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3. Isolation, antimicrobial activity of myxobacterial crude extracts and identification of the most potent strains

Author

Charousová Ivana, Medo Juraj, and Javoreková Soňa

Subjects
Myxococcus sp., Corallococcus sp., antimicrobial activity, multidrug-resistant organisms, Biology (General), QH301-705.5
Abstract

Broad spectrum antimicrobial agents are urgently needed to fight frequently occurring multidrug-resistant pathogens. Myxobacteria have been regarded as “microbe factories” for active secondary metabolites, and therefore, this study was performed to isolate two bacteriolytic genera of myxobacteria, Myxococcus sp. and Corallococcus sp., from 10 soil/sand samples using two conventional methods followed by purification with the aim of determining the antimicrobial activity of methanol extracts against 11 test microorganisms (four Gram-positive, four Gram-negative, two yeasts and one fungus). Out of thirty-nine directly observed strains, 23 were purified and analyzed for antimicrobial activities. Based on the broth microdilution method, a total of 19 crude extracts showed antimicrobial activity. The range of inhibited wells was more important in the case of anti-Gram-positive-bacterial activity in comparison with the anti-Gram-negative-bacterial and antifungal activity. In light of the established degree and range of antimicrobial activity, two of the most active isolates (BNEM1 and SFEC2) were selected for further characterization. Morphological parameters and a sequence similarity search by BLAST revealed that they showed 99% sequence similarity to Myxococcus xanthus − BNEM1 (accession no. KX669224) and Corallococcus coralloides - SFEC2 (accession no. KX669225). As these isolates had antimicrobial activity, they could be considered for use in the development of antibiotics for pharmaceutical use.

Published
2017
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5. Antifungal and synergistic activities of some selected essential oils on the growth of significant indoor fungi of the genusAspergillus

Author

Juraj Čuboň, Lukáš Hleba, Viktória Uzsáková, Miroslava Hlebová, Charousová Ivana, Juraj Medo, Anton Kováčik, Pavel Klouček, and Matej Božik

Subjects
Antifungal, Aspergillus, Fungal growth, Antifungal Agents, Environmental Engineering, biology, medicine.drug_class, Chemistry, Broth microdilution, Fungi, Microbial Sensitivity Tests, General Medicine, biology.organism_classification, Gas Chromatography-Mass Spectrometry, law.invention, Fungicide, law, Oils, Volatile, medicine, Flame ionization detector, Gas chromatography, Food science
Abstract

The study aimed to assess the antifungal activity of twenty-five essential oils (EOs) and the potential synergistic activity of the most effective EOs against significant indoor fungi of the genus Aspergillus [A. fumigatus (KBio-122), A. flavus (KBio-134), A. terreus (KBio-145) and A. niger (KBio-202)]. The chemical composition of all EOs was evaluated by the gas chromatography coupled with mass spectrometry (GC/MS) and gas chromatography with flame ionization detector (GC-FID) analysis. The antifungal susceptibility of EOs was evaluated by using the broth microdilution method. The most effective EOs were selected to determine the minimum inhibitory concentrations (MICs) and minimum fungicidal concentrations (MFCs) at a concentration range from 256 to 0.125 μg/mL. For the synergistic activities, the most effective EOs were tested using the chessboard pattern. The most sensitive strain to treatments with essential oils alone and in the combination of EOs was A. flavus (KBio-134). The chessboard assay showed that combinations of lemongrass and thyme EOs proved the most potent synergistic antifungal activity (FICI = 0.1875) against A. fumigatus (KBio-122). The synergy displayed by a combination of some EOs may be used to control fungal growth or increasing resistance to available synthetic antifungals, consequently permitting the reduction of their most active doses.

Published
2021

7. THE IN VITRO ANTIFUNGAL AND SYNERGISTIC ACTIVITY OF SELECTED PLANT EXTRACTS AND ESSENTIAL OILS AGAINST CANDIDA SPP.

Author

Hlebová, Miroslava, Vešeléniyová, Dominika, Šramková, Zuzana, Vojčíková, Veronika, Kováčik, Anton, Čuboň, Juraj, Charousová, Ivana, Hutárová, Lenka, and Hleba, Lukáš

Subjects
GAS chromatography/Mass spectrometry (GC-MS), PLANT extracts, ESSENTIAL oils, FLAME ionization detectors, ANTIFUNGAL agents, CANDIDA, RED clover
Abstract

This study aimed to evaluate the antifungal activity and the potential synergistic activity of ten plant extracts (EXs) and ten essential oils (EOs) obtained from the same plant species, namely Agrimonia eupatoria L., Arctium lappa L., Arnica montana L., Bidens cernua L., Corylus avellana L., Echinacea purpurea L. (Moench), Menta piperita L., Trifolium pratense L., Syzygium aromaticum L., and Angelica sinensis (Oliv.) Diels were used. Three species of the genus Candida, namely C. albicans, C. glabrata, and C. tropicalis, isolated from polluted estuarine water, were used in this study. The chemical composition of all EOs was evaluated by gas chromatography coupled with mass spectrometry (GC–MS) and gas chromatography with flame ionization detector (GC–FID) analysis. Firstly, the susceptibility of Candida spp. strains to EXs and EOs was screened by the agar disc diffusion method. Then the minimum inhibitory concentration (MIC) of the tested EXs and EOs was determined in a concentration range from 256 to 0.125 µL/mL. Following that, the effective EXs and EOs were divided into two groups (strong effect and weak effect) and tested in combination (strong + weak) at various concentrations depending on the tested strain to determine their potential synergistic effect using the chessboard pattern. The most sensitive strain to treatments with EXs and EOs tested alone and in mixtures was C. albicans. The most potent combinations with other EXs/EOs were observed with Syzygium aromaticum L., for both plant extract and essential oil, respectively. In this study the EOs and EXs tested in combination were much more effective than when acting on the growth of Candida spp. separately. So, the synergistic effect of the combinations of the EXs and EOs can lead to a reduction in the doses of commercial antifungals and can provide an effective way to reduce the infections caused by yeasts of the genus Candida. [ABSTRACT FROM AUTHOR]

Published
2023
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10. THE IN VITRO ANTAGONISTIC EFFECTS OF SOME BACILLUS SPP. ON THE GROWTH AND MYCOTOXIN PRODUCTION OF TOXIGENIC ASPERGILLI.

Author

Hlebová, Miroslava, Uzsáková, Viktória, Charousová, Ivana, and Hleba, Lukáš

Subjects
TOXIGENIC fungi, POSTHARVEST diseases, MICROFUNGI, FILAMENTOUS fungi, FOOD contamination, ASPERGILLUS flavus, FUNGAL cultures, TRICHODERMA
Abstract

This study aimed to investigate the antagonistic effect of three strains of the genus Bacillus on toxigenic fungi contaminating food commodities, with emphasis on two ochratoxigenic isolates (A. ochraceus, A. westerdijkiae) and two aflatoxigenic isolates (A. flavus and A. parasiticus). In vitro studies were carried out using two different methods for cultivation tested bacilli with fungal isolates - coincubation and dual culture method. The most sensitive isolate was A. ochraceus by both used methods. B. mycoides (5.72 ± 6.4 mm) and B. subtilis (5.08 ± 2.84 mm) were able to inhibit its growth and sporulation during ten days of cultivation (both inhibited the sporulation of A. ochraceus 100%) in coincubation. The most effective in the dual culture method were B. subtilis and B. thuringiensis against A. ochraceus (growth inhibition rate 84.40%; 90.55%) and A. flavus (growth inhibition rate 91.54%; 92.43%). The most effective sporulation inhibitors were B. subtilis and B. thuringiensis, which completely inhibited the sporulation of A. ochraceus and A. parasiticus after ten days of coincubation. Likewise, all tested bacterial strains showed complete inhibition of ochratoxin A synthesis in A. ochraceus and A. westerdijkiae exposure to bacterial volatiles. So, the current study illustrated that strains of the genus Bacillus could significantly inhibit the growth, sporulation, and mycotoxin production of toxigenic aspergilli and showing the great potential as a biocontrol agent of preand post-harvest food diseases caused by microscopic filamentous fungi. [ABSTRACT FROM AUTHOR]

Published
2021
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15. Mineral Nitrogen Retention, Nitrogen Availability and Plant Growth in the Soil Influenced by Addition of Organic and Mineral Fertilizers – Lysimetric Experiment

Author

Lukáš Plošek, Hynšt, Jaroslav, Záhora, Jaroslav, Elbl, Jakub, Kintl, Antonín, Charousová, Ivana, and Kovácsová, Silvia

Subjects
Biomass production, Nitrogen, fungi, food and beverages, Compost, complex mixtures, Lysimeter
Abstract

Compost can influence soil fertility and plant health. At the same time compost can play an important role in the nitrogen cycle and it can influence leaching of mineral nitrogen from soil to underground water. This paper deals with the influence of compost addition and mineral nitrogen fertilizer on leaching of mineral nitrogen, nitrogen availability in microbial biomass and plant biomass production in the lysimetric experiment. Twenty one lysimeters were filed with topsoil and subsoil collected in the area of protection zone of underground source of drinking water - Březová nad Svitavou. The highest leaching of mineral nitrogen was detected in the variant fertilized only mineral nitrogen fertilizer (624.58 mg m-2), the lowest leaching was recorded in the variant with high addition of compost (315.51 mg m-2). On the other hand, losses of mineral nitrogen are not in connection with the losses of available form of nitrogen in microbial biomass. Because lost of mineral nitrogen was detected in variant with the least change in the availability of N in microbial biomass. The leaching of mineral nitrogen, yields as well as the results concerning nitrogen availability from the first year of long term experiment suggest that compost can positive influence the leaching of nitrogen into underground water., {"references":["F. Amlinger, B. Götz, P. Dreher, J. Geszti and Ch. Weissteiner, \"Nitrogen in biowaste and yard waste compost: dynamics of mobilisation and availability—a review\", Europien Journal of Soil Biology, vol. 39, pp. 107-116, 2003.","D. Binkley, S. C. Hart, \"The Components of Nitrogen Availability Assessments in Forest Soils\", Advances in Soil Science, vol. 10, pp. 57-112, 1989.","L. G. Bundy and J. J. Meisinger, \"Nitrogen availability indice\", in Methods of soil analysis. Part 2. Microbiological and biochemical properties, R. W. Weaver, S. Angle, P. Bottomley, D. Bezdicek, S. Smith, A. Tabatabai, A.Wollum, Eds. Madison: Soil Society of Amerika, 1994, pp. 951-984.","A. Cellier, C. Francou, S. Houot, Ch. Ballini, T. Gauquelin and V. Baldy. \"Use of urban composts for the regeneration of a burnt Mediterranean soil: A laboratory approach\", Journal of Environmental Management, vol. 95, pp. 238-244, 2012.","P. Cosser (Ed.), Nutrients in Marine and Estuarine Environments. Australia: State of the Environment Technical Paper Series (Estuaries and the Sea), Central Queensland University, Canberra. 1997. T.","D´Hose, M. Cougnon, A. De Vliegher, B. Vandecasteele, N. Viaene, W. Cornelis, E. Van Bockstaele and D. Reheul, \"The positive relationship between soil quality and crop production: A case study on the effect of farm compost application\", Applied Soil Ecology, vol. 75, pp. 189-198, 2014.","L. F. Diaz, M. de Bertoldi, W. Bidlingmaier, E. Stentiford, Compost science and technology. Boston: MA Elsevier, 2007.","J. Elbl, J. K. Friedel, J. Záhora, L. Plošek, A. Kintl, J. Přichystalová, J. Hynšt, L. Dostálová and. K. Zákoutská, \"Leaching of Mineral Nitrogen and Phosphate from Rhizosphere Soil Stressed by Drought and Intensive Rainfall\", International Journal of Agricultural Biosystems Science and Engineering, vol. 7, no. 11, pp. 182-187, 2013.","E. Erhart, W. Hartl and B. Putz, \"Biowaste compost affects yield, nitrogen supply during the vegetation period and crop quality of agricultural crops\", European Journal of Agronomy, vol. 23, pp. 305-314, 2005.\n[10]\tE. Erhart, F. Feichtinger, W. Hartl, \"Nitrogen leaching losses under crops fertilized with biowaste compost compared with mineral fertilization\", Journal of Plant Nutrition and Soil Science, vol. 170, pp. 608-614, 2007.\n[11]\tJ. G. Fuchs, A. Berner, J. Mayer, E. Smidt and K. Schleiss, \"Influence of compost and digestates on plant growth and health: potentials and limits\", in Proceedings of the international congress CODIS 2008, Frick, pp.101-110.\n[12]\tJ. Galloway and E. B. Cowling, \"Reactive nitrogen and the world\", A Journal of the Human Environment, vol. 31, no. 2, pp. 64-71, 2002.\n[13]\tM. Chalhoub, P. Garnier, Y. Coquet, B. Mary, F. Lafolie and S. Houot, \"Increased nitrogen availability in soil after repeated compost applications: Use of the PASTIS model to separate short and long-term effects\", Soil Biology & Biochemistry, vol. 65, pp. 144-157, 2013.\n[14]\tA. Kintl, J. Hynst, J. Zahora, J. Elbl, L. Plosek, L. Halada, I. Tůma, F. Kohút, \"Contrasting effect of nitrogen and phosphorus addition on soil microbial activities at alpine meadow\", in Proceedings of International Masaryk conference for Ph.D. students and young researchers, Hradec Králové, pp. 3693-3702, 2012.\n[15]\t F. Kirschenmann, Alternative agriculture in an energy- and resource-depleting future. Reneable. Agriculture Food Systems, vol. 25, pp. 85–89. 2010.\n[16]\tM. B. Peoples, A. W. Faizah, B. Rerkasem and D. F. Herridge, Methods for evaluating nitrogen fixation by modulated legumes in the field. Canberra: Australian Centre for International Agricultural Research, 1989.\n[17]\tL. Plošek, F. Nsanganwimana, B. Pourrut, J. Elbl, J. Hynšt, A. Kintl, D. Kubná and J. Záhora, \"The Effect oc Compost Addition on Chemical and Nitrogen Characteristics, Respiration Activity and Biomass Production in Prepared Reclamation Substrates\", International Journal of Environmental, Earth Science and Engineering, vol. 7, no. 11, pp. 167-172, 2013.\n[18]\tC. Romero, P. Ramos, C. Costa and M. C. Márquez, \"Raw and digested municipal waste compost leachate as potential fertilizer: comparison with a commercial fertilizer\", Journal of Cleaner Production, vol. 59, pp. 73-78, 2013.\n[19]\tM. A. Sutton, The European nitrogen assessment: sources, effects and policy perspectives. New York: Cambridge University Press, 2011, cha. 1, 5. \n[20]\tM. Tejada, I. Gomez, T. Hernandez and C. Garcia,\"Utilization of Vermicompost in Soil Restoration: Effect on Soil Biological Properties\", Soil Science Society of America Journal, vol. 74, pp. 525-532, 2010.\n[21]\tS. M. Vaughan, R. C. Dalal, S. M. Harper and N. W. Menzies, \"Effect of fresh green waste and green waste compost on mineral nitrogen, nitrous oxide and carbon dioxide from a Vertisol\", Waste Management, vol. 31, pp. 1720-1728, 2011.\n[22]\tP. M. Vitousek, J. D. Aber, R. W. Howarth, G. E. Likens, P. A. Matson, D. W. Schindler, W. H. Schlesinger, G. D. Tilman, 1997. \"Human alteration of the global nitrogen cycle: Sources and consequences\", Ecological Applications, vol. 7, pp. 737-750, 1997\n[23]\tJ. Záhora and L. Mejzlík, \"The leaching of mineral nitrogen into underground water from soil environment of different ecosystems\", Ekológia Travného Porastu, no. 7, pp. 170-174, 2007."]}

Published
2014
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23. Effect of vermicompost on changes in the bacterial community in maize rhizosphere.

Author

HALENÁROVÁ, Eva, MEDO, Juraj, KOVÁCSOVÁ, Silvia, CHAROUSOVÁ, Ivana, MAKOVÁ, Jana, ELBL, Jakub, ZÁHORA, Jaroslav, and JAVOREKOVÁ, Soňa

Subjects
VERMICOMPOSTING, BACTERIAL communities, RHIZOSPHERE, FERTILIZERS for corn, POLYMERASE chain reaction
Abstract

Copyright of Journal of Central European Agriculture is the property of Journal of Central European Agriculture and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Published
2016
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24. Effect of fertilization on biological activity of community of soil streptomycetes.

Author

CHAROUSOVÁ, Ivana, MEDO, Juraj, HALENÁROVÁ, Eva, MAKOVÁ, Jana, and JAVOREKOVÁ, Soňa

Subjects
FERTILIZATION (Biology), SOIL fertility, ORGANIC fertilizers, CLAVIBACTER michiganensis, ACID phosphatase, PHOSPHATASES, BIOPESTICIDES, ANTIFUNGAL agents
Abstract

Copyright of Journal of Central European Agriculture is the property of Journal of Central European Agriculture and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Published
2016
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25. EFFECT OF BIOFERTILIZERS ON MICROBIAL COMMUNITY OF ARABLE SOIL.

Author

Charousová, Ivana, Javoreková, Soňa, Medo, Juraj, Maková, Jana, and Kovácsová, Silvia

Subjects
*SOIL science, *SOIL testing, *BIODIVERSITY, *POLYMERASE chain reaction, *BIOFERTILIZERS
Abstract

Three different organic substrates (compost, bottom sediment and biosludge) were applied to the arable soil in the laboratory conditions (incubation time 105 days at 25°C) in doses of 20 t.ha-1 and 80 t.ha-1. The main objective of the present study was to observe the changes in numbers of bacterial and fungal colonies using classic plate dilution method, changes in genetic microbial biodiversity using Polymerase Chain Reaction - Denaturing Gradient Gel Electrophoresis (PCR-DGGE) and changes in the microbial enzymatic activity (dehydrogenase, FDA - hydrolase and phosphatase activity) at the beginning (1st day) and at the end (105th day) of the experiment. The statistically significant differences in amount of bacteria (ANOVA, Tukey test, P ≤ 0.05) were mostly demonstrated on the 1st day of the experiment. All three tested substrate inputs rapidly increased percentage representation of the monitored bacteria. In case of microscopic fungi, the most significant changes were determined after the addition of bottom sediment, which in both used doses increased number of fungal colonies, particularly the number representation of microscopic fungus Penicillium simplicissimum at the end of the experiment. According to the results of the cluster analysis of PCR-DGGE, the diversity of bacteria from enriched soil was at least influenced by the addition 20 as well as 80 t.ha-1 of compost (similarity from 48 to 62 %). On the other hand, the addition of biosludge (both doses) influenced bacterial diversity at the most (similarity 33 %). In case of microscopic fungi, all three tested substrates affected fungal diversity approximately equally (similarity from 30 to 50 %). According to the results of regression analysis, P< 0.05, high relative abundance of bacteria as well as fungi determined by classic plate dilution method positively correlated with microbial diversity determined by PCR-DGGE and also with dehydrogenase activity. [ABSTRACT FROM AUTHOR]

Published
2014

26. Antimicrobial and enzymatic activity of actinomycetes isolated from soils of coastal islands.

Author

Charousová I, Medo J, Halenárová E, and Javoreková S

Abstract

Microbiological investigation of unexplored ecosystems is crucial for discovering of antibiotic producing actinomycetes. The present study was conducted to determine antimicrobial activity and identify the most active strains. Actinomycetes were isolated using the spread plate technique following by serial dilution of samples on starch casein agar. The screening method consists of primary and secondary testing. The most active isolates were identified based on molecular and cultural methods. 42 out of 66 isolates displayed antimicrobial potential. 63% exhibited antibacterial activity, 16% antifungal activity, and 16% displayed both activities. Identified isolates, Streptomyces scabrisporus , Streptomyces sparsogenes , Streptomyces misakiensis , Streptomyces cirratus , Streptomyces lincolnensis , Streptomyces endophyticus , Streptomyces chartreusis , and Streptomyces alboniger showed a broad spectrum of enzymatic activities. The results indicated that these isolates may serve as antibiotic and enzyme-producing microbes., Competing Interests: There are no conflicts of interest.

Published
2017
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