Ecotoxicology and Environmental Safety 109 (2014) 32–37

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Ecotoxicology and Environmental Safety

journal homepage: www.elsevier.com/locate/ecoenv

Review Effect of trophic status in on fungal species diversity and abundance

A. Pietryczuk a,n, A. Cudowski a, T. Hauschild b a University of Białystok, Institute of Biology, Department of , 15-950 Białystok, Świerkowa 20B, Poland b University of Białystok, Institute of Biology, Department of Microbiology, 15-950 Białystok, Świerkowa 20B, Poland article info abstract

Article history: The objective of this study was to determine the species diversity and abundance of fungi in relation to Received 12 May 2014 the hydrochemical conditions, with special emphasis on the trophic status and degree of of Received in revised form lakes. The study was conducted in 14 lakes of the Augustów Lakeland (central , NE Poland) with 24 July 2014 different hydrological conditions, type of stratification and trophic status. The analyses were performed Accepted 24 July 2014 in the hydrological year 2013. In the waters of the studied lakes, the mean abundance of fungi was Available online 21 August 2014 560073600 CFU/mL. The minimum value (800 CFU/mL) was recorded for the mesotrophic Płaskie , Keywords: and the maximum value (14,000 CFU/mL) was recorded for the eutrophic Pobojno Lake. A total of 38 Aquatic fungi species of fungi were identified, including 11 belonging to the aquatic hyphomycetes; up to 14 species Lakes were potentially pathogenic fungi. The potentially pathogenic fungi, particularly Candida albicans and Organic matter Scopulariopsis fusca, were found in lakes with increased concentrations of chloride and sulphate(VI) ions ITS-PCR and may thus serve as indicators of the degree of water pollution. This paper illustrates that the species RFLP diversity and abundance of fungi in limnic waters depend on the concentration of organic matter, chlorophyll a concentration, and the degree of water pollution. The results suggest that aquatic fungi can be a valuable indicator of the degree of pollution and the sanitary quality of the water. Crown Copyright & 2014 Published by Elsevier Inc. All rights reserved.

Contents

1. Introduction ...... 33 2. Materials and methods ...... 33 2.1. Studyarea...... 33 2.2. Methods of analysis...... 33 2.2.1. Collection of material for analysis...... 33 2.2.2. Water chemistry ...... 33 2.2.3. Fungal cultures...... 33 2.2.4. DNA extraction...... 33 2.2.5. PCR procedure ...... 33 2.2.6. Restriction fragment length polymorphism (RFPL) analysis ...... 34 2.2.7. Statistical analysis ...... 34 3. Results...... 34 4. Discussion ...... 35 5. Conclusion...... 37 Acknowledgements ...... 37 References...... 37

n Corresponding author. E-mail address: [email protected] (A. Pietryczuk). http://dx.doi.org/10.1016/j.ecoenv.2014.07.032 0147-6513/Crown Copyright & 2014 Published by Elsevier Inc. All rights reserved. A. Pietryczuk et al. / Ecotoxicology and Environmental Safety 109 (2014) 32–37 33

1. Introduction (central Europe, NE Poland). Monitoring in water , particularly those used for economic or tourist purposes, which Aquatic fungi are a phylogenetically diverse group of organ- focuses on the abundance and species diversity of microfungi isms. Their occurrence has been recorded in almost all types of appears to be highly desirable. aquatic environments throughout the world. Aquatic fungi can be of autochthonic or allochthonous origin. Microfungi directly enter water through the of soils or are of anthropogenic 2. Materials and methods origin. The role of fungi in aquatic ecosystems mainly involves their participation in the decomposition of organic matter, parti- 2.1. Study area cularly of plant origin (Krauss et al., 2011; Pascoal et al., 2005). The study area covered a group of 14 natural lakes located in central Europe in Romani et al. (2006) suggest that fungi supply bacteria with the the northeastern area of Poland, the Augustów Lakeland (Fig. 1). The monitoring of organic resources necessary for their survival and functioning, limnic waters was performed in the hydrological year 2013 during the occurrence which they could not obtain by themselves. Due to the secretion of of favourable meteorological conditions to facilitate reliable data collection. The enzymes from the oxidase group, fungi are able to decompose studied lakes had varied hydrological conditions. phenol compounds which are less available to other microorgan- isms, such as humic substances, and various xenobiotics (Augustin 2.2. Methods of analysis et al., 2006; Baldrian, 2006; Jain et al., 2005; Junghanns et al., 2005; Steinberg, 2008). Moreover, fungi inhabiting water ecosys- 2.2.1. Collection of material for analysis tems actively participate in the synthesis of autochthonic humic Water samples were collected four times in different hydrological seasons in January, May, August, and October of the hydrological year 2013 using a Limnos substances (Damare and Raghukumar, 2008). The content of sampler from a depth of 0.5 m (). Water temperature, conductivity (EC), organic matter in water, which as suggest by Dunalska (2011) saturation, dissolved oxygen concentration and pH were measured in the contributes to can also considerably affect the field using a HQ40D Hach Lange meter. species structure and abundance of . In addition, decomposition of organic matter may result in the release of 2.2.2. Water chemistry and , which intensifies the eutrophication Analysis of the physical and chemical water parameters was conducted process and may result in algal blooms (Carlson, 1977). immediately after sample collection, determining sulphate(VI), and chloride ion concentrations (APHA, 1992). Chlorophyll a concentration was determined accord- Research the role of fungi in the waters have not been ing to Polish Standard (1986) (PN-86/C-05560/02), the total phosphorus (TP) and conducted for large-scale. Knowledge on the role of fungi in dissolved reactive phosphorus (DRP) concentrations were determined using the waters is fragmentary. After several years of intensive research molybdenian spectrophotometric method (Standard methods for the examination on leaves decomposing in water, a fair amount is known concern- of water and waste water, 1999). The concentration of dissolved organic carbon (DOC) was determined by the high temperature catalytic method of in ing the taxonomic diversity of aquatic hyphomycetes (Krauss et al., a TOC-5050A analyser (Shimadzu) and particulate organic carbon (POC) was 2011; Orłowska et al., 2009; Solé et al., 2008; Sridhar et al., 2001; determined by the chromate method (Bowman, 1998). Wurzbacher et al., 2010), commonly recognised as predominant in many water ecosystems. In addition, the astonishing species 2.2.3. Fungal cultures diversity of fungi belonging to other groups (often typical patho- To estimate fungal abundance, 250 μLofunfiltered water, diluted to ratios of gens) has been demonstrated, but their ecology remains largely 1:10 and 1:100, were placed on Sabouraud agar plates enriched in chloramphenicol 1 1 1 unknown (Nikolcheva and Bärlocher, 2005). It is estimated that (0.5 g/L) and incubated for 5 days at either 37 C, 25 Cor5 C. Each time, this analysis were conducted in 3 replicates. After incubation, the numbers of colonies only approximately 7 percent of the total number of species of and different morphotypes of fungal colonies were determined (Descals, 2007). aquatic fungi have been identified and described thus far. Analyses Fungal abundance was expressed as CFU/mL. on the taxonomic diversity of aquatic hyphomycetes, yeast-like and zoosporic fungi in the waters of Poland and throughout the 2.2.4. DNA extraction world have thus far been performed using microscopic methods A representative colony of each morphotype was subcultured, and its DNA was (Cressa and Smith, 2007; Jobard et al., 2010; Orłowska et al., 2009). isolated using Genomic Mini AX Yeast and Bead-Beat Micro Gravity DNA Isolation It is increasingly emphasised that only the application of mole- Kit according to the manufacturer's instructions. Three hundred and seven DNA samples were isolated, representing 38 morphotypes of fungi. cular methods, such as the fingerprinting (restriction fragment length polymorphism—RFLP) of internal non-coding regions of fi 2.2.5. PCR procedure rDNA (ITS fragments), which show high interspeci c variability, Following Gaitanis et al. (2002) with modifications, the RFLP bands were used permits the determination of the species diversity of fungi and to identify the isolates. PCR reactions were performed in 0.2 mL Eppendorf tubes in their ecological function in various types of waters. a reaction mixture containing 2 μL of isolated DNA, 10 pmol of ITS1 primer (50- 0 0 The objective of this study was to determine the species TCCGTAGGTGAACCTGCGG-3 ), 10 pmol of ITS4 primer (5 -TCCTCCGCTTATTGA- TATGC-30)(Gupta et al., 2000), 11.75 μL of nuclease-free water (A&A Biotechnology, diversity and abundance of aquatic fungi in relation to the μ fi Poland), and 12.5 L of PCR Master MixPlus (A&A Biotechnology, Poland). The PCR hydrochemical conditions, type of strati cation, trophic status, mixtures were first incubated for 3 min at 95 1C, followed by 40 cycles at 95 1C, and degree of pollution of the lakes of the Augustów Lakeland 52 1C and 72 1C for 1 min at each temperature. The last cycle was performed for

Fig. 1. Map of the distribution of the study sites in the Augustów Lakeland in Polish territory in the northern part of the Podlasie region. 34 A. Pietryczuk et al. / Ecotoxicology and Environmental Safety 109 (2014) 32–37

10 min at 72 1C. The presence of a PCR product was confirmed by 1 percent (w/v) 35.6718.4 mg/L and 14.4 74.34 mg/L, respectively (Table 1). The agarose gel electrophoresis and visualised with ethidium bromide. minimum concentrations of both ions were recorded in Płaskie Lake. 2 The SO4 and Cl values were 9.28 mg/L and 5.00 mg/L, respectively. 2.2.6. Restriction fragment length polymorphism (RFPL) analysis The maximum values of SO2 and Cl ions were recorded in Digestions were performed with 10 μL of PCR product in a total volume of 15 μL 4 with 1 reaction buffer and 10 U of EcoRI endonuclease (Sigma-Aldrich, Poland) for 2 h Pobojno Lake (71.3 mg/L and 31.5 mg/L, respectively) (Table 1). at 37 1C. The resulting fragments were separated by electrophoresis on a 2 percent The waters of the studied ecosystems were distinguished by agarose gel and visualised under UV light after ethidium bromide staining (Gaitanis et the mean abundance of fungi at a level of 560073600 CFU/mL. al., 2002). The lengths of the products were evaluated using a 1000 pb DNA ladder. The The minimum value (800 CFU/mL) was recorded for Płaskie Lake results were archived using GelDoc2000 equipment (Bio-Rad) and the software and the maximum value (14,000 CFU/mL) for Pobojno Lake (Fig. 2). program Quantity One. The lengths of the DNA fragments separated on a gel after digestion were compared to the fragment lengths of the reference strains in GenBank. The highest species diversity was observed in 2 lakes: Gorczyckie and Pobojno, where 27 out of the 38 species found in the 14 2.2.7. Statistical analysis studied lakes were identified. The lowest species diversity was To investigate the relationships between environmental and biological data, a recorded for Serwy, Orle, and Płaskie Lake (an average of 10 out of redundancy analysis (RDA) was carried out. RDA is an enhancement of the commonly 38 species) (Table 2). In total, 11 out of the 38 identified species applied principal component analysis (PCA); however, unlike PCA, RDA allows direct were typical aquatic fungi from the hyphomycetes, 14 species were analysis of biotic-environmental components (terBraak, 1994; Van Wijngaarden et al., 1995). To test whether RDA analysis was appropriate for the dataset, the data were potentially pathogenic fungi (Table 2). We found potentially first tested for normality (Kolmogorov–Smirnov test). DCA was used first to determine pathogenic fungi, particularly Candida albicans and Scopulariopsis the character of variability in the studied assemblages: if the length of the first fusca, in lakes with increased organic matter concentrations. The gradient was greater than 2 standard deviations, we assumed a unimodal variation; a other potentially pathogenic fungus, Arthroderma insingulare,was length less than 2 SD indicates a linear variation (LešpandŠmilauer, 2003). The length not detected in these types of lakes (Table 2). Moreover, in the two of the first gradient for the fungi communities was 1.71 SD, indicating linear variation and justifying redundancy analysis. Relationships between nominal and quantitative eutrophic lakes (Gorczyckie and Pobojno), a typical fungus of variables were calculated with V Cramera ranks, but relationships between quantita- origin was identified, namely Leptomitus lacteus (Table 2). tive variables were calculated with Pearson ranks. The analysis of existing differences Only two representatives of the aquatic hyphomycetes, i.e., Angu- fi among dams involved multi-dimensional analysis; speci cally, cluster analysis, where illospora crassa and Flagellospora curvula, were found in all the the Euclidean distance was adopted as the probability measure and the Ward's ł method as the clustering procedure. Statistical calculations were performed using studied lakes, with the exception of Bia e Augustowskie Lake Statistica version 7 and CANOCO for Windows 4.5 software. The map was developed (Table 2). with ArcGIS software version 9.3.1.

3. Results

The conductivity of the waters varied from 222 μS/cm in Płaskie Lake to 501 μS/cm in Pobojno Lake (Table 1). The pH of the lakes varied from slightly acidic, at 6.66 in Płaskie Lake, to alkaline, at 9.03 in Pobojno Lake (Table 1). The chlorophyll a concentration, varied from 1.21 to 57.3 mg/L. The minimum values of this parameter were recorded for Rospuda Lake and the maximum values for Pobojno Lake (Fig. 2). The minimum values of both dissolved organic carbon (DOC) and particular organic carbon (POC) concentrations were recorded for Płaskie Lake and amounted to 2.11 and 0.13 mgC/L, respectively (Table 1). The dissolved organic carbon reached a maximum value of 35.6 mgC/L in Pobojno Lake; the particulate carbon reached a maximum value of 6.13 mgC/L in Kalejty Lake. The lowest value of the total phosphorus (TP) concentration was recorded in Płaskie Lake, at 26.2 μg/L, and the maximum value of TP was recorded in Pobojno Lake, at 201.3 μg/L (Table 1). The dissolved reactive phosphorus (DRP) concentration μ varied from 0.6 (Kalejty Lake) to 64.5 g/L (Serwy Lake) (Table 1). The Fig. 2. Average concentrations and standard deviations of chlorophyll a and mean concentrations of sulphate(VI) and chloride ions were abundance of aquatic fungi in the selected lakes of the Augustów Lakeland.

Table 1 Average value and standard deviations of the primary physico-chemical parameters of in the selected lakes of the Augustów Lakeland.

2 Lakes pH EC [μS/cm] DOC [mgC/L] POC [mgC/L] TP [μgP/L] DRP [μgP/L] SO4 [mg/L] Cl [mg/L]

Sajno (I) 7.5570.07 27476 3.7670.9 0.6970.10 72.070.6 40.877.8 20.770.7 10.070.2 Necko (II) 8.0270.05 345741 7.5870.3 0.9470.08 58.775.6 28.473.1 30.872.3 14.870.9 Rospuda (III) 8.3070.08 324741 11.8 70.7 3.3370.10 77.572.1 13.970.9 37.271.5 16.770.2 Białe Augustowskie (IV) 8.1870.03 33275 9.0870.8 1.2070.09 73.272.3 19.673.9 25.470.8 16.170.1 Długie Augustowskie (V) 8.7070.17 38673 24.171.9 5.2770.56 148715 10.5 78.0 63.573.1 18.870.3 Studzieniczne (VI) 7.7270.03 31674 6.5670.7 0.5070.10 51.5 72.9 31.570.3 23.970.4 11.270.2 Serwy (VII) 7.4070.05 24777 3.3470.8 0.2970.08 68.772.3 60.574.3 17.072.0 8.8270.9 Gorczyckie (VIII) 8.6370.13 35073 19.172.0 3.6870.12 88.174.1 12.976.5 42.972.1 17.070.1 Orle (IX) 7.91 70.08 293718 4.3571.7 0.5370.06 70.1713.4 50.775.9 21.970.5 10.570.2 Pobojno (X) 8.9070.13 442743 31.673.0 4.3670.12 188710 2.18 70.81 69.971.4 23.575.5 Paniewo (XI) 7.8470.03 30277 5.0470.5 0.5670.06 45.972.3 35.873.5 22.870.3 13.370.1 Mikaszewo (XII) 8.4270.17 37774 13.470.5 4.0870.04 96.672.7 8.4672.18 58.971.3 17.7 70.2 Mikaszówek (XIII) 8.4470.26 37073 16.470.8 3.8970.10 119712 11.0 70.6 52.675.2 17.370.1 Płaskie (XIV) 6.8470.31 23378 3.2971.2 0.16 70.03 41.8715.2 37.1716.8 11.471.8 5.19 70.2 A. Pietryczuk et al. / Ecotoxicology and Environmental Safety 109 (2014) 32–37 35

Table 2 Taxonomic diversity of aquatic fungi in the selected lakes of the Augustów Lakeland (accession numbers of the reference strains in GenBank are provided in brackets; study sites are as indicated in Fig. 1).

No Position species I II III IV V VI VII VIII IX X XI XII XIII XIV

1. Acremonium implicatum (JQ910163.1) xx x 2. Alternaria alternata (AY354228.1) xx x x x 3. Anguillospora crassa (AY148106.1) xxx xx x x xxx x x x 4. Anguillospora filiformis (JX089472.1) xxxxx 5. Arthroderma insingulare (AJ877213.1) xx 6. Arthroderma vanbreuseghemii (EU683894.1) xx x x 7. Aspergillus fumigatus (FJ867935.1) xxxxxx xxxx 8. Aspergillus niger (JQ929761.1) xxx x xxxx 9. Aspergillus tubingensis (KC796398.1) xxx x x xxx 10. Aureobasidium pullulans (DQ683019.1) xx x x xx 11. Candida albicans (EF192231.1) xxxxx 12. Chrysosporium queenslandicum (AB219228.1) xx x x x x x x 13. Cryptococcus laurenti (JN627015.2) xx x x x 14. Cryptococcus neoformans (KJ175193.1) xx x 15. Exophiala dermatitidis (KF996500.1) xx x x x x x x 16. Flagellospora curvula (KC834050.1) xxx xx x x xxx x x x 17. Fusarium oxysporum (KC999985.1) xx 18. Fusarium proliferatum (JQ693101.1) xxxx 19. Fusarium verticillioides (KF999013.1) xxxx 20. Geotrichum candidum (KF669518.1) xx x x x x x 21. Hansenula polymorpha (FJ914914.1) xx x x xx 22. Heliscus lugdunensis (AY148103.1) xxx xx x x xxx x x 23. Leptomitus lacteus (AF119597.1) xx 24. Lunulospora curvula (JX089535.1) xxx xx x x xxx x x 25. Microascus cinereus (HG380428.1) xx x x 26. Mucor fragilis (AF474242.1) xx x 27. Penicillium funiculosum (HQ637359.1) xxx x x 28. Rhizophus microsporus (HM999970.1) xxx 29. Rhodotorula rubra (AB916512.1) xx x x xx 30. Scopulariopsis brevicaulis (HG964310.1) xx x xx 31. Tetracladium breve (EU883431.1) xxxxxx 32. Tetracladium marchalianum (EU883423.1) xx x x x x x x 33. Torulopsis glabrata (KF747753.1) xx x x x x x x x x 34. Trichoderma harzianum (AF469188.1) xx x x x x x 35. Trichophyton violaceum (JQ322678.1) xx x x x x x x 36. Tricladium splendens (DQ202511.1) xxx xx x x xxx x x 37. Varicosporium delicatum (JQ412864.1) xxx xx x xxx x x 38. Varicosporium elodeae (JX981463.1) xxx xx x x xxx x x

studies have demonstrated that pH, conductivity, temperature, the concentration of , and particularly nitrogen, phosphorus and organic carbon are important factors affecting the abundance and of mycoplankton (Goh and Hyde, 1996; Krauss et al., 2011; Pascoal and Cassio, 2004; Pietryczuk et al., 2013a; 2013b; 2013c). Moreover, research regarding the effect of temperature on aquatic fungi showed the existence of species occurring through- out the year, and those typical of a particular season of the year (Sharma and Parveen, 2011). The highest abundance and species diversity of fungi among the studied lakes was recorded in Pobojno, Mikaszówek, and Gorczyckie Lake, particularly in autumn. This most likely results from the high concentration of organic matter due to leaf accumulation. The organic matter is mainly decomposed by aquatic hyphomycetes; leaves sub- merged in water serve as a habitat for their development and functioning (Krauss et al., 2011; Nikolcheva and Bärlocher et al., 2005; Pascoal et al., 2005). It also appears important that lakes with the highest abundance and species diversity of fungi accord- Fig. 3. The results of the cluster analysis (on the basis of physico-chemical ing to the cluster analysis (Fig. 3) belong to the group of eutrophic parameters) of the selected lakes of the Augustów Lakeland. lakes (Group I) with a large amount of organic matter. Moreover, they are usually shallow polymictic lakes. This can result in a 4. Discussion higher species diversity of fungi in these lakes because fungal species inhabiting bottom are supplied to the surface Studies on the abundance, species diversity, and ecological water layers. The research conductedalong the Palk Strait of function of yeast-like and filamentous fungi in the limnic waters of South India by Ravikumar et al. (2013) showed that bottom Poland and throughout the world have been exceptionally rare sediments contain a much higher abundance of fungi (100000- (Baldy et al., 2002; Niewolak et al., 2009). However, previous 1200000 CFU/mL) (105–12 105 CFU/mL) than . The 36 A. Pietryczuk et al. / Ecotoxicology and Environmental Safety 109 (2014) 32–37 reason for thisis that a number of fungi in the water are expected water. Moreover, in lakes with a high content of organic matter, the to colonise decaying plant material and the surface of bottom presence of potentially pathogenic fungi was also recorded and sediments. The research presented in this paper shows that included the following: Torulopsis glabrata, Geotrichum candidum, polymictic Płaskie Lake is an exception, where a lower number Candida albicans, Cryptococcus laurentis,andExophiala dermatitidis. of fungal species were identified. This may be because Płaskie Lake These species are allochthonous organisms and usually of anthro- is a mesotrophic lake without outflow. In the remaining meso- pogenic origin. The water is not their natural environ- trophic lakes, such as Necko, Białe Augustowskie, and Studzie- ment, and they do not participate in organic matter decomposition. niczne Lake (Fig. 3, Group II), the abundance of fungi and their Research has shown that there is a strong relationship between species diversity is also considerably lower. They are also deep aquatic fungi and organic matter (Fig. 4), which is responsible for lakes with clearly distinguishable temperature-oxidation layers the progressive eutrophication process of water (Dunalska, 2011), where bottom sediments have no contact with the epilimnion. resulting in algal blooms. On the one hand, there are species of Our study showed statistically significant correlations between fungi that have evolved mechanisms that allow them to function in the abundance of fungi and the dissolved organic carbon (DOC) eutrophic environments (Medeiros et al., 2009; Pascoal and Cásio, concentration (r¼0.88, po0.05) and the particulate organic carbon 2004; Solé et al., 2008). On the other hand, the negative correlation (POC) concentration (r¼0.81, po0.05) (Fig. 4). This finding suggests between the chlorophyll a concentration and the abundance of that it is a highly important parameter in determining the occur- aquatic fungi (r¼0.85, po0.001) may suggest a negative impact of rence of some fungal species and the complete disappearance of one group of organisms on another group. This may be confirmed other fungal species. The majority of dystrophic lakes rich in humic by negative correlation between the abundance of fungi and the substances and peat material, as well as those with a naturally dissolved reactive phosphorus—DRP (r¼0.87, po0.05). As noted in acidic pH, are distinguished by the high species diversity of fungi previous studies, high concentrations of phosphorus in the water belonging to the Chytridiales class (Thormann, 2006; Thormann increase the growth of (Carlson, 1977); however, as men- et al., 2007). Moreover, aquatic fungi, particularly those inhabiting tioned by Cudowski (2014),onlyDRPcanbedirectlyusedbythe sediments, are able to produce their own humic substances. They . In addition, the state of the biotic balance of the surface resemble humic substances naturally occurring in water, although water is expressed as a function of the pH and of they contain more aliphatic (carbohydrates, peptides) and fewer water (Neverova-Dziopak, 2006), as confirmed by the directly aromatic structures (Damare and Raghukumar, 2008). In earlier proportional correlation between the pH and fungal abundance studies (Pietryczuk et al., 2013a, 2013b, 2013c), the biomass of (r¼0.93, po0.01). In lakes with alkaline water (pH of approxi- aquatic fungi, which determinant is a ergosterol concentrations, mately 8.5–9.0), a higher abundance and species diversity of fungi varied from 0.1 to 6.1 μg/L and depended on the type of water and was recorded in comparison with the slightly acidic waters of the DOC and POC concentrations. In lakes with the highest DOC and Płaskie Lake and Serwy Lake. Moreover, in alkaline waters domi- POC concentrations, the highest number of fungal species was nated by allochthonous organisms, potentially pathogenic fungi identified. The majority of the species belonged to the aquatic such as Trichophyton violaceum, Scopulariopsis fusca, Exophiala hyphomycetes and included Varicosporium elodeae, Tricladium dermatitidis, Chrysosporium queenslandicum, Acremonium implica- splendens, Lunulospora curvula,andHeliscus lugdunensis.Their tum,orCandida albicans were recorded. These results support primary role is the decomposition of organic matter present in previous studies that found that the alkalisation of the environment favours the development of fungi that cause diseases in animals and humans (Vylkova et al., 2011). In addition to typically aquatic species, particularly belonging to the hyphomycetes, 14 species that are potentially pathogenic to animals and humans were identified in the waters of the studied lakes. We noted the presence of dermatophytes and yeast-like fungi with Candida albicans predominating. The highest taxonomic diversity of potentially pathogenic fungi, most likely of anthro- pogenic origin, was recorded in Sajno, Necko, Studzieniczne, Białe Augustowskie, Serwy, and Rospuda Lake. These lakes are inten- sively used for tourism in the summer season. High amounts of potentially pathogenic fungi were also recorded in lakes with increased concentrations of chloride (r¼0.80, po0.01) and sul- phate(VI) ions (r¼0.89, po0.01) and conductivity (r¼0.81, po0.05), i.e., parameters indicating water pollution (Fig. 4). Water pollution with chloride and sulphate(VI) ions has no substantial effect on the presence of aquatic hyphomycetes (Fig. 4). In Rospuda, Gorczyckie, Pobojno, Mikaszewo, and Mikaszówek Lake, where the highest concentrations of chloride and sulphate(VI) ions were recorded, the presence of Candida albicans and Rhodotorula Fig. 4. Diplot of redundancy analysis (RDA) results evaluating the relationships rubra was also recorded. According to the literature (Klaerner et between fungi of aquatic hyphomycetes (numbers correspond with the item al., 1997; Kosíková and Sláviková, 1996), these fungi are typical in numbers of a given species from Table 2) and environmental variables. The polluted waters. Moreover, in Gorczyckie and Pobojno Lake, the measured environmental variables and fungal species are shown as arrows. The occurrence of the sewage fungus Leptomitus lacteus, typical in vector orientations represent the direction of strongest change; vector lengths represent relative importance. The distribution and direction of the strongest waters polluted with organic compounds, was recorded. The abundance changes for a fungal species in response to the environment are shown fungus is probably supplied to the waters together with domestic by the corresponding species vectors (grey arrows). In the RDA analysis, a positive sewage from households and the camping site located in the correlation between two environmental factors is expressed by relatively long vicinity of the lakes. Therefore, aquatic fungi appear to be a vectors pointing approximately in the same direction, whereas a negative correla- reliable indicator of water quality. The high abundance of fungi, tion is indicated by arrows pointing in opposite directions. The longer the environmental cline, the stronger the relationship of that parameter with the particularly potentially pathogenic species, suggests pollution of community. Perpendicular arrows indicate no correlation. anthropogenic origin, such as sewage dumping. A. Pietryczuk et al. / Ecotoxicology and Environmental Safety 109 (2014) 32–37 37

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