Czech Journal of Animal Science - In Press
Lactoferrin – the protective component of goat colostrum and milkOriginal Paper
Pavlína Navrátilová, Klára Bartáková, Jan Pospíšil, Lenka Vorlová, Tomáš Kopec, Oto Hanuš, Hana Nejeschlebová, Eva Samková, Josef Kučera, Radoslava Jedelská
Lactoferrin (LF) is an important bioactive component of colostrum, is critical for the development of immunity in the newborns, and an important component of the mammary gland defence system. LF is also an important biomolecule in terms of promoting and restoring human health. The aim of our study was to monitor the dynamics of changes in lactoferrin concentration in goat milk at varying stages of lactation and its correlation with selected components and physicochemical parameters. Colostrum (n = 24) and milk (n = 120) samples were obtained from 12 goats by hand milking. Lactoferrin was determined using reversed-phase high-performance liquid chromatography with an ion-pairing reagent equipped with a Photodiode Array Detector. The LF concentration in colostrum ranged from 206 to 1 228 mg/l, and showed a statistically significant decrease in concentration (P ˂ 0.05) over the colostral period. Significant correlation coefficients (P < 0.001) were found between crude protein and LF (r = 0.896), lactose and LF (r = -0.754), as well as between non-fat solids and LF (r = 0.853). The LF content in milk ranged within a relatively wide range of 94 to 1 115 mg/l although the values were highly variable (vx= 57.0%). Significant correlations were found between fat content and LF in milk (r = 0.429, P < 0.001), crude protein and LF (r = 0.376), non-fat solids and LF (r = 0.361), somatic cell count (SCC) and LF (r = 0.330), as well as log SCC and LF (r = 0.348, P < 0.01).
Lactoferrin content determined in bovine milk by HPLC and mid-infrared spectrometry - relation to udder health and potential for detection of milk adulterationOriginal Paper
Hana Nejeschlebová, Oto Hanuš, Klára Bartáková, Craig Parsons, Eva Samková, Lenka Vorlová, Gavin Thompson, Lucie Hasoňová, Roman Konečný, Pavlína Navrátilová, Marcela Klimešová
Lactoferrin (LF) is a multifunctional glycoprotein associated with the udder health of dairy cows. The aim of this study was to develop a calibration model for LF quantification using mid-infrared spectrometry (MIR-FT; LF MIR-FT). Ion-pair reversed-phase high-performance liquid chromatography (HPLC; LF HPLC) was employed as the reference method. Two sets of individual milk samples, set A (n = 120) and set B (n = 91), were collected from five dairy farms in the Czech Republic. The milk samples in set A were selected so that approximately two-thirds exhibited a somatic cell count (SCC) above 300 000 cells/ml, which proved effective in ensuring greater LF variability. The final calibration set (set C; n = 205) was formed by merging sets A and B, excluding six outliers. For the developed calibration, a cross-validated coefficient of determination of 0.588 7 and a standard error of cross-validation of 67.33 mg/l were achieved. Correlation analysis indicated that several milk parameters in set C correlated with LF MIR-FT in ways consistent with changes typically observed during mastitis (most notably SCC: r = 0.450; lactose monohydrate: r = –0.364; free fatty acids: r = 0.621 and electrical conductivity: r = 0.442). This suggests that the MIR-FT method may have potential for assessing mammary gland health in dairy cows. Furthermore, we aimed to investigate the feasibility of using LF content in milk as an indicator of milk adulteration by artificially reducing SCC through milk centrifugation. For this purpose, two sets of milk samples (n = 20 and n = 68) were collected, both consisting of normal bulk tank milk and bulk tank milk supplemented with abnormal milk from dairy cows in the Czech Republic. Centrifugation of milk resulted in only minimal changes in both LF HPLC and LF MIR-FT (maximum 3.27%), while SCC decreased by nearly 50%. Therefore, LF could serve as a marker for the detection of artificial SCC reduction in milk by centrifugation. The practical application of the MIR-FT method for LF determination is limited by the achieved validation parameters (R²CV, SECV) and the associated estimated expanded combined uncertainty (114.7 mg/l). The MIR-FT method is therefore more suitable for monitoring relative LF changes in milk than for determining exact LF values (e.g., those associated with cow health status, the course of lactation or milk processing).
Colostrum Quality in Northern Italy: The Role of Dry Cow NutritionOriginal Paper
Martina Pavesi, Giulia Gislon, Serena Bonizzi, Stefania Colombini, Nicola Palladini, Maddalena Zucali
This study investigates the effects of dry cow feeding on colostrum quality. A survey was performed, and samples of colostrum and feed ingredients were collected in 25 dairy farms in Northern Italy. Colostrum was analysed for quality in terms of Brix and gamma globulins; diets and feed ingredients were analysed for chemical constituents. The average colostrum quality (n=163) was 24.3±4.41˚Brix and 54.2±20.8 g/l gamma globulin. The refractometer method effectively assessed colostrum quality, correlating accurately with laboratory gamma globulins measurements (R = 0.729,). As resulted by Factor analysis, key nutrients such as protein (13.5 ± 1.49% DM) and fat (4.15 ± 1.58% DM) of dry cow diets influenced colostrum immunoglobulin G Furthermore, Selenium (3.19 ± 3.15 mg/day), Vitamin A (93657±86882 IU/day), Vitamin B12 (0.17±0.35) mg/day), and vitamin E (955 ± 1138 mg/day) supplementation also improved colostrum quality. Colostrum feeding occurred on average 5.87 ± 2.58 hours post-calving with a quantity of 2.74 ± 0.55 L per meal, below the recommended 10% of calf body weight; however, the maximum feeding time was 12 hours, and the minimum colostrum quantity was 1.25 L, highlighting wide improvement possibilities.
Sperm quality improvement of cryopreserved boar semen through colloidal centrifugation gradientOriginal Paper
Francisco Sevilla, Ignacio Araya-Zúñiga, Patricia Cervantes-Acosta, Antonio Hernández-Beltrán, Manuel Barrientos, Belisario Domínguez, Anthony Valverde
Boar semen cryopreservation is limited due to lower sperm cell thermal shock resistance. A colloidal centrifugation gradient is an enhancement to cryopreservation that could improve frozen boar semen quality. Evaluate cryopreserved boar semen quality using a commercial colloidal centrifugation gradient. A total of 15 ejaculates from 5 boars were evaluated across two treatments: a control without colloidal centrifugation and a treatment with commercial colloidal centrifugation. A manual freezing method used two freezing curves in liquid nitrogen. Sperm motility was assessed with Computer-Assisted Semen Analysis, as well as sperm capacitation-like membrane destabilization at 30 min and 150 min post-thaw using Merocyanine 540 (M540) for samples incubated at 37 °C. Spectrophotometry measured lipid peroxidation indirectly by the amount of malondialdehyde; reactive oxygen species production was also determined for sperm samples incubated for 30 min. Gradient centrifugation treatment improved ejaculate sperm motility and membrane destabilization. The proportion of morphologically normal sperm was higher in gradient than control. A total of 74% of spermatozoa retained normal morphology. Lipid peroxidation was lower in the colloidal centrifugation treatment. MDA was lower with gradient (16.4 ± 2.5 vs. 22.3 ± 2.5 µmol/30 × 106 sperm; p < 0.05). Intracellular reactive oxygen species (ROS) and M540‑positivity did not differ significantly between treatments at either time point. The centrifugation process improved quality parameters of frozen boar semen post-thaw.
Effects of Silymarin Supplementation During Late Gestation on Reproductive Performance, Haematological Parameters, Antioxidant Status, and Gut Microbiota in SowsOriginal Paper
Guanglei Cong, Chunxue Liu, Shuangshuang Xia, Junbo Li, Ifen Hung
This study aimed to evaluate the effects of silymarin supplementation during late gestation on reproductive performance, haematological parameters, antioxidant capacity, and gut microbiota composition in sows. Twenty parity-4 crossbred sows (Landrace × Yorkshire) were enrolled and randomly allocated in parity blocks to either a control group (CG; n = 10, basal diet) or a silymarin-supplemented group (SIL; n = 10, basal diet + 200 mg/kg silymarin). The experimental period extended from day 85 of gestation to the completion of farrowing. The results demonstrated that dietary silymarin significantly reduced the number of stillbirths (P < 0.05), without exerting a significant effect on the total number and proportion of live-born piglets (P > 0.05). No notable differences were observed in haematological parameters between the two groups (P > 0.05). However, catalase (CAT) activity and total antioxidant capacity (T-AOC) were significantly elevated in the silymarin group (P < 0.05), and superoxide dismutase (SOD) showed a tendency to increase (P = 0.078). High-throughput 16S rRNA sequencing revealed 1,671 unique feature sequences in the silymarin group and 1,073 in the control group, with 1,600 sequences shared by both groups the two groups. A trend towards increased dominance was observed in the silymarin group (P = 0.082), while both the Shannon and Simpson indices tended to decline (P = 0.087; P = 0.082), suggesting a possible reduction in microbial diversity. Principal coordinate analysis (PCoA) of β-diversity revealed significant structural differences in gut microbiota between the two groups. SIMPER analysis identified Terrisporobacter as the principal genus contributing to these differences. In conclusion, silymarin supplementation during late gestation may enhance reproductive outcomes in sows, potentially through modulation of gut microbial composition and enhancement of systemic antioxidant status.