ABSTRACT
BACKGROUND: The gastrointestinal (GI) microbiota has a strong impact on the health of cats and these populations can be altered in GI disease. Little research has been done to associate improvement in diarrhea with changes in GI microbiota. OBJECTIVE: To evaluate GI microbiota changes associated with diet change and related improvement in diarrhea in cats with chronic naturally occurring diarrhea. ANIMALS: Fifteen adult Domestic Shorthair cats with naturally occurring chronic diarrhea. METHODS: Controlled crossover dietary trial for management of diarrhea. Fecal microbiome was assessed using 454-pyrosequencing. Relationships among fecal score (FS), diet, and microbiome were explored using partial least square method, partial least square method - discriminant analysis, and orthogonal partial least square method with discriminant analysis (OPLS-DA). RESULTS: Dominant bacterial phyla included the Firmicutes and Bacteroidetes, followed by Fusobacteria, Proteobacteria, Tenericutes, and Actinobacteria. Orthogonal partial least squares (OPLS-DA) clustering showed significant microbial differences within cats when fed Diet X versus Diet Y, and with Diet Y versus baseline. Significant correlations were found between the microbiome and FSs. Those bacteria with the strongest correlation with FS included Coriobacteriaceae Slackia spp., Campylobacter upsaliensis, Enterobacteriaceae Raoultella spp., Coriobacteriaceae Collinsella spp., and bacteria of unidentified genera within the families of Clostridiales Lachnospiracea and Aeromonadales Succinivibrionacease, suggesting that increased numbers of these organisms may be important to gut health. CONCLUSIONS AND CLINICAL IMPORTANCE: Alterations in intestinal microbiota were associated with improvement in diarrhea, but, from our data we cannot conclude if changes in the microbiome caused the improvement in diarrhea, or vice versa.
Subject(s)
Cat Diseases/microbiology , Diarrhea/veterinary , Gene Expression Regulation, Bacterial/genetics , Microbiota/genetics , Animals , Cat Diseases/diet therapy , Cat Diseases/genetics , Cats , Cross-Over Studies , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Diarrhea/diet therapy , Diarrhea/genetics , Diarrhea/microbiology , Feces/microbiology , Female , Least-Squares Analysis , Male , Principal Component Analysis , RNA, Ribosomal, 16S/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNAABSTRACT
Closed microwave digestion and a high-pressure asher have been evaluated for wet-oxidation and extraction of lead, cadmium, chromium, and mercury from a range of typical packaging materials used for food products. For the high-pressure asher a combination of nitric and sulfuric acids was efficient for destruction of a range of packaging materials; for polystyrene, however, nitric acid alone was more efficient. For microwave digestion, a reagent containing nitric acid, sulfuric acid, and hydrogen peroxide was used for all materials except polystyrene. Use of the high-pressure asher resulted in the highest recoveries of spiked lead (median 92%), cadmium (median 92%), chromium (median 97%), and mercury (median 83%). All samples were spiked before digestion with 40 microg L(-1) Cd, Cr, and Pb and 8 microg L(-1) Hg in solution. The use of indium as internal standard improved the accuracy of results from both ICP-MS and ICP-AES. Average recovery of the four elements from spiked packaging materials was 92 +/- 14% by ICP-MS and 87 +/- 15% (except for mercury) by ICP-AES. For mercury analysis by CVAAS, use of tin(II) chloride as reducing agent resulted in considerably better accuracy than use of sodium borohydride reagent.