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Abstract

Yeast infections such as otitis externa and seborrheic dermatitis in dogs and cats are frequently associated with Malassezia pachydermatis secondary infection. It is part of the normal cutaneous microflora of most warm-blooded vertebrates, however, under certain conditions, it can become a causative agent of infection that needs to be treated pharmacologically. Azole derivatives are the drugs of the first choice. An interesting trend in developing resistance is the use of natural substances, which include manuka honey with confirmed antimicrobial properties. The main intention of this research was to evaluate the mutual effect of manuka honey in combination with four conventional azole antifungals – clotrimazole, fluconazole, itraconazole, and miconazole – on 14 Malassezia pachydermatis isolates obtained from dogs and 1 reference strain. A slightly modified M27–A3 method (CLSI 2008) and the checkerboard test (Nikolić et al. 2017) were used for this purpose. Our results show an additive effect of all 4 antifungals with manuka honey concurrent use. Based on the determined values of fractional inhibitory concentration index (FICI – 0.74±0.03 when manuka honey combined with clotrimazole, 0.96±0.08 with fluconazole, 1.0±0 with miconazole and 1.16±0.26 with itraconazole), it was found in all cases that the effect of substances used is more pronounced in mutual combination than when used separately.
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Authors and Affiliations

P. Váczi
1
E. Čonková
1
Z. Malinovská
1

  1. Department of Pharmacology and Toxicology, University of Veterinary Medicine and Pharmacy, Komenského 73, 041 81 Košice, Slovakia
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Abstract

A current problem in candidiasis treatment is increasing resistance to azoles, which are often prescribed to patients. The study underlines the high resistance of yeasts to fluconazole, which achieved high MIC (minimal inhibitory concentration) values both alone and in combination with essential oils (EOs). Antifungal activity of Hyssopus officinalis, Thymus vulgaris, Salvia officinalis and Rosmarinus officinalis EOs was determined against 13 clinical isolates of Candida albicans and reference strain Candida albicans ATCC 10231. The synergistic effect was investigated for the combination of itraconazole and fluconazole with Hyssopus officinalis and Thymus vulgaris EOs. Based on the fractional inhibitory concentration index, the synergistic effect was achieved in all of the samples exposed to itraconazole with Hyssopus officinalis (FICI 0.3±0.06). On the other side, the additive effect was proven in use of itraconazole with Thymus vulgaris (FICI 0.75±0.35) and fluconazole with both EOs tested (FICI 0.81±0.19; 0.88±0.57) This study shows the importance of monitoring the synergistic effect of antifungals combined with EOs, because it is a possible solution for reducing the resistance and improving the disease prognosis.
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Bibliography


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Baj T, Biernasiuk A, Wróbel R, Malm A (2020) Chemical composition and in vitro activity of Origanum vulgare L., Satureja hortensis L., Thymus serpyllum L. and Thymus vulgaris L. essential oils towards oral isolates of Candida albicans and Candida glabrata. Open Chem 18: 108-118.
Bolatchiev A, Baturin V, Bazikov I, Maltsev A, Kunitsina E (2020) Effect of antimicrobial peptides HNP-1 and hBD-1 on Staphylococcus aureus strains in vitro and in vivo. Fundam Clin Pharmacol 34: 102-108.
Bona E, Cantamessa S, Pavan M, Novello G, Massa N, Rocchetti A, Berta G, Gamalero E (2016) Sensitivity of Candida albicans to essential oils: are they an alter native to antifungal agents? J Appl Microbiol 121: 1530-1545.
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Cavalcanti YW, Almeida LF, Padilha WW (2011) Anti adherent activity of Rosmarinus officinalis essential oil on Candida albicans: an SEM analysis. Rev Odonto Ciênc 26: 139-144.
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Martins N, Ferreira IC, Barros L, Silva S, Henriques M (2014) Candidiasis: predisposing factors, prevention, diagnosis and alternative treat-ment. Mycopathologia 177: 223-240.
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Raut JS, Karuppayil SM (2014) A status review on the medi cinal properties of essential oils. Ind Crops Prod 62: 250-264.
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Authors and Affiliations

M. Proškovcová
1
E. Čonková
1
P. Váczi
1
D. Marcinčáková
1
M. Harčárová
2

  1. Department of Pharmacology and Toxicology, The University of Veterinary Medicine and Pharmacy, Komenského 73, 041 81 Košice, Slovakia
  2. Department of Animal Nutrition and Husbandry, The University of Veterinary Medicine and Pharmacy, Komenského 73, 041 81 Košice, Slovakia
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Abstract

The aim of this study was to reveal the potentially genotoxic effect of zearalenone on bovine lymphocytes by comet assay in vitro. The bovine lymphocytes were exposed to various zearalenone concetrations (50; 10; 2; 0.4 and 0.08 ppm). The viability and DNA damage of lymphocytes was monitored after 2 h, 24 h, 48 h and 72 h. After 2 hours of zearalenone exposure, statistically significant DNA damage occurred at all tested concentrations of 0.08 ppm (12.2±1.25; p<0.05), 0.4 ppm (12.7±0.88; p<0.01), 2 ppm (12.0±0.51; p<0.01), 10 ppm (11.2±0.47; p<0.01) and at 50 ppm (14.2±0 61; p<0.001). Significantly greater DNA damage was also found after 24 h, 48 h and 72 h. The obtained results showed that zearalenone may induce DNA damage of the bovine lymphocytes.
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Bibliography


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Authors and Affiliations

M. Harcarova
1
E. Conkova
2
S. Kolenicova
3
B. Holeckova
3
M. Proskovcova
2

  1. Department of Animal Nutrition and Husbandry, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 041 81 Kosice, Slovak Republic
  2. Department of Pharmacology and Toxicology, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 041 81 Kosice, Slovak Republic
  3. Department of Biology and Physiology, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 041 81 Kosice, Slovak Republic
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Abstract

The aim of this study was to investigate interactions between conventional antifungal drug and essential oils against isolates of Malassezia pachydermatis. Antifungal activity of Cinnamomum cassia, Melaleuca alternifolia, Mentha piperita, Origanum vulgare and Syzygium aromaticum essential oils were tested against 19 strains of M. pachydermatis isolated from healthy dogs and reference strain M. pachydermatis CBS 1879. The checkerboard assay was used to search for in- teractions. Synergism was observed for the combination of clotrimazole with Melaleuca alternifolia essential oil, Mentha piperita and Origanum vulgare. The combinations of Cinnamomum cassia and Syzygium aromaticum essential oils with clotrimazole showed indifferent effect. Additive antimicrobial activity was observed for the combination of clotrimazole with Syzygium aromaticum and Melaleuca alternifolia essential oils against reference strain. The obtained results showed synergistic interactions between essential oils and clotrimazole which could improve effectiveness of this antifungal drug.

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Authors and Affiliations

E. Bohmova
E. Conkova
M. Harcarova
Z. Sihelska
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Abstract

Candida albicans, a polymorphic yeast, is a physiological component of the human and animal commensal microbiome. It is an etiological factor of candidiasis, which is treated by azole antifungals. Growing resistance to azoles is a reason to look for other alternative treatment options. The pharmacotherapeutic use of plant extracts and essential oils has become increasingly important. In our experiment, C. albicans showed susceptibility to four observed plant extracts and essential oils from peppermint ( Mentha piperita), thyme ( Thymus vulgaris), sage ( Salvia officinalis), and oregano ( Origanum vulgare). Oregano plant extract and essential oil showed the highest antifungal activity, at MIC values of 4.9 mg/mL and 0.4 mg/mL respectively. Therefore, it was subjected to further research on the influence of virulence factors – biofilm formation, extracellular phospholipase production and germ tube formation. Oregano plant extract and essential oil showed an inhibitory effect on the observed C. albicans virulence factors at relatively low concentrations. The extract inhibited the adherence of cells at MIC 12.5 mg/mL and essential oil at MIC 0.25 mg/mL. Degradation of the formed biofilm was detected at MIC 14.1 mg/mL for plant extract and at MIC 0.4 mg/mL for essential oil. Extracellular phospholipase production was most effectively inhibited by the essential oil. In particular, the number of isolates with intensive extracellular phospholipase production decreased significantly. Of the 12 isolates intensively producing extracellular phospholipase, only 1 isolate (4.5%) retained intense production. Essential oil caused up to a 100 % reduction in germ tubes formation and plant extract reduced their formation depending on the concentration as follows: 2.6% (0.8 mg/mL), 21.2 % (6.25 mg/mL), and 64.5 % (12.5 mg/mL) compared to the control.
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Authors and Affiliations

P. Váczi
1
M. Proškovcová
1
E. Čonková
1
D. Marcinčáková
1
M. Bačkorová
2
M. Harčárová
3

  1. Department of Pharmacology and Toxicology
  2. Department of Pharmaceutical Technology, Pharmacognosy and Botany
  3. Department of Animal Nutrition and Husbandry, University of Veterinary Medicine and Pharmacy, Komenského 73, Košice, 041 81, Slovakia

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