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The effect of a multi-species synbiotic on the fecal microbiota of healthy cats (n = 12) and dogs (n = 12) was evaluated. The synbiotic (containing 5 × 10(9)  CFU of a mixture of seven probiotic strains, and a blend of fructooligosaccharides and arabinogalactans) was administered daily for 21 days. Fecal and serum samples were collected before, during, and up to 3 weeks after administration. Changes in the fecal microbiota were analyzed using denaturing gradient gel electrophoresis, 16S rRNA gene libraries, quantitative real-time PCR, and 16S rRNA gene 454-pyrosequencing. Probiotic species were detectable in 10/12 dogs and 11/12 cats during product administration. Abundances of Enterococcus and Streptococcus spp. were significantly increased in at least one time point during administration, and returned to baseline abundance after treatment was discontinued. No changes in the major bacterial phyla were identified on 454-pyrosequencing. No adverse gastrointestinal effects were recorded and no significant changes in gastrointestinal function or immune markers were observed during the study period. This study shows that while the ingestion of probiotics and prebiotics does not appear to alter the predominant bacterial phyla present in feces, supplementation with the investigated synbiotic leads to an increased abundance of probiotic bacteria in the feces of healthy cats and dogs.

-PI for 3 sampling days of > 13.9 µg/g or >21.0 µg/g, respectively, should be considered abnormal in dogs >1 year of age. Fecal cα 1 -PI concentrations in dogs <1 year of age were significantly higher and should be carefully interpreted in this age group.Key words: Alpha 1 -proteinase inhibitor; canine; fecal; immunoassay; puppies.Radioimmunoassay for fecal canine α 1 -proteinase inhibitor 477 methods usually are of higher analytical sensitivity, RIAs are usually easier to control and standardize than ELISAs. Although RIAs require special equipment and the use of a radioactive tracer, they have the advantage of not requiring purification and labeling of large amounts of antibodies from ideally 2 different individuals of a heterologous species, but usually only require a small volume of a heterologous antiserum. An ELISA for the measurement of cα 1 -PI concentration in fecal samples from dogs has previously been developed and analytically validated, 10 but due to the large amount of anti-cα 1 -PI antibodies needed, this smallscale manufacturing methodology was difficult to maintain long-term and has been discontinued, as the original source material was no longer available. Therefore, the current study was aimed at the development and analytical validation of a new RIA suitable for quantifying Fcα 1 -PI (as a clinical marker for GI protein loss in dogs) in a large number of samples, the de novo determination of a reference interval from a large number of healthy dogs, and to compare Fcα 1 -PI concentrations measured in healthy dogs above 1 year of age to that in healthy puppies, as it had been previously noticed that Fcα 1 -PI concentrations are occasionally increased in puppies, yet Fcα 1 -PI has never been studied in healthy puppies. Materials and methods Collection of fecal specimens from dogsSamples of naturally passed feces were collected from 73 healthy adult pet dogs of various breeds (median age: 3.8 years, range: 1.0-17.3 years), and from 28 healthy pet dog puppies of various breeds (mean age: 6.1 months; range: 1.8-10.7 months). All dogs had been vaccinated and dewormed regularly prior to fecal sample collection, did not show clinical signs of GI disease, and did not receive any medications or had any medical conditions known to affect the GI tract. As a high intra-individual variability of cα 1 -PI in fecal samples has been reported (Steiner JM, Ruaux CG, Miller MD, et al.: 2003, Intra-individual variability of fecal α 1 -proteinase inhibitor concentration in clinically healthy dogs. J Vet Intern Med 17:445. Abstract), samples were collected from 3 bowel movements from 3 consecutive days for each dog, placed into pre-weighed polypropylene tubes, a and immediately frozen until extraction. For extraction, feces were then thawed and extracted as previously described. 10 Briefly, aliquots of approximately 1.0 g wet weight were diluted 1:5 in phosphate buffered saline b (PBS) supplemented with 5% (v/v) newborn calf serum, c 1% (v/v) Triton X-100, d and 0.25 mM thimerosal. After homogenization by vortexing...