Veterynarna biotehnologija – Veterinary biotechnology, 2016, 28, 20-28 [in Ukrainian].
Institute of Veterinary Medicine of NAAS
CLOSTRIDIUM DIFFICILE ISOLATION FROM CLINICAL MATERIAL OF PIGS
Introduction. Clostridium difficile – gram-positive anaerobic microorganism that in major cases causes of intestinal (hospital) diarrhea, colitis or pseudomembranous colitis. In humans infection caused by C. difficile is called CDI. The major danger of CDI infection development cause antimicrobials agents that are sensitive to anaerobic microorganism sand also it is important to consider other drugs capable of destroying intestine microbiocenosis. C. difficile causes disease in mammals with the same clinical symptoms in humans. Different types of animals are susceptible to the disease, especially young animals. C. difficile is the causative agent in newborn piglets and can be related to enteritis in calves. The main virulence factors are C. difficile toxin: enterotoxin (type A) and cytotoxin (type B). Virulent strains produce two toxins, and some strains also encode binary toxin (Pa Loc), the role of which is not fully studied in the pathogenesis of the disease.
The goal of the work. To isolate C. difficile pathogen from clinical material of healthy and sick pigs with signs of diarrhea.
Materials and methods. 20 samples from pigs after the act of defecation were collected from one farm of Kyiv oblast. Cultivation was carried out under anaerobic conditions in Wilson-Blair, Kitty-Tarotstsi media, blood agar and selective-differential blood agar with Cefoxitin, Cycloserine and fructose. Identification of C. difficile cultures was carried out by cultural-morphological characteristics and microscopy of microorganism cells after Gram staining and by their ability to survive under aerobic conditions.
Results of the study and discussion. As a result of investigations, we identified three isolates of C. difficile with typical characteristics by bacteriological method. Two positive samples were from pigs with symptoms of diarrhea and one from clinically healthy animal. In addition, we identified C. perfringens (60% of samples) with a clear zone of hemolysis on blood agar and E. coli, St.spp. and other microorganisms as well. Recent studies have shown a high degree of affinity between human pathogen strains and strains isolated from animals and animal products. Given the stability of the pathogen, we can assume that it has worldwide distribution in the environment that creates risks of this zoonosis toxic infection. Data on the isolation of C. difficile from animals in Ukraine is almost absent and there is no known connection of antibiotic therapy with diarrhea in animals. We believe that the only continuous monitoring of biological material from animals, samples of food products and of the environment, will make possible determine the level of threat, sources and ways of transmission of this pathogen in Ukraine.
Conclusions and prospects for further research. In the faeces of pigs three isolates of C. difficile were identified (15% of animals). Perspective direction is defined toxigenicity of isolates, which determine the final diagnosis, and to assess the epidemiological situation in Ukraine.
Keywords: Clostridium difficile, bacteria display, antibiotic therapy, toxins.