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. 2021 Mar 25;11(1):6830.
doi: 10.1038/s41598-021-85988-8.

Comparative proteomic profiling of newly acquired, virulent and attenuated Neoparamoeba perurans proteins associated with amoebic gill disease

Kerrie Ní Dhufaigh  1 , Eugene Dillon  2 , Natasha Botwright  3 , Victor Birlanga  4 , Anita Talbot  5 , Ian O'Connor  5 , Eugene MacCarthy  5 , Orla Slattery  6
Affiliations

Comparative proteomic profiling of newly acquired, virulent and attenuated Neoparamoeba perurans proteins associated with amoebic gill disease

Kerrie Ní Dhufaigh et al. Sci Rep. .

Erratum in

Abstract

The causative agent of amoebic gill disease, Neoparamoeba perurans is reported to lose virulence during prolonged in vitro maintenance. In this study, the impact of prolonged culture on N. perurans virulence and its proteome was investigated. Two isolates, attenuated and virulent, had their virulence assessed in an experimental trial using Atlantic salmon smolts and their bacterial community composition was evaluated by 16S rRNA Illumina MiSeq sequencing. Soluble proteins were isolated from three isolates: a newly acquired, virulent and attenuated N. perurans culture. Proteins were analysed using two-dimensional electrophoresis coupled with liquid chromatography tandem mass spectrometry (LC-MS/MS). The challenge trial using naïve smolts confirmed a loss in virulence in the attenuated N. perurans culture. A greater diversity of bacterial communities was found in the microbiome of the virulent isolate in contrast to a reduction in microbial community richness in the attenuated microbiome. A collated proteome database of N. perurans, Amoebozoa and four bacterial genera resulted in 24 proteins differentially expressed between the three cultures. The present LC-MS/MS results indicate protein synthesis, oxidative stress and immunomodulation are upregulated in a newly acquired N. perurans culture and future studies may exploit these protein identifications for therapeutic purposes in infected farmed fish.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
An example of H&E stained gill samples taken from non-AGD and AGD infected fish from the challenge trial. Scale bar = 100 µm. (A) Gill sample from the attenuated cohort. (B) Gill sample from the virulent cohort.
Figure 2
Figure 2
Relative abundance of the prokaryotic community in the attenuated and virulent cultures by 16S rRNA gene sequencing.
Figure 3:
Figure 3:
2D gel of the reference gel from the newly acquired culture of N. perurans. 11 spots displayed differential expression and were excised for LC MS/MS.
See this image and copyright information in PMC

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