Long-Term Study of Physical, Haematological, and Biochemical Parameters in Cattle with Different Embryo Origins

Abstract

Assisted reproductive technologies are vital in cattle breeding to improve genetic selection and productivity. While early-life differences between artificially inseminated (AI) and in vitro-produced (IVP) cattle have been studied, long-term physiological, haematological, and biochemical effects remain unclear. This observational study assessed AI and IVP cattle from 1.5 to 5 years of age to determine if early differences persist. IVP cattle were produced after the transfer of the embryo produced by supplementing (RF-IVP group) or not supplementing (C-IVP) the embryo culture with oviductal and uterine fluids. Physical evaluations showed body mass index increased until 3.5 years, while temperature and respiratory rate declined with age, with no significant differences between AI and IVP groups. Haematological analysis revealed age-related changes, including decreased red and white blood cell counts and increased mean corpuscular volume and haemoglobin. AI cattle had higher white blood cell counts than IVP groups. Sex significantly influenced many haematological variables. Biochemical analysis showed age-related increases in total protein, creatinine, and urea, and decreases in glucose and alkaline phosphatase. AI cattle had lower cholesterol and creatinine than IVP groups. Despite group differences, all values remained within normal ranges. Sex affected albumin, cholesterol, triglycerides, and creatine kinase. This study provides the first long-term haematological and biochemical reference values for cattle from different reproductive methods, showing that age is the main influencing factor and supporting IVP cattle as a viable alternative to AI in breeding programs.

Keywords: assisted reproductive technologies; biochemical profile; cattle; haematological profile; long-term study.

Figure 1

Evolution of physical parameters across age in cattle born by artificial insemination (AI) and transfer of in vitro embryo produced under a standard protocol (C-IVP), and including reproductive fluids (RF-IVP). Data are expressed as mean ± SEM. (A) Body mass index (BMI), (B) body temperature, (C) respiratory rate (RPM, respirations per minute), and (D) heart rate (beats per minute). Asterisk shows statistical difference at * p < 0.05, ** p < 0.01, and *** p < 0.001; NS, not significant. Horizontal lines denote that all encompassed time points are significantly different. When the vertical lines are added, only the marked time points are significantly different.

Figure 2

Evolution of haematological parameters (red blood cells) across age obtained in cattle born by artificial insemination (AI) and transfer of in vitro embryo produced under a standard protocol (C-IVP), and including reproductive fluids (RF-IVP). Data are expressed as mean ± SEM. (A) Red blood cells (RBC; ×10⁶ cells/μL), (B) mean corpuscular volume (MCV; fL), (C) mean corpuscular haemoglobin (MCH; pg), (D) mean corpuscular haemoglobin concentration (MCHC; g/dL), (E) red blood cell distribution width (RDW, %), (F) haemoglobin content (CH; pg), (G) haemoglobin distribution width (HDW; g/dL), and (H) cell haemoglobin distribution width (CHDW; pg). Asterisk shows statistical difference at * p < 0.05, ** p < 0.01, and *** p < 0.001; NS, not significant. Horizontal lines denote that all encompassed time points are significantly different. When the vertical lines are added, only the marked time points are significantly different.

Figure 3

Evolution of haematological parameters (white blood cells) across age obtained in cattle born by artificial insemination (AI) and transfer of in vitro embryo produced under a standard protocol (C-IVP), and including reproductive fluids (RF-IVP). Data are expressed as mean ± SEM. (A) White blood cells (WBC; ×10³; cells/μL), (B) neutrophils (%), (C) lymphocytes (%), (D) neutrophils (×10³ cells/μL), (E) lymphocytes (×10³ cells/μL), (F) monocytes (%), (G) monocytes (×10³ cells/μL), (H) eosinophils (×10³ cells/μL), (I) basophils (%), and (J) basophils (×10³ cells/μL). Asterisk shows statistical difference at * p < 0.05, ** p < 0.01, and *** p < 0.001; NS, not significant. Horizontal lines denote that all encompassed time points are significantly different. When the vertical lines are added, only the marked time points are significantly different.

Figure 4

Haematological parameters (platelets and reticulocytes) across age obtained in cattle born by artificial insemination (AI) and transfer of in vitro embryo produced under a standard protocol (C-IVP), and including reproductive fluids (RF-IVP). Data are expressed as mean ± SEM. (A) Platelets (PLT; ×10³; cells/μL), (B) mean platelet volume (MPV; fL), (C) plateletcrit (PCT; %), (D) platelet distribution width (PDW; %), (E) reticulocytes (%), and (F) reticulocytes (×10³ cells/μL). Asterisk shows statistical difference at * p < 0.05, ** p < 0.01, and *** p < 0.001; NS, not significant. Horizontal lines denote that all encompassed time points are significantly different. When the vertical lines are added, only the marked time points are significantly different.

Figure 5

Biochemical parameters (protein, glycaemic and kidney biomarkers) across age obtained in cattle born by artificial insemination (AI) and transfer of in vitro embryo produced under a standard protocol (C-IVP), and including reproductive fluids (RF-IVP). Data are expressed as mean ± SEM. (A) Total proteins (TP; g/dL), (B) albumin (ALB; g/dL), (C) globulin (GLOB; g/dL), (D) creatinine (CREA; mg/dL), (E) urea (mg/dL), (F) glucose (GLUC; mg/dL), (G) cholesterol (CHOL; mg/dL), and (H) triglycerides (TRIG; mg/dL). Asterisk shows statistical difference at * p < 0.05, ** p < 0.01, and *** p < 0.001; NS, not significant. Horizontal lines denote that all encompassed time points are significantly different. However, when the vertical lines are added, only the marked time points are significantly different.

Figure 6

Biochemical parameters (hepatic, biliary, and gastrointestinal biomarkers) across age obtained in cattle born by artificial insemination (AI) and transfer of in vitro embryo produced under a standard protocol (C-IVP), and including reproductive fluids (RF-IVP). Data are expressed as mean ± SEM. (A) Amylase (AMS; UI/L), (B) lipase (LIP; UI/L), (C) creatinine kinase (CK; UI/L), (D) alkaline phosphatase (ALP; UI/L), (E) gamma-glutamyl transpeptidase (GGT; UI/L), (F) aspartate aminotransferase (AST; UI/L), (G) alanine aminotransferase (ALT; UI/L), and (H) total bilirubin (TB; mg/dL). Asterisk shows statistical difference at * p < 0.05, and *** p < 0.001; NS, not significant. Horizontal lines denote that all encompassed time points are significantly different. When the vertical lines are added, only the marked time points are significantly different.