Identification of Plasma Fatty Acids in Four Lipid Classes to Understand Energy Metabolism at Different Levels of Ketonemia in Dairy Cows Using Thin Layer Chromatography and Gas Chromatographic Techniques (TLC-GC)

Abstract

Excessive mobilization of adipose tissue in high milk producing dairy cows predisposes to metabolic diseases. The aim of this research was to identify the plasma fatty acids in four lipid classes as biomarkers for the diagnosis of hyperketonemia in bovines using thin layer chromatography and gas chromatographic techniques (TLC-GC). Sixty multiparous Holstein–Friesian dairy cows were enrolled in the study. Blood samples from the coccygeal vein were collected and β-hydroxybutyrate (BHB) was evaluated. All animals were divided into three groups on the basis of ketonemia: BHB < 0.50 mmol/L, 0.50 < BHB < 1.0 mmol/L, and BHB > 1.0 mmol/L. Plasma fatty acid concentrations were evaluated in four lipid classes: Free Fatty Acids (FFA), Triglycerides (TG), Cholesterol Esters (CE) And Phospholipids (PL). The concentration of fatty acids was analyzed using TLC-GC. The results showed the following significance in the lipid classes: 19 fatty acids were significant (p < 0.053) in FFA, nine fatty acids were significant (p < 0.050) in TG, eight fatty acids were significant (p < 0.050) in CE and three fatty acids were significant (p < 0.049) in PL. Eleven parameters were considered as predictive fatty acids related to animals in hyperketonemia. The FFA increased simultaneously with blood BHB levels, although the identified predictive fatty acids related to the TG and CE lipid classes decreased, meanwhile the BHB values increased. In the PL lipid class, no fatty acids were predictive.

Changes of Plasma Fatty Acids in Four Lipid Classes to Understand Energy Metabolism at Different Levels of Non-Esterified Fatty Acid (NEFA) in Dairy Cows

“…The data of this research belong to the “BovineOmics Project” (supported by University of Padua, Padova, Italy), based on the analysis through TLC-GC of the profile of plasma lipids in bovine during the transition period. In the study of Fiore et al, 2020, the data were previously analyzed evaluating blood BHB value whereas, in this study, the plasma concentrations of fatty acids were processed focused on the NEFA blood [ 22 , 23 ]. According to the literature, both NEFA and BHB are recognized as indicators of the risk of developing metabolic disorder in lactating period of dairy cows [ 12 , 24 ].…”

“…Sixty Holstein–Frisian dairy cows were enrolled in early post-partum (26.5 ± 1.5 days in milk). The total mixed ration (TMR) and the chemical composition of lactation diet was previously reported in the study of Fiore et al [ 22 ]. Blood samples, analyzed for this research, were the same collected in the study of Fiore et al, taken from the high producing dairy farm located in Padua, Italy (45°36′ N. 11°40′ E. 23 m above sea-level) [ 18 , 22 , 23 ].…”

“…For each animal, 35 FA were obtained per four lipid classes: free fatty acids (FFA), phospholipids (PL), triglycerides (TG) and cholesterol esters (CE). The extraction of lipids from plasma, the separation of lipid classes by TLC, the methylation of the carbon chain of fatty acids and GC were performed as in accordance with the study of Fiore et al [ 22 ].…”

“…Subsequently, the Receiver Operating Characteristic (ROC) analysis was performed using MedCalc Software (Ostend, Belgium) to establish the cut-off on the plasma fatty acids considered predictive/diagnostic. Statistical analysis was further explained in the study by Fiore et al [ 22 ].…”

Ketosis and its role in Bos taurus reproductive impairment

“…Это согласуется с более свежими исследованиями Фьоре и др. [26], в которых концентрация триглицеридов, фосфолипидов и эфиров холестерина у животных с уровнем β-оксимасляной кислоты меньше 1 ммоль/л была ниже, тогда как при уровне больше 1 ммоль/л концентрация НЭЖК оказывалась выше. Причем произошло значительное увеличение плазменных концентраций пальмитиновой (C16), стеариновой (C18), C18:1 и C18:2 кислот.…”