Differential Exoproteome and Biochemical Characterisation of <i>Neoparamoeba perurans</i>
Infection with the protozoan ectoparasite <i>Neoparamoeba perurans</i>, the causative agent of AGD, remains a global threat to salmonid farming. This study aimed to analyse the exoproteome of both an attenuated and virulent <i>N. perurans</i> isolate using proteomics and cyto...
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MDPI AG
2021-06-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/9/6/1258 |
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| author | Kerrie Ní Dhufaigh Natasha Botwright Eugene Dillon Ian O’Connor Eugene MacCarthy Orla Slattery |
| author_facet | Kerrie Ní Dhufaigh Natasha Botwright Eugene Dillon Ian O’Connor Eugene MacCarthy Orla Slattery |
| author_sort | Kerrie Ní Dhufaigh |
| collection | DOAJ |
| description | Infection with the protozoan ectoparasite <i>Neoparamoeba perurans</i>, the causative agent of AGD, remains a global threat to salmonid farming. This study aimed to analyse the exoproteome of both an attenuated and virulent <i>N. perurans</i> isolate using proteomics and cytotoxicity testing. A disproportionate presence of proteins from the co-cultured microbiota of <i>N. perurans</i> was revealed on searching an amalgamated database of bacterial, <i>N. perurans</i> and Amoebozoa proteins. LC-MS/MS identified 33 differentially expressed proteins, the majority of which were upregulated in the attenuated exoproteome. Proteins of putative interest found in both exoproteomes were maltoporin, ferrichrome-iron receptor, and putative ferric enterobactin receptor. Protease activity remained significantly elevated in the attenuated exoproteome compared with the virulent exoproteome. Similarly, the attenuated exoproteome had a significantly higher cytotoxic effect on rainbow trout gill cell line (RTgill W1) cells compared with the virulent exoproteome. The presence of a phosphatase and serine protease in the virulent exoproteome may facilitate AGD infection but do not appear to be key players in causing cytotoxicity. Altogether, this study reveals prolonged culture of <i>N. perurans</i> affects the exoproteome composition in favour of nutritional acquisition, and that the current culturing protocol for virulent <i>N. perurans</i> does not facilitate the secretion of virulence factors. |
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| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-10T10:34:04Z |
| publishDate | 2021-06-01 |
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| series | Microorganisms |
| spelling | doaj.art-565fca03c9cd4fa5a7136fbe8b65fae32023-11-21T23:26:10ZengMDPI AGMicroorganisms2076-26072021-06-0196125810.3390/microorganisms9061258Differential Exoproteome and Biochemical Characterisation of <i>Neoparamoeba perurans</i>Kerrie Ní Dhufaigh0Natasha Botwright1Eugene Dillon2Ian O’Connor3Eugene MacCarthy4Orla Slattery5Marine and Freshwater Research Centre, Galway-Mayo Institute of Technology, Co. Galway, H91 T8NW Eircode, IrelandCSIRO Agriculture and Food, Livestock & Aquaculture, Queensland Biosciences Precinct, 306 Carmody Road, Brisbane, QLD 4067, AustraliaConway Institute of Biomolecular & Biomedical Research, University College Dublin, Co. Dublin, D04 V1W8 Eircode, IrelandMarine and Freshwater Research Centre, Galway-Mayo Institute of Technology, Co. Galway, H91 T8NW Eircode, IrelandMarine and Freshwater Research Centre, Galway-Mayo Institute of Technology, Co. Galway, H91 T8NW Eircode, IrelandDepartment of Biopharmaceutical and Medical Science, Galway-Mayo Institute of Technology, Co. Galway, H91 T8NW Eircode, IrelandInfection with the protozoan ectoparasite <i>Neoparamoeba perurans</i>, the causative agent of AGD, remains a global threat to salmonid farming. This study aimed to analyse the exoproteome of both an attenuated and virulent <i>N. perurans</i> isolate using proteomics and cytotoxicity testing. A disproportionate presence of proteins from the co-cultured microbiota of <i>N. perurans</i> was revealed on searching an amalgamated database of bacterial, <i>N. perurans</i> and Amoebozoa proteins. LC-MS/MS identified 33 differentially expressed proteins, the majority of which were upregulated in the attenuated exoproteome. Proteins of putative interest found in both exoproteomes were maltoporin, ferrichrome-iron receptor, and putative ferric enterobactin receptor. Protease activity remained significantly elevated in the attenuated exoproteome compared with the virulent exoproteome. Similarly, the attenuated exoproteome had a significantly higher cytotoxic effect on rainbow trout gill cell line (RTgill W1) cells compared with the virulent exoproteome. The presence of a phosphatase and serine protease in the virulent exoproteome may facilitate AGD infection but do not appear to be key players in causing cytotoxicity. Altogether, this study reveals prolonged culture of <i>N. perurans</i> affects the exoproteome composition in favour of nutritional acquisition, and that the current culturing protocol for virulent <i>N. perurans</i> does not facilitate the secretion of virulence factors.https://www.mdpi.com/2076-2607/9/6/1258amoebic gill diseaseexoproteomeLC-MS/MSvirulence factorsprotease activitycytotoxicity |
| spellingShingle | Kerrie Ní Dhufaigh Natasha Botwright Eugene Dillon Ian O’Connor Eugene MacCarthy Orla Slattery Differential Exoproteome and Biochemical Characterisation of <i>Neoparamoeba perurans</i> Microorganisms amoebic gill disease exoproteome LC-MS/MS virulence factors protease activity cytotoxicity |
| title | Differential Exoproteome and Biochemical Characterisation of <i>Neoparamoeba perurans</i> |
| title_full | Differential Exoproteome and Biochemical Characterisation of <i>Neoparamoeba perurans</i> |
| title_fullStr | Differential Exoproteome and Biochemical Characterisation of <i>Neoparamoeba perurans</i> |
| title_full_unstemmed | Differential Exoproteome and Biochemical Characterisation of <i>Neoparamoeba perurans</i> |
| title_short | Differential Exoproteome and Biochemical Characterisation of <i>Neoparamoeba perurans</i> |
| title_sort | differential exoproteome and biochemical characterisation of i neoparamoeba perurans i |
| topic | amoebic gill disease exoproteome LC-MS/MS virulence factors protease activity cytotoxicity |
| url | https://www.mdpi.com/2076-2607/9/6/1258 |
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