Characterization and quantification of feline fecal microbiota using cpn60 sequence-based methods and investigation of animal-to-animal variation in microbial population structure

Improvement of the Representation of Bifidobacteria in Fecal Microbiota Metagenomic Libraries by Application of the cpn60 Universal Primer Cocktail

“…This target has been exploited in microbial ecology studies involving clone libraries of PCR products (6,10,11,17,18,20,28), pyrosequencing of PCR amplicons (31), and quantitative real-time PCR (4,9). Recently, we published a modification of the cpn60 universal primer PCR protocol which includes the use of a cocktail of degenerate primers optimized to give proportional amplifications of sequences across a broad range of GϩC contents (21).…”

“…Recently, we published a modification of the cpn60 universal primer PCR protocol which includes the use of a cocktail of degenerate primers optimized to give proportional amplifications of sequences across a broad range of GϩC contents (21). This approach has subsequently been used in the characterization of human vaginal microbiota (31) and feline fecal microbiota (6). Although in the feline study a large proportion of sequences observed corresponded to bifidobacteria, there is no corresponding 16S rRNA-based study of the samples for comparison.…”

The Microbiome

“…5,17,20,38,55,88,89,115,117,119,126 These microbial changes are accompanied by underlying susceptibilities in the innate immune system of dogs and cats with chronic enteropathies, such as idiopathic IBD, further demonstrating the relationship between gut microbiota and host health. 55,62,63,78,86 It is been shown that the canine microbiota changes with diarrhea, with the most profound changes in dogs with acute hemorrhagic diarrhea (AHD), 117 and characterized by significant decreases in Blautia, Ruminococcaceae including Faecalibacterium, and Turicibacter spp., and significant increases in genus Sutterella and Clostridium perfringens compared to healthy dogs (Fig.…”

Transient response of microbial communities in a water well field to application of an impressed current

“…The primers targeting nirS, dsrB and Geo genes were used to assess potential nitrate-, sulphite-and iron-reducing bacterial activity, respectively. The qPCR assays were performed on a Bio-Rad CFX96 realtime PCR detection system (Biorad Laboratories, Mississauga, ON) according to the method followed by Desai et al (2009). Following thermocycling, the qPCR products were subjected to melting curve analysis to confirm that the fluorescence signal originated from specific PCR product.…”