УДК 637.4.082.474:637.412 ZOOHYGIENIC CHARACTERISTICS OF BIOTECHNOLOGICAL METHODS OF METABOLISM REGULATING OF CHICKENS EMBRYO IN THE PROCESS OF INCUBATION

The research presents the results of experimental studies of the influence of various physicochemical factors (phonophoresis, electrophoresis, electrospray, diffusion, etc.) on the rate of transport of biologically active substances through the protective layer of hatchig eggs shell of crossbred Haysex Brown. The aim of the study was to compare the effectiveness of the use of physicochemical methods of transportation of biologically active substances through the shell of hatching eggs of chickens. For the experiment, three batches of eggs were formed, which were obtained from layer hens Haysex Brown, 144 pieces in each experimental group. The kinetic parameters of BAS transport through bioceramic protective barriers of eggs were calculated based on the determination of BAS concentration on the surface and inside the egg by mass spectrometric method (mass spectrometer with ionization of 252Cf fission fragments "MSBH", (Ltd "SELMI", Sumy, Ukraine). The degree of permeability of bioceramic layers of the shell relative to the model gas mixture which is identical to the atmospheric air was studied by the method of V. Breslavets et other and the mass spectrometric method (gas mass spectrometer "MX 7304A"). Electron microscopic studies were performed on a scanning electron microscope REMMA-102; Visilog 6.11 (Noesis, Belgium) was used to process the obtained digital images to determine the number of shell microdefects per unit area of the digital image. It has been experimentally proved that to increase the efficiency of transport of biologically active substances (BAS) through the bioceramic layers of the shell of hatching eggs of Loman Brown hens, it is advisable to use ultrasonic treatment (phonophoresis, sonophoresis), substances - enhancers, including plant terpenes (L-menthol), DMSO and cyclodextrin. Phonophoretic treatment of hatching eggs increases the hatchability of eggs by 7.6%. At the same time, the gas permeability of the bioceramic layer increases significantly (by 0.43. 10-4 m3 / m2 s).

The problem of delivery of BAS to the area embryo development is achieved by fundamentally different methods [5,6,9,25]. One of them is the introduction of BAS and vaccines in the middle of the hatching egg ("in ovo") using a precision needle microdose (technology Embrex Inc., USA). However, this technology involves a local violation of the integrity of the bioc-eramic layer [4,11]. Also known are technologies for regulating the metabolism of avian embryos that use passive and active transport of BAS through the bioceramic layer. The main driving factor of the first is the free diffusion of substances that are part of the BAS, through the calcite layer of the shell [12,15,18,19].
Factors of active transport include accelerated diffusion and transfer by means of "transporter molecules" and so-called "enhancers" -substances that enhance the transfer processes [13,24,26]. Peculiarities of both approaches to transshell transfer are combined in the technology of "artificial cuticle" for hatching eggs [16]. We note that due to the complex, multicomponent and heterogeneous structure of bioceramic and glycoprotein protective layers of the shell and supra-and subshell membranes, detailed studies of the mechanisms and kinetic parameters of transport of organic substances that are part of BAS, different molecular weight, shape and charge 20,22,23,27].
The aim of the study was a comparative study of the efficiency of transport of biologically active substances through the shell of hatching eggs of Haysex Brown chickens.
The degree of permeability of bioceramic layers of the shell relative to the model gas mixture which is identical to the atmospheric air was studied by the method of V. Breslavets et al. [3] and mass spectrometric method (gas mass spectrometer "MX 7304A"). Electron microscopic studies were performed on a scanning electron microscope REMMA-102, when processing the obtained digital images to determine the number of microdefects of the shell per unit area of the digital image (Y; number of channels,%; table), used the program Visilog 6.11 (Noesis, Belgium). Experimental results (repetition not less than n = 5) were processed statistically using the package Statistica 5.1.
Results of research and discussion. The table shows the results of a series of experiments comparing the degree of efficiency of methods of transporting BAS through the biocrystalline layer of the shell. The control was taken as the amount of BAS received by conventional diffusion for 12 hours at a humidity level of 95-98% and a temperature of 1800,2 0 C; in the middle of the egg at equal temperatures outside and inside the egg. At the end of the experiment, a drop of BAS on the outer surface of the shell was dried, then the egg was carefully retrieved from the protein and yolk, dried again, then selected 0.5 cm2 shell with a layer of BAS applied to the outer surface, broken into 2 equal parts, fixed on the disk surface to keep the sample of the device "IASB" (one part of the outer surface, the other inner). Thus, the analysis of the organic component of the outer and inner surfaces of the shell allowed a comparison in a certain approximation of the number of BAS received during incubation in the middle of the egg. Thus, in the case of normal diffusion in the middle of the egg received 0.03 ± 0.009% of the amount of BAS contained on the surface.
The table shows that the most promising factors for nondestructive transfer of BAS through the biocrystalline layer are ultrasonic treatment (phonophoresis, sonophoresis), plant terpenes (L-menthol), which are already used in biotechnology as enhancers of BAS transport through natural protective structures (leather, bioceramics, etc.), DMSO and CD.
In particular, phonophoretic treatment of hatching eggs of Lohmann Brown crosses increases the hatchability from the initial value of 80.4% to 88.0% (+7.6). This significantly increases the gas permeability (up to 1.91 from the initial value of 1.48. 10 -4 m3 / m2 s) (+ 0.43) under conditions of transfer to

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Серія «Тваринництво», випуск 4 (43), 2020 the middle of the egg 79.2% of the BAS of the total number of eggs applied to the surface (Table 1). It is noteworthy that there is no unambiguous correlation between the level of morphological "ordering" of bioceramic layers of the shell, which in our work is expressed by Y (the number of microdefects in the shell per unit area of digital image of the shell), and the level of gas permeability of the bioceramic layer. Thus, the number of microdefects of the bioceramic layer per unit area of the shell in comparison with the control significantly increases (+ 15.9%) under the influence of "hydraulic shock", as well as with the use of phonophoretic techniques (+ 8.8% relative to control). However, the corresponding increase in the number of BAS received in the middle of the egg is + 22.07% and 79.17% compared to the control. These results provide grounds for assuming the existence of different mechanisms of transfer of organic compounds through the bioceramic layer in the first and second cases. In particular, phonophoretic technology is based on the use of ultrasonic radiation, which is characterized by the so-called "sonocapillary effect", which consists in multiple (sometimes orders of magnitude) enhancement of capillary transport of liquid phase substances in heterogeneous solid phase media.
At the same time, the technique of "hydraulic shock", which has proven itself well in devices for non-destructive transfer of liquid substances through the membrane systems of living tissues, in the case of solid-phase structures is not effective enough. In this aspect, the electrospray technique proved to be more effective -as can be seen from the table, the number of microdefects of the bioceramic layer per unit area of the shell does not differ significantly from the control. However, the efficiency of trans shell transfer exceeds the corresponding figure inherent in the technique of "hydraulic shock" by 58.1%.
According to our data, very promising are the classic substances-"enhancers" -DMSO and plant terpenes (Lmenthol). These substances significantly increase the rate of trans shell shell transfer (+ 65.07% and 58.17%, respectively) and at the same time have little effect on the degree of ordering of heterogeneous layers of bioceramics (table). DMSO and Lmenthol also have a positive effect on the egg hatchability, although they are not able to increase the gas permeability of the bioceramic layer.
Conclusions. It has been experimentally proved that to increase the efficiency of transportation of biologically active substances (BAS) through the bioceramic layers of the shell of hatching eggs of Loman Brown chickens, it is advisable to use ultrasonic treatment (phonophoresis, sonophoresis), enhancers, including plant terpenes (L-menthol), DMSO and cyclodextrin.
Phonophoretic treatment of hatching eggs increases the hatchability of eggs by 7.6%. At the same time, the gas permeability of the bioceramic layer increases significantly (by 0.43. 10-4 m3 / m2  s).
Acknowledgment. The work has been performed under the financial support of the Ministry of Education and Science of Ukraine (state registration number 0119U100551).