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We examined age-related biochemical and histological changes in the fat bodies and hemolymph of Osmia rufa males and females. We analysed solitary bees during diapause, in October and in April; as well as the flying insects following diapause, in May and June. The trophocyte sizes, as well as the numbers of lipid droplets were the greatest at the beginning of diapause. Subsequently, they decreased along with age. Triglyceride and glucose concentrations systematically decreased in fat body cells but increased in the hemolymph from October to June. Concentrations/activities of (enzymatic and non-enzymatic) antioxidant and proteolytic systems, as well as phenoloxidase, aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase levels were constant during the diapause, usually lower in the males than the females. Prior to the diapause/overwintering, the concentrations/activities of all the compounds were higher in the fat bodies than in the hemolymph. Later in the spring and in the summer, they increased in the hemolymph and on the body surfaces, while decreasing in the fat bodies. The global DNA methylation levels increased with age. Higher levels were always observed in the males than in the females. The study will promote better understanding of bee evolution and will be useful for the protection and management of solitary bees, with benefits to the environment and agriculture.

The Varroa destructor mite has recently displayed an ever increasing resistance to new drugs, contributing to CCD proliferation. This work was aimed at determining new viable methods for identifying the pyrethroid resistance of V. destructor and DNA methylation in resistant and sensitive mites. DNA was extracted from Varroa mites. Nucleotide changes in the DNA of pyrethroid-resistant, pyrethroid-sensitive, and control mites were identified with polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) in the case of five mitochondrial gene fragments. More bands were observed in the drug-resistant mites than in the other two groups. Sequencing confirmed these observations. Decreased global DNA methylation levels were observed in the pyrethroid-resistant mites. There exists a previously undescribed mechanism of pyrethroid resistance development in Varroa mites. The PCR-SSCP methods can be considered and further developed as useful tools for detecting V. destructor resistance.

We examined the influence of curcumin-supplemented feeding on worker lifespan, Nosema resistance, key enzyme activities, metabolic compound concentrations and percentage of the global DNA methylation. Two worker groups (Apis mellifera) were set up: 1) control group; workers were fed ad libitum with sucrose syrup; 2) workers were fed with the syrup with the addition of curcumin. Dead workers were removed every two days and the Nosema spp. infection levels were assessed. Hemolymph was taken from living workers for biochemical analyses. The global DNA methylation level was analysed using DNA from worker heads and thoraces. The bees that consumed curcumin lived longer and were less infested with Nosema spp. The curcumin-treated workers had higher concentrations of proteins, non-enzymatic biomarkers (triglycerides, glucose, cholesterol, Mg2+ and Ca2+), uric acid and creatinine, as well as elevated activities of antioxidant enzymes (SOD , GPx, CAT , GST ), neutral proteases, protease inhibitors, enzymatic biomarkers (AST , ALT , ALP ). The concentrations of albumin and urea, and the activities of acidic and alkaline proteases were higher in the control group. Curcumin decreased global DNA methylation levels especially in older bees in which the natural, age-related level increase was observed. Most of the parameters increased over the apian youth and adulthood, and decreased in older bees. The decrease was markedly delayed in the bees fed with curcumin. Curcumin appeared to be an unexpectedly effective natural bio-stimulator, improving apian health and vitality. This multifactorial effect is caused by the activation of many biochemical processes involved in the formation of apian resistance.

Trzem grupom pszczół w klatkach podawano syrop cukrowy (grupa kontrolna), syrop cukrowy z dodatkiem amfoterycyny B (AmB) w dawce 0,5 mg/ml oraz syrop cukrowy z AmB w dawce 0,25 mg/ml. AmB wpływała na skrócenie długości życia pszczół oraz obniżała poziom globalnej metylacji DNA w porównaniu do grupy kontrolnej, jednocześnie zwiększała stężenie białek na powierzchni ciała. AmB odkładała się w jelicie tylnym pszczół. Pszczoła miodna okazała się użytecznym modelem dla badań efektów ubocznych terapii antygrzybiczej z zastosowaniem AmB. Między innymi, za efekty uboczne takiej terapii uważa się zmiany epigenetyczne i przyspieszenie procesów starzenia.

We examined the influence of bromfenvinphos, a commonly used acaricide, on activities of many metabolic enzymes affecting the biochemical defences/physiology of the western honeybee, Apis mellifera L. (Hymenoptera: Apidae), as well as on some metabolic compound concentrations, percentage of global DNA methylation, and Nosema spp. infection levels. Bromfenvinphos-treated workers had decreased haemolymph volumes and higher protein concentrations on their cuticle but lower protein concentrations in the haemolymph. They had higher global DNA methylation levels independent of the age-related variants. Bromfenvinphos decreased the activities of antioxidant enzymes ( SOD, GPx, CAT, GST), acidic, neutral, and alkaline protease inhibitors and enzymatic physiological markers ( AST, ALT, ALP), and concentrations of urea, uric acid, creatinine, cholesterol, glucose, Mg2+, and Ca2+ in worker haemolymph, depending on the age of the bees. Protease activities were higher only in the haemolymph of young bromfenvinphos-treated bees in comparison with untreated bees. This compound decreased the activities of alkaline proteases and neutral protease inhibitors on the cuticle. Unexpectedly, in the treated bees, the activities of acidic and neutral proteases, and acidic and alkaline protease inhibitors, were higher in the young bees and lower in the older workers in comparison to the untreated group. The bromfenvinphos-treated workers were more heavily infested with Nosema spp. Thus, bromfenvinphos not only supressed many levels of biochemical defences, and therefore stress-resistance-related biochemical pathways but also visibly increased the Nosema spp. infection levels. [ABSTRACT FROM AUTHOR]

Simple Summary: The queens and sterile workers arise from genetically identical eggs but as imagoes, they differ in their life span, DNA methylation, and their functions. In the absence of the queen, the larvae develop into rebels, i.e., workers with increased reproductive potential. We assumed that since rebels are similar to the queen in many anatomical and behavioral features, they live longer and have lower levels of global DNA methylation, even when infected, e.g., by Nosema spp. Rebels always lived longer in comparison in normal workers and unexpectedly extended longevity of normal workers when they were together, similarly as the presence of a queen did. Rebels became infected more easily but tolerated the infection better. They also had lower level of global DNA methylation than normal workers. These features expand possibilities of the use of honeybees as a model for studies on senescence, nosemosis, eusocial evolution, and epigenetics. The widely accepted hypothesis in life history evolution about the trade-off between fecundity and longevity is not confirmed by long-living and highly fecund queens in eusocial insects. The fact that the queens and facultatively sterile workers usually arise from genetically identical eggs but differ in DNA methylation makes them a good model for studies on senescence, eusocial evolution, and epigenetics. Therefore, honeybees seem to be especially useful here because of long living rebel-workers (RW) with high reproductive potential recently described. Longevity, ovariole number, nosema tolerance, and global DNA methylation have been assayed in normal workers (NW) versus RW in hives and cages. RW always lived longer than NW and unexpectedly extended longevity of NW when they were together, similarly as the presence of a queen did. RW lived longer despite the fact that they had higher Nosema spore load; surprisingly they became infected more easily but tolerated the infection better. Global DNA methylation increased with age, being lower in RW than in NW. Therefore, RW are queen-like considering global DNA methylation and the link between fecundity, longevity, and body maintenance. Presented features of RW expands possibilities of the use of honeybees as a model for studies on senescence, nosemosis, eusocial evolution, and epigenetics. [ABSTRACT FROM AUTHOR]