Results T-RFLP analysis of the impact of cage type on intestinal microbiota The microbiota in ileal and caecal samples from the first experiment were characterised by creating individual
T-RFLP fingerprint profiles for each sample. Profiles were generated on the basis of the number of Terminal Restriction Fragments (T-RFs) in the range of 60 – 850 bp. The relationship AUY-922 concentration between two profiles could then be Tideglusib ic50 calculated by pair wise comparisons as a Dice similarity coefficient (SD), however to compensate for the variation between individual comparisons, the mean of the SD values was calculated and used to compare cage groups. The Dice coefficients from the first experimental study are shown in Table 1. In ileum, the highest Dice score was found between samples within same cage, and especially CC and AV diverged clearly from each other (SD 54.3 ± 9.6) with FC being in between, sharing profiles with both the other cages (CC SD 67.4
± 9.9 and AV 66.8 ± 11.4). When sampling was done 4 weeks later, higher SD values were calculated within cage, while values between cages were in the range 65.5-67.5. This shows that layers sharing the same environment also had comparable ileal microbiota, and this similarity increased over time. The height of the T-RF peaks reflected BTK inhibitor cost the prevalence of individual species in the microbiota. Ileum was characterized by having the same 3-4 dominating T-RFs in all cage groups, but other individual T-RFs were also present. Before
inoculation 10.5 ± 1.7 different T-RFs were detected in CC, while FC had 6.5 ± 2.7 and AV 7.3 ± 3.5. These were maintained throughout the study, although an increase was found in AV (10.7 ± 2.7). The four most dominating T-RFs in all samples were 393 bp, 406 bp, 597 bp, and 550 bp. These T-RFLP fragments could be equated with by different Lactobacillus species by in silico digest of 16S rDNA. Although the total number of detectable T-RFs remained constant in the ileum, an inverted relationship was found between one group of T-RFs: 406 bp, 606 bp and 550 bp which decreased 6-phosphogluconolactonase in height, whereas as a new and unidentified T-RF 813 bp emerged. This shift was primarily found in layers from FC and a few layers from other cages, and this may explain some of the differences observed in SD between cages. Table 1 Comparisons of T-RFLP profiles of microbiota in the ileum and caecum of layers housed in different cage systems Before Inoculation Mean SD Location Cage n T-RF Conventional Furnished Aviary Ileum Conventional 4 10.5 ± 1.7 70.5 ± 12.4 – - Furnished 4 6.5 ± 2.7 67.4 ± 9.9 65.9 ± 7.5 – Aviary 4 7.3 ± 3.5 54.3 ± 9.6 66.8 ± 11.4 72.3 ± 7.0 Caecum Conventional 4 39.5 ± 6.6 66.4 ± 6.0 – - Furnished 4 39.8 ± 4.2 60.8 ± 3.5 75.1 ± 6.0 – Aviary 4 52.7 ± 23.5 38.6 ± 6.3 38.5 ± 4.8 45.4 ± 14.
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