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African swine fever is one of the most serious threats to the world pig industry due to high infectivity and mortality rates among pigs. To date, no effective means of active prevention of the infection have been developed. The only effective method of control is passive monitoring of the spread of the pathogen among the population of domestic and wild pigs, detection of infected animals and their depopulation. The study analyzes the spread of African swine fever in Ukraine from 2012 to 2024 and compares it with the countries that share a common border – Poland, Romania, Hungary, Slovakia and Moldova. In Ukraine, the first outbreaks were recorded in 2012 in Zaporizhzhia region among domestic pigs. In total, 619 outbreaks were detected during the study period: 482 among domestic pigs and 137 among wild pigs. The largest number of outbreaks was recorded in Odesa (64), Poltava (54), Mykolaiv (52) and Kyiv (46) regions. In Eastern European countries, African swine fever was detected later: in Poland – since 2014 (1304 among domestic and 17871 among wild pigs), Romania – since 2017 (6729 and 3649, respectively), Hungary and Slovakia – since 2018 (0 and 7875; 72 and 3645). In Moldova, the first outbreak was in 2020 (39 among domestic and 45 among wild pigs). The highest total number of outbreaks was recorded in Poland (19175), mainly among wild boars (93.2%). In Hungary, all detected cases involved wild animals. The analysis revealed a statistically significant difference in the number of African swine fever outbreaks between the analyzed countries. There is also a difference in the number of outbreaks within the analyzed time period. If we analyze the number of cases since 2018, there is no statistically significant difference. Prevention and control of African swine fever are complicated by the circulation of the pathogen among wild boars, non-compliance with biosecurity measures by owners of small pig farms and the movement of infected animals. Comprehensive monitoring with early detection of outbreaks and timely destruction of infected animals plays a key role.

Coagulase-positive staphylococci are an important infectious agentcausing numerous infections in animals. Staphylococcus aureus andStaphylococcus pseudintermedius share a number of similar cultural andbiochemical characteristics, which makes their differentiation difficult.Since these species have different zoonotic potential, it is advisable todevelop rapid and specific schemes for species differentiation of pathogens.We have studied the cultural and biochemical characteristics ofStaphylococcus spp. isolated from dogs, cats and cows. In total, 103halophilic coccal cultures were isolated from companion animals and45 from cows, of which 55 and 30 cultures were coagulase-positivestaphylococci, respectively. The reactions that can be used to differentiateS. pseudintermedius and S. aureus were studied. Growth inhibition zonesaround the disk with polymyxin B antibiotic for S. pseudintermediuswere statistically higher (p

The results of optimization of the Staphylococcus spp. identification protocol by polymerase chain reaction with agarose gel detection and approbation of the protocol with wild strains selected from dogs are presented. Determination of the parameters of specificity and sensitivity of the method was performed on museum strains of cocci S. epidermidis ATCC 14990, S. aureus ATCC 25923, S. aureus subsp. aureus UKM B-918, S. pneumoniae ATCC 49619 and E. faecalis ATCC 194433. DNA extraction was performed using the IndiSpin Pathogen Kit. The ready PCR mix NEB OneTaq® 2X Master Mix with Standard Buffer was used to prepare the reaction mixture. Primers targeted to the tuf gene region using an amplification product of 370 bp were used for the study. The reaction results were recorded in a 2% agronomic gel with the addition of ethidium bromide at a concentration of 0.5%. The optimal annealing temperature was determined by the temperature gradient method. In a study of the specificity of the method, three museum strains of staphylococci were identified as positive, while strains of other cocci did not give reaction products. The sensitivity study of the method was to detect the amplification product in seven dilutions of bacterial suspension that meet McFarland turbidity standards, the lowest concentration was further diluted 10, 100 and 1,000 times. The last dilution, which showed the presence of the amplification product corresponds to 2×106 CFU in 200 μl of saline used for DNA isolation. PCR protocol was tested on wild staphylococcal strains. Ear and nasal swabs of dogs, as well as washes from the transfer cage were selected for the study. The primary inoculation of the material was carried out on mannitol salt agar, on this medium only the growth of halophilic microorganisms is possible. Growth was found on 17 Petri dishes. The PCR washings of these cups indicated the presence of staphylococci in the test materials. The results of in-laboratory PCR testing indicate that the primer we used gives high indicators of specificity and sensitivity. Our tested technique can be used to confirm the presence of Staphylococcus spp. bacteria in the primary culture of smears taken from dogs.