GENETIC ANALYSIS OF BRAZILIAN ISOLATES AND DEVELOPMENT OF A PCR-BASED DIAGNOSTIC METHOD FOR DETECTION OF Cyniclomyces guttulatus IN DOG FEEDS
Cyniclomyces guttulatus; Diagnostic; Dogs;
The frequency of emerging infections in man and animals caused by yeast has increased in recent years and thus the need of greater efficiency in the diagnosis and characterization of these pathogens. Reports of yeast associated with diarrhea in dogs, or any other animal, are scarce. However, data produced over the past decade by researchers in Europe, Asia and North America has indicated that the yeast Cyniclomyces guttulatus may represent an emerging and opportunistic pathogen of dogs. More recently, this hypothesis has been supported by reports of this species of yeast in association with gastrointestinal disorders in Brazilian dogs. Cyniclomyces guttulatus is widely recognized as a natural inhabitant of the digestive tract of rabbits and guinea pigs. However, the difficulties associated with the culture of this yeast have served to limit research on the biology and potential pathogenicity of this microorganism. At present, the diagnosis of the infection with this yeast requires isolation from clinical material as a pure culture, followed by nucleotide sequence analysis of a variable region (D1 / D2 domain) of the gene encoding the large ribosomal rRNA subunit (26S). The study is based developed a PCR-based diagnostic system for the detection of C. guttulatus, and its association with gastrointestinal disorders in dogs. Firstly, a PCR-RFLP-based approach to facilitate the definitive identification of cultures of C. guttulatus, recovered from clinical material, without the need sequencing. Second, a PCR-based assays for the specific detection of C. guttulatus directly from stool samples, without the need for culture, with the objective of developing rapid diagnosis that would guarantee the rapid application of effective treatment and consequently, improvements in animal health and welfare.