Standardizing sugar probe-based techniques to measure intestinal permeability in broilers

A concern for intestinal health of broilers, that includes intestinal barrier function, has been one of the main focuses that poultry researchers and the industry want to improve. Yet, there is not a practical and standardized technique to non-invasively measure intestinal permeability of broilers. The available techniques, such as ELISA, qRT-PCR, Ussing chambers, and histological approaches are laborious, indirect, and terminal. Understanding this critical need, we tried to standardize a sugar probe-based assay using lactulose (Lac) and mannitol (Man) to measure intestinal permeability in male and female Ross 308 broilers by performing 4 sequential trials. In trial 1, 21-d old broilers orally received 2 mL/kg body weight (BW) of solution containing 25 g Lac and 5 g Man dissolved in 100 mL of pure water. Bloods samples from each bird were collected at 60-, 90-, and 120-min after Lac and Man (LacMan) dosing using serum and EDTA-treated plasma tubes. The Lac value of plasma collected at 90 minutes post dosing had the least variation and this collection time-point was selected for subsequent trials. In trial 2, birds were dosed with different amounts of Lac and Man with the lowest amount (5 g Lac and 1 g Man dissolved in pure water to reach 100 mL of final solution) still allowing Lac and Man detection from EDTA-treated blood samples by liquid chromatography (LC), which was then used in trial 4. Unfortunately, glucose was found to severely interfere with Lac quantification in trial 3 in which samples were collected using Na-Heparin-treated tubes. However, after applying a skimming technique when evaluating LC chromatograms, the Lac values of groups subjected to 24-h feed withdrawal (FW) prior to LacMan dosing were higher (P < 0.01) than groups provided free access to feed. Use of the Lac:Man ratio to predict intestinal permeability was not achieved in trial 3 due to possible inaccuracy of Man in capturing total absorptive capacity. Hence, trial 3 was followed by trial 4 where adjustment in the LacMan solution was made (15 g Lac and either 3 or 5 g Man dissolved in pure water to reach 100 mL of final solution). Yet, glucose interference still existed causing Lac undetectability in several samples. Throughout all trials, we demonstrated that Lac and Man still pose various limitations to be reliable markers for intestinal permeability in broilers, whether coming from the intestinal stress model, sample handling and processing, or the analytical side. High concentration of glucose in broilers that interfere formation of Lac peak in the chromatogram is the major hurdle and requires urgent attention before proceeding. Hence, caution should be used when predicting intestinal permeability of broilers using Lac and Man, unless it has been fully standardized.

Name thesis:degree_name

M.S.

Level thesis:degree_level

Thesis

Discipline thesis:degree_discipline

Nutritional Sciences

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2023

Author dc:creator

• Sujiyanto, -

Contributors dc:contributor

• Dilger, Ryan N.
• Emmert, Jason L.
• Stein, Hans H.

• Broiler
• Lactulose
• Mannitol
• Intestinal Permeability

Statement dc:rights

• Copyright 2023 Sujiyanto

Language dc:language

en, eng

Handle dc:identifier

https://hdl.handle.net/2142/122100

OAI record oai:identifier

https://www.ideals.illinois.edu/oai-pmh?verb=GetRecord&metadataPrefix=etdms&identifier=oai%3Awww.ideals.illinois.edu%3A2142%2F122100