Indoor Air Mycological Survey and Occupational Exposure in Libraries in Mato Grosso-Central Region—Brazil

Background: Indoor air quality in environments where there is great circulation of people, posing risks to the health of its occupants, including allergic problems, infections and contaminations, can be aided by climatic factors, chemicals and biological agents housed in these environments, influencing the location and providing favorable conditions for the degradation of bibliographic collections. The present study investigated the presence of fungi in indoor environments in seven public and private libraries in the central region of Brazil, Mato Grosso, and verified the impact on occupational health. Results: A total of 26,194 fungal specimens were isolated from 342 dust samples collected using three techniques: Andersen’s sampler (12.3%), exposure plate dish (25.1%) and sterile swab (62.6%). A total of 184 fungal species were identified: 156 (84.8%) mycelial fungi and 28 (15.2%) yeast fungi, belonging 54 fungal genera, 43 (79.6%) mycelial fungi and 11 (20.4%) yeast fungi. The genus Aspergillus (40.6%) was one of the main fungi present in indoor air. Aspergillus niger (12.3%) was identified as the most prevalent species in literary environments, followed by Cryptococcus spp. (7.1%) and Cladosporium cladosporioides (7.0%). In relation to seasonal distribution, there was a greater fungal isolation in the dry season (54%); followed by the rainy season (46%). Conclusion: These results suggest the substrates researched in the evaluated environments presented in the form of documents, books and papers associated with dust and air humidity become suitable for microbiological proliferation. These findings highlight the importance of minimizing the risk of exposure to fungal agents, identified in pathogenic and toxigenic microenvironments in library collections.

• Biological Hazards
• Library Collections
• Anemophilous Fungi
• Indoor Air Quality

1. Introduction

Recent studies suggested that there may be around 1.5 to 5.1 million extant fungal species and about 1200 new species are being described in each year [1] . However, the authors caution that these numbers refer to inferences and estimates for each country.

The great biological diversity of Brazilian regions makes Brazil a country with high biodiversity, coupled with the endemism of some species. This richness is distributed in the national territory in six continental biomes: Atlantic Rainforest, Pantanal, Amazon, Caatinga, Cerrado and Pampas, which are characterized by certain uniformity in their environmental regions such as climate, temperature, soil and characteristic physiognomy [2] .

Fungal microorganisms can affect all types of archives, regardless of their constitution. Their propagules are dispersed in the environment and over the surface of objects in most places. In this way, careful attention must be paid to fungal contamination in books, periodicals and documents, since this exposes such archives to the risk of damage [3] . Contamination can also provoke allergic processes, with aggravated clinical symptoms occurring among those that handle these archives and become exposed to the action of these fungi [4] .

Diseases related to the quality of indoor air have been classified by the World Health Organization [5] as Sick Building Syndrome (SBS). Currently symptoms related to SBS are described by those who remain in enclosed spaces 90% of the time. Diagnosing SBS is eminently epidemiological [4] [6] .

More complex health effects occur when characterizing SBS (Sick Building Syndrome) or BRI (Building Related Illness). The symptoms can include irritation of mucous membranes, skin and eyes, fatigue, headache, malaise, lethargy, difficulty concentrating, sensitivity to odors and flu-like symptoms, but no causative agent is usually identified. In almost all cases, BRI is an advanced stage of SBS [6] .

The objective of this work was to evaluate the fungal biodiversity in different environments of seven libraries of Mato Grosso-Central West region of Brazil, and to establish a possible correlation with the occupational health of the visitors.

2. Materials and Methods

2.1. Climatic Characterization

The climate is characteristic of the semiarid region-Savana Tropical (AW), with a mean annual rainfall of 1469.4 mm and average annual temperature of 24˚C to 26˚C. The municipality is surrounded by three large ecosystems: the Amazon, the cerrado and the pantanal. The city of Cuiabá is the capital of the State of Mato Grosso, its location presents the coordinates 15˚35' - 56˚07' and altitude of 151 meters above sea level [7] (Figure 1).

2.2. Description of Sample Collection

Sample collections were divided into two seasonal periods: two conducted in the rainy season in January and March (2014), and another in the dry season in July to September (2014), totaling 342 samples using three methodologies. Samples were collected on two random days within each climatic period, ranging from 15 to 20 days between the first collection and the second collection for each climatic period (two in the dry season and two in the rainy season).

The temperature (T) and relative humidity (RH) were evaluated inside these repositories at each sampling point at the time the microbiological collection was being carried out using two digital thermocouples with reading capacity −10 + 60˚C (Model 7429.02.0.00 Brand Incoterm).

One-third the samples collected were achieved by sedimentation in petri dishes containing Sabouraud agar (Difco^TMBD^TM, France) added with cloranphenicol (100 mg/mL) [8] and 114 samples were collected using the Andersen’s

Sampler technique and another 114 samples by rubbing a sterile swab on the surfaces of books, periodicals and other documents in libraries.

The petri dishes were opened inside the selected locations, mainly placed on surfaces parallel to the floor, such as countertops, cabinets and shelves, for about 30 min, at a height of 1.20 cm to 1.50 cm, as described by Gambale [9] and Reis-Menezes [10] . As the exposure time elapsed, the other collections by the Swab method on books and the sampler method of Andersen were applied. The places chosen for exposure were standardized so that each of the petri dishes ware also exposed equally during the four collections.

For collection, the swab was moistened in 20% saline with chloramphenicol. The collection area was delimited using a sterile template, 10 cm × 10 cm (100 cm²). The swabs were pressure-rubbed at an angle of 30˚ with the test surface, twenty times zig-zag shape, in the diagonal directions, in the area of surface collection, in the space delimited by the mold. Other sections of the books and documents that are arranged on the shelves were also rubbed for substrate collection [11] [12] .

The dust samples were packed in sterilized polyethylene boxes and bags and transported to the Laboratory of Investigation in Micology (UFMT). In the laboratory, each swab was seeded on a 150 × 15 mm petri dish containing Sabouraud agar with chloramphenicol (100 mg/mL), which was then incubated in a BOD incubator at 27˚C for 5 - 7 days, until colony forming units (CFUs) were observed [8] .

2.3. Fungal Isolation and Species Identification

Observation of the macro morphological characteristics of the colonies was achieved using specific media to observe and identify the characteristic macro morphology of the colonies: CYA 25 (Czapeck Agar Yeast Extract 25), CYA 37 (Czapeck Yeast Extract Agar at 37), CY 20S (Czapeck Yeast Extract Agar with 20% sucrose) and MEA (Malt Extract Agar); as previously established [13] [14] [15] were carnation leaf agar (CLA), banana leaf agar (BLA) and potato dextrose agar (PDA) [16] .

Riddel’s technique [8] was used to assess the microscopic structures of yeast and filamentous fungi on slides stained with lactophenol cotton blue. Colonies with yeast-like forms were purified and seeded on CHROMagar^TMCandida, according to the characteristics of the colonies using specific morphophysiological evidence. Biochemical tests were performed, urease technique, CGB (L-canavanine-glycine-bromothymol blue), production of phenol-oxidase in ágar seeds niger (Guizotia abyssinica) and auxanogram, which included the assimilation of carbon sources and nitrogen source [17] . These were used as complementary tests for differentiation, identification and confirmation at the species level.

Identification was carried out by specific mycological methods for each group of isolated fungi, as well as with specific bibliographic [8] [14] [15] [17] - [23] .

2.4. Data Analysis

Were analysed the data using Sörensen’s similarity index (S_S), Fisher’s (α), Shannon-Wiener’s diversity index (H'), Simpson’s diversity index (1/D), Margalef’s richness index (I_Mg), Menhinick’s richness index (1/d), diversity indices were calculated using software Microsoft Excel^TM 2010.

3. Results

3.1. Study Areas Characterization

3.1.1. Diversity Samples

A total 26,194 colonies were isolated and 54 taxa and 183 species of fungal microorganisms were identified, of these species, 156 (85.2%) characterized as mycelial fungi and 27 (14.7%) as yeast fungi. The fungal genera isolated were mycelial fungi (43; 79.6%) and yeasts (11; 20.4%). A total of 342 samples were collected in the following sections: periodicals, study areas, rare books, archive storage and display areas of seven libraries, four located in the city of Cuiabá and three located in the city of Várzea Grande, Mato Grosso, Brazil (Figure 1).

Differences were observed among the three collection techniques applied in the acquisition of fungal propagules. The swab technique proved to be more efficient in capturing the samples 16,387 (62.6%), followed by the sedimentation technique in petri dishes 6578 (25.1%) and the Andersen’s sampler technique 3229 (12.3%) (Table 1).

The incidence of fungal propagules was also verified for rainy season (January to March/2014) and dry season (July to September/2014) collections in the seven libraries a, b, c, d, e, f, g evaluated (Figure 1). The choice of sampling sites was randomized [24] , have been observed 26,194 fungal CFU’s grown in the petri dishes exposed (gravitational method), collected from Andersen’s sampler and seeded by swabs (Figure 2). The number of Colony Forming Units (CFU) for the dry season 13,991 (54.3%) presented a higher index isolating compared to the rainy season 12,062 (46.6%) (Table 1).

The number of Colony Forming Units (CFU) for the dry season 13,991 (54.3%) presented a higher index isolating compared to the rainy season 12,062 (46.6%). Despite this high isolation rate of colonies in the dry period, this period presented a lower number of isolates of fungic genera (35 taxa) than the rainy season (45 taxa).

Fifty-four taxa were identified from samples obtained in the four collections conducted, two during the rainy season and the two during the dry season. Twenty-seven genera (50%) were identified from samples collected in both seasons: Acremonium, Alternaria, Aspergillus, Aureobasidium, Bipolaris, Candida, Chrysonilia, Chrysosporium, Cladophialophora, Cladosporium, Cryptococcus, Curvularia, Fonsecaea, Fusarium, Mucor, Mycelia sterilia, Neocytalidium, Paecilomyces, Penicillium, Pestalotiopsis, Phaeacremonium, Rhizopus, Rhodotorula, Saccharomyces, Sporobolomyces, Talaromyces, Trichoderma.

Table 1

Frequency of colony-forming units (CFU) of filamentous fungi and yeasts isolated from dust seven libraries, applying three collection methodology: Andersen’ sampler (AS); exposure in petri dish (PD) and sterile swab (SW), during the rainy season and dry season in 2014, in the municipalities of Cuiabá and Várzea Grande―MT, Brazil

ENVIROMENTAL SPECIMENS			SAMPLES
			AS	PD	SW	TOTAL
SEQ.	Filo/Families	Isolated species	N	N	N	N	%
ASCOMYCOTA
01	Aspergillaceae	Aspergillus aculeatus	40	143	48	231	0.9
02	Aspergillaceae	Aspergillus amstelodami	0	11	4	15	0.1
03	Aspergillaceae	Aspergillus awamorii	118	280	679	1077	4.1
04	Aspergillaceae	Aspergillus candidus	5	26	5	36	0.1
05	Aspergillaceae	Aspergillus carbonarius	98	143	189	430	1.6
06	Aspergillaceae	Aspergillus carneus	0	12	3	15	0.1
07	Aspergillaceae	Aspergillus chevalieri	0	12	2	14	0.1
08	Aspergillaceae	Aspergillus clavatus	59	21	194	274	1.0
09	Aspergillaceae	Aspergillus clavatus-nanicus	0	26	2	28	0.1
10	Aspergillaceae	Aspergillus conicus	0	0	8	8	0.0
11	Aspergillaceae	Aspergillus fischerianus	6	27	3	36	0.1
12	Aspergillaceae	Aspergillus flavipes	0	24	3	27	0.1
13	Aspergillaceae	Aspergillus flavus	285	532	801	1618	6.2
14	Aspergillaceae	Aspergillus flavus-furcatis	0	10	0	10	0.0
15	Aspergillaceae	Aspergillus fumigatus	39	50	93	182	0.7
16	Aspergillaceae	Aspergillus giganteus	12	10	0	22	0.1
17	Aspergillaceae	Aspergillus glaucus	0	35	0	35	0.1
18	Aspergillaceae	Aspergillus japonicus	114	207	308	629	2.4
19	Aspergillaceae	Aspergillus melleus	0	2	7	9	0.0
20	Aspergillaceae	Aspergillus minesclerotigenes	0	0	8	8	0.0
21	Aspergillaceae	Aspergillus nidulans	71	74	661	806	3.1
22	Aspergillaceae	Aspergillus niger	464	783	1972	3219	12.3
23	Aspergillaceae	Aspergillus nomius	0	2	0	2	0.0
24	Aspergillaceae	Aspergillus ochraceus	17	15	123	155	0.6
25	Aspergillaceae	Aspergillus ornatulus	3	0	0	3	0.0
26	Aspergillaceae	Aspergillus oryzae	19	24	144	187	0.7
27	Aspergillaceae	Aspergillus parasiticus	90	110	141	341	1.3
28	Aspergillaceae	Aspergillus penicillioides	0	3	11	14	0.1
29	Aspergillaceae	Aspergillus quadrillineatus	0	0	25	25	0.1
30	Aspergillaceae	Aspergillus reptans	2	0	9	11	0.0
31	Aspergillaceae	Aspergillus rubrum	0	17	0	17	0.1

32	Aspergillaceae	Aspergillus sclerotiioniger	4	0	0	4	0.0
33	Aspergillaceae	Aspergillus sclerotiorum	10	8	3	21	0.1
34	Aspergillaceae	Aspergillus sojae	5	10	8	23	0.1
35	Aspergillaceae	Aspergillus sulphureus	0	0	3	3	0.0
36	Aspergillaceae	Aspergillus sydowii	9	6	8	23	0.1
37	Aspergillaceae	Aspergillus tamarii	197	252	232	681	2.6
38	Aspergillaceae	Aspergillus terreus	16	49	64	129	0.5
39	Aspergillaceae	Aspergillus unguis	17	31	99	147	0.6
40	Aspergillaceae	Aspergillus ustus	0	10	0	10	0.0
41	Aspergillaceae	Aspergillus versicolor	37	16	49	102	0.4
42	Aspergillaceae	Aspergillus violaceofuscus	0	8	12	20	0.1
43	Aspergillaceae	Monascus ruber	0	9	0	9	0.0
44	Aspergillaceae	Penicillium chrysogenum	6	12	21	39	0.1
45	Aspergillaceae	Penicillium citreonigrum	7	3	12	22	0.1
46	Aspergillaceae	Penicillium citrinum	174	145	331	650	2.5
47	Aspergillaceae	Penicillium corylophilum	0	1	6	7	0.0
48	Aspergillaceae	Penicillium digitatum	4	0	4	8	0.0
49	Aspergillaceae	Penicillium expansum	17	13	0	30	0.1
50	Aspergillaceae	Penicillium glabrum	63	142	87	292	1.1
51	Aspergillaceae	Penicillium implicatum	2	2	0	4	0.0
52	Aspergillaceae	Penicillium italicum	0	8	10	18	0.1
53	Aspergillaceae	Penicillium oxalicum	0	3	28	31	0.1
54	Aspergillaceae	Penicillium restrictum	0	25	11	36	0.1
55	Aspergillaceae	Penicillium sclerotiorum	0	2	0	2	0.0
56	Aspergillaceae	Penicillium simplicissimum	0	2	37	39	0.1
57	Aspergillaceae	Penicillium spinulosum	7	12	0	19	0.1
58	Aspergillaceae	Penicillium variabilis	0	0	63	63	0.2
59	Botryosphaeriaceae	Macrophomina phaseolina	7	0	0	7	0.0
60	Chaetomiaceae	Chaetomium globosum	5	0	0	5	0.0
61	Clavicipitaceae	Mariannaea elegans	0	3	0	3	0.0
62	Clavicipitaceae	Tritirachium oryzae	5	6	0	11	0.0
63	Davidiellaeae	Cladosporium cladosporioides	194	357	1274	1825	7.0
64	Davidiellaeae	Cladosporium colocasiae	0	0	5	5	0.0
65	Davidiellaeae	Cladosporium delicatulum	0	0	31	31	0.1
66	Davidiellaeae	Cladosporium elatum	0	13	0	13	0.0
67	Davidiellaeae	Cladosporium herbarum	48	8	0	56	0.2
68	Davidiellaeae	Cladosporium macrocarpum	2	0	0	2	0.0

69	Davidiellaeae	Cladosporium sphaerospermum	5	148	126	279	1.1
70	Davidiellaeae	Cladosporium tenuissimum	0	33	0	33	0.1
71	Dothioraceae	Aureobasidium melanogenum	6	55	54	115	0.4
72	Dothioraceae	Aureobasidium pullulans	25	78	71	174	0.7
73	Dothioraceae	Hormonema dematioides	0	0	5	5	0.0
74	Endomycetaceae	Geotrichum candidum	0	0	2	2	0.0
75	Endomycetaceae	Geotrichum clavatum	0	0	5	5	0.0
76	Herpotrichiellaceae	Cladophialophora bantiana	25	89	0	114	0.4
77	Herpotrichiellaceae	Cladophialophora carrionii	43	15	12	70	0.3
78	Herpotrichiellaceae	Fonsecaea compacta	0	0	12	12	0.0
79	Herpotrichiellaceae	Fonsecaea pedrosoi	0	0	3	3	0.0
80	Herpotrichiellaceae	Rinocladiella aquaspersa	12	0	0	12	0.0
81	Microascaceae	Graphium putredinis	0	2	3	5	0.0
82	Microascaceae	Scopulariopsis brevicaulis	0	0	2	2	0.0
83	Nectriaceae	Fusarium avenaceum	1	0	8	9	0.0
84	Nectriaceae	Fusarium chlamydosporum	5	0	0	5	0.0
85	Nectriaceae	Fusarium dimerum	0	5	0	5	0.0
86	Nectriaceae	Fusarium graminearum	9	97	190	296	1.1
87	Nectriaceae	Fusarium oxysporum	2	160	433	595	2.3
88	Nectriaceae	Fusarium proliferatum	0	67	23	90	0.3
89	Nectriaceae	Fusarium roseum	0	1	0	1	0.0
90	Nectriaceae	Fusarium solani	10	92	8	110	0.4
91	Nectriaceae	Fusarium sporotrichoides	6	0	8	14	0.1
92	Nectriaceae	Fusarium verticillioides	37	93	115	245	0.9
93	Saccharomycetaceae	Bispora bertulina	0	10	0	10	0.0
94	Saccharomycetaceae	Candida albicans	3	2	17	22	0.1
95	Saccharomycetaceae	Candida glabrata	9	0	10	19	0.1
96	Saccharomycetaceae	Candida intermedia	3	3	0	6	0.0
97	Saccharomycetaceae	Candida kefyr	0	15	14	29	0.1
98	Saccharomycetaceae	Candida krusei	2	5	21	28	0.1
99	Saccharomycetaceae	Candida parapsilosis	0	11	29	40	0.2
100	Saccharomycetaceae	Candida tropicalis	0	2	43	45	0.2
101	Saccharomycetaceae	Candida viswanathii	0	0	3	3	0.0
102	Saccharomycetaceae	Candida zeylanoides	0	3	0	3	0.0
103	Saccharomycetaceae	Hanseniaspora apiculata	0	6	0	6	0.0
104	Saccharomycetaceae	Hansenula anomala	0	9	0	9	0.0
105	Saccharomycetaceae	Saccharomyces cerevisae	1	23	58	82	0.3

106	Saccharomycetaceae	Saccharomyces ellipsoideus	3	2	0	5	0.0
107	Trichocomaceae	Talaromyces purpurogenus	25	28	2	55	0.2
108	Trichocomaceae	Talaromyces rugulosus	2	0	11	13	0.0
109	Trichocomaceae	Talaromyces verruculosus	6	0	0	6	0.0
BASIDIOMYCOTA
110	Cryptococaceae	Cryptococcus spp.	53	272	1540	1865	7.1
111	Cryptococaceae	Rhodotorula aurantiaca	0	0	63	63	0.2
112	Cryptococaceae	Rhodotorula glutinis	29	24	985	1038	4.0
113	Cryptococaceae	Rhodotorula minuta	17	16	220	253	1.0
114	Cryptococaceae	Rhodotorula mucilaginosa	6	75	1193	1274	4.9
115	Sporodiobolaceae	Sporobolomyces roseus	4	0	0	4	0.0
116	Sporodiobolaceae	Sporobolomyces salmonicolor	0	3	0	3	0.0
117	Trichosporonaceae	Trichosporon spp.	4	0	8	12	0.0
DEUTEROMYCOTA
118	Amphisphariaceae	Pestalotiopsis funerea	0	0	0	0	0.0
119	Amphisphariaceae	Pestalotiopsis microspora	2	14	5	21	0.1
120	Botryosphaeriaceae	Neoscytalidium dimidiatum	0	12	53	65	0.2
121	Botryosphaeriaceae	Neoscytalidium hialinum	17	84	23	124	0.5
122	Botryosphaeriaceae	Phaeoacremonium parasiticum	2	19	60	81	0.3
123	Clavicipitaceae	Paecilomyces carneus	9	8	64	81	0.3
124	Clavicipitaceae	Paecilomyces farinosus	0	0	72	72	0.3
125	Clavicipitaceae	Paecilomyces fumosoroseus	0	0	8	8	0.0
126	Clavicipitaceae	Paecilomyces lilacinus	32	14	401	447	1.7
127	Clavicipitaceae	Paecilomyces penicillatus	0	2	0	2	0.0
128	Clavicipitaceae	Paecilomyces variotii	38	157	34	229	0.9
129	Clavicipitaceae	Paecilomyces viride	40	29	21	90	0.3
130	Herpotrichiellaceae	Phialophora richardsiae	0	13	0	13	0.0
131	Hypocreaceae	Acremonium chrysogenum	0	8	1	9	0.0
132	Hypocreaceae	Acremonium falciforme	0	22	14	36	0.1
133	Hypocreaceae	Acremonium kiliensi	0	15	103	118	0.5
134	Hypocreaceae	Alternaria alternata	52	110	323	485	1.9
135	Hypocreaceae	Alternaria chlamydospora	0	2	0	2	0.0
136	Hypocreaceae	Alternaria citri	0	2	26	28	0.1
137	Hypocreaceae	Alternaria geophila	0	6	41	47	0.2

138	Hypocreaceae	Alternaria infectoria	12	0	239	251	1.0
139	Hypocreaceae	Alternaria longissima	0	7	0	7	0.0
140	Hypocreaceae	Alternaria tenuissima	0	12	0	12	0.0
141	Hypocreaceae	Trichoderma longibrachiatum	0	21	0	21	0.1
142	Hypocreaceae	Trichoderma roseum	0	3	0	3	0.0
143	Hypocreaceae	Trichoderma viride	45	123	344	512	2.0
144	Microascaceae	Scedosporium apiospermum	11	2	0	13	0.0
145	Microascaceae	Scedosporium prolificans	0	6	0	6	0.0
146	Nectriaceae	Cylindrocarpon destructans	0	0	6	6	0.0
147	Onygenaceae	Chrysosporium fastidium	18	6	0	24	0.1
148	Onygenaceae	Chrysosporium keratinophilum	0	11	0	11	0.0
149	Onygenaceae	Chrysosporium zonatum	0	0	0	0	0.0
150	Pleosporaceae	Bipolaris australiensis	2	0	81	83	0.3
151	Pleosporaceae	Bipolaris hawaiiensis	5	0	0	5	0.0
152	Pleosporaceae	Bipolaris spicifera	6	22	24	52	0.2
153	Pleosporaceae	Epicoccum nigrum	0	12	0	12	0.0
154	Pleosporaceae	Epicoccum purpurascens	24	0	0	24	0.1
155	Pleosporaceae	Exserohilum rostratum	6	3	0	9	0.0
156	Pleosporaceae	Ulocladium alternariae	0	1	0	1	0.0
157	Pleosporaceae	Ulocladium botrytis	0	19	10	29	0.1
158	Pleosporaceae	Stemphylium botryosum	5	0	0	5	0.0
159	Pleosporales	Curvularia aerea	6	9	39	54	0.2
160	Pleosporales	Curvularia brachyspora	7	0	0	7	0.0
161	Pleosporales	Curvularia clavata	3	7	0	10	0.0
162	Pleosporales	Curvularia eragostridis	0	0	12	12	0.0
163	Pleosporales	Curvularia lunata	29	137	619	785	3.0
164	Pleosporales	Curvularia ovoide	0	2	3	5	0.0
165	Pleosporales	Curvularia pallescens	10	0	0	10	0.0
166	Sordariaceae	Chrysonilia sitophila	59	151	27	237	0.9
167	Stachybotryaceae	Stacybotrys equinata	5	0	0	5	0.0
168	Trichosphaeriaceae	Nigrospora sphaerica	0	0	5	5	0.0
ZIGOMYCOTA
169	Cunninghamellaceae	Cunninghamela bertholetiae	3	12	31	46	0.2
170	Mucoraceae	Absidia corymbifera	0	3	0	3	0.0
171	Mucoraceae	Circinella muscae	0	0	2	2	0.0
172	Mucoraceae	Mucor circinelloides	2	39	38	79	0.3
173	Mucoraceae	Mucor hiemalis	8	14	109	131	0.5

174	Mucoraceae	Mucor mucedo	0	0	17	17	0.1
175	Mucoraceae	Mucor racemosus	0	0	3	3	0.0
176	Mucoraceae	Rhizopus chlamydosporus	1	0	0	1	0.0
177	Mucoraceae	Rhizopus microsporus	0	0	7	7	0.0
178	Mucoraceae	Rhizopus rhizopodiformis	0	3	0	3	0.0
179	Mucoraceae	Rhizopus oryzae	35	55	66	156	0.6
180	Mucoraceae	Rhizopus schipperae	4	0	0	4	0.0
181	Mucoraceae	Rhizopus stolonifer	55	77	224	356	1.4
182	Syncephalastraceae	Scyncephalastrum racemosum	0	0	54	54	0.2
183	AGNOMICETES	Mycelia sterilia	45	105	40	190	0.7
TOTAL			3229	6578	16387	26194	100

Of these, 45 taxa were isolated in the rainy season, of which 18 (33.3%) were exclusive to this season: Absidia, Chaetomium, Circinella, Cunninghamela, Cylindrocarpon, Exserohilum, Geotrichum, Graphium, Hanseniospora, Hormonema, Macrophomina, Mariannae, Nigrospora, Scopulariopsis, Scedosporium, Trichosporon, Tritirachium, Ulocladium. In the dry season, 35 taxa were isolated, of which 9 (16.7%) were exclusive to this season: Bispora, Epicocum, Hansenula, Monascus, Phialophora, Rinocladiela, Scyncephalastrum, Stacybotrys, Stemphylium.

Isolates that did not sporulate were identified as morphotypes (Mycelia sterilia), based on their morphological characteristics. Were identified 190 CFU (0.7%) as Mycelia sterilia (Agonomycetales) an order of imperfect fungi. The genera with a higher dominance in the total fungi isolated, according to the number of species were: Aspergillus, 42 species, 10,628 isolates (40.6%); Rhodotorula, 4 species, 2,628 isolates (10%); Cladosporium, 8 species, 2,244 isolates (8.6%); Followed by Cryptococcus 1,865 isolates (7.1%); Fusarium 10 species, 1370 isolates (5.2%); Penicillium, 15 species, 1,260 isolates (4.8%); Paecilomyces, 7 species, 929 isolates (3.5%); also by Curvularia, 7 species; 883 isolates (3.4%); Alternaria, 7 species, 832 isolates (3.2%) and Trichoderma, 3 species; 536 isolates (2.0%).

Regarding the sociological structure of a species in its totality in the community, indicated that prevalence of the species isolated in this study; we can cite the “Black Aspergilli”, Aspergillus niger 3.219 (12.3%) isolated colonies, followed by lignolytic yeast Cryptococcus complex 1865 (7.1%) colonies and the pigmented fungus Cladosporium cladosporioides 1825 (7.0%) colonies, as the most commonly isolated in the four collections performed (Table 1).

When we evaluated the prevalence of these fungal entities in relation to each collection, that is, two in the humid season (1^st and 2^nd collection) and two in the dry season (3^rd and 4^th collection), we can infer that the three most prevalent species. In the first collect of the wet season, Aspergillus niger mycelial fungi predominated 971 (3.7%) CFU, followed by another “Black Aspergilli”, Aspergillus awamori 625 (2.4%) and Aspergillus flavus 623 (2.4%) isolates. In the second collect of the wet season, again the predominance of Aspergillus niger, this time 878 (3.3%) isolates, followed by the carotenoid yeast Rhodotorula mucilaginosa 504 (1.9%) and the anamorph fungus Penicillium citrinum 424 (1.6%) isolated.

In the third collect, already destined to the dry season, we observed the presence of the capsulated yeast Cryptococcus complex 1129 (4.3%) CFUs, followed by Aspergillus niger mycelial fungus 783 (3%) colonies and Cladosporium cladosporioides 735 (2.8%) of the isolates and finally in the fourth and final collect, Rhodotorula glutinis was predominant 738 (2.8%), followed by mycelial fungi, Aspergillus niger 587 (2.2%) isolates and by the darkly pigmented mold Cladosporium cladosporioides 574 (2.2%) fungal isolates (Table 1).

3.1.2. Species Diversity, Richness and Dominance

The calculated diversity indexes (Table 2) were divided into three groups, according to the diversity component they express: those that express richness and diversity (S, I_Mg, M_n), those that analyze the equitability (H’) and the one expressing dominance (D’).

Among the genera isolated in the dry and rainy season there was a statistically significant difference (p = 0.023). The Sörensen similarity index (S_S) was 93.2% for the mycotas isolated in both seasons (dry/rainy) showed a high species similarity between the mycotas of the two periods surveyed (dry/rainy). These indices may be considered high, indicating the existence of qualitative differences in the composition of the mycotas isolated, despite the quantitative similarities in the number of species isolated in both seasons.

There was little difference in the richness of fungal species (S), except in the sedimentation technique by petri dish in the rainy season (103), followed by swab in the rainy season (92) (Table 2). It was observed that the greatest number of fungal isolates (CFU) occurred in the dry period.

The diversity and richness of fungi species were characterized by the following indices: The diversity index of Shannon-Wiener (H’) was calculated by representing the number of individuals as well as the number of taxa in each site collected. The maximum value of H’ was observed by the swab technique in the dry period (H’ = 1.47) and in the wet period (H’ = 1.31), respectively (Table 2). These results may be indicative that variations in temperature, rainfall and humidity may affect the distribution of species over the periods, since the Shannon-Wiener index expresses species heterogeneity and this heterogeneity seems not to be related only to with different climatic variations.

Simpson’s diversity index (1/D) was also studied because it measures the dominant species. Greater dominance was observed, again by the Swab technique (0.0080) during the dry season, while during the wet season it was found (0.0043) (Table 2).

Dominance (D) indicates the dominance of a given species. The value was higher in the Andersen sampler technique (0.0147) during the rainy season and also lower in the rainy season (0.0097) by the sedimentation technique in petri dish. However, the lowest value of (D) was observed in the total values of the rainy season (0.0071). In addition, Fisher’s alpha diversity indicated a higher value in the dust samples collected by the technique of exposure in petri dishes in the rainy season (2.66), followed by the swab technique in the rainy season (2.61) (Table 2).

The dominance index of Berger Parker (1/d) expresses the proportion of the total parcel that is due to the dominant species. Compared to sample components at sampling sites with respect to CFU size, the Berger-Parker index of dominance expressed a higher number in the wet period by the Andersen sampler (0.1603) and species samples sharing communities in the plate dish technique in the rainy period and dry period swab (0.1530), respectively.

Margalef (I_Mg) is index of diversity that also expresses species richness, but weighted by the sample size. Margalef index, which evaluates the diversity of communities based on the numerical distribution of individuals (CFU) of different genera, depending on the total number of individuals (CFU), our results gave an index of 14.77 in the rainy season and 13.09 in the dry season (Table 2). The richness of species was studied, besides the diversity of Margalef, the Menhinick index was used. The results indicated that the maximum species richness was reported in the dry period by the Andersen’s sampler technique and the minimum swab technique, also in the dry period (Table 2).

Table 2

Diversity index of fungi found in the libraries of Cuiabá and Várzea Grande-MT, found in the dust collection of three techniques (Andersen’s sampler, exposition of petri dish and sterile swab); during the dry and rainy seasons in 2014

Diversity index	Andersen’s sample		Plate dish		Swab		Total
	dry	rainy	dry	rainy	dry	rainy	dry	rainy
Species (S)	72	68	84	103	75	92	126	140
Individuals (N)	1401	1828	3572	3006	9018	7369	13991	12203
Dominance D	0.0139	0.0147	0.0119	0.0097	0.0133	0.0109	0.0079	0.0071
Shannon H’	0.3491	0.4164	0.7805	0.6660	1.4713	1.3150	2.2682	2.0532
Simpson 1/D	0.0001	0.0003	0.0007	0.0007	0.0080	0.0043	0.0140	0.0106
Margaleff (IMg)	9.7999	8.9203	10.1456	12.7367	8.1256	10.2189	13.0943	14.7724
Menhinick (Mn)	4.5953	1.5905	1.4055	1.8786	0.7898	1.0717	1.0652	1.2673
Berger-Parker (1/d)	0.1221	0.1603	0.0904	0.1530	0.1530	0.1487	0.1173	0.1515
Fisher (α)	2.4884	2.4600	2.5649	2.6664	2.5086	2.6102	2.7667	2.8192

3.1.3. Occupational Questionnaire

During the collection, an occupational survey was carried out on the staff of the libraries where the parameters related to the condition and wellbeing of employees in the workplace was observed. 71 employees distributed in seven libraries and collections of two cities surveyed participated in the research; (40/56.3%) belonging to the male gender and (31/43.7%) belonging to the female gender, ranging from 15 to 69 years old (mean = 44.7, SD = 15.2, 95% CI % (±3.6)).

Were obtained (50/70.4%) of librarians and collections held a workload equal to 8 hours, and they occupied positions as sector managers, librarians, analysts and administrative assistants. However, (18/25.4%) of the employees had a workload of 6 hours per day, and they were employed by clerks, administrative assistants, security guards, secretaries and trainees. Finally, (3/4.2%) were registered with a workload of 4 hours, occupying the roles of trainees and apprentices.

When analyzing the responses of the 71 employees of these literary establishments in the research, it was possible to observe that (18/25.4%) did not report complaints and (53/74.6%) referred to complaints resulting from allergic and respiratory reactions. These were especially reported by employees who worked 8 hours a day (37/52.1%) followed by (16/22.5%) by those who worked 6 hours daily. Of these (32/45.1%) were male and 21/29.5% female.

When asked what type of allergic reaction it was possible to verify that the complaints reported by the employees were mixed and 195 complaints of clinical manifestations were reported; and (85/43.6%) complaints from male individuals and (110/56.4%) complaints from female subjects. The most frequent were: sneezing (29/14.9%); cough (21/10.8%), irritation/clearing (16/8.2%), dry mouth (8/4.1%), coryza (12/6.2%), pruritus (15/7.7%), burning and irritation (14/7.2%), erythema (9/4.6%) and considering the eyes: pruritus (28/14.4%), lacrimation (11/5.6%), burning (19/9.7%), redness (13/6.7%).

3.1.4. Internal Climatic Condition

The mean values of the values found for temperature (˚C) within the libraries reached fluctuations in the wet period of (27.6˚C) - SD = 2.1˚C, while in the dry period the recorded temperature was (27.2˚C) - SD = 1.3˚C. Measurements were made in the external areas of these libraries, with the values found in the averages of these temperatures in the wet period (29.1˚C) with SD = 1.3˚C, and in the dry period (26.9˚C) - SD = 1.7˚C (Figure 3).

In relation to the relative humidity of the air (RH%) the internal average observing the parameters recommended by the technical norms. In the wet period the unit was equal to 44.5% - SD = 5.7%, while in the dry period 40.3% - SD = 5.4%. In the external areas of the libraries, the mean value for humidity was 58.7% - SD = 4.4% in the wet period and 44.3% - SD = 7.1% in the dry period (Figure 3).

4. Discussion

4.1. Fungal Collection

The results obtained showed that the indoor environment of libraries consists of a variety of fungal microrganisms capable of causing deterioration of literary archives, allergic reactions and sensitization in atopic individuals, and possible fungal infections of various etiologies. These findings corroborate studies on airborne mycobiota conducted by several research groups and their casuistic [3] [4] [9] [10] [25] [26] .

According [6] [8] [27] modern humans spend an average of 87% of the day indoors; inhaling air contained in these environments. Considering this fact, studies of indoor air quality (IAQ) are important and have been highlighted by several authors in their series, such as those performed in: residences [28] , educational institutions [29] , hospitals [25] [26] , wards [30] , laboratory [31] , airports [32] , railway stations [33] ; caves [34] ; huts [35] ; theaters [36] ; wineries [37] ; vivarium [38] , avian [39] , shopping centers, buindings and hotels [40] [41] , carpets [42] various internal environmentals [43] and libraries [10] [44] [45] .

Fungi are ubiquitous in all environments and the abundance of other fungi varies with time and place. In general, both outer and inner environments are colonized by species of Cladosporium, Penicillium, Aspergillus and Alternaria and by yeast and also Mycelia sterilia. Cladosporium is always the dominant fungus in outdoor environments, but in indoor environments in suitable conditions Aspergillus and Penicillium are generally dominant [45] [46] .

In this work, Aspergillus spp. (10,628; 40.6%), one of the main airborne fungi isolated the world over [21] was isolated in all the collect, being the species Aspergillus niger (3219; 12.3%) demonstrating that it is constantly present in numerous locations. According to Prince & Meyer [47] and Miller [48] Aspergillus and Penicillium are genera that are usually found inside environments, while the concentrations of spores of Alternaria spp next to Cladosporium spp in the open air are generally more prevalent in the peaks during summer and fall in temperate locations. Other researchers in their casuistic also reported the existence of these fungal entities predominating in the literature records [49] [50] . Aleksic [51] , conducting studies on the health risk of mycotoxins produced by fungi growing indoors, have shown numerous light spores that can be easily aerosolized and inhaled along with mycotoxins.

Some studies report the inverse for isolation of the genera Aspergillus and Penicillium. Sautour [31] conducted a survey of fungal contamination in hospitalar unit, they isolated Penicillium spp. (27% - 38%, while Faure [52] conducted a study on airborne microbiota in a French hospital and identified this genus as the most frequent from 1992 to 1999, with a percentage of 28.4%. According to Lacaz [8] find the optimum temperature for greater propagation of the genus Penicillium is 20˚C - 25˚C, and this in many places, such as decaying air, soil and organic matter.

According to Pitt [15] , this inversion occurs because the species of both genera are found almost everywhere on the planet. Penicillium is a common dominant species in cold climates, particularly in cold temperate zones, while Aspergillus is most common in the tropics and warm locations. The state of Mato Grosso has a tropical climate and this explains the high rate of isolation of this fungal genre detected in this casuistic.

A number of airborne fungal species are considered important to medical pathologies, including those belonging to the genera Alternaria spp., Aspergillus spp., Cladosporium spp., Fusarium spp., Mucor spp., Penicillium spp. and Rhizopus spp. It has become apparent that these are especially allergenic elements, a factor that is particularly disturbing to clinicians, since these microorganisms are abundantly dispersed in the environment. This is the reason that investigations concerning the occurrence of environmental fungi (usually opportunistic and contaminants) have become important in the prevention of allergic diseases caused by pathogens that are potentially harmful to humans [33] .

Saprophytic fungi considered common in storage areas, like Acremonium spp, Aspergillus spp, Cladosporium spp, Curvularia spp, Fusarium spp, Penicillium spp and Rhizopus spp, presented an expressive quantity of spores in the environments evaluated. These microorganisms are responsible for numerous opportunistic diseases, including onychomycosis, keratitis, ear infections, allergic conditions, mycotoxicosis, as well as urinary, pulmonary and systemic infections.

Dunlap [53] reported that fungi as well as other species of microorganisms on the planet have a circadian clock, and these species differences are more likely to occur due to environmental changes where the temperature may be slightly warmer in the places or days where they occurred collections, such as during the summer. The findings of Silveira [33] report that there is influence of temperature on the occurrence of species in different climatic periods. Amend [54] ; still the humidity can be a differential factor as also the length of the day also changes with the climatic period being able to contribute to the variability and diversity of isolated species. These statements corroborate the results found in this study, indicating that geographically isolated species may belong to the local biota.

It was possible to verify that the most commonly isolated species belonged to the genus Aspergillus, Penicillium, Cladosporium, Fusarium, Paecilomyces, Rhodotorula and Cryptococcus, evaluating of cities of Cuiabá and Várzea Grande and the same substrate of the internal dust of libraries of these cities. This fact can signal even with the changes that have taken place in the city’s distinctive phytophysiognomy that the species can remain predominant in the place. This citation also complies with Mezzari [55] who report that the anemophilic fungal microbiota may be similar or different in each city or region.

Situations observed in the collection period were the urban works that occurred in both cities due to the advent of the World Cup (2014). The cities Cuiabá and Várzea Grande underwent several changes in infrastructure, such as structural works and construction of road networks, excavations, tunnel construction and a series of changes and transformations, which contributed greatly to the dispersion of dust contained in the soil of the cities surveyed, contributing possibly to increase the isolation of the fungal microorganisms isolated in this work.

Temperate climates favor the appearance of soil fungi, possibly justifying the detection of Cryptococcus (1865; 7.1%) samples in this study. According to Kidd [56] [57] , these basidiomycetic yeasts occur at 15 cm from the soil, in trees, wood chips, straw and other “natural reservoirs”. This occurs with many other fungi, such as Aspergillus fumigatus 182 (0.7%) where their spores are so small, and other species Aspergillus niger 3219 (12.3%) and Aspergillus flavus 1,618 (6.2%) that any disturbance can dislodge and disperse them via air currents easily aerosolized by soil disturbances [58] .

In Brazil, among the main anthropogenic activities that pollute atmospheric air in urban and industrialized regions are the burning of fossil fuels, industrial processes, incineration of urban waste, mining, civil construction and associated inadequate agricultural practices [59] .

There is still to be made an inference about the burnings carried out during the dry months in Mato Grosso (August, September and October) in Cerrado areas, which are routinely becoming an event that is already part of the region’s phytophysiognomy. This fact is capable of generating constant uncomfortable visual and aggravating health problems common to this period and contributing to the increase of drought and thermal sensation of excessive heat faced by the population of Mato Grosso state.

In view of the above, these events may have contributed greatly to the capture of fungal spores carried by atmospheric air of the region, facilitating the dispersion of various microorganisms (some considered uncommon in isolation and others considered of extreme importance) in the etiology of respiratory and systemic diseases.

In relation to the yeasts isolated in this study we can highlight those of the genus Cryptococcus spp. (1865, 7.1%) which deserve due attention due to possible pathogenicity in immunocompromised and immunocompetent individuals. Fewer are the records of this fungal entity in dust, highlighting the work carried out by Leite-Jr [44] , which isolated seven different species of this basidiomycetic yeast being Cryptococcus neoformans, C. gatti, C. albidus, C. luteolus, C. uniguttulatus, C. humiculus and C. terreus in the dust of libraries of the Midwest region of the country, highlighting the species C. gatti (36.6%), as the agent thinned with this substrate.

A British study established a relationship between the presence of symptoms and an air conditioning system [40] ; however, it was in the 1990s that SBS became a common concept in the scientific literature [60] . In Brazil, it only began to receive greater attention from health authorities following the death of the Minister of Communications, Sérgio Motta, in 1998, because his clinical status was aggravated by the presence of fungi growing in the air conditioning system of his cabinet offices.

It is estimated that human inhale about 200 filamentous spores daily containing mycotoxins and fungal volatiles, and some of these compounds react to lung alveoli [58] . The interviews conducted by questions on health and users of mental state allows somehow draw conclusions if the symptoms proposed by SED tend to decrease or disappear when users are away from the building in which they work.

In the seven libraries evaluated, 30 environments were investigated: (10; 33.3%) classified as open environments with air circulation and presence of people constantly; (10, 33.3%) being classified as closed environments, type 1, that maintained air circulation and presence of people; followed by (3; 10%) of closed environments, type 2, with little circulation of air and people and still (7, 23,3%) classified as unhealthy environment, that is, closed and without air circulation and with flow rare of people. It was not possible to make inferences and comparisons between the libraries, due to their size and different dimensions, capacity of support, specific rooms, number of employees, different environments and also the disposition of the environments offered to the public and visitors.

It is important to highlight that when working in libraries, the biological risk of the presence of fungi, bacteria and mites is ever-present, especially on paper, leather, fabric and wood, substrates that these organisms use as their habitat. These risks may be higher or lower depending on the volume and the general environment, and on the level of monitoring of temperature and relative humidity [9] [61] . Libraries present suitable substrates in the form of old paper, fabrics, dust and glue, in association with the humidity of air conditioners. Workers in these establishments who suffer from allergic rhinitis, asthma and bronchitis can be affected when handling old books, periodicals and documents [3] .

Among the mycelial species, four airborne fungi are considered important triggers of respiratory allergies: Alternaria spp., Aspergillus spp., Cladosporium spp. and Penicillium spp. The literature describes the importance of these microorganisms and the allergic complications they cause [9] [62] [63] . Several Brazilian research groups have reported, in their respective casuistics, symptoms that these agents trigger in people who are prone to allergies [25] [26] [45] .

The sensitization of individuals occurs through exposure to fungi and due to their high molecular weight, these microorganisms possess an antigenic capacity caused by different substances embedded within their structure that promote a variety of reactions [55] [64] .

According Gioda and Aquino Neto [4] , the causes of SBS can be explained by a number of factors, including a lack of fresh air, poor air distribution, poor temperature control, inadequate design and irregular air conditioning system maintenance. Thus, not only is it problematic to characterize SBS in a particular building, it is also difficult to identify the primary cause.

Several researchers have conducted surveys concerning the health and mental state of occupants and any symptoms manifested while remaining inside and while outside these buildings as an effective way to identify SBS [4] . It is known that the impact of remaining inside establishments with high concentrations of microorganisms and poor moisture control, allied with work, physical fatigue and stress factors are perceived by the body and mind, leading to symptoms known as allergies or skin problems. The importance of microbiota in libraries worldwide was reviewed by Gallo [62] . In his research, he affirmed that the cause of this problem could be dust or components that affix to the books and shelving.

Allergic rhinitis at present represents a global public health problem and affects 10% to 25% of the population in Western countries. The prevalence of allergic rhinitis in Brazil is not known, although studies conducted in the 80% decade indicate an estimate of 15%. In the 1990s, studies (International Study of Asthma and Allergies in Childhood-ISAAC) showed a prevalence of 30% in children and adolescents. In adults, it is possible that this prevalence reaches 20% or more [65] . In 2011, in a study conducted by Indian researchers, led by Sathavahana Chowdary [66] found a prevalence of 44% in its casuistry.

According WHO [5] , occupants of buildings with moisture problems and containing fungal growth are at higher risk of contracting respiratory diseases, including infectious diseases. This information suggests that in these conditions considered to be often unhealthy, the risk of developing diseases such as allergic rhinitis and asthma can be increased. Kleinheinz & Campbell [67] in their studies also stated that allergic reactions may be related to the presence of certain fungi or portions of fungi in indoor air or associated with extracellular compounds produced. French researcher [51] affirm the presence of mycotoxins in indoors should be taken into consideration as an important parameter of air quality, that part of the toxic load was found on very small particles―dust or tiny fragments, that could be easily inhaled.

In this research, when the employees were questioned about the clinical and epidemiological investigation of the work environment, it was possible to verify that (20/28.2%) of the interviewees reported complaints due to rhinitis in the work environment. Asthma (5/7%) referred to the effects of this reaction and (8/11.2%) as a consequence of sinusitis and finally, (20/28.2%) referred to another type of infection.

Since 2002, the Occupational Safety and Health Administration (OSHA) in the United States has been delivering documented results on the effects of asthma on workers in commercial and institutional environments, revealing the presence of fungi in these workplaces. OSHA research has shown that fungi can cause skin irritation and have effects on the cardiovascular, pulmonary, nervous, reproductive, and carcinogenic effects [68] .

However, according to Hess-Kosa [69] SED is defined only if the presence of these symptoms occurs in a significant number of people. The presence of similar symptoms among the occupants of the building is of extreme importance for the detection of SED [70] .

In the city of New Delhi [71] and Brazil [4] cited polls with users of buildings as an effective way of identifying SED. According to WHO standards, more than half of the indoor environments of buildings such as schools, hospitals, residences, movies, theaters, shopping centers, show poor air quality and suggest that occupants of 30% of new or refurbished buildings around the world presented complaints compatible with poor indoor air quality [5] .

Based on the results of the individual surveys applied to Mato Grosso state library staff, it was possible to verify the most prevalent symptoms. In this case the percentage of individuals is equal to or greater than 20%, this will be a strong indicator of the existence of SBS among the occupants of the building, as suggested [5] .

Few studies show the association between SBS and the age of individuals in 2013 [72] . In China, demonstrated that Adults’ SBS symptoms are associated with a history of asthma, allergic rhinitis or eczema. This fact proves that the manifestations provoked by inhalation of bioaerosols, provoking asthma and rhinitis can affect any age. These results are in accordance with the findings [73] [74] .

Our results showing considering the employees, that (18/25.4%) reported no complaints regarding allergic manifestations and (53/74.6%) reported some allergic manifestations during the working day. Among the 53 employees who reported allergic discomfort (34/64.2%) were enrolled in the age group below 44 years of age, reflecting those who reported more problems related to occupational activity and irritations due to their stay in work environments; (19/35.8%) remained in the age group above 44 years.

Evaluated the internal air [75] of environments, it was found that females (44.3%) were more susceptible to inhaled infections than males (26.2%). It was possible to verify in this study that, although women (31/43.7%) represented a lower index than men (40/56.3%), they reported more complaints of symptomatology (110/56.4%) compatible with SBS compared to men (85/43.6%). This finding confirms previous reports which have indicated the female gender as the most susceptible to this problem, corroborating with results found in this study with the reports [76] .

In 2005 [75] also revealed that women are seen as strong candidates to present significant risks of perceiving eye symptoms. Other researchers consider IAQ as one of the main determinants of SBS [76] .

Studies showed [76] [77] [78] [79] [80] that considering gender, there is a greater impact factor on women’s health. According Kinman & Griffin [81] the results are not unequivocal and many studies include the possibilities that women may be more sensitive to environmental factors.

Others researchers [77] reported differences in the occurrence of SBS symptoms between women and men resulting in factors external to the internal environment. However [79] suggested when evaluating 1,024 adolescents of both genders that higher endogenous estrogen levels in sexually mature adolescents appear to protect them against the effects of aeroallergens, although their overall prevalence is considered to be higher. Bell [82] also point out in their research that women are more susceptible to SBS due to higher estrogen/progesterone ratios. Hormone levels play an important role in neural sensitization because of prolonged and repeated exposure to external stimuli.

The Brazilian Association of Technical Standards (1980) cites that the relative humidity of the air recommended for the deposit of manuscripts and rare works is between 40% to 50% and the temperature is between 21˚C to 23˚C and values above this can cause damage to the collection. The resolution RE n˚9, January 16, 2003 [83] advocates these same parameters [84] . Additionally [84] reports that for good chemical and physical paper conservation, it is advisable to maintain a temperature between 18˚C and 22˚C and relative humidity between 45 and 55%.

Cassares & Moi [85] reported that it is more recommended to keep the archives and libraries at a temperature closer to 20˚C and relative humidity at 45% to 50%, avoiding in any way the oscillations of 3˚C temperature and 10% relative humidity.

Arundel [86] have suggested that high RU increases the occurrence of allergic diseases and respiratory infections. Humidity between 40% and 70% minimizes bacterial and fungal infections, and most fungal species do not grow in RU less than 60%.

There seems to be a divergence between the authors regarding the levels of temperature and humidity fluctuations reported by several authors. In this way, it is possible to conclude that the desirable values according to the mentioned authors, can be between 18˚C and 23˚C considering the temperature and 40 to 60% for humidity.

Costa [84] emphasizes that the imbalance of temperature and relative humidity causes in the collection a dynamics of contraction and elongation of the elements that compose the paper besides favoring the proliferation of biological agents, such as: fungi, bacteria, insects and rodents. The lower the temperature, the longer the paper stays and the longer it lasts. Moisture also greatly affects the paper as follows if very high, accelerates acid degradation and if it is too low, facilitates the attack of biological agents.

The differences in the results between this and other studies can be attributed to geographical location, climatic period, abiotic factors and the time and methodology of collection, since the techniques used in this study may differ greatly compared with other techniques. It is also clear that abiotic factors, such as temperature and relative humidity, exert an important influence on CFUs. This observation is fully corroborated Samson [14] , who pointed out that fungi only occur under favorable conditions, which vary for each species, and that adaptability is a determining factor.

5. Conclusions

These results show that the air conditioning systems of the libraries were not able to maintain the temperature and the relative humidity of the air in the evaluated environments causing high temperature oscillation inside the same ones and thus providing risk factors for the collection favoring the proliferation of fungi which can trigger allergic and respiratory problems.

The internal conditions found in the studied libraries (relative humidity, temperature, conservation of the collections, dust, books, atmospheric air pollution, etc.) in this study, found non ideal values, in the matter of conservation of the stocked materials. These researched libraries need to be applied conservation and cleaning methodologies to optimize the conditions of internal environments. In particular, with regard to questions of relative humidity and internal temperature fluctuations, special care should be taken to avoid the proliferation of fungal agents.

Hygiene, conservation and air quality care are effective conditions for resol- ving an IAQ problem, especially where the sources are opportunistic microorga- nisms that settle in those places where heat, darkness and humidity coexist, and these known sources can be controlled. Periodic cleaning, maintenance of air conditioners and air conditioning in the interiors, as well as the use of dehumidifiers help to provide adequate conditions in these environments and all can be performed individually or at the same time.

Internal air quality management and control programs, aided in the orientation and education of users, employees and technical part of these study and knowledge establishments; become important tools in the prevention and proper functioning of these sites and consequently minimization of problems related to Sick Building Syndrome.

In summary, this study showed clear evidence of fungal diversity in the libraries evaluated and indicated that more effective methods of disinfection and sterilization are required to combat these potentially pathogenic fungal organisms.

Acknowledgements

Financial support for this study was provided by the Research Foundation of the State of Mato Grosso (Fundação de Amparo à Pesquisa no Estado de Mato Grosso, FAPEMAT) and Coordination of Improvement of Higher Level Personnel (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, CAPES).

Author Contributions

Diniz Pereira Leite Júnior was the researcher-in-chief, and designed and performed the study. Diniz Pereira Leite Júnior and Rosane Christine Hahn drafted the study. All authors collected and examined the biological samples, performed the experiments and all authors contributed to data analysis. Diniz Pereira Leite Júnior wrote the paper, and all authors made critical revisions and approved the final manuscript.

Conflicts of Interest

The authors declare no conflict of interest. The founding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results. The authors declare there are no conflicts of interest.

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