Methionine ( Met ), an essential amino acid in poultry diets, when overdosed may cause hyperhomocysteinemia, which is mainly a trigger for cardiovascular diseases in humans. Homocysteine is neutralized (remethylated) in the presence of folic acid ( FA ), which also plays an important role in hematopoiesis and participates in the synthesis of DNA, and its deficiencies may result in the development of neural tube defects. One of the basic tools in studying the impact of both xenobiotics and nutrients on the animal organism is hematological analysis. Thus, the aim of this study was to determine the effect of in ovo supplementation with Met and FA on the hematological parameters of broiler chickens. On the 17th day of incubation, embryonated eggs (Ross 308) were injected with 5 or 25 mg of Met per egg (M5 and M25), 3 and 15 mg of FA per egg (F3 and F15), or a mixture of these 2 compounds (M5/F3 and M25/F15). The broilers were reared in accordance with welfare regulations and fed with commercial diets ad libitum . Blood samples were collected on the first, seventh, and 35th day of rearing (D1, D7, and D35), and complete hematological analysis was performed. The observed changes in red blood cell parameters probably result from physiological changes occurring during bird growth. Mean erythrocyte volume decreased with the age of chickens in the control, M5, and M25 groups, but not in those supplied with FA. Among supplemented groups, the number of white blood cells on D1 was lower only in group M5 than in the sham (C) group. The analysis of leukograms showed no significant differences between the groups. Comparing D1 with D7 in the group injected with a higher dose of Met and FA (MF25/15), a statistically significant increase in the percentage of lymphocytes and a significant decrease in the percentage of heterophils were observed. In addition, in the group injected with a higher FA dose (F15), there was statistically significant reduction in the percentage of eosinophils and a significant increase in the percentage of monocytes at day 7 compared with day 1. It seems that Met supplementation led to temporary immunosuppression in the animals.
Introduction Riboflavin (7,3S,4R)-2,3,4,5-tetrahydr oxypentyl]benzo[g]pteridine-2,4-dione, C 17 H 20 N 4 O 6 , vitamin B2) plays a key role in energy metabolism as a source of flavin mononucleotide (riboflavin-5′-phosphate, FMN) and flavin adenine dinucleotide (FAD). Moreover, it is a component of the flavin enzymes and coenzymes, which carry electrons in oxidation and reduction reactions, and it also plays a key role in the respiratory chain and the oxidation of fatty acids and amino acids. Furthermore, riboflavin is involved in the Krebs cycle and the metabolism of folate, vitamin B6, (pyridoxine), and vitamin B12 (cobalamin) (1). Riboflavin plays an important role in the innate immunity of both plants and animals (1-3). The immunomodulatory role of riboflavin in avian immunity is particularly highly appreciated (3).In vertebrates, flavin deficiencies lead to diseases such as glossitis, cheilosis, and organic acidurias (4). A deficiency of riboflavin is very rare in adult birds; however, its disappearance from the organism is associated with the overall hypovitaminosis (5-7). In avian species, riboflavin is required for proper embryonic development; therefore, it is accumulated in the developing chicken egg at an amount of about 300 µg per egg (8). Riboflavin deficiency is often related to mutations in an Rd gene encoding a riboflavin-binding protein (RfBP), which is responsible for deposition of riboflavin in eggs (9). In rd/rd mutants, effects of deficiency become apparent after day 10 of incubation (5). Embryo death occurs suddenly around day 13 of incubation and is the result of inhibition of activity of flavin-dependent enzymes, hypoglycemia, and impaired fatty acid oxidation. The adverse effects of the Rd gene mutation can be abolished by in ovo administration of free riboflavin or FMN but not RfBP (10). Riboflavinsupplemented embryos survived and developed properly, but those injected with RfBP died. This indicates that the unbound riboflavin that is injected into egg whites can be used by the developing embryo to ensure its proper
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