9bb85a0271821215ab6b3c1402a71bf3 ANS_a2025v15p02810.pdf 7f1482ce43d41a3d265bfe42aaab6efef9fb3833 ANS_a2025v15p02810.pdf 9f4a23ef676b43c3749f16e87e2114220a5ec27dc58571c6af87c7a685b7d032 ANS_a2025v15p02810.pdf Title: Exploratory Metabolomic Fingerprinting of Aqueous Humor in Healthy Horses and Donkeys, and in Horses with Ocular Pathologies Subject: This study aims to generate foundational metabolomic data of aqueous humor (AH) in healthy horses and donkeys, and to investigate potential changes or trends in the metabolomic profile associated with age, sex or ocular pathology in horses. The AH metabolomic fingerprint from 5 donkeys and 35 equine eyes (17 controls, 8 with cataracts, 6 with retinal disease and 4 with anterior chamber disease (ACD)) were analyzed using nuclear magnetic resonance (NMR) spectroscopy. A linear mixed-effects model, with individual horse as a random effect and group as a fixed effect, with multiple testing correction using the Benjamini–Hochberg false discovery rate (FDR) method was used to compare groups. The metabolomic profile of the donkeys and horse’s AH is very similar to that of other mammals. Threonine was higher in young horses (p = 0.04), and creatinine was elevated in males (p = 0.04). Compared with control groups, dimethyl sulfone was higher in the retina (p < 0.00) and cataract (p = 0.05) groups. Arginine (p = 0.05) and valine (p = 0.03) were lower in the retina group compared to controls. This study successfully characterized the AH metabolomic profile in healthy horses and donkeys and identified several metabolites that could be associated with ocular pathology, warranting further investigation to determine their potential as biomarkers of ocular disease. Keywords: metabolomics; fingerprinting; aqueous humor; horses; donkeys; oculomics Author: Ignacio Corradini, Eduard Jose-Cunilleras, Pau Nolis, María Mar López-Murcia and Aloma Mayordomo-Febrer Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25; modified using OpenPDF 1.4.2 CreationDate: Fri Sep 26 12:40:56 2025 CEST ModDate: Fri Sep 26 13:08:51 2025 CEST Custom Metadata: yes Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 16 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1349523 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- JCGAPA+URWPalladioL-Ital Type 1 Custom yes yes yes 122 0 SSXPJJ+VnURWPalladioL Type 1 Custom yes yes yes 123 0 IFUYKG+URWPalladioL-Bold Type 1 Custom yes yes yes 124 0 OKZIYA+URWPalladioL-Roma Type 1 Custom yes yes yes 125 0 WTPQBM+CMSY10 Type 1 Builtin yes yes yes 179 0 OVNEXW+EURM10 Type 1 Builtin yes yes yes 180 0 WBVKDJ+URWPalladioL-BoldItal Type 1 Custom yes yes yes 228 0 MJKFDC+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 213 0 MJKFBB+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 214 0 MJKFCB+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 215 0 MJKFDC+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 254 0 MJKFBB+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 255 0 MJKFCB+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 256 0 MJKFDC+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 261 0 MJKFBB+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 262 0 MJKFCB+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 263 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-10-29 04:37:11 CET RepresentationInformation: ANS_a2025v15p02810.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-10-28 09:40:42 CET Size: 1349523 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 531 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Item: Title: Materials and Methods Children: Item: Title: Animals and Study Design Item: Title: Ophthalmic Examination Item: Title: Sample Collection and Preparation Item: Title: Data Acquisition and Analysis Item: Title: Quantification of Metabolites Item: Title: Comparative Analysis Item: Title: Results Children: Item: Title: Samples Item: Title: Characterization of the Metabolomic Profile of the AH of the Horse and Donkey Healthy Eyes Item: Title: Characterization of the Metabolomic Profile of AH According to Age and Sex Groups of Normal Equine Eyes Item: Title: Characterization of the Metabolomic Profile of the AH of the Equine Eyes with Ocular Disease Item: Title: Discussion Children: Item: Title: Age-Related Variations in AH Composition Item: Title: Significant Metabolite Changes in Horse AH of Eyes with Cataracts and Retinal Disease Item: Title: Metabolomic Trends in Horse AH Associated with Ocular Disease Item: Title: Conclusions Item: Title: References Info: Title: Exploratory Metabolomic Fingerprinting of Aqueous Humor in Healthy Horses and Donkeys, and in Horses with Ocular Pathologies Author: Ignacio Corradini, Eduard Jose-Cunilleras, Pau Nolis, María Mar López-Murcia and Aloma Mayordomo-Febrer Subject: This study aims to generate foundational metabolomic data of aqueous humor (AH) in healthy horses and donkeys, and to investigate potential changes or trends in the metabolomic profile associated with age, sex or ocular pathology in horses. The AH metabolomic fingerprint from 5 donkeys and 35 equine eyes (17 controls, 8 with cataracts, 6 with retinal disease and 4 with anterior chamber disease (ACD)) were analyzed using nuclear magnetic resonance (NMR) spectroscopy. A linear mixed-effects model, with individual horse as a random effect and group as a fixed effect, with multiple testing correction using the Benjamini–Hochberg false discovery rate (FDR) method was used to compare groups. The metabolomic profile of the donkeys and horse’s AH is very similar to that of other mammals. Threonine was higher in young horses (p = 0.04), and creatinine was elevated in males (p = 0.04). Compared with control groups, dimethyl sulfone was higher in the retina (p < 0.00) and cataract (p = 0.05) groups. Arginine (p = 0.05) and valine (p = 0.03) were lower in the retina group compared to controls. This study successfully characterized the AH metabolomic profile in healthy horses and donkeys and identified several metabolites that could be associated with ocular pathology, warranting further investigation to determine their potential as biomarkers of ocular disease. 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