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In vitro culture of the zoonotic nematode Anisakis pegreffii (Nematoda, Anisakidae)
BACKGROUND: Anisakiasis is a foodborne disease caused by the third-stage larvae (L3) of two species belonging to the genus Anisakis: Anisakis pegreffii and Anisakis simplex sensu stricto. Both species have been the subject of different -omics studies undertaken in the past decade, but a reliable in...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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BioMed Central
2023
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9896804/ https://www.ncbi.nlm.nih.gov/pubmed/36732837 http://dx.doi.org/10.1186/s13071-022-05629-5 |
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author | Mladineo, Ivona Charouli, Artemis Jelić, Filip Chakroborty, Anand Hrabar, Jerko |
author_facet | Mladineo, Ivona Charouli, Artemis Jelić, Filip Chakroborty, Anand Hrabar, Jerko |
author_sort | Mladineo, Ivona |
collection | PubMed |
description | BACKGROUND: Anisakiasis is a foodborne disease caused by the third-stage larvae (L3) of two species belonging to the genus Anisakis: Anisakis pegreffii and Anisakis simplex sensu stricto. Both species have been the subject of different -omics studies undertaken in the past decade, but a reliable in vitro culture protocol that would enable a more versatile approach to functional studies has never been devised. In nature, A. pegreffii shows a polyxenous life-cycle. It reproduces in toothed whales (final host) and disseminates embryonated eggs via cetacean faeces in the water column. In the environment, a first- (L1) and second-stage larva (L2) develops inside the egg, and subsequently hatched L2 is ingested by a planktonic crustacean or small fish (intermediate host). In the crustacean pseudocoelom, the larva moults to the third stage (L3) and grows until the host is eaten by a fish or cephalopod (paratenic host). Infective L3 migrates into the visceral cavity of its paratenic host and remains in the state of paratenesis until a final host preys on the former. Once in the final host’s gastric chambers, L3 attaches to mucosa, moults in the fourth stage (L4) and closes its life-cycle by becoming reproductively mature. METHODS: Testing two commercially available media (RPMI 1640, Schneider’s Drosophila) in combination with each of the six different heat-inactivated sera, namely foetal bovine, rabbit, chicken, donkey, porcine and human serum, we have obtained the first reliable, fast and simple in vitro cultivation protocol for A. pegreffii. RESULTS: Schneider’s Drosophila insect media supplemented with 10% chicken serum allowed high reproducibility and survival of adult A. pegreffii. The maturity was reached already at the beginning of the third week in culture. From collected eggs, hatched L2 were maintained in culture for 2 weeks. The protocol also enabled the description of undocumented morphological and ultrastructural features of the parasite developmental stages. CONCLUSIONS: Closing of the A. pegreffii life-cycle from L3 to reproducing adults is an important step from many research perspectives (e.g., vaccine and drug–target research, transgenesis, pathogenesis), but further effort is necessary to optimise the efficient moulting of L2 to infective L3. GRAPHICAL ABSTRACT: [Image: see text] |
format | Online Article Text |
id | pubmed-9896804 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-98968042023-02-04 In vitro culture of the zoonotic nematode Anisakis pegreffii (Nematoda, Anisakidae) Mladineo, Ivona Charouli, Artemis Jelić, Filip Chakroborty, Anand Hrabar, Jerko Parasit Vectors Research BACKGROUND: Anisakiasis is a foodborne disease caused by the third-stage larvae (L3) of two species belonging to the genus Anisakis: Anisakis pegreffii and Anisakis simplex sensu stricto. Both species have been the subject of different -omics studies undertaken in the past decade, but a reliable in vitro culture protocol that would enable a more versatile approach to functional studies has never been devised. In nature, A. pegreffii shows a polyxenous life-cycle. It reproduces in toothed whales (final host) and disseminates embryonated eggs via cetacean faeces in the water column. In the environment, a first- (L1) and second-stage larva (L2) develops inside the egg, and subsequently hatched L2 is ingested by a planktonic crustacean or small fish (intermediate host). In the crustacean pseudocoelom, the larva moults to the third stage (L3) and grows until the host is eaten by a fish or cephalopod (paratenic host). Infective L3 migrates into the visceral cavity of its paratenic host and remains in the state of paratenesis until a final host preys on the former. Once in the final host’s gastric chambers, L3 attaches to mucosa, moults in the fourth stage (L4) and closes its life-cycle by becoming reproductively mature. METHODS: Testing two commercially available media (RPMI 1640, Schneider’s Drosophila) in combination with each of the six different heat-inactivated sera, namely foetal bovine, rabbit, chicken, donkey, porcine and human serum, we have obtained the first reliable, fast and simple in vitro cultivation protocol for A. pegreffii. RESULTS: Schneider’s Drosophila insect media supplemented with 10% chicken serum allowed high reproducibility and survival of adult A. pegreffii. The maturity was reached already at the beginning of the third week in culture. From collected eggs, hatched L2 were maintained in culture for 2 weeks. The protocol also enabled the description of undocumented morphological and ultrastructural features of the parasite developmental stages. CONCLUSIONS: Closing of the A. pegreffii life-cycle from L3 to reproducing adults is an important step from many research perspectives (e.g., vaccine and drug–target research, transgenesis, pathogenesis), but further effort is necessary to optimise the efficient moulting of L2 to infective L3. GRAPHICAL ABSTRACT: [Image: see text] BioMed Central 2023-02-02 /pmc/articles/PMC9896804/ /pubmed/36732837 http://dx.doi.org/10.1186/s13071-022-05629-5 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Mladineo, Ivona Charouli, Artemis Jelić, Filip Chakroborty, Anand Hrabar, Jerko In vitro culture of the zoonotic nematode Anisakis pegreffii (Nematoda, Anisakidae) |
title | In vitro culture of the zoonotic nematode Anisakis pegreffii (Nematoda, Anisakidae) |
title_full | In vitro culture of the zoonotic nematode Anisakis pegreffii (Nematoda, Anisakidae) |
title_fullStr | In vitro culture of the zoonotic nematode Anisakis pegreffii (Nematoda, Anisakidae) |
title_full_unstemmed | In vitro culture of the zoonotic nematode Anisakis pegreffii (Nematoda, Anisakidae) |
title_short | In vitro culture of the zoonotic nematode Anisakis pegreffii (Nematoda, Anisakidae) |
title_sort | in vitro culture of the zoonotic nematode anisakis pegreffii (nematoda, anisakidae) |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9896804/ https://www.ncbi.nlm.nih.gov/pubmed/36732837 http://dx.doi.org/10.1186/s13071-022-05629-5 |
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