Detection of the macrolide phosphorylase gene in the intestinal contents of pigs

The growth and development of agricultural production is restricted by infectious diseases and, as a consequence, leads to the associated economic losses. Antibiotics are widely used to prevent the occurrence of infectious diseases. Their uncontrolled use entails the emergence of new antibiotic-resistant bacterial strains. The determining factor of bacterial resistance to the macrolide group of antibiotics is the macrolide phosphorylase (MphA) gene. It is found in the genome of bacteria of the Enterobacteriaceae. The presence of the MphA gene in food in the long term can adversely affect the health of the nation due to the development of resistance to macrolide antibiotics in people. In this regard, modern agricultural production requires simple and massive methods for determining the antibiotic resistance of microorganisms. In this regard, the aim of the study was to develop a real-time polymerase chain reaction (RT-PCR) for the possibility of monitoring the MphA gene in the intestinal contents of pigs and poultry.Samples of feces (80 samples) from pigs aged 40-50 days served as the material for the study. Isolation of DNA by traditional silico-sorption method were used. The selection of oligonucleotide primers was performed using the IDT Oligo Analyzer software. RT-PCR was set up on a BioRad CFX96 Real-Time PCR-System. As a result of the research, 30% of positive samples were identified. At the same time, the threshold reaction cycle varied from 36 Q to 42С1 This fact proves the high distribution of the macrolide phosphorylase gene in the intestinal contents of pigs and poultry at agricultur al enterprises in the Russian Federation. However, according to our data, the number of copies of the desired gene was not very high.Based on the data obtained and the analysis of the literature, we believe that the problem of antibiotic resistance must be solved in a complex way through specific and non-specific prevention of infectious diseases. Specific prophylaxis includes the widespread use of bacteriophages and autogenous vaccines. Non-specific prophylaxis includes the use of prebiotics, probiotics, and silver-containing preparations.

pig breeding, poultry farming, antibiotic resistance, macrolide phosphorylase, polymerase chain reaction

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