Serum pregnancy-associated glycoprotein profiles during early gestation in Karya and Konya Merino sheep.

BACKGROUND: Reported as being expressed by mono- and binucleate placental cells, pregnancy-associated glycoproteins (PAGs) are released into the blood circulation from the ruminant placenta. Circulating gestational PAGs levels may differ between sheep breeds. OBJECTIVE: This study was aimed at the close monitoring of the serum PAGs profiles of Karya and Konya Merino sheep during early pregnancy. METHODS: Fifteen Karya and 15 Konya Merino ewes were synchronized by a 12-day treatment with progesterone-impregnated intravaginal sponges. After the sponges were withdrawn, the ewes were administered 400 IU of equine chorionic gonadotropin. The ewes were allowed to mate naturally, and all animals were sampled for blood as of the day of mating (day 0) at weeks 1, 2, 3, 4 and 5. Pregnancy diagnoses were made by transabdominal ultrasonography at week 5. At weeks 6, 7, 8, 9 and 11, blood samples were collected only from the pregnant ewes. The blood samples were centrifuged at 3000 × g, and extracted sera were stored at -20°C until being used for laboratory analyses. Serum PAGs levels were determined with the aid of a commercial PAG-enzyme-linked immunosorbent assay test originally developed for pregnancy diagnosis in cattle. Differences in the between the PAGs levels throughout pregnancy and the group effect (Karya and Konya Merino) were determined with a two-way mixed analysis of variance. Pairwise comparisons were made using a Bonferroni adjustment. RESULTS: PAGs levels showed a linear increase with the advance of pregnancy in both Karya and Konya Merino sheep. No difference was detected between the breeds for serum PAGs levels. The serum PAGs levels of the pregnant and non-pregnant ewes differed as of the fourth week. CONCLUSION: The serum PAGs levels of the Karya and Konya Merino ewes were similar during the first 11 weeks of gestation, and pregnancy diagnosis could be made based on serum PAGs levels as of the 4th week in both breeds.

Evaluation of refractometry methods for estimating passive immunity status in neonatal foals.

The objective of this cross-sectional study was to determine the accuracy of the digital Brix and serum total protein (TP) refractometers for estimating different passive immunity status in neonatal foals. In total, 18- to 40-h old purebred Arabian foals (n = 185) were used. Serum TP concentrations, total solid percentages and IgG concentrations were measured with a digital serum TP refractometer, digital Brix refractometer and the gold standard radial immunodiffusion (RID) assay, respectively. Correlation coefficients were calculated between the refractometer and RID assay results. A receiver operating characteristic analysis was used to select the optimal cut-offs for both refractometers. Test performance and agreement were evaluated using diagnostic test characteristics at optimal thresholds and areas under the ROC curve, and by calculating Cohen's kappa coefficient. The sensitivity and specificity of the digital Brix refractometer at optimal cut-offs (≤7.8%, ≤7.9%, ≤8.2%, ≤8.3%, ≤9.0%) were 100 and 69.3%; 100% and 68.5%; 70.5% and 71.0%; 88.3% and 85.5%; 88.1% and 76% to estimate RID-IgG of <400 mg/dL, <800 mg/dL, <1500 mg/dL, <2500 mg/dL and < 3000 mg/dL, respectively. The sensitivity and specificity of the digital serum TP refractometer at optimal cut-off (≤4.6 g/dL, ≤4.6 g/dL, ≤4.8 g/dL, ≤5.0 g/dL, ≤5.4 g/dL) were 100 and 69.3%; 100% and 72.8%; 90% and 72.8%; 72.9% and 83.9%; 84.4% and 88% to estimate RID-IgG of <400 mg/dL, <800 mg/dL, <1500 mg/dL, <2500 mg/dL and < 3000 mg/dL, respectively. In conclusion, the refractometers showed a good potential as screening tools for the estimation of different IgG concentrations in neonatal foals.

Diagnostic accuracy of digital Brix and serum total protein refractometers in estimating different passive immunity levels in dairy calves.

The aim of this study was to determine the diagnostic accuracy of digital serum total protein (TP) and digital Brix refractometers in estimating different passive immunity levels (<10, <18, <25 mg/mL) in dairy calves. Blood samples were collected from 260 apparently healthy Holstein calves, aged 2-7 days. Serum IgG concentrations were measured using digital Brix and TP refractometers and the radial immunodiffusion (RID) assay, as the gold standard. Data were analyzed by a receiver operating characteristics (ROC) analysis, the area under the ROC curves (AUC) and Cohen's kappa (κ). Optimal thresholds were determined as < 8.4, < 9.0 and < 9.4% for the digital Brix refractometer, and < 5.0, < 5.4 and < 5.8 g/dL for the serum TP refractometer in estimating IgG concentrations of < 10, < 18, < 25 mg/mL, respectively. The sensitivity (Se) and specificity (Sp) of the Brix refractometer were 96.3% and 88.8% for < 8.4% Brix, 97.0% and 83.4% for < 9.0% Brix, and 85.5% and 77.8% for < 9.4% Brix, respectively. The Se and Sp of the serum TP refractometer were 96.3% and 90.1% for < 5.0 g/dL, 91.0% and 89.6% for < 5.4 g/dL, 79.6% and 85.2% for < 5.8 g/dL, respectively. The discriminant ability of the refractometers was moderately accurate in estimating IgG concentrations of < 10 and < 18 mg/mL, and highly accurate in estimating IgG concentrations of < 25 mg/mL. Both refractometers substantially agreed with RID-IgG results and almost perfectly agreed with each other. In conclusion, the digital Brix and digital serum TP refractometers offer a good utility for determining different passive immunity levels in dairy calves.

Effect of the interval from follicle aspiration to initiation of lengthened FSH treatment on follicular superstimulatory and superovulatory responses and embryo production in lactating Simmental cows.

The present study evaluated follicular superstimulatory (FSS) and superovulatory (SOV) responses and in vivo embryo production in lactating Simmental cows treated with FSH starting 1 or 2 days after follicle aspiration (FA). The performance of a lengthened superovulation program, named 6dFSH-P36-hCG60, is described. At random stages of the estrous cycle, cows (n = 52) were subjected to ultrasound-guided transvaginal aspiration of all follicles ≥ 5 mm. After FA, cows were randomly assigned to one of two groups in which FSH treatments started 1 or 2 days after FA (groups FA-1D and FA-2D, respectively). Cows were superstimulated with a total of 500 µg pFSH over 6 days on a decreasing dose schedule and were pre-treated with a single dose of 400 IU of eCG 24 h before the start of FSH treatments. Follicular superstimulatory (the mean numbers of follicles ≥ 8 mm on the day of hCG treatment) and SOV responses (the mean numbers of CL and cows with ≥ 3 CL at the time of collection) were similar in FA-1D and FA-2D groups. However, when compared to FA-1D group, the number of unfertilized ova tended to decrease (0.4 vs 1.7; P = 0.065) and percentage of fertilized ova tended to increase (95.8% vs 84.6%; P = 0.066) in FA-2D group. Moreover, the mean numbers and percentages of both transferable embryos (8.0 and 77.6% vs 6.4 and 57.7%) and freezable embryos (5.3 and 51.5% vs 3.5 and 31.1%) were numerically higher in FA-2D group than FA-1D group. The results of the study suggest that starting a lengthened superovulation programs in Simmental cows 2 days after FA has potential to increase percentage of fertilized ova and the number of transferable and freezable embryos, although new studies may be needed to confirm this findings.