Widespread infection, diversification and old host associations of Nosema Microsporidia in European freshwater gammarids (Amphipoda)

PLoS Pathog. 2023 Aug 21;19(8):e1011560. doi: 10.1371/journal.ppat.1011560. eCollection 2023 Aug.

Abstract

The microsporidian genus Nosema is primarily known to infect insects of economic importance stimulating high research interest, while other hosts remain understudied. Nosema granulosis is one of the formally described Nosema species infecting amphipod crustaceans, being known to infect only two host species. Our first aim was to characterize Nosema spp. infections in different amphipod species from various European localities using the small subunit ribosomal DNA (SSU) marker. Second, we aimed to assess the phylogenetic diversity, host specificity and to explore the evolutionary history that may explain the diversity of gammarid-infecting Nosema lineages by performing a phylogenetic reconstruction based on RNA polymerase II subunit B1 (RPB1) gene sequences. For the host species Gammarus balcanicus, we also analyzed whether parasites were in excess in females to test for sex ratio distortion in relation with Nosema infection. We identified Nosema spp. in 316 individuals from nine amphipod species being widespread in Europe. The RPB1-based phylogenetic reconstruction using newly reported sequences and available data from other invertebrates identified 39 haplogroups being associated with amphipods. These haplogroups clustered into five clades (A-E) that did not form a single amphipod-infecting monophyletic group. Closely related sister clades C and D correspond to Nosema granulosis. Clades A, B and E might represent unknown Nosema species infecting amphipods. Host specificity seemed to be variable with some clades being restricted to single hosts, and some that could be found in several host species. We show that Nosema parasite richness in gammarid hosts is much higher than expected, illustrating the advantage of the use of RPB1 marker over SSU. Finally, we found no hint of sex ratio distortion in Nosema clade A infecting G. balcanicus. This study shows that Nosema spp. are abundant, widespread and diverse in European gammarids. Thus, Nosema is as diverse in aquatic as in terrestrial hosts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amphipoda* / genetics
  • Animals
  • Female
  • Fresh Water
  • Humans
  • Nosema* / genetics
  • Phylogeny

Supplementary concepts

  • Nosema granulosis

Grants and funding

We acknowledge the l’Agence Nationale de la Recherche (ANR) (Grant # ANR-15-CE32-0006) for paying AQ salary and molecular analysis; as well as Narodowa Agencja Wymiany Akademickiej (Poland) for funding travel between France and Poland in terms of analysing the results and writing the manuscript within ‘Polonium’ exchange program (PPN/BIL/2018/1/00138) to KBS, RW, AQ, MG, TR. The sampling was performed during a series of scientific expeditions in the years 2004–2021 supported by Ministerstwo Nauki i Szkolnictwa Wyższego, Poland (N N303 579439) to MG, as well as French: l’Agence Nationale de la Recherche (Recolnat ANR-11-INBS-0004), Conseil Regional Bourgogne Franche-Comté (Excellence BFC “PIA‐Recolnat‐BFC”), Centre national de la recherche scientifique (CNRS-INSU OSU-THETA Franche-Comté Bourgogne SRO 2022) to RW.