy and
outdoor counts of mold spores in Ankara atmosphere,
Turkey. The Journal of Asthma, 43, 421–426. doi:
10.1080/02770900600710706.
Bousquet, P. J., Chinn, S., Janson, C., Kogevinas, M., Burney,
P., & Jarvis, D. (2007). Geographical variation in the
prevalence of positive skin tests to environmental aeroallergens in the European community respiratory health
survey. Allergy, 62, 301–309. doi:10.1111/j.1398-9995.
2006.01293.x.
Bustos Delgado, I., Angulo Romero, J., Dominguez Vilches,
E., & Galan Soldevilla, C. (2008). Preliminary survey of
fungal spore biodiversity in the nature reserve of ‘‘Sierra
de Hornachuelos’’ in Andalucia (Spain). Aspects of
Applied Biology 89, 59–60.
Corbaz, R. (1972). Etudes des spores fongiques captées dans
l’air I. Dans un vignoble. Phytopathologische Zeitschrift,
74, 318–328. doi:10.1111/j.1439-0434.1972.tb02587.x.
Diaz, M. R., Iglesias, I., & Jato, M. V. (1998). Seasonal variation of airborne fungal spore concentrations in a vineyard
of North–West Spain. Aerobiologia, 14, 221–227. doi:
10.1007/BF02694210.
Dighton, J. (1998). Analysis of micromycete communities in
soil: A critique of methods. Mycological Research, 7,
796–798.
Farr, D. F., & Rossman, A. Y. (2008). Fungal Databases.
Systematic Mycology and Microbiology Laboratory,
ARS, USDA. http://nt.ars-grin.gov/fungaldatabases/.
Gioulekas, D., Damialis, A., Papakosta, D., Spieksma, F.,
Giouleka, P., & Patakas, D. (2004). Allergenic fungi spore
records (15 years) and sensitization in patients with
respiratory allergy in Thessaloniki-Greece. Journal of
Aerobiologia (2009) 25:99–109
Investigational Allergology and Clinical Immunology, 14,
225–231.
Gönczöl, J., & Révay, Á. (2004). Fungal spores in rainwater:
stemflow, throughfall and gutter conidial assemblages.
Fungal Diversity, 16, 67–86.
Gonianakis, M. I., Neonakis, I. K., Gonianakis, I. M., Baritaki,
M. A., Bouros, D., Potamias, G., et al. (2006). Mold
allergy in the Mediterranean island Crete, Greece: A 10year volumetric, aerobiological study with dermal sensitization correlations. Allergy and Asthma Proceedings, 27,
354–362. doi:10.2500/aap.2006.27.2918.
Hirst, J. M. (1952). An automatic volumetric spore trap. The
Annals of Applied Biology, 39, 257–265. doi:10.1111/
j.1744-7348.1952.tb00904.x.
Járai-Komlódi, M., & Tóth, S. (1993). Studies of rare airborne
fungal spores and conidia in Hungary. Acta Botanica
Hungarica, 38, 383–399.
Jones, A. M., & Harrison, R. M. (2004). The effects of
meteorological factors on atmospheric bioaerosol
concentrations—a review. The Science of the Total
Environment, 326, 151–180. doi:10.1016/j.scitotenv.2003.
11.021.
Kendrick, B. (1990). Fungal allergens. In E. G. Smith (Ed.),
Sampling and identifying allergenic pollens and moulds
(pp. 134–164). San Antonio: Blewstone Press.
Kurup, V., Shen, H., & Banerjee, B. (2000). Respiratory fungal
allergy. Microbes and infection, 2, 1101–1110. doi:
10.1016/S1286-4579(00)01264-8.
Lacey, M., & West, J. (2006). The air spora. Dordrecht:
Springer.
Magurran, A. E. (1988). Ecological diversity and its measurement. New South Wales: Croom Helm.
Magyar, D. (2007). Aeromycological aspects of mycotechnology. In M. K. Rai (Ed.), Mycotechnology: Current
trends and future prospects (pp. 226–263). New Delhi:
I.K. International Publishing House.
Magyar, D. (2008). The tree bark: A natural spore trap. Aspects
of Applied Biology 89, 7–16.
109
Magyar, D., Gönczöl, J., Révay, Á., Grillenzoni, F., & SeijoCoello, M. D. C. (2005). Stauro- and scolecoconidia in
floral and honeydew honeys. Fungal Diversity, 20, 103–
120.
Patil, G. P., & Taillie, C. (1979). An overview of diversity. In J.
F. Grassle, G. P. Patil, W. Smith, & C. Taillie (Eds.),
Ecological diversity in theory and practice (pp. 3–27).
Maryland: International Publishing House.
Pérez-Atanes, S., Seijo-Coello, M. D. C., & Méndez-Álvarez,
J. (2001). Contribution to the study of fungal spores in
honeys of Galicia (NW Spain). Grana, 40, 217–222. doi:
10.1080/001731301317223240.
Picco, A. M. (1992). Presence in the atmosphere of vine and
tomato pathogens. Aerobiologia, 8, 459–463. doi:10.1007/
BF02272916.
Pyrri, I., & Kapsanaki-Gotsi, E. (2007). A comparative study
on the airborne fungi in Athens, Greece, by viable and
non-viable sampling methods. Aerobiologia, 23, 3–15.
doi:10.1007/s10453-006-9039-6.
Shapiro, S. S., Wilk, M. B., & Chen, H. J. (1968). Comparative
study of various tests for normality. Journal of the
American Statistical Association, 63, 1343–1372. doi:
10.2307/2285889.
Shoemaker, R. A., LeClair, P. M., & Smith, J. D. (1974). Some
parasites of Bromus inermis and airborne spores trapped
over brome grass crops. Canadian Journal of Botany, 52,
2415–2421. doi:10.1139/b74-313.
Tilak, S. T., & Pande, B. N. (2005). Current trends in aeromycological research. In M. K. Rai & S. K. Deshmukh
(Eds.), Fungi: Diversity and biotechnology (pp. 281–510).
Jogpur: Scientific Publishers.
Troutt, C., & Levetin, E. (2001). Correlation of spring spore
concentrations and meteorological conditions in Tulsa,
Oklahoma. International Journal of Biometeorology, 45,
64–74. doi:10.1007/s004840100087.
Wilken-Jensen, K., & Gravesen, J. (1984). Atlas of moulds in
Europe causing respiratory allergy. Copenhagen: ASK
Publishing, Foundation for Allergy Research in Europe.
123
Emma Tedeschini