Extracts with Nutritional Potential and Their Influence on the Rheological Properties of Dough and Quality Parameters of Bread.

Formulating basic food to improve its nutritional profile is one potential method for food innovation. One option in formulating basic food such as bread is to supplement flours with specified amounts of non-bakery raw materials with high nutritional benefits. In the research presented here, we studied the influence of the addition of curcumin and quercetin extracts in amounts of 2.5% and 5% to wheat flour (2.5:97.5; 5:95). The analysis of the rheological properties of dough was carried out using a Mixolab 2. A Rheofermentometer F4 was used to assess the dough's fermentation, and a Volscan was used to evaluate the baking trials. The effect of the extracts on the rheological properties of dough was measured and found to be statistically significant, with curcumin shortening both dough development time and dough stability. Doughs made with greater quantities of extract had a greater tendency to early starch retrogradation, which negatively affects the shelf life of the end products. The addition of extracts did not significantly affect either the ability to form gas during fermentation or its retention, which is important because this gas is prerequisite to forming a final product with the required volume and porosity of crumb. Less favourable results were found on sensory evaluation, wherein the trial bread was significantly worse than the control wheat bread. The panel's decision-making might have been influenced by the atypical colour of the bread made with additives, and in case of a trial bread made with quercetin, by a bitter taste. From the technological point of view, the results confirmed that the composite flours prepared with the addition of extracts of curcumin and quercetin in amounts of 2.5% and 5% can be processed according to standard procedures. The final product will be bread with improved nutritional profile and specific sensory properties, specifically an unconventional and attractive colour.

The Vapor Phase of Selected Essential Oils and Their Antifungal Activity In Vitro and In Situ against Penicillium commune, a Common Contaminant of Cheese.

This study aimed to determine the in vitro and in situ antifungal activity of (14) selected essential oils (EOS), namely clove, thyme, red thyme, litsea, eucalyptus, niaouli, fennel, anise, cumin, basil, rosemary, sage, bergamot mint, and marjoram, by vapor contact against the growth of two strains of Penicillium commune (KMi-183 and KMi-402). Furthermore, to exclude the negative effect of EOs on the lactic acid bacteria (LABs) (Streptococcus spp.) on cheeses, their influence was monitored. Next, the sensory evaluation of cheese treated by EOs was evaluated. The results show that litsea and clove EOs were the most effective in the vapor phase against both tested strains. These EOs were characterized by the highest amount of α- (40.00%) and ß-Citral (34.35%) in litsea and eugenol (85.23%) in clove. The antitoxicogenic activity of less effective (in growth inhibition) EOs on cyclopiazonic acid (CPA) production by the tested strains was also observed. The growth of Streptococcus spp. (ranging from 8.11 to 9.69 log CFU/g) was not affected by the EOs in treated cheese. Even though the evaluators recognized some EOs in sensory evaluation by the triangle test, they did not have a negative effect on the taste and smell of the treated cheeses and were evaluated as edible. The antifungal activity of EOs against several types of microscopic fungi and their effect on the sensory properties of treated foods needs to be further tested to achieve the most effective protection of foods from their direct contaminants.

Antifungal Activities of Essential Oils in Vapor Phase against Botrytis cinerea and Their Potential to Control Postharvest Strawberry Gray Mold.

Essential oils (EOs) from aromatic plants seem to have the potential to control several fungal pathogens and food contaminants. Botrytis cinerea is the main strawberry fruit contaminant causing high losses during storage. Here, thirteen EOs applied in the vapor phase were evaluated for their potential to inhibit the growth of three different strains of B. cinerea isolated from strawberry fruits. Eight EOs (lemongrass, litsea, lavender, peppermint, mint, petitgrain, sage, and thyme) were able to completely inhibit the growth of B. cinerea for 7 days when applied at a concentration of 625 µL·L-1. Four EOs with the lowest minimal inhibition concentrations (thyme, peppermint, lemongrass, and litsea) have been tested on strawberry fruits intentionally inoculated by B. cinerea. All four EOs showed high inhibition at a concentration of 250 or 500 µL·L-1, but only peppermint EO was able to completely inhibit B. cinerea lesion development at a concentration of 125 µL·L-1. The sensory evaluation of strawberries treated by EOs at a concentration 125 µL·L-1 resulted in a statistically significant decrease in taste, aftertaste, aroma, and overall quality. Lemongrass and litsea EOs scored better than thyme and peppermint ones, thus forming two viable methods for B. cinerea suppression and the extension of packed strawberries' shelf life.

Phenotypic and genotypic diversity of airborne fungal spores in Demänovská Ice Cave (Low Tatras, Slovakia).

This paper is the first aero-mycological report from Demänovská Ice Cave. Fungal spores were sampled from the internal and external air of the cave in June, 2014, using the impact method with a microbiological air sampler. Airborne fungi cultured on PDA medium were identified using a combination of classical phenotypic and molecular methods. Altogether, the presence of 18 different fungal spores, belonging to 3 phyla, 9 orders and 14 genera, was detected in the air of the cave. All of them were isolated from the indoor samples, and only 9 were obtained from the outdoor samples. Overall, airborne fungal spores belonging to the genus Cladosporium dominated in this study. However, the spores of Trametes hirsuta were most commonly found in the indoor air samples of the cave and the spores of C. herbarum in the outdoor air samples. On the other hand, the spores of Alternaria abundans, Arthrinium kogelbergense, Cryptococcus curvatus, Discosia sp., Fomes fomentarius, Microdochium seminicola and T. hirsuta were discovered for the first time in the air of natural and artificial underground sites. The external air of the cave contains more culturable airborne fungal spores (755 colony-forming units (CFU) per 1 m3 of air) than the internal air (from 47 to 273 CFU in 1 m3), and these levels of airborne spore concentration do not pose a threat to the health of tourists. Probably, the specific microclimate in the cave, including the constant presence of ice caps and low temperature, as well as the location and surrounding environment, contributes to the unique species composition of aeromycota and their spores in the cave. Thus, aero-mycological monitoring of underground sites seems to be very important for their ecosystems, and it may help reduce the risk of fungal infections in humans and other mammals that may arise in particular due to climate change.

The in vitro effect of selected essential oils on the growth and mycotoxin production of Aspergillus species.

The aim of the present study was to assess the antifungal and anti-toxinogenic activity of 15 essential oils (EOs) against three fungi of the genus Aspergillus (A. parasiticus KMi-227-LR, A. parasiticus KMi-220-LR and A. flavus KMi-202-LR). The minimum inhibitory doses (MIDs) of the tested essential oils and their antifungal activity were determined using the micro-atmosphere method. The original commercial essential oil samples of Jasminum officinale L., Thymus vulgaris L., Syzygium aromaticum (L.) Merrill & Perry, Rosmarinus officinalis L., Ocimum basilicum L., Eucalyptus globulus Labill., Salvia officinalis L., Citrus limon (L.) Burm, Origanum vulgare L., Lavandula angustifolia Mill., Carum carvi L., Citrus sinensis (L.) Osbeck., Zingiber officinalis Rosc., Mentha piperita L. and Cinnamomum zeylanicum Nees. (C. verum J.S.Presl.) were produced in Slovakia (Calendula a.s., Nová Lubovna, Slovakia). All essential oils exhibited activity against all tested strains of fungi. After 14 days of incubation, A. flavus (KMi-202-LR) showed the highest susceptibility with a growth inhibition percentage (GIP) of 18.70% to C. limon and 5.92% to C. sinensis, while A. parasiticus (KMi-220-LR) exhibited a GIP of 20.56% to J. officinale. The minimum inhibitory doses (MIDs) of EOs with the most significant activity were recorded. The best antifungal activity, using the micro-atmosphere method was found in S. aromaticum with an MID of 62.5 µL L-1 air, T. vulgaris (MID of 62.5 µL L-1 air) and O. vulgare (MID of 31.5 µL L-1 air) against all tested strains. Mycotoxin production of the tested strains was evaluated by the thin layer chromatography (TLC) method. Mycotoxin production of AFB1 and AFG1 was inhibited following all treatments with C. carvi, R. officinale and S. officinale, Eucalyptus globulus L. and O. basilicum L. Essential oils exhibited a potential inhibition activity against toxic fungi, although, these affected only the production of AFB1.

Mycobiota of Underground Habitats: Case Study of Harmanecká Cave in Slovakia.

Harmanecká Cave is located in the Harmanec Valley to the northwest of Banská Bystrica city, in the southern part of the Great Fatra Mountains, Slovakia. This cave is the most important underground locality of bat occurrence in Slovakia (population of 1000 to 1500 individuals). The study aimed at mycological evaluation of the air, the water, and the rock surface of Harmanecká Cave in Slovakia. The samples were taken on 24 July 2014. To examine the air, the Air Ideal 3P sampler was used. Microbiological evaluation of the rock surface was performed using swab sampling and the water by using the serial dilution technique. The authors observed a relationship between air temperature and the concentration of fungi. The concentration of airborne fungi increased with the increase in the air temperature and decreased with distance from the entrance to the cave. The density of airborne fungi isolated from the outdoor air samples was 810.5 colony-forming units (CFU) per 1 m3 of air and from 27.4 to 128.5 CFU for the indoor air samples. From the rock surface inside the cave, 45.0 to 106.6 CFU per 1 cm2 were isolated, whereas from the water, 29.9 CFU per 1 ml were isolated. Seven species of filamentous fungi were isolated from the external air samples and 12 species of filamentous fungi and 3 species of yeast-like fungi from the internal air samples. From the surface of the rocks inside the cave, 5 species of filamentous fungi and 1 species of yeast-like fungi were cultured, whereas from the water samples, 6 species of filamentous fungi were cultured. Cladosporium spp. were the fungi most frequently isolated from the external air; from the internal air, Penicillium urticae was most frequently isolated; from the rock surface, it was Gliocladium roseum; and from the water, it was P. chrysogenum. The species found in the cave can be pathogenic for humans and animals, especially for immunocompromised persons, and they can also cause biodegradation of the rocks. However, the concentration of airborne fungi inside the cave did not exceed official limits and norms stated as dangerous for the health of tourists.

Fungi on wheat bran and their toxinogenity.

To survey the overall fungal contamination and toxicological potential of isolated fungi, a mycological investigation of 56 samples of wheat bran was carried out during March-June 2004 in Slovakia. Despite of low values of water activity found in the samples reaching an average of 0.54, the individual samples were contaminated with fungi in a range from 1.82 x 10(1) - 3.42 x 10(4) colonies forming units per gram of sample (CFU/g). A total number of 65 fungal species pertaining to 23 genera were recovered. The most dominant and diverse genus was Penicillium (20 spp.) followed by Aspergillus (10 spp.) and Cladosporium (3 spp.) with 100 percentages, 89 percentages and 72 percentages frequency, respectively. Isolates of potentially toxinogenic species recovered from the samples were found to produce various mycotoxins, namely citrinin (23 isolates), cyclopiazonic acid (43 isolates), griseofulvin (23 isolates), ochratoxin A (14 isolates), patulin (30 isolates), penitrem A (18 isolates) and sterigmatocystin (7 isolates). Furthermore, fumonisin B1 as well as moniliformin producing Fusarium strains, were recovered. It is evident that this type of cereal commodity is a significant source of toxicologically relevant species of which majority are true toxinogenic.

Fungi recovered from Slovakian poultry feed mixtures and their toxinogenity.

To contribute towards the knowledge of microbiology of feeds, more than 100 samples of poultry feed mixtures from Slovakia were mycologically investigated in terms of the overall fungal diversity and toxicological potential of isolated fungi. The study revealed that out of 22 genera recovered, Penicillium was the most frequent and diverse genus, followed by Aspergillus and Mucor being found in 89 % (34 spp.), 69 % (11 spp.) and 50 % (4 spp.), respectively. The most frequently encountered taxa were Fusarium proliferatum, followed by Penicillium aurantiogriseum, Mucor racemosus, Penicillium crustosum and Aspergillus flavus . In addition, the following genera were recorded (in descending order) Rhizopus (44 %, 3 spp.), Eurotium (42 %, 5 spp.), Fusarium (42 %, 3 spp.), Cladosporium (31 %, 1 sp.), Alternaria (22 %, 3 spp.), Absidia (16 %, 3 spp.), Acremonium (12 %, 2 spp.), Scopulariopsis (10 %, 2 spp.), Paecilomyces (4 %, 1 sp.), Ulocladium (3 %, 1 sp.), Trichoderma (2 %, 1 sp.), Zygorrhynchus (2 %, 1 sp.), and finally Emericella, Epicoccum, Geosmithia, Monascus, Stachybotrys, Syncephalastrum and Wardomyces , all were encountered in 1 % of the samples being represented by a single species. The mean value counts of total fungi ranged from 1 x 10 (3) to 200 x 10(5) cfu/g. Outcomes from mycotoxin screening within the appropriate potentially toxinogenic species showed a number of mycotoxin producers, namely those forming aflatoxin B (1) (n=3), citrinin (17), cyclopiazonic acid (76), fumonisin B(1) (86), griseofulvin (42), moniliformin (18), ochratoxin A (5), patulin (56), penitrem A (30) and sterigmatocystin (10).

Incidence of trichothecenes and zearalenone in poultry feed mixtures from Slovakia.

A total of 50 samples of poultry feed mixtures of Slovakian origin were analyzed for eight toxicologically significant Fusarium mycotoxins, namely zearalenone (ZON), A-trichothecenes: diacetoxyscirpenol (DAS), T-2 toxin (T-2) and HT-2 toxin (HT-2) and B-trichothecenes: deoxynivalenol (DON), 3-acetyl-deoxynivalenol (3-ADON), 15-acetyl-deoxynivalenol (15-ADON) and nivalenol (NIV). The A-trichothecenes and the B-trichothecenes were detected by means of high pressure liquid chromatography with tandem mass spectrometry detection (HPLC-MS/MS) and gas chromatography electron capture detection (GC-ECD), respectively. Reversed phase-high performance liquid chromatography with a fluorescence detector (RP-HPLC-FLD) was used for ZON detection. The most frequent mycotoxin detected was T-2, which was found in 45 samples (90%) in relatively low concentrations ranging from 1 to 130 microg kg(-1) (average 13 microg kg(-1)), followed by ZON that was found in 44 samples (88%) in concentrations ranging from 3 to 86 microg kg(-1) (average 21 microg kg(-1)). HT-2 and DON were detected in 38 (76%) and 28 (56%) samples, respectively, in concentrations of 2 to 173 (average 18 microg kg(-1)) for HT-2 and 64 to 1230 microg kg(-1) sample (average 303 microg kg(-1)) for DON. The acetyl-derivatives of DON were in just four samples, while NIV was not detected in any of the samples investigated. In as many as 22 samples (44%), a combination of four simultaneously co-occurring mycotoxins, i.e. T-2, HT-2, ZON and DON, was revealed. Despite the limited number of samples investigated during this study poultry feed mixtures may represent a risk from a toxicological point of view and should be regarded as a potential source of the Fusarium mycotoxins in Central Europe. This is the first reported study dealing with zearalenone and trichothecene contamination of poultry mixed feeds from Slovakia.

Incidence of fumonisins, moniliformin and Fusarium species in poultry feed mixtures from Slovakia.

A total of 50 samples of poultry feed mixtures of Slovak origin were analysed for fumonisin B(1) and B(2) (FB(1), FB(2)) and moniliformin (MON) using SAX-clean up procedure being detected by high pressure liquid chromatography with mass spectrometry (HPLC-MS) and diode array detection (HPLC-DAD), respectively. The samples were also simultaneously investigated for Fusarium species occurrence, and for the capability of Fusarium isolates recovered to produce FB(1) and MON in vitro. FB1 was detected in 49 samples (98 %) in concentrations ranging from 43 to 798 microg x kg(-1), and FB(2) in 42 samples (84 %) in concentrations ranging from 26 to 362 microg x kg(-1). MON was detected in 26 samples (52 %) in concentrations that ranged from 42 to 1,214 microg x kg(-1). Only two Fusarium populations were encountered, namely F. proliferatum and F. subglutinans, of which the former was the most dominant and frequent. All 86 F. proliferatum isolates tested for FB1-production ability proved to be producers of the toxin although none of them produced MON. On the contrary, MON production was observed in a half out of 16 F. subglutinans isolates tested, yet no FB1 production was detected in this case. Despite the limited number of samples investigated during this study, it is obvious that poultry feed mixtures may represent a risk from a toxicological point of view and should be regarded as a potential source of the Fusarium mycotoxins in central Europe. This is the first reported study dealing with fumonisin and moniliformin contamination of poultry feeds from Slovakia.

Mycological survey of ripped service tree fruits (Sorbus domestica L.) with an emphasis on toxinogenic fungi.

To investigate a possible incidence of microscopic fungi in ripped service tree (Sorbus domestica L.) fruits, a mycological survey was carried out during October-November 2003 in Slovakia. This rare kind of fruit is renowned for its significant curative actions in various human and animal diseases. The study revealed that all 24 surface sterilized fruits analysed were contaminated with fungi. The most dominant endogenous contaminant encountered was Cladosporium cladosporioides (Fres.) de Vries followed by Alternaria alternata (Fr.) Keissler and Penicillium expansum Link with 88%, 63% and 54% frequency, respectively. Furthermore, 24 other fungal species were associated with the fruits as well. P. expansum (40), P. carneum (Frisvad) Frisvad/Penicillium paneum Frisvad (35) and P. griseofulvum Dierckx (6) isolates recovered from the fruits were screened by an agar plug method for production of mycotoxin patulin, all with positive results. In addition, citrinin, griseofulvin and zearalenon production by appropriate species isolates were detected. Despite of a limited number of samples of the service tree fruits examined during this study, it was concluded that the ripening of service tree fruits is accompanied with the presence of typical rotting-fungi as well as of some others, which have not previously been reported in connection with fruit. Furthermore, it was suggested that the ripped service tree fruits should be considered as a potential source of significant fungal secondary metabolites including mycotoxins. Secondary metabolite profiles of the species identified during the study were included here.

Identification and enumeration of Fusarium species in poultry feed mixtures from Slovakia.

Thirty-three samples of poultry mixed feeds collected in the region of Nitra (Slovakia) from August 2001-April 2002 were assayed for the incidence of Fusarium Link: Fr. species. In tested samples, the total frequency of isolated fusaria was determined to be 48 % (16 samples) and their counts ranged from 0.2 x 10(2) to 2.4 x 10( 4 ) CFU per g of the sample on Dichloran Chloramphenicol Peptone Agar (DCPA). Of the total amount of Fusarium isolates (609), the highest part (i.e. 584 isolates) was represented by Fusarium proliferatum (Matsushima) Nirenberg, being isolated in all the samples tested. Fusarium subglutinans (Wollenw. et Reinking) Nelson, Tousson et Marasas (in total 24 isolates) was found in 3 positive samples (9%) and Fusarium oxysporum Schlecht. Fr. (a single isolate) was found in one positive sample only (3%). Data of these significant mycotoxin producers found in examined samples with particular mycotoxins as well as their habitats were summarised briefly. The results refer to a large incidence of the potentially toxinogenic Fusarium species, mainly F. proliferatum, in the feeds of Slovakian origin which represent entry components of the food chain. The study also points out a potential risk of feed contamination with hazardous toxic compounds, especially by carcinogenic fumonisins and cardiotoxic moniliformin. From the hygienic point of view, it will be necessary to continue monitoring and evaluating this occurrence.