We examined the influence of caffeine on honeybee lifespan, Nosema resistance, key enzyme activities, metabolic compound concentrations, and total DNA methylation levels. Caffeine slowed age-related metabolic tendencies. Bees that consumed caffeine lived longer and were not infested with Nosema spp. Caffeine-treated workers had higher protein concentrations. The levels increased with aging but they then decreased in older bees. Caffeine increased the activities of antioxidant enzymes (SOD, GPx, CAT, GST), AST, ALT, ALP, neutral proteases, and protease inhibitors, and the concentrations of uric acid, triglycerides, cholesterol, glucose, and Ca2+. Acidic and alkaline protease activities were lower in the bees treated with caffeine. Creatinine and Mg2+ concentrations were higher in the caffeine-treated workers but only up to 14 days of age. Caffeine significantly decreased DNA methylation levels in older bees. The compound could be considered as a natural diet supplement increasing apian resistance to stress factors. Our studies will enhance possibilities of using Apis mellifera as a model organism in gerontological studies.

Natural bioactive preparations that will boost apian resistance, aid body detoxification, or fight crucial bee diseases are in demand. Therefore, we examined the influence of coenzyme Q10 (CoQ10, 2,3-dimethoxy, 5-methyl, 6-decaprenyl benzoquinone) treatment on honeybee lifespan, Nosema resistance, the activity/concentration of antioxidants, proteases and protease inhibitors, and biomarkers. CoQ10 slows age-related metabolic processes. Workers that consumed CoQ10 lived longer than untreated controls and were less infested with Nosema spp. Relative to controls, the CoQ10-treated workers had higher protein concentrations that increased with age but then they decreased in older bees. CoQ10 treatments increased the activities of antioxidant enzymes (superoxide dismutase, GPx, catalase, glutathione S-transferase), protease inhibitors, biomarkers (aspartate aminotransferase, alkaline phosphatase, alanine aminotransferase), the total antioxidant potential level, and concentrations of uric acid and creatinine. The activities of acidic, neutral, and alkaline proteases, and concentrations of albumin and urea were lower in the bees that were administered CoQ10. CoQ10 could be taken into consideration as a natural diet supplement in early spring before pollen sources become available in the temperate Central European climate. A response to CoQ10 administration that is similar to mammals supports our view that Apis mellifera is a model organism for biochemical gerontology.

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: It was postulated that a phytobiotic preparation containing cinnamon oil and citric acid added to drinking water for chickens in a suitable amount and for a suitable time would beneficially modify the microbiota composition and morphology of the small intestine, thereby improving immunity and growth performance without inducing metabolic disorders. The aim of the study was to establish the dosage and time of administration of such a phytobiotic that would have the most beneficial effect on the intestinal histology and microbiota, production results, and immune and metabolic status of broiler chickens. The experiment was carried out on 980 one-day-old male chickens until the age of 42 days. The chickens were assigned to seven experimental groups of 140 birds each (seven replications of 20 individuals each). The control group (G-C) did not receive the phytobiotic. Groups CT-0.05, CT-0.1, and CT-0.25 received the phytobiotic in their drinking water in the amount of 0.05, 0.1, and 0.2 mL/L, respectively, at days 1–42 of life (continuous application, CT). The birds in groups PT-0.05, PT-0.5, and PT-0.25 received the phytobiotic in the same amounts, but only at days 1–7, 15–21, and 29–35 of life (periodic application, PT). Selected antioxidant and biochemical parameters were determined in the blood of the chickens, as well as parameters of immune status and redox status. The morphology of the intestinal epithelium, composition of the microbiome, and production parameters of chickens receiving the phytobiotic in their drinking water were determined as well. The addition of a phytobiotic containing cinnamon oil and citric acid to the drinking water of broiler chickens at a suitable dosage and for a suitable time can beneficially modify the microbiome composition and morphometry of the small intestine (total number of fungi p < 0.001, total number of aerobic bacteria p < 0.001; and total number of coliform bacteria p < 0.001 was decreased) improving the immunity and growth performance of the chickens (there occurred a villi lengthening p = 0.002 and crypts deepening p = 0.003). Among the three tested dosages (0.05, 0.1, and 0.25 mL/L of water) of the preparation containing cinnamon oil, the dosage of 0.25 mL/L of water administered for 42 days proved to be most beneficial. Chickens receiving the phytobiotic in the amount of 0.25 mL/L had better growth performance, which was linked to the beneficial effect of the preparation on the microbiome of the small intestine, metabolism (the HDL level p = 0.017 was increased; and a decreased level of total cholesterol (TC) p = 0.018 and nonesterified fatty acids (NEFA) p = 0.007, LDL p = 0.041, as well as triacylglycerols (TAG) p = 0.014), and immune (the level of lysozyme p = 0.041 was increased, as well as the percentage of phagocytic cells p = 0.034, phagocytosis index p = 0.038, and Ig-A level p = 0.031) and antioxidant system (the level of LOOH p < 0.001, MDA p = 0.002, and the activity of Catalase (CAT) p < 0.001 were decreased, but the level of ferric reducing ability of plasma (FRAP) p = 0.029, glutathione p = 0.045 and vitamin C p = 0.021 were increased).

The proteolytic system on the body surface of the honey bee has been insufficiently researched. In this study the body surface proteolytic activity was examined in queens at various developmental stages (eggs, larvae, pupae and imagines) in different seasons (spring, summer, autumn, winter). Extracts of the body surface material with water and detergent were used for an in vitro analysis of the proteolytic activity and protease inhibitor level assaying, as well as for an electrophoretic separation of the extracts in polyacrylamide gels. The following methods were used: protein content testing by the Lowry method (modified by Schacterle-Pollack), protease activity testing by the Anson method and protease inhibitor activity testing by the Lee and Lin method. Our studies revealed a high protease activity in an acidic environment (pH = 2.4; the material rinsed with detergent), as well as in neutral (pH = 7) and alkaline (pH = 11.2) environments (the material rinsed with water in both cases). The highest protein concentration values were observed in the imagines from summer. The lowest activities of the proteases and protease inhibitors were determined in the eggs from summer. The highest activities of the acidic, neutral and alkaline proteases were observed in the pupae from spring. The highest number of protease activity bands in PAGE zymography was obtained for the neutral and alkaline activities in the queens for all the seasons. In the queens all the catalytic protease types were present: asparagine and cysteine proteases at pH = 2.4; cysteine proteases and metalloproteases at pH = 7 and serine proteases at pH = 11.2. These results were crucial for the analysis of immunity mechanisms on the body surface of the honey bee.

The aim of the study was to test whether the use of probiotic bacteria Bacillus subtilis or Enterococcus faecium or a phytobiotic containing cinnamon oil can improve the metabolic parameters, immune status, gut microbiota and histology, and growth performance of broiler chickens. The experiment was carried out on 560 one-day-old male Ross 308 broiler chickens raised until the age of 42 days. The broiler chickens were assigned to 4 experimental groups of 140 birds each (7 replications of 20 individuals each). The control group (Control) did not receive additives. A probiotic preparation containing live bacterial cultures of Enterococcus faecium (EF, in the amount of 0.25 g/l) or Bacillus subtilis (BS, 0.25 g/l) or a phytobiotic preparation containing cinnamon oil (OC, 0.25 ml/l) was administered to the broiler chickens with their drinking water throughout the rearing period. The most important results indicate that the use of BS and OC resulted in: a significant (P≤0.05) increase in the level of ferric reducing ability of plasma (FRAP), high-density cholesterol (HDL) and glutathione (GSH + GSSH) and a significant (P≤0.05) decrease in the level of malondialdehyde (MDA), lipid hydroperoxides (LOOH), total cholesterol (TC), triacylglycerols (TAG), nonesterified fatty acids (NEFA) and interleukin 6 (IL-6), a ratio of heterophils : leukocytes (H:L) and alkaline phosphatase (ALP) activity, alanine aminotransferase (ALT), acidic phosphatase (AC) and creatinine kinase (CK), relative to the C group. In the blood of broiler chickens from the OC treatment, aspartate aminotransferase (AST), lactate dehydrogenase activity and 3-hydroxy-butyrate dehydrogenase (HBDH) significantly (P≤0.05) decreased in relation to the C group, and in broiler chickens from EF and BS treatments there was an increase (P≤0.05) in haemoglobin (Hb) content. Compared with group C, in the broiler chickens’ nutritional content from EF, BS and OC treatments, the total number of coliforms and number of fungi significantly (P≤0.05) dropped and the number of aerobic bacteria increased (P≤0.05) in the length of the villus and the depth of the crypt. It has been found that Bacillus subtilis, Enterococcus faecium and phytobiotic containing cinnamon oil can improve the microbiological and histological appearance of broiler chicken intestine. The addition of probiotic bacteria Bacillus subtilis or phytobiotic containing cinnamon oil to drinking water is more preferable than Enterococcus faecium regarding stimulation of the immune system, blood redox status parameters, parameters of metabolic changes and the gut microbiome and morphometry.

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________________________________________________________________________________ AbstractThe research focused on the effect of a prebiotic additive, mannanoligosaccharides (Bio-Mos), in the diet of turkey hens on their growth performance and measurements of pro-oxidation and antioxidation systems in their blood. The investigation was performed on 240 six-week-old turkey hens of the heavy Big-6 breed, randomly divided into two groups. Group I was the control group, whereas the birds in group II were fed the control diet with a 0.5% addition of Bio-Mos. Bio-Mos did not increase the concentration of lipid peroxidation products, peroxide H 2 O 2 and malone dialdehyde. However, it contributed to the increased concentration of some antioxidation parameters such as vitamin C, iron and zinc in the blood. Moreover, it led to an improved growth performance. The study suggested that mannanoligosaccharides can be used in practice as a dietary additive for turkeys, stimulating the mechanisms of the birds' antioxidation defence and improving their growth performance. ________________________________________________________________________________