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

Abstract

Matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI IMS) is a powerful technology used to investigate the spatio-temporal distribution of a huge number of molecules throughout a body/tissue section. In this paper, we report the use of MALDI IMS to follow the molecular impact of an experimental infection of Apis mellifera with the microsporidia Nosema ceranae. We performed representative molecular mass fingerprints of selected tissues obtained by dissection. This was followed by MALDI IMS workflows optimization including specimen embedding and positioning as well as washing and matrix application. We recorded the local distribution of peptides/proteins within different tissues from experimentally infected versus non infected honeybees. As expected, a distinction in these molecular profiles between the two conditions was recorded from different anatomical sections of the gut tissue.More importantly, we observed differences in the molecular profiles in the brain, thoracic ganglia, hypopharyngeal glands, and hemolymph. We introduced MALDI IMS as an effective approach to monitor the impact of N. ceranae infection on A. mellifera. This opens perspectives for the discovery of molecular changes in peptides/proteins markers that could contribute to a better understanding of the impact of stressors and toxicity on different tissues of a bee in a single experiment.

Phagocytosis Is the Sole Arm of Drosophila melanogaster Known Host Defenses That Provides Some Protection Against Microsporidia Infection

“…Several other host defenses were either not relevant or unexpectedly promoted the infection by the parasite ( Table 2 ). Our work therefore provides a novel perspective on insect host defenses against one obligate intracellular parasite that differs from the picture gained through descriptive transcriptomic or proteomic analyses ( 51 , 52 ).…”

“…As the impact of microsporidia on economically important invertebrates ( e.g ., silkworm, shrimps) and broadly on insect populations ( e.g ., honeybees, mosquitoes) has been increasing over the years, using Drosophila melanogaster and its genetic tools and knowledge will be helpful to investigate insect host-defenses to the parasite. Indeed, a current limitation of the study of these defenses is that they rely essentially on descriptive transcriptomic and proteomic studies ( 51 , 52 ) but are rarely followed up by experimental characterization. In Drosophila , it is easier to directly test the functional relevance of specific host defenses by genetic loss-of-function approaches.…”

Proteomics and Immune Response Differences in Apis mellifera and Apis cerana Inoculated with Three Nosema ceranae Isolates

“…In adult honeybees, Nosema infection induces several behavioral and molecular alterations such as (i) reduction of worker lifespan; 17 (ii) cognitive changes including acceleration of the behavior-age polyethism, 18 alteration of olfactory learning and memory; 17,19 (iii) dysbiosis of the endogenous gut microbial communities and impairment of the host defense system; 20−22 (iv) alteration of the hemolymph carbohydrate homeostasis, 23 lipid loss; 24 and (v) dysfunction of the gut tissue. 25 Like other insects, honeybees have evolved defense mechanisms against pathogens that help them survive under biological stress, including Nosema invasion. 26 The mechanisms of the insect immune system against parasites and pathogens rely mostly on both humoral and cellular responses.…”

“…These obligate intracellular parasites infect the digestive tract of adult honeybees where they can develop and multiply before being transmitted horizontally by ingestion of spores from contaminated food, fecal-oral transmission, or ingestion of contaminated water. ,, A visual symptom of Nosema infection is diarrhea, while microscopic observations reveal tissue lesions, spore accumulation, and colonization by Nosema mycelium in the gut tissue and Malpighian tubule system. In adult honeybees, Nosema infection induces several behavioral and molecular alterations such as (i) reduction of worker lifespan; (ii) cognitive changes including acceleration of the behavior-age polyethism, alteration of olfactory learning and memory; , (iii) dysbiosis of the endogenous gut microbial communities and impairment of the host defense system; − (iv) alteration of the hemolymph carbohydrate homeostasis, lipid loss; and (v) dysfunction of the gut tissue …”

Occurrence of Nosemosis in honey bee,Apis melliferaL. at the apiaries of East Kazakhstan

“…In light of the fact, that many studies prove negative effects of nosemosis at the level of a bee family ( Farrar, 1947 ; Fries, 1998 ; Anderson & Giacon, 1992 ; Higes et al, 2009 ; Mayack & Naug, 2009 ; Botías et al, 2013 ; Tlak Gajger et al, 2015 ; Ostroverhova, 2018 ; Golubeva, 2018 ; Bajgazanov, Pashayan & Tikhomirova, 2019 ; Bajgazanov & Tikhomirova, 2020 ; Kartal et al, 2021 ; Salkova et al, 2022 ; Houdelet et al, 2022 ), the need arises to proceed with monitoring studies of honey bee at the apiaries of East Kazakhstan Region.…”