Research Article

Modulation of Rumen Microbial Communities in Tropical Holstein Dairy Cows through Lemon Peel Essential Oil Supplementation: A Metagenomic Analysis

Number: 1 February 18, 2026
Yuzuncu Yıl University Journal of Agricultural Sciences Cover
Yuzuncu Yıl University Journal of Agricultural Sciences
PDF Download

Modulation of Rumen Microbial Communities in Tropical Holstein Dairy Cows through Lemon Peel Essential Oil Supplementation: A Metagenomic Analysis

Abstract

Lemon peel essential oil (LPEO) is a promising natural additive for modulating the rumen microbiota in dairy cattle. This study evaluated the effects of LPEO supplementation on fecal microbial diversity and composition in tropical Holstein cows, with focus on methanogenic archaea and fiber-degrading bacteria. Cows received 0, 2.5, or 5.0 mL/head/day of LPEO. Overall microbial diversity remained stable across treatments (Shannon index: P = 0.4838; Simpson index: P = 0.497), and species richness increased (Control: 28.0, High LPEO: 31.0; P = 0.0572), indicating preserved ecosystem stability. The Firmicutes/Bacteroidetes (F/B) ratio remained optimal (>3.0) and increased with LPEO dose (Control: 3.57; High LPEO: 3.94), suggesting enhanced fiber degradation. Despite non-significant p-values, strong dose-dependent trends were observed: Bacteroidota declined (slope = –0.819, R² = 0.702), Firmicutes increased (slope = 0.557, R² = 0.467), and the Archaea/Bacteria ratio decreased by 8.9% (0.00981→0.00891), indicating reduced methanogenic potential. Inter-phylum ecological relationships were preserved, including a perfect negative correlation between Bacteroidota and Firmicutes (ρ = –1.000, P < 0.0001), and network connectivity indices remained stable (0.980–0.982). These results highlight LPEO’s selective antimicrobial activity—modulating microbial structure without broad-spectrum disruption. Limonene (40.60% of LPEO) likely mediated the selective suppression of archaea and enrichment of fiber degraders. The strong predictive capacity of dose-response models supports the biological relevance of observed trends. LPEO supplementation at 5 mL/head/day offers potential for improved rumen function and environmental sustainability in tropical dairy systems, warranting larger-scale validation.

Keywords

References

  1. Bi, Y., Zeng, S., Zhang, R., Diao, Q., & Tu, Y. (2018). Effects of dietary energy levels on rumen bacterial community composition in Holstein heifers under the same forage to concentrate ratio condition. BMC Microbiology, 18(1), 1–11. https://doi.org/10.1186/s12866-018-1213-9
  2. Fadillah, M., Widodo, E., & Djunaidi, I. (2024). Effect of Ultrasound-Assisted Maceration on Yield Percentage, Chemical Compound and Antimicrobial Activity of Citrus nobilis Peel Essential Oil. Yuzuncu Yil University Journal of Agricultural Sciences, 34(4), 656-668. https://doi.org/10.29133/yyutbd.1499319
  3. Hidayet, H. M., & Mustafa, K. N. (2025). Impact of Diet Supplementation with Fenugreek Seeds on Digestibility and Some Rumen and Blood Parameters in Lambs. Yuzuncu Yil University Journal of Agricultural Sciences, 35(3), 381-391. https://doi.org/10.29133/yyutbd.1621030
  4. Jami, E., White, B. A., & Mizrahi, I. (2014). Potential role of the bovine rumen microbiome in modulating milk composition and feed efficiency. PLoS ONE, 9(1), e85423. https://doi.org/10.1371/journal.pone.0085423
  5. Kibegwa, F. M., Bett, R. C., Gachuiri, C. K., Machuka, E., Stomeo, F., & Mujibi, F. D. (2023). Diversity and functional analysis of rumen and fecal microbial communities associated with dietary changes in crossbreed dairy cattle. PLoS ONE, 18(1), e0274371. https://doi.org/10.1371/journal.pone.0274371
  6. Kim, J. N., Méndez–García, C., Geier, R. R., Iakiviak, M., Chang, J., Cann, I., & Mackie, R. I. (2017). Metabolic networks for nitrogen utilization in Prevotella ruminicola 23. Scientific Reports, 7(1), 7851. https://doi.org/10.1038/s41598-017-08463-3
  7. Melkina, O. E., Plyuta, V. A., Khmel, I. A., & Zavilgelsky, G. B. (2021). The mode of action of cyclic monoterpenes (−)-limonene and (+)-α-pinene on bacterial cells. Biomolecules, 11(6), 806. https://doi.org/10.3390/biom11060806
  8. National Research Council. (2001). Nutrient requirements of dairy cattle: Seventh revised edition. The National Academies Press. https://doi.org/10.17226/9825

Details

Primary Language

English

Subjects

Animal Biotechnology in Agriculture

Journal Section

Research Article

Authors

Mustofa Hilmi This is me
0000-0002-4045-2576
Indonesia

Novita Dewi Kristanti This is me
0000-0003-1117-4383
Indonesia

Publication Date

February 18, 2026

Submission Date

June 24, 2025

Acceptance Date

January 1, 2026

Published in Issue

Year 2026 Number: 1

DOI

https://doi.org/10.29133/yyutbd.1725613

IZ

https://izlik.org/JA42PK98GK

Cite

RIS / Bibtex
APA
Daning, D. R. A., Aji, I., Rahayu, S., Hilmi, M., & Kristanti, N. D. (2026). Modulation of Rumen Microbial Communities in Tropical Holstein Dairy Cows through Lemon Peel Essential Oil Supplementation: A Metagenomic Analysis. Yuzuncu Yıl University Journal of Agricultural Sciences, 1, 1725613. https://doi.org/10.29133/yyutbd.1725613
Creative Commons License
Yuzuncu Yil University Journal of Agricultural Sciences by Van Yuzuncu Yil University Faculty of Agriculture is licensed under a Creative Commons Attribution 4.0 International License.