Cultivation of Protozoa Parasites In Vitro: Growth Potential in Conventional Culture Media versus RPMI-PY Medium
The in vitro cultivation of Leishmania and Trypanosoma parasites plays an important role in the diagnosis and treatment of parasitic diseases. Although Evans’s modified Tobie and Novy–MacNeal–Nicolle media, for <i>Leishmania spp.</i> and <i>Trypanosoma cruzi</i>, respectively...
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MDPI AG
2023-03-01
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| Series: | Veterinary Sciences |
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| Online Access: | https://www.mdpi.com/2306-7381/10/4/252 |
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| author | Germano Castelli Eugenia Oliveri Viviana Valenza Susanna Giardina Flavia Facciponte Francesco La Russa Fabrizio Vitale Federica Bruno |
| author_facet | Germano Castelli Eugenia Oliveri Viviana Valenza Susanna Giardina Flavia Facciponte Francesco La Russa Fabrizio Vitale Federica Bruno |
| author_sort | Germano Castelli |
| collection | DOAJ |
| description | The in vitro cultivation of Leishmania and Trypanosoma parasites plays an important role in the diagnosis and treatment of parasitic diseases. Although Evans’s modified Tobie and Novy–MacNeal–Nicolle media, for <i>Leishmania spp.</i> and <i>Trypanosoma cruzi</i>, respectively, are the two commonly used media for both isolation and maintenance of strains in vitro, their preparation is expensive and laborious and requires fresh rabbit blood from housed animals. The purpose of this study was to evaluate the in vitro growth of both parasites with an alternative monophasic, blood-free, easy, and affordable medium called RPMI-PY, which was previously demonstrated suitable for the in vitro growth of <i>Leishmania infantum</i>. The potential growth of different Leishmania species and <i>Trypanosoma cruzi</i> was evaluated in traditional culture media versus RPMI-PY medium, and we recorded the protozoa parasites’ morphology via orange acridine–ethidium bromide staining. The results of our study show that RPMI-PY medium can be used for <i>Trypanosoma cruzi</i>, <i>Leishmania amazonensis</i>, <i>Leishmania major</i>, <i>and Leishmania tropica</i> species since in all the species except <i>Leishmania braziliensis</i>, the exponential growth of the parasite was observed, in many cases higher than conventional media. The staining confirmed not only their growth during the 72 h investigation but also the optimal morphology and viability of the protozoa in the RPMI-PY medium. |
| first_indexed | 2024-03-11T04:27:29Z |
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| language | English |
| last_indexed | 2024-03-11T04:27:29Z |
| publishDate | 2023-03-01 |
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| record_format | Article |
| series | Veterinary Sciences |
| spelling | doaj.art-c95b3ca58f11451eaa03498b03ca45632023-11-17T21:43:17ZengMDPI AGVeterinary Sciences2306-73812023-03-0110425210.3390/vetsci10040252Cultivation of Protozoa Parasites In Vitro: Growth Potential in Conventional Culture Media versus RPMI-PY MediumGermano Castelli0Eugenia Oliveri1Viviana Valenza2Susanna Giardina3Flavia Facciponte4Francesco La Russa5Fabrizio Vitale6Federica Bruno7Centro di Referenza Nazionale per le Leishmaniosi (C.Re.Na.L.), WOAH Leishmania Reference Laboratory, Istituto Zooprofilattico Sperimentale della Sicilia, Via Gino Marinuzzi 3, 90129 Palermo, ItalyCentro di Referenza Nazionale per le Leishmaniosi (C.Re.Na.L.), WOAH Leishmania Reference Laboratory, Istituto Zooprofilattico Sperimentale della Sicilia, Via Gino Marinuzzi 3, 90129 Palermo, ItalyCentro di Referenza Nazionale per le Leishmaniosi (C.Re.Na.L.), WOAH Leishmania Reference Laboratory, Istituto Zooprofilattico Sperimentale della Sicilia, Via Gino Marinuzzi 3, 90129 Palermo, ItalyCentro di Referenza Nazionale per le Leishmaniosi (C.Re.Na.L.), WOAH Leishmania Reference Laboratory, Istituto Zooprofilattico Sperimentale della Sicilia, Via Gino Marinuzzi 3, 90129 Palermo, ItalyCentro di Referenza Nazionale per le Leishmaniosi (C.Re.Na.L.), WOAH Leishmania Reference Laboratory, Istituto Zooprofilattico Sperimentale della Sicilia, Via Gino Marinuzzi 3, 90129 Palermo, ItalyCentro di Referenza Nazionale per le Leishmaniosi (C.Re.Na.L.), WOAH Leishmania Reference Laboratory, Istituto Zooprofilattico Sperimentale della Sicilia, Via Gino Marinuzzi 3, 90129 Palermo, ItalyCentro di Referenza Nazionale per le Leishmaniosi (C.Re.Na.L.), WOAH Leishmania Reference Laboratory, Istituto Zooprofilattico Sperimentale della Sicilia, Via Gino Marinuzzi 3, 90129 Palermo, ItalyCentro di Referenza Nazionale per le Leishmaniosi (C.Re.Na.L.), WOAH Leishmania Reference Laboratory, Istituto Zooprofilattico Sperimentale della Sicilia, Via Gino Marinuzzi 3, 90129 Palermo, ItalyThe in vitro cultivation of Leishmania and Trypanosoma parasites plays an important role in the diagnosis and treatment of parasitic diseases. Although Evans’s modified Tobie and Novy–MacNeal–Nicolle media, for <i>Leishmania spp.</i> and <i>Trypanosoma cruzi</i>, respectively, are the two commonly used media for both isolation and maintenance of strains in vitro, their preparation is expensive and laborious and requires fresh rabbit blood from housed animals. The purpose of this study was to evaluate the in vitro growth of both parasites with an alternative monophasic, blood-free, easy, and affordable medium called RPMI-PY, which was previously demonstrated suitable for the in vitro growth of <i>Leishmania infantum</i>. The potential growth of different Leishmania species and <i>Trypanosoma cruzi</i> was evaluated in traditional culture media versus RPMI-PY medium, and we recorded the protozoa parasites’ morphology via orange acridine–ethidium bromide staining. The results of our study show that RPMI-PY medium can be used for <i>Trypanosoma cruzi</i>, <i>Leishmania amazonensis</i>, <i>Leishmania major</i>, <i>and Leishmania tropica</i> species since in all the species except <i>Leishmania braziliensis</i>, the exponential growth of the parasite was observed, in many cases higher than conventional media. The staining confirmed not only their growth during the 72 h investigation but also the optimal morphology and viability of the protozoa in the RPMI-PY medium.https://www.mdpi.com/2306-7381/10/4/252protozoa parasitesculture mediaRPMI-PY medium<i>Leishmania</i> spp.<i>Trypanosoma cruzi</i> |
| spellingShingle | Germano Castelli Eugenia Oliveri Viviana Valenza Susanna Giardina Flavia Facciponte Francesco La Russa Fabrizio Vitale Federica Bruno Cultivation of Protozoa Parasites In Vitro: Growth Potential in Conventional Culture Media versus RPMI-PY Medium Veterinary Sciences protozoa parasites culture media RPMI-PY medium <i>Leishmania</i> spp. <i>Trypanosoma cruzi</i> |
| title | Cultivation of Protozoa Parasites In Vitro: Growth Potential in Conventional Culture Media versus RPMI-PY Medium |
| title_full | Cultivation of Protozoa Parasites In Vitro: Growth Potential in Conventional Culture Media versus RPMI-PY Medium |
| title_fullStr | Cultivation of Protozoa Parasites In Vitro: Growth Potential in Conventional Culture Media versus RPMI-PY Medium |
| title_full_unstemmed | Cultivation of Protozoa Parasites In Vitro: Growth Potential in Conventional Culture Media versus RPMI-PY Medium |
| title_short | Cultivation of Protozoa Parasites In Vitro: Growth Potential in Conventional Culture Media versus RPMI-PY Medium |
| title_sort | cultivation of protozoa parasites in vitro growth potential in conventional culture media versus rpmi py medium |
| topic | protozoa parasites culture media RPMI-PY medium <i>Leishmania</i> spp. <i>Trypanosoma cruzi</i> |
| url | https://www.mdpi.com/2306-7381/10/4/252 |
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