Metabolomics reveals early pregnancy biomarkers in sows: a non-invasive diagnostic approach.

In an effort to enhance reproductive management and reduce non-productive periods in swine breeding, this study presents a novel, non-invasive metabolomics approach for the identification of early pregnancy biomarkers in sows. Utilizing an untargeted metabolomics approach with mass spectrometry analysis, we examined saliva samples from pregnant (n = 6) and non-pregnant control sows (n = 6, artificially inseminated with non-viable sperm). Our analysis revealed 286 differentially expressed metabolites, with 152 being up-regulated and 134 down-regulated in the pregnant group. Among these, three metabolites, namely Hyodeoxycholic acid, 2'-deoxyguanosine, and Thymidine, emerged as potential early pregnancy biomarkers. These biomarkers were further evaluated using targeted LC-MS/MS quantification and qualification, accompanied by ROC curve analysis. The study confirmed Hyodeoxycholic acid and 2'-deoxyguanosine as promising biomarkers for early pregnancy detection, offering potential for future implementation in swine production environments. This research establishes a robust theoretical foundation for the development of innovative molecular diagnostic techniques and explores new avenues for molecular genetic breeding and non-invasive diagnostics, ultimately enhancing fertility and productivity in sow herds.

Development of a fluorometric Cuboni test for the semi-quantitative measurement of urinary estrogen levels and pregnancy detection in mares.

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-17ß 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.

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.

A simple, inexpensive and portable on-farm test for pregnancy diagnosis and ovary status in cows via chemical analysis of urine.

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.

Interferon-Stimulated Gene Expression in Peripheral Blood Leucocytes as a Convenient Prediction Marker for Embryo Status in Embryo-Transferred Japanese Black Cows during the Peri-Implantation Period.

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.

Assessment of diagnostic accuracy and effectiveness of trans-abdominal real-time ultrasound imaging for pregnancy diagnosis in breeding sows under intensive management.

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.

Embryonic resorption rates at canine pregnancy diagnoses: A retrospective evaluation.

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.

Transcriptomic analysis reveals gene expression changes in peripheral white blood cells of cows after embryo transfer: Implications for pregnancy tolerance.

Most embryo losses occur in the first trimester of pregnancy in cows and include losses following embryo transfer. There is a resulting negative economic impact on cattle production systems when this occurs. Cellular and molecular mechanisms behind the maternal immune response to the growing embryo have not been fully characterized. The objective of this study was to examine the gene expression profiles of peripheral white blood cells (PWBCs) from pregnant cows 21 days after an embryo was transferred, and cows that were treated equally but lost the embryo. Specifically, we obtained and compared the transcriptome of PWBC from heifers that became pregnant at day 21 (N = 5) or failed to become pregnant after the embryo transfer (N = 5). Sequencing data can be accessed by Gene Expression Omnibus (GEO) with the accession number GSE210665. A total of 13,167 genes were evaluated for differential expression between groups. A total of 682 genes showed differential expression (p-value <.01), 302 genes were up-regulated while 380 were down-regulated due to pregnancy. The most significant genes were COL1A2, H2AC18, HTRA1, MMP14, CD5L, ADAMDEC1, MYO1A and RPL39, among others. Most of the significant genes are related to the up-regulation of inflammatory chemokine activity and immune defence response. Our findings extend the current knowledge that pregnancy alters the PWBC by promoting immune tolerance, cell chemotaxis, blood coagulation, angiogenesis, inflammatory response, cell adhesion and cytokine secretion. Our data suggest that pregnancy and ectoparasites could trigger poorly described genes in PWBC of cows, and a few previously escribed genes, such as IFI44. These results could shed light on the genes and mechanisms that promote tolerance to pregnancy and allow survival of the developing embryo.

An intelligent method for pregnancy diagnosis in breeding sows according to ultrasonography algorithms.

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.

Overview of ectopic pregnancy diagnosis, management, and innovation.

Ectopic pregnancies are the leading cause of maternal mortality in the first trimester, with an incidence of 5%-10% of all pregnancy-related deaths. Diagnosis of ectopic pregnancies is difficult due to clinical mimics and non-specific symptoms of abdominal pain and vaginal bleeding. The current standard for ectopic pregnancy diagnosis includes ultrasound imaging and ß-human chorionic gonadotropin (ß-hCG) monitoring. In addition to ß-hCG, serum markers are being explored as a potential for diagnosis, with activin-AB and pregnancy-associated plasma protein A specifically showing promise. Other diagnostic methods include endometrial sampling, with dilation and curettage showing the highest specificity; however, frozen section reduces the diagnostic timeline which may improve outcomes. Treatment options for confirmed ectopic pregnancies include medical, surgical, and expectant management. Chosen treatment methodology is based on ß-hCG levels, hematologic stability, and risk of ectopic pregnancy rupture. Current innovations in ectopic pregnancy management aim to preserve fertility and include laparoscopic partial tubal resection with end-to-end anastomosis and uterine artery embolization with intrauterine infusion of methotrexate. Psychological interventions to improve patient mental health surrounding ectopic pregnancy diagnosis and treatment are also valuable innovations. This literature review aims to bring light to current ectopic pregnancy diagnostics, treatments, and future directions.

Evaluation of RET-He values as an early indicator of iron deficiency anemia in pregnant women

Abstract Introduction During pregnancy, women are at an increased risk of developing iron-deficiency anemia. Objective The objective of this study was to assess the diagnostic performance of the reticulocyte hemoglobin equivalent (RET-He) in the early detection of iron-deficiency anemia in a group of pregnant women and to establish a reference range for this parameter in a group of control individuals. Method: A total of 60 patients and 130 control subjects were included in the study. Blood samples collected from the subjects were submitted to a complete blood count and a serum ferritin test and the data were analyzed by comparing the groups and ROC curves. Results The reference range found for the RET-He was between 29.75pg and 38.24pg, with a median of 35pg. The receiver operating characteristic (ROC) curve analysis for the ferritin parameter showed an area under the curve of 0.732 for the RET-He, 0.586 for hemoglobin, 0.551 for the mean corpuscular hemoglobin concentration and 0.482 for the mean corpuscular volume. Conclusion Early diagnosis of iron deficiency anemia in pregnancy is essential to prevent damage to both maternal and fetal health. The RET-He presents an excellent potential as an auxiliary tool for the diagnosis of iron deficiency in pregnant women.

Comparative evaluation of two commercial pregnancy-associated glycoproteins tests for early detection of pregnancy in dairy cattle.

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.

Comparison of ultrasonography, progesterone assay, ballottement, and non-return-to-heat with digital rectal palpation technique for early pregnancy diagnosis in Red Sokoto goat.

The objective of the study was to evaluate the effectiveness of the digital rectal palpation (DRP) technique for early pregnancy diagnosis and to compare the results of experiments I and II in the Red Sokoto goat. Experiment I had 68 goats with 52 does and 16 bucks. Does were divided into prostaglandin F2 -alpha (PGF2 α; n = 18), progesterone pessaries (P4 P; n = 18), and Control (n = 16) groups as oestrus synchronizaton methods. Bucks were used for oestrus detection (n = 6) and breeding (n = 10). Comparative pregnancy diagnosis was carried out on day 21 post-breeding using ultrasonography (US), DRP, progesterone assay (PA), ballottement (BL), and non-return-to-heat (NRH). Experiment II was a repeat but had 51 animals with 42 does and nine bucks; three bucks for oestrus detection and six for breeding. Does were divided into recto-vaginal artificial insemination (AI) by DRP (n = 14), vaginal speculum AI (n = 14), and Control-natural service (n = 14) groups, indicating breeding methods. Oestrus was synchronized with PGF2 α and pregnancy diagnosis was carried out on day 21 using US, DRP, PA, and NRH. The results for both experiments were similar. Pregnancy rates for PGF2 α group were 66.2%, 66.7%, 64.8%, 62.1%, and 63.0% for US, DRP, PA, BL, and NRH, respectively; P4 P had 81.5%, 81.5%, 42.6%, 20.3%, and 42.6% for US, DRP, PA, BL, and NRH, respectively; while Control was 73.6%, 79.2%, 70.9%, 73.6%, and 73.6% for US, DRP, PA, BL, and NRH, respectively. Proportions are significantly (p < .000) different in the P4 P group. It was concluded that the DRP technique was effective and consistent in early pregnancy diagnosis on day 21 and comparable to US, PA, BL, and non-return-to-oestrus in both experiments in Red Sokoto goat does.

Evaluation of RET-He values as an early indicator of iron deficiency anemia in pregnant women.

INTRODUCTION: During pregnancy, women are at an increased risk of developing iron-deficiency anemia. OBJECTIVE: The objective of this study was to assess the diagnostic performance of the reticulocyte hemoglobin equivalent (RET-He) in the early detection of iron-deficiency anemia in a group of pregnant women and to establish a reference range for this parameter in a group of control individuals. METHOD: A total of 60 patients and 130 control subjects were included in the study. Blood samples collected from the subjects were submitted to a complete blood count and a serum ferritin test and the data were analyzed by comparing the groups and ROC curves. RESULTS: The reference range found for the RET-He was between 29.75pg and 38.24pg, with a median of 35pg. The receiver operating characteristic (ROC) curve analysis for the ferritin parameter showed an area under the curve of 0.732 for the RET-He, 0.586 for hemoglobin, 0.551 for the mean corpuscular hemoglobin concentration and 0.482 for the mean corpuscular volume. CONCLUSION: Early diagnosis of iron deficiency anemia in pregnancy is essential to prevent damage to both maternal and fetal health. The RET-He presents an excellent potential as an auxiliary tool for the diagnosis of iron deficiency in pregnant women.

Patterns of Reproductive Management in Sheep and Goat Farms in Greece.

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.

Validation of pregnancy associated glycoproteins-based ELISA kits to determine early pregnancy status in lactating Nili-Ravi buffaloes.

The objective of the current study was to evaluate pregnancy-associated glycoproteins (PAGs) based enzyme-linked immunosorbent assay (ELISA) kits utilizing whole blood, serum, or milk samples for diagnosis of early pregnancy status in lactating Nili-Ravi buffaloes. Dairy buffaloes (n = 174) of mixed parity, 4-6 years of age, having mean (± standard deviation) days in milk 165 ± 87, and body condition score of 3.26 ± 0.34 were randomly enrolled in this study. Buffaloes were exposed to penile deviated bulls with 12 h interval for estrus detection during peak breeding season and eventually bred naturally at their respective standing estrus (day 0). Blood and milk samples were collected at days 24, 28, and 35 post-breeding to run a rapid visual pregnancy test® (RVPT) as a buffalo-side test or ELISA-based assay in the laboratory to detect early pregnancy status. Transrectal B-mode ultrasonography was performed to diagnose pregnancy at day 35 post-breeding and used as a gold standard to validate results of RVPT or ELISA-based tests. The RVPT is a visual readout test for pregnancy detection and had sensitivity (77.9 vs. 89.7 vs. 93.3%), specificity (77.9 vs. 89.7 vs. 93.3%), and accuracy (84.5 vs. 90.1 vs. 94.2%) at days 24, 28, and 35 post-breeding, respectively. The PAGs were assayed using ELISA kits in serum and had sensitivity (77.9 vs. 89.7 vs. 93.3%), specificity (84.2 vs. 87.7 vs. 93.9%), and accuracy (82.1 vs. 88.4 vs. 93.7%) at days 24, 28, and 35 post-breeding, respectively. Similarly, PAGs were also analysed using ELISA kits in milk samples and had sensitivity (77.6 vs 89.5 vs 95.0%), specificity (89.1 vs 91.9, vs 93.9%), and accuracy (85.1 vs 91.1 vs 94.3%) at days 24, 28, and 35 post-breeding, respectively. Overall, the Kappa values in this study exceeded 0.85 at day 35 post-breeding using RVPT or ELISA-based test kits in serum or milk samples, indicating a high level of agreement between PAGs detection method and gold standard for pregnancy diagnosis. The pregnancy outcomes based on ELISA-based PAGs detection at day ≥28 post-breeding had a high negative predictive value, indicating that the probability of incorrectly administering prostaglandins to pregnant buffaloes would be low if these tests were implemented on a commercial dairy herd. Taken together, it is concluded that PAGs-based determination of pregnancy using RVPT or ELISA in blood, serum, or milk samples can be used effectively for pregnancy diagnosis at ≥28 days post-breeding with more than 90% accuracy in Nili-Ravi buffaloes.

Advances in the ultrasound diagnosis in equine reproductive medicine: New approaches.

Ultrasound technology has led to new lines of research in equine reproduction, and it has helped to greatly improve clinical diagnosis and reproductive outcomes in equine practice. This review aims to discuss the potential clinical uses and new approaches of ultrasonography in equine reproduction. Doppler modalities are usually used to evaluate the vascularization of the follicles, corpus luteum (CL), and the uterus in the mare for diagnostic purposes. Inclusion of Doppler ultrasound in artificial insemination and embryo transfer programs could improve the reproductive outcome of these techniques. Better selection of recipients based on CL functionality, early pregnancy diagnosis 7-8 days postovulation of the donor before flushing or diagnosis of mares with endometritis with pathological increases of blood flow are examples of clinical applications in the mare. In the stallion, colour Doppler ultrasound has improved the diagnostic potential of B-mode ultrasound, improving the differential diagnosis of pathologies such as testicular torsion (decrease or absence of blood flow in the cord) and orchitis (increased blood flow in the cord). The incorporation of pulsed Doppler ultrasound into the reproductive evaluation of the stallion has enabled early identification of stallions with testicular dysfunction, thus allowing administration of timely treatment and subsequent improvements of the fertility prognosis for these animals. In addition, this technique has been used in the monitoring of patients undergoing medical and surgical treatments, thus verifying their efficacy. Recently, computer-assisted pixel analysis using specific software has been performed in research work in order to semi-quantitatively evaluate the vascularization (colour and power Doppler) and echotexture of different organs. These softwares are now being developed for clinical purposes, as is the case with Ecotext, a computer program developed for the evaluation of testicular echotexture, providing information on testicular functionality.

Selection of early pregnancy specific proteins and development a rapid immunochromatographic test strip in cows.

The most of embryo losses occur before the day 16 after artificial insemination, but there is no low cost and easy operation that can detect pregnancy with high accuracy within three weeks post-insemination in cattle. In this study, blood samples were collected at day 18 of the estrous cycle, and days 18, 25 and 35 of pregnancy, and relative levels of interferon stimulated genes (ISGs), Toll-like receptor (TLRs), complement components, early pregnancy factor (EPF) and pregnancy-associated plasma protein A (PAPPA) proteins were analyzed through Western blot. In addition, a colloidal gold immunochromatographic test strip was developed using the selected antibody, and the test was used for early pregnancy diagnosis. The results showed that there were changes in relative levels of plasma ISGs, TLRs, complement components, EPF and PAPPA proteins during early pregnancy in cattle, and complement component 1q (C1q) could be used as an ideal marker to develop a colloidal gold immunochromatographic test strip for early pregnancy diagnosis. In addition, the accuracy of pregnancy diagnosis by this test strip was 91.67% (11/12) for pregnant cows and 80% (8/10) nonpregnant cows at day 18 after insemination. In conclusion, the changes in plasma ISGs, TLRs, complement components, EPF and PAPPA proteins may be related to the maternal systemic immune modulation during early pregnancy, and a colloidal gold immunochromatographic test strip was developed for early pregnancy diagnosis using C1q as the ideal marker in cows. However, this colloidal gold immunochromatographic test strip needs further studies to improve the accuracy.

Economics of Rebreeding Nonpregnant Dairy Cows Diagnosed by Transrectal Ultrasonography on Day 25 after Artificial Insemination.

Pregnancy rates of Holstein cows showed a substantial decline in the past years, which caused intensive TAI programs for nonpregnant cows to shorten the period between unsuccessful insemination and the next attempt on the same cow. Although many studies examined the improvement in pregnancy rates following TAI, only a few examined the economic impact of such programs. In this study, we look at the feasibility of reproductive programs that included early pregnancy diagnosis performed by transrectal ultrasonography 25 days after artificial insemination (AI) and TAI of nonpregnant cows. This resulted in the following two TAI programs: a modified OvSynch program with a second PGF2α treatment at 24 h interval (GPPG, n = 100) and a modified OvSynch program with an intravaginal progesterone-release device inserted between days 0−7 (PRID + GPPG, n = 100). Cows included in the TAI programs recorded an improvement in the cumulative pregnancy rate (67% vs. 53%; 69% vs. 53%) compared to those in which this strategy was not applied (p < 0.05). An economic analysis was performed using a decision-support tool to estimate the net present value (NPV; USD/cow/year). The analysis revealed a difference in NPV of 89.6 USD/cow/year between the programs (rebreeding the nonpregnant cows following the TAI program vs. AI at detected estrus). In summary, rebreeding the nonpregnant cows after early negative pregnancy diagnosis (25 days after AI) using this strategy can improve the cumulative pregnancy rate and profitability of dairy farms.

Progress on Infrared Imaging Technology in Animal Production: A Review.

Infrared thermography (IRT) imaging technology, as a convenient, efficient, and contactless temperature measurement technology, has been widely applied to animal production. In this review, we systematically summarized the principles and influencing parameters of IRT imaging technology. In addition, we also summed up recent advances of IRT imaging technology in monitoring the temperature of animal surfaces and core anatomical areas, diagnosing early disease and inflammation, monitoring animal stress levels, identifying estrus and ovulation, and diagnosing pregnancy and animal welfare. Finally, we made prospective forecast for future research directions, offering more theoretical references for related research in this field.