# Antiviral and anti-inflammatory evaluation of herbal extracts: Implications for the management of calf diarrheal diseases

**Authors:** Xi-Rui Xiang, Eun-Seo Lee, Junho Lee, Su Min Kyung, Han Sang Yoo, Tofazzal Md Rakib, Tofazzal Md Rakib, Tofazzal Md Rakib, Tofazzal Md Rakib

PMC · DOI: 10.1371/journal.pone.0342013 · PLOS One · 2026-02-06

## TL;DR

This study explores how herbal extracts can fight viruses and reduce inflammation in calves, potentially offering new treatments for calf diarrhea.

## Contribution

The study systematically evaluates antiviral and immunomodulatory effects of three herbal extracts in vitro.

## Key findings

- All three herbal extracts inhibited bovine rotavirus and bovine coronavirus in vitro.
- The extracts reduced inflammatory mediator expression in macrophage models.
- Extract C modulated T helper cell responses and suppressed regulatory T-cell transcription in bovine PBMCs.

## Abstract

Traditional herbal extracts are attracting attention in the context of animal disease control because of their low side effects, diverse bioactive compounds, and low antimicrobial resistance risk. However, the underlying mechanisms remain inadequately understood. To characterize the multifaceted biological activities that underlie their therapeutic potential, this study systematically evaluated the antiviral and complex immunomodulatory properties of three distinct herbal combinations (designated Extracts A, B, and C) in vitro. The antiviral activities of the extracts were tested against bovine rotavirus and bovine coronavirus, two major pathogens of neonatal calf diarrhea, and their regulation of inflammatory mediators was assessed in a murine macrophage model (RAW 264.7 cells) stimulated with lipopolysaccharide from Escherichia coli by measuring nitric oxide production and the gene expression of inflammation related enzymes. Immunomodulatory effects were investigated by analyzing the gene expression of T helper cell-associated cytokines in both RAW 264.7 cells and bovine peripheral blood mononuclear cells (PBMCs). All three extracts exhibited inhibitory activity against both viruses and reduced the expression of specific inflammatory mediators. Furthermore, the extracts demonstrated complex immunomodulatory effects in bovine PBMCs, with Extract C promoting T helper 1 and T helper 17 responses while suppressing the regulatory T-cell transcription factor Forkhead Box P3. In conclusion, this in vitro study demonstrates that these herbal extracts possess antiviral and immunomodulatory potential, providing a basis for future studies to determine their relevance in viral infections associated with calf diarrhea.

## Linked entities

- **Species:** Mus musculus (taxon 10090)

## Full-text entities

- **Genes:** IFNG (interferon gamma) [NCBI Gene 281237], Akt1 (Akt serine/threonine kinase 1) [NCBI Gene 11651] {aka Akt, LTR-akt, PKB, PKB/Akt, PKBalpha, Rac}, TBX21 (T-box transcription factor 21) [NCBI Gene 538933], Actb (actin, beta) [NCBI Gene 11461] {aka Actx, E430023M04Rik, beta-actin}, Sik2 (salt inducible kinase 2) [NCBI Gene 235344] {aka G630080D20Rik, Snf1lk2}, FOXP3 (forkhead box P3) [NCBI Gene 506053], IL22 (interleukin 22) [NCBI Gene 507778] {aka IL-22}, Mapk8 (mitogen-activated protein kinase 8) [NCBI Gene 26419] {aka JNK, JNK1, Prkm8, SAPK1}, Ptgs2 (prostaglandin-endoperoxide synthase 2) [NCBI Gene 19225] {aka COX2, Cox-2, PES-2, PGHS-2, PHS II, PHS-2}, Ifng (interferon gamma) [NCBI Gene 15978] {aka IFN-g, If2f, Ifg}, Tnf (tumor necrosis factor) [NCBI Gene 21926] {aka DIF, TNF-a, TNF-alpha, TNFSF2, TNFalpha, Tnfa}, Il21 (interleukin 21) [NCBI Gene 60505] {aka IL-21}, IL4 (interleukin 4) [NCBI Gene 280824] {aka BSF-1, IL-4}, IL5 (interleukin 5) [NCBI Gene 280825] {aka IL-5, TRF}, Foxp3 (forkhead box P3) [NCBI Gene 20371] {aka JM2, scurfin, sf}, GATA3 (GATA binding protein 3) [NCBI Gene 505169], Pcx (pyruvate carboxylase) [NCBI Gene 18563] {aka Pc, Pcb}, IL10 (interleukin 10) [NCBI Gene 281246] {aka IF2A}, IL17A (interleukin 17A) [NCBI Gene 282863] {aka IL-17, IL17}, Pdk1 (pyruvate dehydrogenase kinase, isoenzyme 1) [NCBI Gene 228026] {aka B830012B01, D530020C15Rik}, IL2 (interleukin 2) [NCBI Gene 280822] {aka IL-2, TCGF}, IL1B (interleukin 1 beta) [NCBI Gene 281251], CD4 (CD4 molecule) [NCBI Gene 407098], CTLA4 (cytotoxic T-lymphocyte associated protein 4) [NCBI Gene 281732] {aka CD152, CTLA-4}, COX2 (cytochrome c oxidase subunit II) [NCBI Gene 17709], Nfkb1 (nuclear factor of kappa light polypeptide gene enhancer in B cells 1, p105) [NCBI Gene 18033] {aka NF-KB1, NF-kappaB, NF-kappaB1, p105, p50, p50/p105}, Il5 (interleukin 5) [NCBI Gene 16191] {aka Il-5}, IL13 (interleukin 13) [NCBI Gene 281247], Nos2 (nitric oxide synthase 2, inducible) [NCBI Gene 18126] {aka MAC-NOS, NOS-II, Nos-2, Nos2a, i-NOS, iNOS}, Il6 (interleukin 6) [NCBI Gene 16193] {aka Il-6}, TNF (tumor necrosis factor) [NCBI Gene 280943] {aka TNF-a, TNF-alpha, TNFa}, RORC (RAR related orphan receptor C) [NCBI Gene 527470]
- **Diseases:** enteritis (MESH:D004751), dehydration (MESH:D003681), Diarrheal Diseases (MESH:D004403), viral diarrhea (MESH:D014777), gastrointestinal (MESH:D005767), Inflammation (MESH:D007249), Diarrhea (MESH:D003967), bacterial infections (MESH:D001424), infection (MESH:D007239), inflammatory bowel disease (MESH:D015212), PC (MESH:C536209), macrophage (MESH:D055501), Cytotoxicity (MESH:D064420)
- **Chemicals:** nitrite (MESH:D009573), chloroform (MESH:D002725), penicillin (MESH:D010406), streptomycin (MESH:D013307), ethanol (MESH:D000431), DMSO (MESH:D004121), Bavachin (MESH:C459212), SYBR Green (MESH:C098022), water (MESH:D014867), LPS (MESH:D008070), MTT (MESH:C070243), CO2 (MESH:D002245), Propolis (MESH:D011429), genipin (MESH:C007834), DEPC (MESH:D004047), baicalin (MESH:C038044), NO (MESH:D009569), B. serrata (-), isopropanol (MESH:D019840)
- **Species:** Mus musculus (house mouse, species) [taxon 10090], Commiphora myrrha (myrrh, species) [taxon 318982], Severe acute respiratory syndrome coronavirus 2 (no rank) [taxon 2697049], Escherichia coli (E. coli, species) [taxon 562], Boswellia serrata (species) [taxon 613112], Bos taurus (bovine, species) [taxon 9913], Betacoronavirus (genus) [taxon 694002], Rotavirus (genus) [taxon 10912], Alpharicinrhavirus blanchseco (species) [taxon 2843852], Homo sapiens (human, species) [taxon 9606], Nypa fruticans (nipa palm, species) [taxon 4718], Gardenia jasminoides (species) [taxon 114476], Scutellaria baicalensis (Baikal skullcap, species) [taxon 65409], Glycyrrhiza uralensis (Chinese licorice, species) [taxon 74613], Bovine coronavirus (no rank) [taxon 11128], Bovine rotavirus (no rank) [taxon 10927]
- **Mutations:** A-C at 6, A-C for 4, C at 100
- **Cell lines:** TF104 — Homo sapiens (Human), Erythroleukemia, Cancer cell line (CVCL_3608), MA-104 monkey — Chlorocebus pygerythrus (Vervet monkey), Spontaneously immortalized cell line (CVCL_3845), RAW 264.7 — Mus musculus (Mouse), Mouse leukemia, Cancer cell line (CVCL_0493), MDBK — Bos taurus (Bovine), Spontaneously immortalized cell line (CVCL_0421)

## Full text

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## Figures

9 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12880661/full.md

## References

71 references — full list in the complete paper: https://tomesphere.com/paper/PMC12880661/full.md

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Source: https://tomesphere.com/paper/PMC12880661