Peculiarities of morphological indicators of quail spleen (Coturnix japonica Temminck et Schlegel, 1849) when using protein-vitamin mineral supplements

O. Dunaievska, I. Sokulskiy, L. Horalskyi, M. L. Radzikhovskyi, B. Gutyj
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The question of increasing the productivity of quail breeding, as a branch of precocious animal husbandry, has always occupied the attention of biologists, morphologists, physiologists, geneticists and technologists. This topical problem has various scientific aspects: morphological, histochemical, physiological, genetic, etc. Special attention in the conducted research is required to study the impact of additives on the body of quails, the absence of negative effects and the production of safe products for humans. We chose the spleen as a biomarker, because the spleen is an important peripheral organ of immune protection, sensitive to the action of various factors, whose morphofunctional indicators determine the magnitude and nature of the impact. The purpose of the research was to determine the effect of modern protein-vitamin mineral supplements, which are advertised and offered by producers in the poultry industry to stimulate the growth and development of the bird, on the quail body. In the studies, the features of the microscopic structure and morphometry of the spleen of control and experimental animals were used as sensitive characteristics of the action of additives. The proposed methods are aimed at researching the impact of protein-vitamin mineral supplements on the relevant industry. So, for 21 days, mineral supplements “Multilife”, “Missy” and “Standard Agro” were added to the main ration of quails. Using macrometric methods, the body weight of quails, the weight of the spleen (absolute and relative) were determined. For histological studies, pieces of the spleen were fixed in a 10 % formalin solution, and after embedding in paraffin, histological sections were prepared from it, which after deparaffinization were stained with hematoxylin and eosin, according to the methods of Brachet and Van Gieson. For electron microscopic examination, the spleen was immediately fixed with a 2.5 % solution of glutaraldehyde in a phosphate buffer with additional fixation in a 1 % solution of osmium tetroxide according to Caulfield. For the first time, the ultrastructural organization of the spleen of Japanese quail was investigated after feeding protein-vitamin mineral supplements, which may indicate the absence of their negative impact on the structural and functional state of the spleen. Important morphological structural indicators of the morphofunctional state of the spleen were determined: the relative area of the white pulp, red pulp, lymphoid nodules, periarterial lymphoid sheaths, and the contractile apparatus. According to the analysis of the morphological results obtained by us, it was investigated that the microscopic structure of the quail spleen is characterized by the presence of a support-contractile apparatus with a relative area of 4.58 ± 0.76 % and single radial trabeculae. The parenchyma is formed by white and red pulp without clear boundaries. Lymphoid nodules without bright centers, periarterial lymphoid sheaths, and ellipsoids were identified as part of the white pulp. The ultrastructural organization of the spleen of quails had peculiarities: the nuclei of leukocytes contain large nucleoli, the endotheliocytes of the central artery of the lymphoid follicle perform a phagocytic function by accumulating autophagosomes in their cytoplasm. The macroscopic, microscopic and ultramicroscopic structure of quails that were added to the main diet with the listed additives did not undergo any changes. As a result, the following supplements led to an increase in the body weight of quails: by 5.78 % when using the Multilife supplement, by 12.51 % when using the Missy supplement, and by 8.12 % when using the Standard Agro supplement, and the weight of the spleen on 7.17 %, 16.04 % and 10.54 %, respectively. The conducted studies showed that the relative weight of the organ practically did not change. At the same time, there is a trend towards an increase in the relative area of the white pulp and the supporting-contractile apparatus. Taking into account the obtained data, it can be concluded that our research confirms the feasibility of using modern protein-vitamin mineral supplements in poultry farming to stimulate the growth and development of poultry. 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Abstract

In most countries of the world, poultry farming is one of the main branches of agriculture, which provides the population with high-quality food products. To date, the positive dynamics of the growth of such a branch of industrial poultry farming as quail breeding has been noted. Quails have a complex of excellent economic and productive advantages compared to other agricultural birds – high body temperature, intensive metabolism in the body, small size, precocity, high egg productivity. The question of increasing the productivity of quail breeding, as a branch of precocious animal husbandry, has always occupied the attention of biologists, morphologists, physiologists, geneticists and technologists. This topical problem has various scientific aspects: morphological, histochemical, physiological, genetic, etc. Special attention in the conducted research is required to study the impact of additives on the body of quails, the absence of negative effects and the production of safe products for humans. We chose the spleen as a biomarker, because the spleen is an important peripheral organ of immune protection, sensitive to the action of various factors, whose morphofunctional indicators determine the magnitude and nature of the impact. The purpose of the research was to determine the effect of modern protein-vitamin mineral supplements, which are advertised and offered by producers in the poultry industry to stimulate the growth and development of the bird, on the quail body. In the studies, the features of the microscopic structure and morphometry of the spleen of control and experimental animals were used as sensitive characteristics of the action of additives. The proposed methods are aimed at researching the impact of protein-vitamin mineral supplements on the relevant industry. So, for 21 days, mineral supplements “Multilife”, “Missy” and “Standard Agro” were added to the main ration of quails. Using macrometric methods, the body weight of quails, the weight of the spleen (absolute and relative) were determined. For histological studies, pieces of the spleen were fixed in a 10 % formalin solution, and after embedding in paraffin, histological sections were prepared from it, which after deparaffinization were stained with hematoxylin and eosin, according to the methods of Brachet and Van Gieson. For electron microscopic examination, the spleen was immediately fixed with a 2.5 % solution of glutaraldehyde in a phosphate buffer with additional fixation in a 1 % solution of osmium tetroxide according to Caulfield. For the first time, the ultrastructural organization of the spleen of Japanese quail was investigated after feeding protein-vitamin mineral supplements, which may indicate the absence of their negative impact on the structural and functional state of the spleen. Important morphological structural indicators of the morphofunctional state of the spleen were determined: the relative area of the white pulp, red pulp, lymphoid nodules, periarterial lymphoid sheaths, and the contractile apparatus. According to the analysis of the morphological results obtained by us, it was investigated that the microscopic structure of the quail spleen is characterized by the presence of a support-contractile apparatus with a relative area of 4.58 ± 0.76 % and single radial trabeculae. The parenchyma is formed by white and red pulp without clear boundaries. Lymphoid nodules without bright centers, periarterial lymphoid sheaths, and ellipsoids were identified as part of the white pulp. The ultrastructural organization of the spleen of quails had peculiarities: the nuclei of leukocytes contain large nucleoli, the endotheliocytes of the central artery of the lymphoid follicle perform a phagocytic function by accumulating autophagosomes in their cytoplasm. The macroscopic, microscopic and ultramicroscopic structure of quails that were added to the main diet with the listed additives did not undergo any changes. As a result, the following supplements led to an increase in the body weight of quails: by 5.78 % when using the Multilife supplement, by 12.51 % when using the Missy supplement, and by 8.12 % when using the Standard Agro supplement, and the weight of the spleen on 7.17 %, 16.04 % and 10.54 %, respectively. The conducted studies showed that the relative weight of the organ practically did not change. At the same time, there is a trend towards an increase in the relative area of the white pulp and the supporting-contractile apparatus. Taking into account the obtained data, it can be concluded that our research confirms the feasibility of using modern protein-vitamin mineral supplements in poultry farming to stimulate the growth and development of poultry. Especially valuable is the absence of a negative effect on the morphofunctional indicators of the spleen, which was used as a biomarker in research.
使用蛋白质-维生素-矿物质补充剂时鹌鹑脾脏(Coturnix japonica Temminck et Schlegel, 1849)形态指标的特殊性
在世界上大多数国家,家禽养殖是农业的主要分支之一,为居民提供优质食品。迄今为止,鹌鹑饲养这一工业化家禽饲养分支的发展势头良好。与其他农禽相比,鹌鹑具有经济和生产方面的综合优势--体温高、体内新陈代谢旺盛、体型小、早熟、产蛋率高。作为早熟畜牧业的一个分支,提高鹌鹑繁殖生产力的问题一直是生物学家、形态学家、生理学家、遗传学家和技术专家关注的焦点。这一热点问题涉及多个科学方面:形态学、组织化学、生理学、遗传学等。在进行研究时,需要特别注意研究添加剂对鹌鹑身体的影响、无负面影响以及生产对人类安全的产品。我们选择脾脏作为生物标志物,因为脾脏是重要的免疫保护外周器官,对各种因素的作用敏感,其形态功能指标决定了影响的大小和性质。研究的目的是确定现代蛋白质-维生素-矿物质补充剂对鹌鹑身体的影响,这些补充剂是家禽业生产商为刺激鸟类生长发育而宣传和提供的。在研究中,对照组和实验组动物脾脏的显微结构和形态特征被用作添加剂作用的敏感特征。所提出的方法旨在研究蛋白质-维生素矿物质补充剂对相关产业的影响。因此,在鹌鹑的主饲料中添加了矿物质补充剂 "Multilife"、"Missy "和 "Standard Agro",为期 21 天。使用宏观测量方法测定了鹌鹑的体重、脾脏重量(绝对重量和相对重量)。为了进行组织学研究,将脾脏切片固定在 10% 福尔马林溶液中,然后用石蜡包埋,制备组织学切片,按照 Brachet 和 Van Gieson 的方法,去石蜡后用苏木精和伊红染色。为了进行电子显微镜检查,脾脏立即用 2.5 % 的戊二醛溶液在磷酸盐缓冲液中进行固定,然后按照考菲尔德的方法用 1 % 的四氧化锇溶液进行固定。这是首次对日本鹌鹑饲喂蛋白质-维生素-矿物质补充剂后的脾脏超微结构组织进行研究,这可能表明补充剂对脾脏的结构和功能状态没有负面影响。测定了脾脏形态功能状态的重要形态结构指标:白髓、红髓、淋巴结、动脉周围淋巴鞘和收缩器的相对面积。根据我们对形态学结果的分析,研究发现鹌鹑脾脏的微观结构特点是存在相对面积为 4.58 ± 0.76 % 的支持-收缩器和单个径向小梁。实质由白色和红色浆液组成,边界不清。白髓中有无亮点的淋巴结、动脉周围淋巴鞘和椭圆体。鹌鹑脾脏的超微结构组织具有特殊性:白细胞的核含有大的核小体,淋巴滤泡中央动脉的内皮细胞通过在其胞质中积累自噬体而发挥吞噬功能。在主食中添加所列添加剂后,鹌鹑的宏观、微观和超微结构没有发生任何变化。因此,以下添加剂导致鹌鹑体重增加:使用 Multilife 添加剂时增加 5.78%,使用 Missy 添加剂时增加 12.51%,使用 Standard Agro 添加剂时增加 8.12%,脾脏重量分别增加 7.17%、16.04% 和 10.54%。研究表明,器官的相对重量实际上没有变化。同时,白浆和支持-收缩器的相对面积有增加的趋势。考虑到所获得的数据,可以得出结论:我们的研究证实了在家禽养殖中使用现代蛋白质-维生素-矿物质补充剂来刺激家禽生长发育的可行性。尤其难能可贵的是,研究中作为生物标志物的脾脏形态功能指标没有受到负面影响。
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