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In vitro anticoccidial activity of thymol, carvacrol, and saponins
The anticoccidial activity of thymol, carvacrol, and saponins was assessed in an in vitro model of coccidiosis. Eimeria spp. sporozoites were collected from field samples, characterized, and used for 2 different invasion assays on Madin-Darby Bovine Kidney cells (MDBK). The cells were challenged wit...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7647770/ https://www.ncbi.nlm.nih.gov/pubmed/33142451 http://dx.doi.org/10.1016/j.psj.2020.07.035 |
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author | Felici, Martina Tugnoli, Benedetta Ghiselli, Federico Massi, Paola Tosi, Giovanni Fiorentini, Laura Piva, Andrea Grilli, Ester |
author_facet | Felici, Martina Tugnoli, Benedetta Ghiselli, Federico Massi, Paola Tosi, Giovanni Fiorentini, Laura Piva, Andrea Grilli, Ester |
author_sort | Felici, Martina |
collection | PubMed |
description | The anticoccidial activity of thymol, carvacrol, and saponins was assessed in an in vitro model of coccidiosis. Eimeria spp. sporozoites were collected from field samples, characterized, and used for 2 different invasion assays on Madin-Darby Bovine Kidney cells (MDBK). The cells were challenged with 5 × 10(4) sporozoites without (control) or with various treatments: saponins (10 ppm), thymol, and carvacrol (7 ppm each) or a combination of saponins, thymol, and carvacrol at 2 doses; MIX 1 (saponins 5 ppm, thymol 3.5 ppm, and carvacrol 3.5 ppm) and MIX 2 (saponins 10 ppm, thymol 7 ppm, and carvacrol 7 ppm). The treated cells were incubated at 37°C for 24 h (invasion assay 1) and for 2, 24, and 48 h (invasion assay 2). The efficiency of invasion was determined by counting the sporozoites left in the supernatant that were not able to invade the cells, whereas intracellular Eimeria DNA was detected by qPCR to confirm the data. Data were analyzed with ANOVA, and differences were considered significant when P value was ≤0.05. Data from invasion assay 1 showed that the thymol and carvacrol-containing blends significantly reduced invasion, especially in combination with saponins at the highest dose. Saponins alone did not have a strong inhibiting activity but acted synergistically with the other molecules. Interestingly, in invasion assay 2, it was found that the effect of the highest dose of the blend of saponins, thymol, and carvacrol was already visible at 2 h postinfection, whereas the other treatments were significantly successful at 24 h postinfection. The invasion assay protocol was designed to screen molecules in vitro starting from field fecal samples, and it can represent a potential tool in Eimeria research. Moreover, this study shows that invasion in MDBK cells by Eimeria sporozoites is inhibited in presence of thymol, carvacrol, and saponins, thus highlighting the anticoccidial potential of these compounds. |
format | Online Article Text |
id | pubmed-7647770 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-76477702020-11-13 In vitro anticoccidial activity of thymol, carvacrol, and saponins Felici, Martina Tugnoli, Benedetta Ghiselli, Federico Massi, Paola Tosi, Giovanni Fiorentini, Laura Piva, Andrea Grilli, Ester Poult Sci Immunology, Health and Disease The anticoccidial activity of thymol, carvacrol, and saponins was assessed in an in vitro model of coccidiosis. Eimeria spp. sporozoites were collected from field samples, characterized, and used for 2 different invasion assays on Madin-Darby Bovine Kidney cells (MDBK). The cells were challenged with 5 × 10(4) sporozoites without (control) or with various treatments: saponins (10 ppm), thymol, and carvacrol (7 ppm each) or a combination of saponins, thymol, and carvacrol at 2 doses; MIX 1 (saponins 5 ppm, thymol 3.5 ppm, and carvacrol 3.5 ppm) and MIX 2 (saponins 10 ppm, thymol 7 ppm, and carvacrol 7 ppm). The treated cells were incubated at 37°C for 24 h (invasion assay 1) and for 2, 24, and 48 h (invasion assay 2). The efficiency of invasion was determined by counting the sporozoites left in the supernatant that were not able to invade the cells, whereas intracellular Eimeria DNA was detected by qPCR to confirm the data. Data were analyzed with ANOVA, and differences were considered significant when P value was ≤0.05. Data from invasion assay 1 showed that the thymol and carvacrol-containing blends significantly reduced invasion, especially in combination with saponins at the highest dose. Saponins alone did not have a strong inhibiting activity but acted synergistically with the other molecules. Interestingly, in invasion assay 2, it was found that the effect of the highest dose of the blend of saponins, thymol, and carvacrol was already visible at 2 h postinfection, whereas the other treatments were significantly successful at 24 h postinfection. The invasion assay protocol was designed to screen molecules in vitro starting from field fecal samples, and it can represent a potential tool in Eimeria research. Moreover, this study shows that invasion in MDBK cells by Eimeria sporozoites is inhibited in presence of thymol, carvacrol, and saponins, thus highlighting the anticoccidial potential of these compounds. Elsevier 2020-08-12 /pmc/articles/PMC7647770/ /pubmed/33142451 http://dx.doi.org/10.1016/j.psj.2020.07.035 Text en © 2020 Published by Elsevier Inc. on behalf of Poultry Science Association Inc. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Immunology, Health and Disease Felici, Martina Tugnoli, Benedetta Ghiselli, Federico Massi, Paola Tosi, Giovanni Fiorentini, Laura Piva, Andrea Grilli, Ester In vitro anticoccidial activity of thymol, carvacrol, and saponins |
title | In vitro anticoccidial activity of thymol, carvacrol, and saponins |
title_full | In vitro anticoccidial activity of thymol, carvacrol, and saponins |
title_fullStr | In vitro anticoccidial activity of thymol, carvacrol, and saponins |
title_full_unstemmed | In vitro anticoccidial activity of thymol, carvacrol, and saponins |
title_short | In vitro anticoccidial activity of thymol, carvacrol, and saponins |
title_sort | in vitro anticoccidial activity of thymol, carvacrol, and saponins |
topic | Immunology, Health and Disease |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7647770/ https://www.ncbi.nlm.nih.gov/pubmed/33142451 http://dx.doi.org/10.1016/j.psj.2020.07.035 |
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