Proteomics of Anatomical Sections of the Gut of Nosema-Infected Western Honeybee (Apis mellifera) Reveals Different Early Responses to Nosema spp. Isolates

Abstract

Honeybees play an important role in pollinating native plants and agricultural crops and produce valuable hive products. Within the last decade, honeybee colonies have been reported to be in decline, due to both biotic and abiotic stress factors including pathogens and pesticides. This study evaluated the impact of different isolates of Nosema spp.[Nosema apis spores (NA), Nosema ceranae from Apis mellifera from France (NF), N. ceranae from Apis cerana from Thailand (NC1), and N. ceranae from A. mellifera from Thailand (NC2)] on the different gut sections of newly emerged adult A. mellifera bees. With an attempt to decipher the early impact of Nosema spp. on the first barrier against Nosema infection, we used off-gel bottom-up proteomics on the different anatomical sections of the gut four days post inoculation. A total of 2185 identified proteins in the esophagus, 2095 in the crop, 1571 in the midgut, 2552 in the ileum, and 3173 in the rectum were obtained. Using label-free quantification, we observed that the response of the host varies according to the Nosema spp. (N. apis versus N. ceranae) and the geographical origin of Nosema. The proteins in the midgut of A. mellifera, orally inoculated with spores of N. ceranae isolated from France, were the most altered, when compared with controls, exhibiting 50 proteins down-regulated and 16 up-regulated. We thereby established the first massspectrometry-based proteomics of different anatomical sections of the gut tissue of Nosema-infected A. mellifera four days post inoculation, following infection by different isolates of Nosema spp. that provoked differential host responses. We reported an alteration of proteins involved in the metabolic pathways and specifically eight proteins of the oxidative phosphorylation pathway. More importantly, we propose that the collagen IV NC1 domain-containing protein may represent an early prognostic marker of the impact of Nosema spores on the A. mellifera health status. Data are available via ProteomeXchange with the identifier PXD021848.

Genetic Variation in Antimicrobial Activity of Honey Bee (Apis mellifera) Seminal Fluid

“…Chitinases are known to be present in the honey bee gut, specifically the acidic mammalian chitinase (GB46749) and the chitinase 5 (GB53565) (Houdelet et al, 2021), the latter being the same chitinase identified in seminal fluid previously (Grassl et al, 2016). Larvae have also shown deregulation of the AmelCht chitinase (GB15116), in response to infection with the fungal pathogen Ascosphaera apis (chalkbrood), and this chitinase shows high sequence similarity to gut-specific chitinases identified in the locust (Locusta migratoria manilensis) and the mosquito (Anopheles gambiae) (Aronstein et al, 2010).…”

In vitro Effects of Prebiotics and Synbiotics on Apis cerana Gut Microbiota

“…However, yeasts have been recognized as dominant fungi in the gut of the Apis genus, but the increased abundance of yeast has been related to diseases and malnutrition or response to antibiotics and insecticides ( Gilliam and Prest 1972 ; Ptaszyńska et al 2016 ). The infection with intestinal fungal pathogens such as Nosema apis and N. ceranae has been recognized to cause shortened lifespan and reduction in colony size of honeybees ( Houdelet et al 2021 ). Our results suggest that IMO combined with selected Lactobacillus strains maintain gut microbiota homeostasis of A. cerana .…”

Molecular histoproteomy by MALDI mass spectrometry imaging to uncover markers of the impact of Nosema on Apis mellifera

“…ways [67]. To pursue our investigations on the underlying molecular mechanisms, MALDI IMS was conducted on the entire body of A. mellifera in this context in order to follow and establish the first comprehensive spatial histo-proteomics picture of the impact of Nosema infection on whole bee sections.…”

“…The impact of Nosema disease at the individual level includes numerous physiological alterations, such as (i) metabolic changes [55,56], (ii) energy stress/management [56], (iii) pheromone and hormone production [57], (iv) inhibition of epithelial cells apoptosis [58], (v) immune function suppression [55,59,60], (vi) life span shortening [56,61,62], (vii) cognitive deficits [63,64], (viii) acceleration of transition to forager activities [65], and (ix) changes of gut microbiota [66]. We recently conducted study with the aim to decipher the impact of a per‐os inoculation of A. mellifera with spores of Nosema on the first off‐gel proteomics on the different anatomical sections of the gut [67]. Upon oral infection, we observed at an early stage of the infection (4 days) that midgut proteins were the most altered (50 down‐regulated, 16 up‐regulated) compared to the control experiment with most of them being involved in metabolic and oxidative phosphorylation pathways [67].…”

“…We recently conducted study with the aim to decipher the impact of a per‐os inoculation of A. mellifera with spores of Nosema on the first off‐gel proteomics on the different anatomical sections of the gut [67]. Upon oral infection, we observed at an early stage of the infection (4 days) that midgut proteins were the most altered (50 down‐regulated, 16 up‐regulated) compared to the control experiment with most of them being involved in metabolic and oxidative phosphorylation pathways [67]. To pursue our investigations on the underlying molecular mechanisms, MALDI IMS was conducted on the entire body of A. mellifera in this context in order to follow and establish the first comprehensive spatial histo‐proteomics picture of the impact of Nosema infection on whole bee sections.…”