Furthermore, future studies with longer follow-up periods than 14 days after

treatment cessation will be useful to evaluate the long-term effect of tylosin on the jejunal microbiota. AZD5363 nmr Result of such studies may indicate the time needed for the microbiota to return to its pre-treatment state. Conclusion In conclusion, using deep massive parallel pyrosequencing we identified additional bacterial phyla and demonstrated the enormous species richness present in the small intestine of healthy dogs. We have demonstrated a profound and pervasive effect of tylosin on microbial diversity and various bacterial groups. These bacterial groups may represent candidates for exploration in clinical studies, and their changes will need to be correlated with clinical outcome, to further understand the effect of tylosin on gastrointestinal health. Methods Animals

Five healthy dogs, each with a pre-existing jejunal fistula inserted approximately 60 cm distal to the pylorus were used in this study [21]. All dogs were considered healthy and had no recent MI-503 ic50 history of gastrointestinal disease. All dogs were unrelated and approximately two years old. Their body weights ranged from 12 to 19 kg, and their body condition scores ranged between 3 and 4 (median 3) on a 5-point scale. The dogs received a commercial dry dog food (Mastery Adult Essential Maintenance, Dog’n Cat International, Vauvert, France) twice a day throughout the study period. According to the manufacturer, the food composition was 28% crude protein, 20% crude fat, 7% crude ash, and 2.5% crude fibre. During the study period, Histamine H2 receptor the dogs were cared for by the same personnel. All dogs were housed at the same laboratory animal unit at the Faculty of Veterinary Medicine, University of Helsinki, Finland. Dogs were housed in separate pens and treated individually. All dogs were fed at the same time each day. Tylosin was administered at 20 to 22 mg/kg q 24 hr for

a period of 14 consecutive days. This is the same dose that has previously been recommended for the treatment of tylosin-responsive diarrhea [34]. Sample collection The study had been approved by the Finnish Ethical Committee with license number ESLH-2007-09833/Ym-23. Mucosal brush samples were collected by advancing a sterile Cell Cycle inhibitor cytology brush through the fistula as described previously [23]. Samples were collected on day 0 (baseline), day 14 (after 14 days of tylosin administration), and day 28 (14 days after withdrawal of tylosin). To ensure consistency in sample collection, the same person collected all the samples during the whole study period. Furthermore, the samples were obtained according to a timetable with each sample collected exactly at the same time after feeding (i.e. dogs were fed consecutively, so that each sample could be collected in each dog at the same time after feeding). Samples were homogenized, properly labeled, and immediately frozen and stored at -80°C until further analysis.