Publication: Long-term biochemical and haematological analysis, with focus on sex effects, in pigs derived from assisted reproductive technologies
Authors
Mikita, Małgorzata ; Romero Aguirregomezcorta, Jon ; Cánovas, Sebastián ; Heras, Sonia ; Ramis, Guillermo ; Gadea, Joaquín ; Coy, Pilar ; Romar Andrés, Raquel
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© 2025 Universidad de Murcia. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/ This document is the Accepted Manuscript, version of a Unpublished Work.
Abstract
Rising livestock demand has increased use of assisted reproductive technologies (ART). Evidence indicates long-term effects of ART on molecular physiology and metabolism. This study explores ART's impact on biochemical and haematological parameters in a colony of pigs from 1 to 2.5 years of age. Animals were born after artificial insemination (AI) and transfer of embryos in vitro produced with (RF-IVP) or without (C-IVP) oviductal and uterine fluids during in vitro fertilization and embryo culture media (París-Oller et al., J Anim Sci and Biotechnol, 2021). Pigs were maintained under identical conditions and sampled every six months. Number of animals in each group decreased throughout life: 33 to 9 (AI); 22 to 10 (C-IVP); and 14 to 4 (RF-IVP). The number ofblood samples was the same as the number of animals and were collected via jugular venipuncture in lithium heparin tubes and immediately transported to laboratory. Plasma was obtained through centrifugation (1200 g, 20 min, 4 ºC, Eppendorf 5810 R) of blood collected via direct jugular venipuncture in lithium heparin tubes. Plasma was assessed using a chemistry analyser (Olympus AU400, Japan) for creatinine (CREA), urea, amylase, creatine kinase (CK), cholesterol, alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), glucose, aspartate aminotransferase (AST), alanine aminotransferase (ALT), lipase, total protein (TP), albumin (ALB), globulin (GLOB), triglycerides (TRIGL), and total bilirubin (TBIL). A haematology analyser (Siemens ADVIA® 120, USA) assessed red blood cell (RBC) count, haemoglobin concentration (HB), haematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), cell haemoglobin concentration mean (CHCM), red blood cell distribution width (RDW), haemoglobin concentration distribution width (HDW), white blood cell (WBC) count; and the differential count of neutrophils, lymphocytes, monocytes, eosinophils, and basophils. Reticulocyte percentage, mean corpuscular volume of reticulocytes (MCVr), haemoglobin content in reticulocytes (CHr) and platelet count were analysed. Data were analysed by mixed-effects model and likelihood-ratio test to determine effects of age and sex on the variables. Post-hoc test for multiple comparisons Tukey's method was used. Statistical significance was set at P < 0.05. Sex significantly influenced biochemical and haematological profiles. In biochemical parameters, females exhibited higher ALBU and urea concentrations, while males demonstrated elevated GLOB levels. Haematological analysis revealed sex-specific differences: females showed increased CH and PCDW values, whereas males presented higher HDW, PDW, and MPC parameters. Age-sex interactions were observed for TP, GLOB, urea, TRIGLI, amylase, and TBIL. Significant age-related changes occurred in RDW, CH, HDW, WBC, neutrophil percentage and count, lymphocyte percentage, PMDW, CHr, and MCVr. The interaction between sex and group was observed in ALT, HCT, and percentage of reticulocytes. ALT showed differences in males between C-IVP and RF-IVP group (p=0.01); HCT also in males between AI and both C-IVP and RF-IVP groups (p=0.01; p=0.02 respectively). A significant interaction was demonstrated between AI and C-IVP in males, as well as in the percentage of reticulocytes. These physiological findings provide essential reference data for veterinary practitioners and livestock producers, demonstrating minor long-term biochemical and haematological variations between naturally and artificially conceived swine. However, observed differences appear clinically insignificant, suggesting comparable physiological development regardless of conception method.
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