Short communication: Gut microbial colonization of the mouse colon using faecal transfer was equally effective when comparing rectal inoculation and oral inoculation based on 16S rRNA sequencing

Ditte Olsen Lützhøft; Lidia Sánchez-Alcoholado; Peter Tougaard; Caroline M. Junker Mentzel; Witold Kot; Dennis S. Nielsen; Axel Kornerup Hansen

doi:10.1016/j.rvsc.2019.09.009

Short communication: Gut microbial colonization of the mouse colon using faecal transfer was equally effective when comparing rectal inoculation and oral inoculation based on 16S rRNA sequencing

Ditte Olsen Lützhøft^*, Lidia Sánchez-Alcoholado, Peter Tougaard, Caroline M. Junker Mentzel, Witold Kot, Dennis S. Nielsen, Axel Kornerup Hansen

^*Corresponding author for this work

Abstract

In the present study we hypothesized that a higher degree of gut microbiota (GM) transfer and colonization could be reached by rectal inoculation compared to oral inoculation, which is commonly used in mouse studies for GM transfer. We treated C57BL/6NTac Specific Pathogen Free (SPF) mice with antibiotics and subsequently we inoculated these with GM from donor mice of the same strain by either the oral or the rectal inoculation method. 16S rRNA gene sequencing of the colon microbiota showed no difference in microbial community on account of inoculation method as determined by unweighted UniFrac distance metrics in C57BL/6NTac SPF mice. In addition, qPCR analysis on colon tissue revealed no difference in mRNA expression between the inoculation methods. Next, the SPF mice were compared to germ-free (GF)-mice to identify differences in inoculation efficacy. Whether the mice were antibiotic treated SPF or GF clearly influenced GM determined by 16S rRNA gene sequencing where the SPF mice experienced up-regulation of S24–7 (p = .0001) and a decrease in Rikenellaceae (p = .016) compared to GF mice. qPCR analysis on colon tissue revealed up-regulation in mRNA gene expression of Il6, Il10, Reg3g and transcription factor RORγt (Rorc) in GF mice compared to SPF mice on a significant level (p < .05). This gene expression profile is consistent with post colonization development of the intestinal barrier in GF mice.

Original language	English
Journal	Research in Veterinary Science
Volume	126
Pages (from-to)	227-232
Number of pages	6
ISSN	0034-5288
DOIs	https://doi.org/10.1016/j.rvsc.2019.09.009
Publication status	Published - 2019

Keywords

16S rRNA sequencing
Conventional mice
Faecal transplantation
Germ-free mice
Oral inoculation
Rectal inoculation

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10.1016/j.rvsc.2019.09.009

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Lützhøft, D. O., Sánchez-Alcoholado, L., Tougaard, P., Junker Mentzel, C. M., Kot, W., Nielsen, D. S., & Hansen, A. K. (2019). Short communication: Gut microbial colonization of the mouse colon using faecal transfer was equally effective when comparing rectal inoculation and oral inoculation based on 16S rRNA sequencing. Research in Veterinary Science, 126, 227-232. https://doi.org/10.1016/j.rvsc.2019.09.009

Short communication : Gut microbial colonization of the mouse colon using faecal transfer was equally effective when comparing rectal inoculation and oral inoculation based on 16S rRNA sequencing. / Lützhøft, Ditte Olsen; Sánchez-Alcoholado, Lidia; Tougaard, Peter et al.

In: Research in Veterinary Science, Vol. 126, 2019, p. 227-232.

Research output: Contribution to journal › Journal article › Research › peer-review

Lützhøft, DO, Sánchez-Alcoholado, L, Tougaard, P, Junker Mentzel, CM , Kot, W , Nielsen, DS & Hansen, AK 2019, 'Short communication: Gut microbial colonization of the mouse colon using faecal transfer was equally effective when comparing rectal inoculation and oral inoculation based on 16S rRNA sequencing', Research in Veterinary Science, vol. 126, pp. 227-232. https://doi.org/10.1016/j.rvsc.2019.09.009

Lützhøft DO, Sánchez-Alcoholado L, Tougaard P, Junker Mentzel CM , Kot W , Nielsen DS et al. Short communication: Gut microbial colonization of the mouse colon using faecal transfer was equally effective when comparing rectal inoculation and oral inoculation based on 16S rRNA sequencing. Research in Veterinary Science. 2019;126:227-232. doi: 10.1016/j.rvsc.2019.09.009

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title = "Short communication: Gut microbial colonization of the mouse colon using faecal transfer was equally effective when comparing rectal inoculation and oral inoculation based on 16S rRNA sequencing",

abstract = "In the present study we hypothesized that a higher degree of gut microbiota (GM) transfer and colonization could be reached by rectal inoculation compared to oral inoculation, which is commonly used in mouse studies for GM transfer. We treated C57BL/6NTac Specific Pathogen Free (SPF) mice with antibiotics and subsequently we inoculated these with GM from donor mice of the same strain by either the oral or the rectal inoculation method. 16S rRNA gene sequencing of the colon microbiota showed no difference in microbial community on account of inoculation method as determined by unweighted UniFrac distance metrics in C57BL/6NTac SPF mice. In addition, qPCR analysis on colon tissue revealed no difference in mRNA expression between the inoculation methods. Next, the SPF mice were compared to germ-free (GF)-mice to identify differences in inoculation efficacy. Whether the mice were antibiotic treated SPF or GF clearly influenced GM determined by 16S rRNA gene sequencing where the SPF mice experienced up-regulation of S24–7 (p = .0001) and a decrease in Rikenellaceae (p = .016) compared to GF mice. qPCR analysis on colon tissue revealed up-regulation in mRNA gene expression of Il6, Il10, Reg3g and transcription factor RORγt (Rorc) in GF mice compared to SPF mice on a significant level (p < .05). This gene expression profile is consistent with post colonization development of the intestinal barrier in GF mice.",

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T2 - Gut microbial colonization of the mouse colon using faecal transfer was equally effective when comparing rectal inoculation and oral inoculation based on 16S rRNA sequencing

AU - Lützhøft, Ditte Olsen

AU - Sánchez-Alcoholado, Lidia

AU - Tougaard, Peter

AU - Junker Mentzel, Caroline M.

AU - Kot, Witold

AU - Nielsen, Dennis S.

AU - Hansen, Axel Kornerup

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N2 - In the present study we hypothesized that a higher degree of gut microbiota (GM) transfer and colonization could be reached by rectal inoculation compared to oral inoculation, which is commonly used in mouse studies for GM transfer. We treated C57BL/6NTac Specific Pathogen Free (SPF) mice with antibiotics and subsequently we inoculated these with GM from donor mice of the same strain by either the oral or the rectal inoculation method. 16S rRNA gene sequencing of the colon microbiota showed no difference in microbial community on account of inoculation method as determined by unweighted UniFrac distance metrics in C57BL/6NTac SPF mice. In addition, qPCR analysis on colon tissue revealed no difference in mRNA expression between the inoculation methods. Next, the SPF mice were compared to germ-free (GF)-mice to identify differences in inoculation efficacy. Whether the mice were antibiotic treated SPF or GF clearly influenced GM determined by 16S rRNA gene sequencing where the SPF mice experienced up-regulation of S24–7 (p = .0001) and a decrease in Rikenellaceae (p = .016) compared to GF mice. qPCR analysis on colon tissue revealed up-regulation in mRNA gene expression of Il6, Il10, Reg3g and transcription factor RORγt (Rorc) in GF mice compared to SPF mice on a significant level (p < .05). This gene expression profile is consistent with post colonization development of the intestinal barrier in GF mice.

AB - In the present study we hypothesized that a higher degree of gut microbiota (GM) transfer and colonization could be reached by rectal inoculation compared to oral inoculation, which is commonly used in mouse studies for GM transfer. We treated C57BL/6NTac Specific Pathogen Free (SPF) mice with antibiotics and subsequently we inoculated these with GM from donor mice of the same strain by either the oral or the rectal inoculation method. 16S rRNA gene sequencing of the colon microbiota showed no difference in microbial community on account of inoculation method as determined by unweighted UniFrac distance metrics in C57BL/6NTac SPF mice. In addition, qPCR analysis on colon tissue revealed no difference in mRNA expression between the inoculation methods. Next, the SPF mice were compared to germ-free (GF)-mice to identify differences in inoculation efficacy. Whether the mice were antibiotic treated SPF or GF clearly influenced GM determined by 16S rRNA gene sequencing where the SPF mice experienced up-regulation of S24–7 (p = .0001) and a decrease in Rikenellaceae (p = .016) compared to GF mice. qPCR analysis on colon tissue revealed up-regulation in mRNA gene expression of Il6, Il10, Reg3g and transcription factor RORγt (Rorc) in GF mice compared to SPF mice on a significant level (p < .05). This gene expression profile is consistent with post colonization development of the intestinal barrier in GF mice.

KW - 16S rRNA sequencing

KW - Conventional mice

KW - Faecal transplantation

KW - Germ-free mice

KW - Oral inoculation

KW - Rectal inoculation

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DO - 10.1016/j.rvsc.2019.09.009

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SN - 0034-5288

VL - 126

SP - 227

EP - 232

JO - Research in Veterinary Science

JF - Research in Veterinary Science

ER -

Short communication: Gut microbial colonization of the mouse colon using faecal transfer was equally effective when comparing rectal inoculation and oral inoculation based on 16S rRNA sequencing

Abstract

Keywords

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