Complexity and temporal stability of the digestive microbiota of captive cheetahs by community fingerprinting

Introduction: Feeding mismanagement has been pointed out as an important factor for the development of gastrointestinal and metabolic diseases in captive cheetahs. Given the crucial role of the host’s intestinal microbiota in feed fermentation and health maintenance, a better understanding of the diversity and temporal stability of the cheetah’s intestinal ecosystem is essential for improvement of current feeding strategies. Materials and Methods: With 2-monthly intervals, fecal samples of two adult captive cheetahs from a Belgian zoo were collected within 15 minutes after defecation and immediately stored at -80°C. The dietary regime (randomly raw beef or whole rabbit) of both animals was recorded daily. Given the general poor culturability of intestinal anaerobic bacteria, a culture-independent approach was applied to gain a first insight in the complexity and stability of the fecal microbiota. DNA extracts obtained from 1g of homogenized fecal sample were used as templates in a community PCR targeting the ribosomal V3-16S rRNA gene and subsequent profiling of the resulting PCR amplicons with Denaturing Gradient Gel Electrophoresis (DGGE). The resulting DGGE fingerprints were analyzed for number, intensity and distribution of bands using the BioNumerics v. 5.10 (Applied Maths, Belgium) software. Results and discussion: At the time of abstract submission, three samples were collected from cheetah 1. Band profile clustering analysis using the Pearson correlation coefficient revealed that the bi-monthly samples shared 67% similarity indicating moderate temporal stability of the fecal microbiota. By scoring individual DGGE bands based on their unique position, 31 different band-classes were found which reflect the bacterial richness within the fecal microbiota of cheetah 1. Interestingly, six band-classes were found to occur in all three samples and might correspond to the dominant core microbiota of cheetah 1. On the other hand, nine band-classes were unique to a single time point and may represent the transient fecal microbiota of this animal. Conclusion: The molecular approach proves to be a powerful tool to gain insights in the phylogenetic complexity of the microbial communities in the intestinal tract of captive cheetahs. By the time of the symposium, more animals and samples will have been included which will enable more in-depth analysis of the fecal microbiota in captive cheetahs