Detection of pregnancy in sheep using an ELISA for pregnancy-specific protein B.

The early detection of pregnancy and the determination of fetal numbers have economic benefits in sheep production because of the seasonal breeding patterns where missing a breeding opportunity means the loss of one productive year. The purpose of this study was to evaluate the efficacy of the B6-HRP ELISA for ovine pregnancy-specific protein B (oPSPB) measurement in the detection of pregnancy and estimation of fetal numbers in different sheep breeds. BioPRYN® ELISA assay kit was used for the detection of pregnancy in the experimental animals. Ninety-three ewes of three breeds (British Milksheep - BM, Lacaune - L and Transylvanian Racka - TR), each from three farms in Hungary, were included in the study. BM and L ewes were artificially inseminated (AI). Thirty-five days after AI, all ewes were examined by transabdominal ultrasound. The TR flock was mated naturally over a six-week period. At the end of the mating period, the ewes were similarly examined by ultrasound. Blood samples were taken from all pregnant ewes twice (35 and 65 days after AI), and serum samples were assayed by the BioPRYN test. It can be concluded that the detection of serum PSPB by ELISA is a much easier, safer, less expensive and highly accurate method for the detection of ovine pregnancy. Although some breed-related differences were detectable at 35 and 65 days post breeding, no differences in oPSPB levels were found in pregnant ewes carrying different numbers of fetuses.

Is twin pregnancy, calving and pregnancy loss predictable by serum pregnancy-specific protein b (pspb) concentration 28-35 days after ai in dairy cows?

Double ovulation occurs more frequently in multiparous cows with high milk production than in primiparous cows and the rate of twin pregnancy/calving is increasing worldwide. Diagnosis of twin pregnancy is possible by ultrasound at the time of early pregnancy examination [28-34 days after artificial insemination (AI)]. Pregnancy proteins are also well-known indicators of gestation. The risk of pregnancy loss during the first trimester of gestation for cows carrying twins is three to nine times higher than for cows carrying singletons. Pregnancy-specific protein B (PSPB) is a good indicator not only of pregnancy but also of pregnancy loss. The aims of this study were (a) to collect calving data in some Hungarian Holstein-Friesian herds (n = 7,300) to compare PSPB serum concentrations (measured 29-35 days post insemination) in twin- and singleton-calving cows (Trial 1), and (b) to check the predictive value of PSPB serum concentration for twin pregnancy and pregnancy loss in high-producing Spanish Holstein-Friesian cows (n = 98; Trial 2). Our results showed almost 7% twin calving rate. Although hormonal treatments are commonly believed to be major causes of twin pregnancies, our data do not support this hypothesis. The only exception is the single PGF injection, which significantly increased twin calving. No effect of milk production on the risk of twin pregnancy was found, and twin pregnancy increased with parity. The AI bull, the bull's sire, the bull's grandfather and the cow's father also affected twin calving (P ≤ 0.02). We found much higher frequency of twin calving in cows diagnosed pregnant with higher than 3 ng/ml serum PSPB concentrations at 29-35 days after insemination. In Trial 2, non-significant but well-marked differences were found in PSPB serum concentration between singleton- and twin-pregnant cow samples (2.1 and 2.9 ng/ml) at different bleeding times. Probably the small size of the study population and the effects of milk production on PSPB values may explain this lack of significance.

Macroscopic and histological characteristics of fluid-filled ovarian structures in dairy cows.

The primary objective of this study was to use macroscopic and histological features of corpora lutea with a cavity and anovulatory cystic ovarian structures, present in 90 pairs of abattoir-derived dairy cow ovaries, as the basis to clarify the nomenclature of ovarian structures. Excluding morphologically normal ovarian fol-licles (antrum < 2 cm, wall < 1 mm), there were 27 fluid-filled ovarian structures. Ovulatory structures > 16 mm in diameter were designated as Group A (cavity ≤ 10 mm and wall > 10 mm) or Group B (cavity > 10 mm and wall < 10 mm). The volume of luteal tissue was less (P < 0.05) in Group B than in Group A, whereas that of a solid corpus luteum (CL) was intermediate (least square means ± SEM: 72 ± 1.92, 11.22 ± 1.57 and 5.84 ± 1.92 cm3, respectively). There was a greater proportion (P < 0.05) of small luteal cells in Group B compared to a solid CL, whereas Group A was intermediate (58.6 ± 5.3, 37.4 ± 5.3 and 44.0 ± 4.4%, respectively). Connective tissue was thicker (P < 0.05) in Group B than in Group A (295.4 ± 46.9 vs. 153.9 ± 38.2 µm). Based on the above-mentioned characteristics and differences, Groups A and B were designated as a CL with a cavity and a cystic CL, respectively. Furthermore, there were three groups of anovulatory ovarian structures. Structures in Group C were termed persistent/anovulatory follicles (overall diameter and wall thickness ≤ 20 and 1-3 mm, respectively). Finally, Groups D and E were designated as a follicle-fibrous cyst and a follicle-luteinised cyst (based on histological structure) for anovulatory structures with an overall diameter and wall thickness of ≥ 20 and ≤ 3 mm, and ≥ 20 and ≥ 3 mm, respectively.

Seasonal fertility differences in synchronised dairy cows: ultrasonic, metabolic and endocrine findings.

The aim of this study was to identify relationships among seasonal differences of fertility, metabolic parameters and appearance of irregular luteal forms in high-yielding dairy cows. Holstein-Friesian cows were put on the Provsynch regimen in winter (n = 10) and in summer (n = 10). Blood sampling (starting 35 days post partum) and rectal ultrasound examinations (starting post insemination) were carried out once a week in each examination period. Metabolic [plasma nonesterified fatty acid (NEFA) and beta-hydroxybutyrate (BHB) levels, ferric reducing ability of plasma (FRAP) and serum beta-carotene] and endocrine parameters [plasma thyroxine (T4), triiodothyronine (T3), insulin-like growth factor (IGF-I) and insulin levels] were measured. In summer, two cows were excluded from the study because of metritis and none of the remaining animals became pregnant, but 6 of the 8 cows had irregular luteal forms (ILF) on their ovaries. In winter, one cow was excluded because of metritis and 6 of the 9 cows became pregnant, while 2 of the 3 open cows had irregular luteal forms. In summer the mean plasma NEFA and BHB concentrations were significantly higher, while serum carotene and plasma IGF-I concentrations were significantly lower than in winter. The high plasma NEFA concentration found in summer seemed to be in association with the lower body condition score (BCS) caused by depressed appetite. In conclusion, statistical analysis supports the hypothesis that increased plasma NEFA and BHB and decreased plasma IGF-I concentrations may result in reduced fertility in summer. These changes may be associated with the more frequent appearance of ILFs and probably have a negative effect on ovarian function and/or oocyte quality.