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.
This study was aimed at the close monitoring of the serum PAGs profiles of Karya and Konya Merino sheep during early pregnancy.
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 After the sponges were withdrawn, the ewes were administered 400 IU of equine chorionic gonadotropin.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.
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.
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.

The aim of the present study was to develop a semi-quantitative urine pregnancy test for mares based on the Cuboni reaction and to verify the reliability of this test. The urine specimens were hydrolyzed by heating in the presence of hydrochloric acid. The resulting free estrogens were extracted from the urine matrix using toluene. Sulfuric acid was added to the toluene extract and the mixture was heated again. The lower layer in the test tube containing sulfuric acid was used for fluorescence measurements with excitation at 355 nm and measurement at 535 nm. The fluorometric Cuboni test revealed that the fluorescence counts in urine samples collected after the second trimester of gestation were significantly higher than those obtained from barren mares. The levels of estrogens, including equilin, estrone and estardiol-17β exhibited a dose-dependent increase in fluorescence counts, whereas other steroids, such as progesterone, testosterone, and cortisol, did not affect fluorescence. Heat treatment of urine samples with hydrochloric acid significantly increased the fluorescence counts in those collected after the second trimester of gestation compared to non-pregnant samples, implying the presence of large amounts of conjugated estrogens in pregnant mare urine. Fluorescence counts in urine samples obtained during pregnancy showed a positive relationship with estrone concentrations as measured by enzyme immunoassay. The results of the present study showed that the fluorometric Cuboni test facilitates urine fluorescence counts depending on the urinary estrogen content and is capable of discriminating between pregnancy and non-pregnancy states beyond the second trimester of gestation in mares.

Primiparous and multiparous dairy cattle were evaluated for pregnancy using both transrectal ultrasonography and a point-of-care pregnancy test (Alertys OnFarm Test), which measures pregnancy-associated glycoproteins through lateral diffusion, between Days 28-34 of pregnancy results were compared. A total of 637 animals were included in this study. Pregnancy was confirmed via manual palpation between Days 57-64. Data on parity, calving, and time of artificial insemination (AI) were also collected and evaluated. Overall the accuracy of the lateral diffusion test was 93.1% with 98.9% sensitivity, 88.7 % specificity, 86.8 % positive predictive value, and 99.1% negative predictive value. In heifers, the Alertys OnFarm Test had 100% sensitivity and 81.6% specificity. In contrast, the test had a sensitivity and specificity of 98.5 and 89.5%, respectively for multiparous cows. The pregnancy loss between early diagnosis and confirmation increased with parity. Heifers suffered losses as low as 2.6%, whereas animals in the third parity had significantly more losses (17.9%). Season also affected losses with spring pregnancy losses being considerably higher than autumn losses. The veterinary workload was also evaluated. Using the combined method of pregnancy testing, animals were first tested with the lateral diffusion test; then, the test-negative animals were scanned again, and the number of scanned animals was reduced. The results ranged between 3274% on farms. We concluded that veterinary workload could be reduced by using the point-of-care test. However, farm-level differences may have affected the results of the present study.

Ovary dysfunction causes an aberrant endocrine surge at various reproductive cycle stages, negatively impacting fertility and economic profit. Optimizing dairy cow performance requires determining ovarian status and detecting early pregnancy. Still, little to no information is available about the diagnosis of the ovarian condition using urine chemical analysis at the field level in Bangladesh. This study aimed to develop a simple, inexpensive and portable on-farm technique for pregnancy diagnosis and ovary status determination in cows via chemical urine analysis. Fifty reproductively healthy cows were recruited from different donor farms. Prior to artificial insemination (AI), all selected cows were placed in a single ovsynch program. TAI (timed artificial insemination) was carried out. Urine was routinely collected from Day 0-55 days at estrus cycle stages for routine chemical analysis using barium chloride (BaCl2), followed by commercially available protein strip tests. The developed techniques for pregnancy and ovary status diagnosis in cows were validated with rectal palpation (RP). Barium chloride (BaCl2) analysis of urine revealed white precipitation corresponding to a mature follicle in the ovary during estrus and colorless precipitation corresponding to the corpus luteum during the diestrus period. Positive pregnancy was indicated by the presence of a colorless precipitate in the BaCl2 test, and a protein value of less than 100 mg/dl was found in the protein strip test. The maximum accuracy (42/50, 84%) was observed between 25 and 35 days, as confirmed by RP. Perplexing results were seen 45-55 days after AI, between pregnancies and luteal cystic disease. In both cases, we discovered that the BaCl2 precipitation was colorless. However, the protein value in the context of luteal cystic disease was found to be higher than 100 mg/dl. The barium chloride test, followed by protein strip tests, is a simple and portable way to diagnose pregnancy and determine ovarian status in cows at the field level.

Pregnancy diagnosis during early gestation is important for cattle reproduction. The expression of interferon-stimulated genes (ISGs) in peripheral blood leukocytes (PBLs) was studied in embryo-transferred (ET) Japanese Black cattle. ISGs in PBLs-ISG15, MX1, MX2, and OAS1-were detected in multiple ovulation ET cattle using a real-time quantitative polymerase chain reaction, and receiver operating characteristic (ROC) curve analysis was performed. Gestational status was predicted using the average ISG levels during the normal estrous cycle (AVE) and the Youden index from the ROC curve analysis as cutoff values. The ISG15, MX1, and MX2 levels were significantly higher in pregnant cattle (n = 10) than in non-pregnant cattle (n = 23) on gestation day 21, whereas the levels of all ISGs were similar between non-pregnant and non-pregnant cattle with late embryonic death (n = 7). ISG15, MX1, and MX2 appropriately predicted the gestational status of ET cows. The statistical evaluation of the diagnostic accuracy in ET cows on day 21 of gestation presented higher values of sensitivity, specificity, accuracy, and positive predictive values of ISG15, MX1, and MX2 using the Youden index than using the AVE. Therefore, ISG15, MX1, and MX2 are excellent biomarkers of gestational status during the peri-implantation period in ET cattle.

Early and accurate determination of pregnancy is critical to optimum reproductive performance in pigs and enables farmers to early rebreed or cull non-pregnant animals. Most of the conventional diagnostic methods are unsuitable for systematic application under practical conditions. The advent of real-time ultrasonography has made it possible to establish relatively more reliable pregnancy diagnosis. The present study was carried out to evaluate the diagnostic accuracy and effectiveness of trans-abdominal real-time ultrasound (RTU) imaging vis-à-vis pregnancy status in sows reared under intensive management. Trans-abdominal ultrasonographic examinations were performed using a mechanical sector array transducer and portable ultrasound system in crossbred sows from 20 days post-insemination for up to next 40 days. Animals were followed up for subsequent reproductive performance with farrowing data used as the definitive test for deriving predictive values. Accuracy for diagnosis was determined by diagnostic accuracy measures like sensitivity, specificity, predictive values, and likelihood ratios. Before 30 days of breeding, RTU imaging had 84.21% sensitivity and 75% specificity. Relatively higher false diagnosis rates were obtained in animals checked at or before 55 days after AI than in animals checked after 55 days (21.73% versus 9.09%). Negative pregnancy rate was low with 29.16% (7/24) false positives. Overall sensitivity and specificity, using farrowing history as the gold standard, were 94.74% and 70.83% respectively. The sensitivity of testing tended to be slightly lower in sows with litter size of less than 8 total born piglets, compared to sows with 8 or more piglets. Overall positive likelihood ratio was 3.25 while negative likelihood ratio was 0.07. The results indicate that pregnancy in swine herds can be reliably detected earlier in gestation by 30 days post-insemination using trans-abdominal RTU imaging. This non-invasive technique with portable imaging system can be used as an integral part of reproductive monitoring and sound management practices for profitable swine production systems.

Pregnancy diagnosis in canines is generally performed during embryonic phase, between 19 and 35 days of gestation. At this stage embryonic resorptions can be observed, which, according to the literature, affects 11-26% of conceptuses and 5-43% of pregnancies. Resorption has been hypothesized as a physiological event in uterine overcrowding, however other factors may be involved, such as infectious or non-infectious diseases. This study aimed to retrospectively evaluate the incidence of embryo resorption at ultrasonographic pregnancy diagnosis in different dog breeds, and to identify the main factors determining the occurrence of the resorption sites. 95 pregnancy diagnoses were performed 21-30 days post-ovulation by ultrasound examination on 74 different animals. Breed, weight, and age of the bitches were recorded, and the reproductive anamnesis was collected from their medical records. The overall pregnancy rate was 91.6%. In 48.3% of pregnancies (42/87), at least one resorption site was visible, and embryonic resorption rate was 14.2% (61 resorption sites/431 total structures). Binary logistic regression showed a significant effect of age (P < 0.001), but not the size of the litter (P = 0.357), nor the size of the mother (P = 0.281) or any previous reproductive problems (P = 0.077). Age was significantly higher in pregnancies with resorptions than in normal ones (60.88 ± 18.24 and 40.27 ± 15.74 months, respectively, P < 0.001). The embryonic resorption rate was in line with previous findings, while the incidence of affected pregnancies was higher. Although resorptions may occur physiologically in pregnancies with large litters, a relationship between embryo resorption and litter size was not identified in our sample group, while aging increased the resorption rates. This, together with the occurrence of repeated embryonic resorptions in some bitches included in the study, suggests how resorptions could also be the result of pathological events. The underlying mechanisms and other factors that may be involved need further clarification.

Pig breeding management directly contributes to the profitability of pig farms, and pregnancy diagnosis is an important factor in breeding management. Therefore, the need to diagnose pregnancy in sows is emphasized, and various studies have been conducted in this area. We propose a computer-aided diagnosis system to assist livestock farmers to diagnose sow pregnancy through ultrasound. Methods for diagnosing pregnancy in sows through ultrasound include the Doppler method, which measures the heart rate and pulse status, and the echo method, which diagnoses by amplitude depth technique. We propose a method that uses deep learning algorithms on ultrasonography, which is part of the echo method. As deep learning-based classification algorithms, Inception-v4, Xception, and EfficientNetV2 were used and compared to find the optimal algorithm for pregnancy diagnosis in sows. Gaussian and speckle noises were added to the ultrasound images according to the characteristics of the ultrasonography, which is easily affected by noise from the surrounding environments. Both the original and noise added ultrasound images of sows were tested together to determine the suitability of the proposed method on farms. The pregnancy diagnosis performance on the original ultrasound images achieved 0.99 in accuracy in the highest case and on the ultrasound images with noises, the performance achieved 0.98 in accuracy. The diagnosis performance achieved 0.96 in accuracy even when the intensity of noise was strong, proving its robustness against noise.

The aim of this study was to determine and compare the diagnostic accuracy of two pregnancy-associated glycoproteins tests (IDEXX on-farm pregnancy test [OFPT] and IDEXX rapid visual pregnancy test [RVPT]) for early pregnancy diagnosis in dairy cattle. Blood samples were collected from Holstein cows (n = 317) by coccygeal venipuncture 28-31 days after artificial insemination (AI). The OFPT and RVPT were performed on the farm within 2 h after blood collection using whole blood or blood serum. Transrectal ultrasonography (USG) was performed for pregnancy diagnosis on day 32 post-AI as a gold standard. One-hundred fourteen cows were diagnosed as pregnant and 203 were determined to be nonpregnant. Furthermore, embryonic mortality was detected in four of the nonpregnant animals, based on fragmented/dispersed embryonic membranes and the absence of heartbeat. Sixteen and 14 false positive results (13 jointly for both tests) were obtained for the RVPT and OFPT, respectively. Three false negative results were obtained for the RVPT. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy for the OFPT were 100%, 93.1%, 89.1%, 100% and 95.6%, and for RVPT were 97.4%, 92.1%, 87.4%, 98.4% and 94.0%, respectively. The ability of both tests to distinguish between pregnant and open cows was very good (AUC of both tests above 0.9). The OFPT and RVPT almost perfectly agreed with each other. According to McNemar\'s analysis, false positive results have been caused difference between the two pregnancy tests and USG. In conclusion, both the RVPT and OFPT proved to be reliable and practical methods for pregnancy diagnosis 28-31 days after AI in dairy cows. However, the results of both pregnancy tests were affected by the occurrence of embryonic mortality around the time of their employment.

This paper reports findings regarding patterns of reproductive management in 325 sheep flocks and 119 goat herds throughout Greece. The objectives were to describe the patterns of reproductive management in sheep and goat farms, to highlight factors that were associated with these management patterns and the clinical application of the various reproductive management approaches in the farms and to establish potential associations with production outcomes. The median months of the start of the mating period for adult sheep and goats were May and June, respectively and for ewe lambs and replacement goats these were August and September, respectively. The median duration of the mating period was 2 months for adults and 1 month for replacements. The median ratio of female to male animals was 22:1 and 25:1 in sheep and goat farms, respectively. Control of reproduction was applied in 33.2% and 16.8% of sheep and goat farms, respectively, mostly by intravaginal progestagen sponges and less often by melatonin implants; multivariable analysis indicated five (availability of milking parlour, number of ewes in a farm, number of daily milkings, daily period spent by farmers at the farm, farming tradition in the family) and two (number of daily milkings, availability of milking parlour) predictors in sheep and goat farms, respectively. Pregnancy diagnosis ultrasonographically was performed in 36.6% and 16.8% of sheep and goat farms, respectively; multivariable analysis indicated as significant three (management system applied in farms, age of farmer, farming tradition in the family) and two (management system applied in farms, availability of milking parlour) predictors in sheep and goat farms, respectively. The control of reproduction and pregnancy diagnosis were associated with a higher number of vaccinations during pregnancy: 2.6 and 2.7, respectively, versus 2.2 and 2.1 in farms where they were not performed. The average per farm number of lambs/kids born per female animal was 1.33 ± 0.01 and 1.30 ± 0.02 for sheep and goat farms, respectively; five (control of reproduction, location of the farm, presence of working staff, collaboration with veterinary practice, breed of ewes) and two (control of reproduction, breed of female goats) predictors were identified in sheep and goat farms, respectively, for high numbers of lambs/kids. Lambs/kids were taken away from their dams at the age of 50 and 65 days, respectively. The average culling age (females, males) was 5.9 and 4.4 years and 6.9 and 4.9 years for sheep and goats, respectively. Farmers sourced animals to be used as replacements for those removed from the farm, mostly from the animals in their own farms, considering criteria related to milk production. Finally, the application of the control of reproduction was associated with increased milk production and the number of newborns, whilst an inverse correlation between the culling age of animals and milk production was seen.