The changes of alkaline and acid phosphatase activity and the indexes of mineral metabolism in laying hens under the influence of selenium and zinc nanochelates and vitamin E

• Published in: Bìoresursi ì prirodokoristuvannâ
• Main Authors: Nischemenko, N., Koziy, V., Trokoz, V., Omelchuk, O., Poroshynska, O., Stovbecka, L., Еmelynenko, A.
• Format: Article in Journal/Newspaper
• Language: Ukrainian
• Published: Національний університет біоресурсів і природокористування України 2019
• Subjects: Alkaline and acid phosphatases; selenium nanoaquachelates; zinc; vitamin E; Calcium; inorganic Phosphorus; laying hens; productivity; лужна і кисла фосфатази; наноаквахелати; Селен; Цинк; вітамін Е; Кальцій; неорганічний Фосфор; кури-несучки; продуктивність; Carbonic acid
• Online Access: http://journals.nubip.edu.ua/index.php/Bio/article/view/13449; https://doi.org/10.31548/bio2019.05.020

In the diet of laying hens, the level of their mineral supply, in particular calcium and phosphorus, is important. They are especially necessary during the period of intensive egg position. These elements are a large part of the shell, and so their performance depends largely on the egg productivity and egg quality. (Suray, 2006; Lemesheva, 2007). A peculiarity of calcium exchange in egg-laying poultry is that the laying hens use a stock of calcium accumulated in the bone marrow, which is sufficient to form 6 to 30 eggs (Bilokon et al., 2012; Shtele, 2017). Inorganic phosphorus an important role in metabolism in the birds body plays. About 80–85 % of phosphorus is found in bone and 15–20 % is in other tissues and fluids. Of great importance in the phosphorus-calcium metabolism regulation belongs to the group of enzymes-phosphatases (Ionov et al., 2011; Melnyk, 2008), in particular alkaline (EC 3.1.3.1) and acidic (EC 3.1.3.2) phosphatases, which take participation in the catalysis of phosphorus esters in blood plasma and in tissues.The purpose of the study was to investigate the effect of selenium and zinc nanoaquachelates with vitamin E on alkaline and acid phosphatase activity, calcium and inorganic phosphorus content in the serum of laying hens.Methods. The experiments were performed on laying hens of the Lohmann-Brown breed, aged 45 weeks. Feeding of the birds was carried out with dry balanced feeds with nutrition in accordance with the rules of feeding. It was formed by the method of analogues of two groups of chickens: control and experimental, 40 heads each. The confinement conditions corresponded to the hygienic parameters, the chickens were kept in cages with free access to feed and water. Layers of the experimental group per head were obtained in the standard diet: Zn 30 ml/kg, SE – 30 ml/kg and vitamin E – 40 mg/kg, and in the control group for each head was added to the diet 30 ml. of distilled water. The experiment lasted 90 days, accounting for egg productivity throughout the experiment for each group separately. During the experiment serum calcium and inorganic phosphorus content were investigated using the method described by Danchuk et al. (2013). The activity of alkaline and acidic phosphatases was determined using the Felicity Diagnostic Kit. Other conditions of the experiment were reported previously (Nischemenko et al., 2019). The obtained research results were processed by statistics conventional methods of using Microsoft Excel.Results and discussion. It is known that enzymes are high molecular weight organic compounds of a protein nature that act as biological catalysts in a living organism. The group of phosphatases is alkaline phosphatase, which is involved in the catalysis of phosphorus esters in blood plasma and tissues. In birds, the value of this enzyme is also that it is actively involved in the metabolism of minerals in the body of laying and transfer of calcium ions in the formation of eggshells (Fisinin, 2016). The activity of alkaline serum phosphatase is due to the intensity of metabolic processes. It has been established that excessive increase of enzyme activity in 2-3 times in comparison with the norm is observed in cholestasis and disturbance of mineral metabolism. The activity of LF increases most often in the animal during its intensive growth and development, and in laying hens – during egg formation and egg position (Fisinin, 2016; Nishchemenko et al., 2016).Significant physiological growth of alkaline phosphatase activity during the period of active egg position is necessary for laying hens, since the enzyme mentioned above is involved, both in metabolic processes and in the transfer of calcium and phosphorus ions. In the laying body, these macronutrients are used to form the eggshell, as indicated by a number of literary messages (Okolelova, 2016; Okolelova, 2016-1).We found that alkaline phosphatase activity increased significantly by the 60-90th day of the experiment by 32.3–38.6 % in the layers of the experimental group compared with the control. Therefore, an increase in the activity of LF in the serum of laying hens during the period of active egg position is a necessary adaptive response of their body. It can be assumed that alkaline phosphatase activity increases during this period due to the bone isoenzyme and is ensured by the intensive functioning of osteoclasts, and the presence in the diets of selenium and zinc chelates with vitamin E contributes to the increase of activity of this enzyme.Acid phosphatase activity also increased on the 30th day of the study, but its probable increase was observed only on the 60th and 90th day of the experiment, respectively, by 16.5–21.0% (p <0.01). Such an increase in the activity of CF is, in our opinion, necessary for the active formation of the eggshell, which provides the necessary concentration of calcium ions and inorganic Phosphorus. Other researchers also point to this fact (Batman, 1968; Apukhovska et al., 2006). It should be noted that the deposition of mineral salts is catalyzed not only by alkaline but also acidic phosphatase. In addition, the enzyme carbonic anhydrase promotes the formation of carbonic acid, which, with Ca2 ions, forms calcium carbonates. In this case, acid phosphatase also promotes the transport of phosphorus-containing compounds that are required for the formation of the mineral base of the shell. There are limited reports in the scientific literature on the effect of nanoaquachelates of different elements on the physiological state and activity of phosphatases (Kaplunenko et al, 2007; Borysevych et al., 2009; 2009-1) in laying hens at industrial plants. Therefore, the study of the effect of selenium and zinc nanoaquachelates with vitamin E on the body of the layers requires careful study, including the effect of these drugs on the activity of enzymes and other indicators. As a result of the 90-day experiment, we determined changes in the concentration of calcium and inorganic Phosphorus in the blood of laying control and experimental groups. Prior to the experiment, the levels of calcium and inorganic Phosphorus in both groups of chickens were almost the same. He underwent some changes in the experimental group of hens after consumption of chelate compounds of selenium, zinc and vitamin E. In particular, the concentration of calcium in the blood of chickens in the experimental groups during the experiment was higher than in the control at 12.8-18.2 %. However, the probable increase in the level of this important macronutrient was established by us only in the 90th day of research. The increase in the content of Ca++ ions in the blood of laying hens during the period of intensive egg position can in our view be explained by its active "leaching" from the bone tissue and intensive absorption from the gastrointestinal tract for the needs of the body and, in particular, the formation of the shell.It should be noted that alkaline phosphatase, which transports calcium ions, plays an important role in the formation of the shell. The thickness of the shell depends largely on mineral nutrition and the presence in the diet of calcium, inorganic Phosphorus and vitamins D and E (Apukhovska et al., 2006). It is known that, with each egg, the laying egg loses about 2.0–2.5 g of calcium from its body (Palii, 2019; Kavtarashvili & Okolelova, 2006). The opinion is also expressed (Kavtarashvili & Okolelova, 2006) that two-thirds of this amount of macronutrients comes from the consumption of calcium feed and a third from the body's reserves.Analyzing changes in the level of inorganic Phosphorus, we note that in the blood of chickens of the experimental group compared with the control group of this element during the experiment was on average more than 3.2–26.3 %, but the probable increase was established only on the 90th day of research. The absorption of phosphorus by the body of a bird depends on the form in which it is included in the diet. In the diet, this trace element is in the form of organic compounds - phosphatides, phosphoproteins, which are well soluble in water and more accessible to the body of laying hens, and contributes to the presence of selenium and zinc chelates in combination with vitamin E. These changes can also be explained by that Zinc and Selenium provide transport processes related to the metal-enzyme transformation of a large number of biochemical compounds, and Vitamin E, in turn, also participates in various biological reactions and facilitates the processes of oxidation, reduction, etc. in the body of a bird.We hypothesize that an increase in the concentration of Phosphorus in the serum of chickens during the period of intensive egg position should be considered as an adaptive response of the organism, which is due to the need for increased synthesis of proteins used to form egg proteins. On the other hand, during ovogenesis, inorganic Phosphorus is used as an integral part of the egg shell (Okolelova, 2016; Liebers et al., 2019). We found a positive correlation between the presence in the diet of chelate compounds of selenium, zinc with vitamin E and the activity of alkaline and acid phosphatase (r = 0,510 and r = 0,390), calcium (r = 0,618) and inorganic phosphorus (r = 0,430) in the serum layer. It was found that egg laying increased in chickens of the experimental group compared to the control by 8.6%, improved morphometric and qualitative indicators of eggs (Nishchemenko et al., 2019).Conclusions. Addition to the diet of laying hens of nanoaquachelates selenium and zinc with vitamin E probably increases the activity of alkaline and acid phosphatase in the serum of laying hens by 32.3–38.6 and 16.5–21.0 %, respectively. A significant increase in calcium concentration of 18.2 % and inorganic phosphorus was found to be 26.3 % in the blood of chickens receiving mineral-vitamin supplement for the 90th day of the experiment. In other research terms, there is a tendency to increase the studied indicators level. The study’s results suggest that chelated forms of selenium, zinc, and vitamin E themselves as synergists in the regulation of mineral metabolism in the layers body show. This effect of nanoaquachelates is based on their biophysical properties and the activation of many biochemical processes according to the Borisovich – Kaplunenko – Kosinov effect (Borysevych et al., 2009, 2010). Сучасне птахівництво України – це галузь сільського господарства, яка найбільш інтенсивно розвивається та її важливим завданням є покращення життєздатності птиці на різних етапах розвитку, а також підвищення продуктивності. Одним із можливих методів збільшення яєчної продуктивності є застосування біологічно активних препаратів, у тому числі й наноаквахелатів біогенних та біоцидних металів, таких як Аргентум, Купрум, Магній, Кобальт, Селен, Цинк тощо. Ці біологічно активні речовини поліпшують фізіологічний стан і обмін речовин, сприяють кращому засвоєнню поживних речовин раціону птиці, а також позитивно впливають на несучість курей. Відомо, що у країнах Європейського союзу та США заборонено використання антибіотиків, які через можливі залишки в продукції мають негативний вплив на організм споживача. На сьогодні в Україні розробляються нові методи та застосовуються альтернативні кормові добавки, що здатні замінити антибіотичні стимулятори продуктивності, росту, розвитку та захисту здоров’я птиці. До речовин, які тут є перспективними відносять наноаквахелати біогенних та біоцидних металів і, зокрема, Селену та Цинку. Для вивчення впливу цих елементів разом із токоферолом на активність важливих ензимів – лужної та кислої фосфатази, а також на показники мінерального обміну проведені досліди на курках-несучках породи Ломан Браун. Встановлено, що згодовування наноаквахелатів Селену і Цинку з вітаміном Е позитивно впливає на активність лужної та кислої фосфатази, яка зростала протягом досліду відповідно на 17,2–38,6 та 16,5–21,0 %. Уміст Кальцію та неорганічного Фосфору в сироватці крові курок-несучок також збільшився. Зокрема, вміст Кальцію у дослідних несучок був вірогідно вищим під час експерименту на 12,8–18,2 % порівняно з контрольною групою. Така ж картина була і стосовно неорганічного Фосфору (10,0–26,3 %). Отримані нами результати свідчать про збільшення активності лужної і кислої фосфатази, стимуляцію фосфорно-кальцієвого обміну, завдяки вираженим властивостям наноаквахелатів біогенних металів Селену, Цинку з вітаміном Е, які засновані на їх біофізичних властивостях і активації багатьох біохімічних процесів згідно з ефектом Борисевича–Каплуненка–Косінова.