The Impact of Pastured Livestock Food Consumption on Fatty Acid and Metabolomic Profiles

This randomized, double-blind, parallel-arm clinical trial investigates the impact of consuming animal products from regenerative/pastured versus conventional farming practices on fatty acid composition, inflammatory markers, and metabolomic profiles in adults aged 30-65 years. The study aims to address gaps in understanding the nutritional and health implications of farming practices by comparing postprandial inflammatory and metabolic responses to these diets.

Participants will be randomly assigned to one of two groups and provided with weekly allocations of animal-source foods. Dietary compliance will be assessed through 24-hour dietary recalls and food logs. Blood, urine, and stool samples will be collected at baseline and end of study to measure the primary and secondary outcomes, including:

Red blood cell omega-3 fatty acid levels as a marker of cardiovascular health. A panel of inflammatory markers, including IL-6, TNF-alpha, CRP.

Plasma metabolites such as amino acids, organic acids, and vitamin derivatives.

Lipid profiles (LDL, HDL, VLDL, cholesterol, and triglycerides). Participants will be instructed to consume only the foods provided by the study to ensure dietary control and reduce confounding factors. Biospecimens will be stored for future analysis to examine broader metabolic impacts.

By addressing how different farming practices influence health markers, this study aims to provide evidence-based insights for public health and inform dietary recommendations for sustainable and health-conscious food systems.

• Pastured Livestock Diet (Dietary supplement): Participants will consume a diet consisting of animal-source foods (chicken, beef, pork, and eggs) from regenerative/pastured farming practices. Foods will be provided weekly in specific quantities for a 16-week intervention. Adherence will be monitored through dietary logs and ASA24 dietary assessments.
• Conventional Livestock Diet (Dietary supplement): Participants will consume a diet consisting of animal-source foods (chicken, ground, pork, and eggs) from conventional farming practices. Foods will be provided weekly in specific quantities for a 16-week intervention. Adherence will be monitored through dietary logs and ASA24 dietary assessments.

Primary outcomes

• Red Blood Cell (RBC) Omega-3 Fatty Acid Levels [Baseline and 16 weeks (end of intervention).]: Assessment of changes in red blood cell omega-3 fatty acid composition as a marker of cardiovascular health. Blood samples will be collected at baseline and after the 16-week intervention to measure levels using gas chromatography-mass spectrometry (GC and LC/MS).

Secondary outcomes

• Metabolomics Profile Analysis [Baseline to Week 16 (End of Study)]: Changes in plasma metabolomic profiles will be assessed to identify variations in metabolites, such as amino acids, organic acids, and vitamin derivatives, associated with the consumption of pastured versus conventional animal products. This analysis will provide insights into the biochemical impacts of dietary interventions on metabolic pathways, inflammation, and overall health. Samples will be analyzed using mass spectrometry for untargeted and/or targeted metabolomics profiling.

Sex: All, ages: 30 Years to 65 Years, accepts healthy volunteers

Inclusion Criteria:

* Age between 30 and 65 years. BMI between 25 and 35 kg/m². Weight stability within the last three months (loss or gain < 4%). Hemoglobin A1C (HbA1C) ≤ 6.4%. Fasting plasma glucose concentration < 126 mg/dL. Ability to understand and communicate in English or Spanish.

Exclusion Criteria:

* Use of medications known to affect study outcomes (e.g., NSAIDs, corticosteroids, anticoagulants) that cannot be discontinued temporarily.

Blood pressure ≥ 140/90 mmHg without physician clearance. Unwillingness to forgo seafood consumption or omega-3 supplement intake during the study.

Alcohol consumption > 14 drinks per week. Tobacco product use within the last three months. Receipt of a COVID vaccine within the last two weeks. Engagement in high-level competitive exercise (e.g., triathlon, marathon, powerlifting).

Diagnosed inflammatory diseases (e.g., autoimmune disorders, coeliac disease, arthritis).

Diagnoses of active malignancy, congestive heart failure, diabetes mellitus, or chronic obstructive pulmonary disease.

Use of antibiotics within the last 60 days. Pregnancy or planning pregnancy within the next month. Lactating women. Allergies to meat or eggs. Inability or unwillingness to comply with the study protocol, including adherence to study visits and dietary requirements.

Dietary preferences that exclude meat or eggs (e.g., vegan, vegetarian, halal). Any condition that, in the opinion of the research team, makes the participant unsuitable for the study.

• McAfee AJ, McSorley EM, Cuskelly GJ, Fearon AM, Moss BW, Beattie JA, Wallace JM, Bonham MP, Strain JJ. Red meat from animals offered a grass diet increases plasma and platelet n-3 PUFA in healthy consumers. Br J Nutr. 2011 Jan;105(1):80-9. doi: 10.1017/S0007114510003090. PMID 20807460