Efficacy of repeated ovum pick-up in Podolic cattle for preservation strategies: a pilot study
ITALIAN JOURNAL OF ANIMAL SCIENCE
Authors: Presicce, Giorgio Antonio; Neglia, Gianluca; Salzano, Angela; Padalino, Barbara; Longobardi, Valentina; Vecchio, Domenico; De Carle, Esterina; Gasparrini, Bianca
The study evaluated the effects of eCG treatment prior to ovum pick up (OPU) on follicular population, oocyte and embryo yields in summer and autumn in Podolic cattle. The effects of repeated OPU on cattle wellbeing was also documented. Twenty-six animals were used, and split into two groups, treatment (OPU; n = 18) and control (CG; n = 8). The OPU cattle were subsequently split into two subgroups (n = 9) and underwent repeated OPU, without and with eCG priming, for a total of 8 sessions (4 sessions/season). Follicular population, oocyte and embryo yields were recorded in those sub-groups. CG was handled in the same manner of OPU, except for epidural anaesthesia and follicular aspiration. Biochemical profile, serum protein electrophoresis and haptoglobin levels were analysed in OPU and CG. Hormonal priming increased the number of medium follicles (1.7 +/- 0.2 vs 0.9 +/- 0.2, p < .05), while it decreased the recovery rate and number of cumulus-enclosed oocytes (COCs) (recovery rate: 38.3 +/- 3.5 vs 60.5 +/- 4.0%; COCs: 2.3 +/- 0.3 vs 3.4 +/- 0.4, respectively; p < .01). However, priming increased cleavage (72.9 +/- 5.7 vs 49.4 +/- 5.4; p < .05) and blastocyst (41.1 +/- 5.7 vs 23.0 +/- 4.2; p = .054) rates. With regard to season's effect, a higher number of COCs was recorded in autumn than in summer (3.1 +/- 0.4 vs 2.6 +/- 0.3; p < .05) without affecting though the number of embryos produced (0.9 on average). Since haematological parameters did not vary between OPU and CG, our preliminary data suggest that repeated OPU may be used as a conservation strategy in Podolic cattle without affecting wellbeing.
Lyophilization as pre-processing for sample storage in the determination of vitamin D-3 and metabolites in serum and plasma
Authors: Castillo-Peinado, L. S.; Calderon-Santiago, M.; Priego-Capote, F.
Determination of vitamin D levels in human biological specimens has gained a high relevance over the last decades, essentially because low levels have been associated with several biological disorders. In fact, vitamin D deficiency has become a worldwide health concern covering all ages and genders. The storage of biofluids has to be considered for determination of vitamin D and metabolites in order to fully preserve matrices status. This study attempts to evaluate lyophilization of serum and plasma as a pre-processing step for sample storage prior to quantitative analysis of vitamin D-3 and its main hydroxylated metabolites-25(OH)D-3, 24,25(OH)(2)D-3 and 1,25 (OH)(2)D-3. The protocol including sample lyophilization was characterized in terms of analytical features and compared to the same method, based on SPE-LC-MS/MS, without lyophilization. Sensitivity, precision and accuracy were not affected when we operated with lyophilized serum and plasma and results provided by a set of twenty-four serum samples from DEQAS (Vitamin D External Quality Assessment Scheme) were in agreement with reported concentrations for 25(OH)D-3 and 1,25(OH)(2)D-3. A stability study programmed for 9 months allowed ensuring that the concentration of vitamin D-3 and metabolites in lyophilized serum and plasma stored at room temperature was not affected during this period. This research has demonstrated that the quantitation of target metabolites is not under the influence of lyophilization. Therefore, including lyophilization prior to analysis could reduce shipment and storage costs, avoid delays of sample processing, and increase the stability of the target analytes due to an effective quenching process.