Effect of oral administration of metronidazole or prednisolone on fecal microbiota in dogs

Abstract

Gastrointestinal microbiota have been implicated in the pathogenesis of various gastrointestinal disorders in dogs, including acute diarrhea and chronic enteropathy. Metronidazole and prednisolone are commonly prescribed for the treatment of these diseases; however, their effects on gastrointestinal microbiota have not been investigated. The objective of this study was to evaluate the effects of these drugs on the gastrointestinal microbiota of dogs. Metronidazole was administered twice daily at 12.5 mg/kg to a group of five healthy dogs, and prednisolone at 1.0 mg/kg daily to a second group of five healthy dogs for 14 days. Fecal samples were collected before and after administration (day 0 and 14), and 14 and 28 days after cessation (day 28 and 42). DNA was extracted, and the bacterial diversity and composition of each sample were determined based on 16S ribosomal RNA (rRNA) gene sequences using next-generation sequencing (Illumina MiSeq). In the group administered metronidazole, bacterial diversity indices significantly decreased at day 14, and recovered after the cessation. Principal coordinates analysis and hierarchical dendrogram construction based on unweighted and weighted UniFrac distance matrices revealed that bacterial composition was also significantly altered by metronidazole at day 14 compared with the other time points. The proportions of Bacteroidaceae, Clostridiaceae, Fusobacteriaceae, Lachnospiraceae, Ruminococcaceae, Turicibacteraceae, and Veillonellaceae decreased, while Bifidobacteriaceae, Enterobacteriaceae, Enterococcaceae, and Streptococcaceae increased at day 14 and returned to their initial proportions by day 42. Conversely, no effect of prednisolone was observed on either the bacterial diversity or composition. Reducing pathogenic bacteria such as Fusobacteria and increasing beneficial bacteria such as Bifidobacterium through the administration of metronidazole may be beneficial for promoting gastrointestinal health; however, further investigations into the effects on diseased dogs are needed.

Figure 1. Average proportion of bacterial phyla identified in dogs at each time point.

Results of dogs administered metronidazole (A) and prednisolone (B). Error bars represent standard error of the mean. Asterisks indicate statistically significant differences from day 0, and daggers indicate statistically significant differences from day 14 (P<0.05).

Figure 2. Rarefraction analysis of V4 16S rRNA gene sequences obtained from fecal samples.

Results from dogs administered metronidazole (A) and prednisolone (B). Lines represent the average of each time point and the error bars represent standard deviations. This analysis was performed using a randomly selected subset of 9,915 (A) or 8,153 (B) sequences per sample. Operational Taxonomical Units (OTUs) in this analysis were defined by 97–100% similarity.

Figure 3. Principal coordinates analysis (PCoA) of V4 16S rRNA genes from canine fecal samples.

Figures were calculated using unweighted UniFrac distances. (A) Result of dogs administered metronidazole. Metronidazole-affected samples (blue, day 14) were separated from the other samples, primarily along PCoA axis 1 (accounting for 33.94% of all variability among samples). (B) Result of dogs administered prednisolone. Prednisolone administration did not induce alteration of bacterial composition.

Figure 4. Hierarchical dendrogram and fecal microbial composition of each sample at the phylum level.

Figures were constructed using unweighted UniFrac distances. (A) Result of dogs administered metronidazole. This dendrogram showed that the samples obtained at day 14 were clustered. (B) Result of dogs administered prednisolone. No clustering was observed at any time points.