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The detection of Campylobacter’s sources contamination in beef
An increased risk of food poisoning in human during consumption of contaminated food and water is registered in Ukraine and worldwide every year. A significant part in the etiology of acute intestinal infections belongs to "non-traditional" bacteria. These microorganisms are most important is Campylobacter genus, which account for 10–15 % of sporadic cases of diarrheal disease, as well as a significant amount of water and food, including milk outbreaks described in foreign literature.
The main natural reservoir of Campylobacter are chickens, turkeys, wild birds, rodents and cattle, sheep, goats and pigs. There are Campylobacter localized mainly in the intestine and excreted in faeces infecting the environment, and during slaughtering and primary processing – products of slaughter in cattle.
This infection is spread with contaminated feeding, drinking bowls, tools, bedding, infected food, especially animal water.
The detection of Campylobacter species DNA in research material under the absence of clinical manifestations of the infection with bacteria’s indicates that in case of breaking rules during primary processing and circulation of animal slaughter provides increase in the number of these microorganisms (Campylobacter jejuni,Campylobacter coli, Campylobacter lari) and risk for infection, which occurs with symptoms of poisoning in consumers.
The aim of the study was to identify the Campylobactergenus bacteria in environmental objects (water, fodder and litter) in dairy farms by polymerase chain reaction.
The research materials were: water bowls, hay and silage mixture of racks and warehouse, litter on the farm.
The preparation of samples for DNA isolation ofCampylobacter research trial was conducted according to the "Methodological guidelines for the selection, handling, storage and sample preparation of biological material for diagnostic PCR".
The DNA isolation from test material was performed by PCR Ampli Sens of test systems "DNA Sorb-B-50." The Detection of PCR amplification products were determined by electrolytic separation of amplificatic mixture in shaded ethidium bromide agarose gel.
After preparation of samples and separation of pure DNA the samples were subjected to electrophoresis research to detect the presence of Campylobacter. Therewith the Ethidium bromide contacted with fragments of double-stranded DNA that appeared in the gel as light bars for UV radiation (λ = 290–330 nm). For visualize these bands were used a special device – transilyuminate and the results documented pictures. As a positive control we used the scale DNA containing Campylobacter DNA fragments of different lengths to estimate the size of PCR reaction products.
Further, we see that water silage and haylage mixture from feeding reacted positively on presence of bacteria of the genus Campylobacter. The DNA fragments were divided by the molecular weight in agarose gels. The specificity of the amplified DNA bands confirmed their placement relative molecular weight to markers and placing fragment amplification positive
Thus, the objects of the farm environment contaminated bacteria of the Campylobacter genus. Failure of proper sanitary-hygienic requirements for slaughter and primary processing of animals are contributes to contamination products of slaughter by Campylobacter.
The results of this study encourage us to conduct researches on the following number and type of Campylobacter in the environment (water, food, litter) and subsequently to detect Campylobacter in raw beef after slaughter of cattle.
The researches have found out that bacteria of Campylobacter is present in the farm’s medium, such as water, hay and silage mixture of feeding and litter that can later enter the animals body.
Key words: campylobacters, cattle, dairy farms, food, water, polymerase chain reaction.
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