Carbohydrate tetrachloride induces injury of liver parenchyma evoking fast and severe liver fibrosis, and is associated with irreversible structural and functional changes in testes, reducing fertility, decreasing potential pregnancy rate, and affecting its development. Liver showed high potential to regenerate, however the self-restoring after liver fibrosis was not accompanied with recovery of the reproductive system.

To edit the duck genome by HDR-directed integration of the EGFP gene into the duck host genome in combination with SMGT using CRISPR/Cas9. Methods. HDR-mediated gene of green fluorescent protein (EGFP) was crried out by the combined action of four plasmids. The pX330 contained the Cas9 gene. Two plasmids contained sgRNA genes: pBR322-sgRNA1 and pBR322-sgRNA2. The pBR322-HDR-EGFP plasmid was constructed to contain the DNA vector with left homologous sequence part(LHP), the EGFP gene coding sequences and the right homologous sequence part(RHP). The DNA sequence data for designing the HDR-EGFPinsert and sgRNA 1 and sgRNA 2 were taken from the genome DNA sequence of Anas platyrhynchos Spindlin 1 (SPIN1) gene. Twenty four ducks (13 males and 11 females) of the Shaoxing breed were used for this experiment. The sperm transfection was performed using Lipofectamine 2000. Results. Thirty one ducks were obtained, 19 of which carried the EGFP gene. F2 analysis revealed that 16 ducks (F1) (14 females and 2 males) transmitted the transgene DNA to their offsprings. Thus 27.6 % (56/203) of F2 descendants were positive for the transgene DNA construct. Conclusions. Exogenous DNA was successfully inserted into the duck genome.

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This article presents an elaboration of the protocol for the method of sexing wild birds based on the polymorphism of the CHD gene using P2/P8 primer for Common Pheasant – Phasianus colchicus (Linnaeus, 1758; Galliformes, Phasianidae); Silver Lofur or Silver Pheasant – Lophura nycthemera (Linnaeus, 1758; Galliformes, Phasianidae), Budgerigar – Melopsittacus undulatus (Shaw, 1805; Psittaciformes, Psittacidae), Herring Gull – Larus argentatus (Pontoppidan, 1763; Charadriiformes, Laridae), and White Stork – Ciconia ciconia (Linnaeus, 1758; Ciconiiformes, Ciconiidae). Blood samples were taken from Common Pheasant, Silver Pheasant and White Stork using the “drop of blood on paper” method. For the Budgerigar and the Herring Gull, DNA was isolated from the feather follicle. To isolate DNA, a commercial NeoPrep 100 DNA reagent kit (Neogen, Ukraine) was used. Primers P2/P8 were used for PCR; PCR was performed using GenPac PCR Core reagents (Neogen, Ukraine). We selected the optimal amount of Tag polymerase, the amount of DNA and primers and, according to the amount of reagents, set acceptable amplification modes and electrophoresis agarose gel percentage. Prior to PCR, additional DNA gel electrophoresis purification is proposed, which increases the percentage of positive sex determination results. It was found that the ideal mixture for the 5 bird species was an amplification mixture (total volume 20 µL, containing 1 U Tag polymerase, 100 ng DNA and 0.6 µM of each primer). The amplified CHD-Z fragment of Common and Silver pheasants is of ~340 n. p., CHD-W ~360 n. p. Herring Gull and Budgerigar have ~350 n. p. of CHD-Z length, and ~400 n. p. of CHD-W length, White Stork has its CHD-Z of ~ 370 n. p. long. It is advisable to investigate the genome of the experimental bird species using horizontal electrophoresis in agar’s gel with the concentration of 5%, which makes it possible to clearly visualize the female genotype. The universal protocol of the method of sex determination based on polymorphism of the CHD gene for the 5 studied bird species is described. These results of the study led to the conclusion that for the simultaneous sexing of several species of birds, it is advisable to develop a unified protocol for determining the status of the CHD gene, with the aim of clarifying the gender, as well as new approaches in ornithology and ecology aimed at determining interspecific differences associated with gene polymorphism. Identification of differences in fragment sizes may be useful for identifying the species in cases when birds form mixed pairings for taxonomic and phylogenetic comparisons.

Aim. Injury using CCl4 triggers a cascade of reactions and conditions specific for systemic lesions caused by most external anthropogenic environmental factors. Conditions for obtaining a model of systemic injury of an organism have been developed for further performing of restoration technologies with the use of stem cells and cytokines. Methods. Tumor predisposed ICR mice were chosen as the model. Systemic damage was caused with multiple intraperitoneal administration of carbon tetrachloride. Results. The optimal doses and load duration, excess above which leads to irreversible changes in the body that are not amenable to self-healing, were found. Conclusions. The model obtained can be used to study various recovery technologies of systemic damages. Keywords: systemic disease, model, carbon tetrachloride.

The paper studies interspecies physiological and genetic differences between the white-collared flycatcher and the pied flycatcher of the genus Ficedula. The fact that the flycatchers are capable of interspecies hybridization is one more reason for particular interest to these species. Using our own-developed method of taking venous blood from the eye’s sinus, we took blood samples from birds in the protected area of Homilshanski Forests, Kharkiv, Ukraine, to study their leukogram patterns. The bird feathers were also collected for genetic analysis -- to identify interspecies differences with application of the ISSR technique. It was revealed that the percentage of heterophiles in the nestlings of the pied flycatcher was lower than in the nestlings of the white-collared flycatcher. The micronucleus test did not reveal any significant difference in the interspecies groups. The spectra of amplification products obtained with the primer (AGC)6 G showed that the white-collared flycatchers had a more heterogeneous structure. The study of the leukogram, micronucleus test, and the ISSR analysis can be especially effective in the study of intra-species genetic differentiation.