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Pregnancy can be detected by several methods, including rectal palpation, measurement of progesterone hormone concentration, and ultrasound (USG). The aim of this study is to diagnose early pregnancy and monitor pregnancy development using ultrasound in Javanese Banteng (Bos javanicus). Three female Banteng from Taman Safari Indonesia, Cisarua, Bogor, Indonesia, were used for this study. Pregnancy diagnosis was performed 11, 18 and 20 days after insemination. Diagnosis was based on the presence of the amniotic sac, heart rate, and embryonic and fetal development. Embryonic and fetal development were monitored until day 60 of gestation, and placental development was monitored from day 60 to day 270. The results showed that on day 20 after insemination, a 0.33-cm chorioallantois membrane with a white spot inside was detected. On day 27, a 0.55 cm embryo and a 1.16 cm amniotic sac were detected. The size of the placenta of the three bantengs was 1.21 ± 0.3 cm on day 60 and continued to increase until it reached 5.57 ± 0.26 cm on day 270. This study concludes that it is possible to diagnose early pregnancy at day 20 after fertilization and placental development in Banteng. [ABSTRACT FROM AUTHOR]

Effective early pregnancy diagnosis is crucial for commercial rabbit breeding. Early pregnancy diagnosis enables the implementation of staged feeding for pregnant does, effectively preventing excessive weight gain and reducing the high mortality rates of kits during the birthing stage. This not only enhances production efficiency but also ensures the health and well-being of the breeding rabbits. The study introduces a method and device, the Rabbit Pregnancy Identification Device (RAPID), for detecting rabbit pregnancies using matrix optical sensing. RAPID comprises eight sensor modules and a central host unit. Each sensor module is equipped with three LEDs emitting light at wavelengths of 660 nm, 850 nm, and 940 nm, along with two photodiodes for data collection. A mobile application was developed to enable flexible control of the device. Signal-to-noise ratio tests were conducted to evaluate the stability of data collection by the device across varying light intensities. The experimental results reveal a direct correlation between light intensity levels and the signal-to-noise ratio of collected data. Notably, under a light intensity level of 4, RAPID achieves a signal-to-noise ratio ranging from 42 to 45 dB, satisfying the necessary criteria for data collection. Different classification models were trained using sample data from 216 does across various batches, and their generalization capabilities were evaluated. The experimental findings indicate that the optimal time for RAPID to diagnose the pregnancy status of does is on the 14th day after insemination, achieving an accuracy of 86.63 % and a recall of 80.49 %. Moreover, the model exhibits a degree of generalization, achieving an accuracy of 78.36 % when classifying another batch of sample data. RAPID achieves an accuracy of 97.25 % for pregnancy diagnosis of older does, which is 7.44 % higher than that of younger does; the accuracy rate for pregnancy diagnosis of does with sparse hair is 86.92 %, which is 4.78 % higher than that of does with dense hair. By comparing the effectiveness of using data from 8 sensor modules and data from a single sensor module for pregnancy diagnosis of different batches of does, it was found that the former exhibits more stable generalization capability in doe pregnancy detection.

This publication is directed to beef producers, farmworkers, and Extension agents with the purpose of providing information regarding current pregnancy diagnosis methods for beef cattle. Written by Maria Camila Lopez Duarte, Kalyn Waters, Nick Simmons, and Angela Gonella and published by the UF/IFAS Department of Animal Sciences, May 2024.

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.

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Abstract 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.

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.

Synchronization of estrus involves handling the estrus cycle so that a large number of cows can be bred at approximately the same time. Proper cow/heifer selection as well as efficient coordination are all key elements for the success of synchronization programs which were lacking in the study area. The study was conducted with the objective to evaluate the effect of Alfaglandin C and fixed factors on estrus synchronization and efficiency of Artificial Insemination (AI) in North Shoa zone of the Oromia region. The study population encompassed indigenous and crossbred cows and heifers found in six purposively selected kebeles (smallest administrative unit within a district) from the three selected districts (Wucale, Dagam and Kuyuu). Multistage sampling procedure was used to select target districts, kebeles and experimental cows/heifers. A total of 180 cows and heifers fulfilling estrus synchronization criteria were selected purposively and injected with synchronizing hormone (Alfaglandin C). The level of significance among categorical variables was determined using the chi-square (ω²) test. About 70.6% of the cows and heifers injected with Alfaglandin C exhibited estrus. Exhibition of estrus following hormone injection was significantly (p<0.05) lower in Wuchale district compared with that of the other two districts. Likewise, the agroecology, farming system, herd group, and breed had a significant (p<0.05) effect on estrus exhibition. Among cows and heifers inseminated following estrus exhibition, 62.2% conceived as confirmed by pregnancy diagnosis (PD) test conducted at 45-day post AI, which dropped to 58.3% as observed by the PD test conducted three months' post AI. Agroecology, farming system, herd group, and genotype of cows and heifers showed significant (p<0.05) influence on the conception rate (CR). The highest CR was observed among cows of Kuyuu district, which was significantly (p<0.05) different from that of the other two districts. The overall calving rate was 91.1%. The semi-intensive part of the study also showed better estrus synchronization response and better reproductive performances which directly associated with the management practices. Treating industrial byproducts, better management, and promoting AI can achieve better estrus synchronization and efficiency of artificial insemination. [ABSTRACT FROM AUTHOR]

Simple Summary: Practically and economically, predicting pregnancy status is crucial for food animal production. Recently, the expression of interferon-stimulated genes (ISGs) in peripheral blood leukocytes has been examined as a biomarker for predicting the gestational status during maternal recognition in ruminants. Although the ISG levels in the blood cells of cattle have been reported, the practical application of this detection method in embryo-transferred (ET) cows has not been evaluated in detail. The levels of three of the four ISGs in this study were significantly higher in pregnant cows than in non-pregnant cows on day 21 of gestation. The diagnostic performance results showed that the three ISGs are appropriate biomarkers for predicting the gestational status of ET cows. The statistical evaluation of diagnostic accuracy in ET cows indicated that the values of sensitivity, specificity, and accuracy were very high and the positive predictive values of the three ISGs were estimated. The results of this study suggest that ISGs are good biomarkers for gestational status during the peri-implantation period in ET cattle. 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. [ABSTRACT FROM AUTHOR]

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Auspicious pulse diagnosis/pregnancy diagnosis in traditional Chinese medicine involves such issues as medical skills, narrative skills, family decency, and ethics. It is an excellent case for the exploration of ethical dilemmas in traditional Chinese medical practice. The early classical medical texts such as Su Wen (Basic Questions) and Ling Shu Jing (Spiritual Pivot Canon) provide a principle-based ethical guide for doctor-patient communication, while popular fiction such as Hong Lou Meng (A Dream of Red Mansions), Yu Mu Xing Xin Bian (Stories: Entertain to Enlighten), and Feng Yue Meng (Courtesans and Opium) in the Ming and Qing dynasties present literary examples for solving ethical dilemmas. This article will analyze these texts from three perspectives. First, the doctors in the text were subject to gender order and other delicate etiquette and customs, therefore were unable to make the diagnosis without embarrassing the patients and jeopardizing family decency. Second, the narrator tends to attribute pregnancy misdiagnosis to three reasons: incomplete patient information, doctors' poor narrative competence, and doctors' corrupted medical ethics. Finally, the Ming-Qing fiction proposes three methods to solve this moral dilemma: clear pulse reading, tactful speech, and taboo challenging. This discussion of moral dilemmas in pregnancy diagnosis in traditional Chinese medical practice can be used as a reference for the localization of narrative medicine. [ABSTRACT FROM AUTHOR]

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.

Simple Summary: The paper presents practices related to reproductive management in sheep and goat farms in Greece. It presents descriptive information regarding the practices followed and the predictors associated with them. The results also indicate that various reproductive practices can have significant effects on the production outcomes (milk production, lamb production) in sheep and goat farms. The correct implementation of reproductive management in sheep and goat farms, based on scientific principles and compliance with established regulations, is important for the improvement in the welfare of farm animals and high production outcomes. 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. [ABSTRACT FROM AUTHOR]

The purpose of the present work was to study effectiveness for early pregnancy diagnosis in cattle of the new enzyme immunoassay (EIA) sandwich kit commercially available based on the measurement of pregnancy-associated glycoproteins (PAGs). 120 Holstein-Friesian cattle of mixed age and parity were comprised from different dairy herds. The pregnant females (n = 68) were diagnosed by ultrasonography at day 35-40 after artificial insemination and confirmed by transrectal exploration at 2-3 months after AI. The non-pregnant females (n = 52) were housed in the absence of males during the experimental period. Blood samples were collected from coccygeal vessels of females into EDTA tubes. The serum was obtained by centrifugation and the serum was stored at - 20 C until assay. The PAG concentrations in pregnant and non-pregnant females were determined in serum by EIA kit. The reproducibility inter- and intra-assay of the PAG-EIA is satisfactory (2.78 and 13.19 %, respectively). The accuracy ( 94.8 %) and the test of parallelism were largely acceptable. No cross-reaction was observed with the different hormones tested at different dilutions. PAG-EIA system gave 100 % sensitivity and negative predictive values. Whereas, specificity and positive predictive value were 91.93 and 71.15 %, respectively. The accuracy of pregnancy diagnosis by PAG-EIA was 87.5 %. In conclusion, the present study shows clearly that the EIA kit can be used to measure PAG in serum cows for the detection of gestation in Algeria. Therefore, this alternative technique could be recommended to replace the radioactive methods in immunoassays to improve the reproductive performances and an efficient tool for reproductive management of dairy cattle.

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.

The use of Doppler ultrasonography to quantify blood vascularization in reproductive organs has increased over the past decade. Doppler technology has predominantly been explored in research settings to evaluate uterine blood flow and to assess follicular and luteal blood perfusion. Recent research has also explored the use of Doppler technology in applied reproductive management for both the beef and dairy industries and has focused on the use of luteal color Doppler ultrasonography to evaluate embryo transfer recipients and perform early pregnancy diagnosis. Although significant progress has been made and current literature indicates a strong potential for the applied use of Doppler ultrasonography to increase reproductive efficiency in the cattle industry, uptake of this technology is still currently limited. This review summarizes the recent developments in the applied use of color Doppler ultrasonography for reproductive management in both beef and dairy cattle herds.

A pregnancy diagnosis is an important standard for control of livestock’s reproduction in paricular dairy cattle. High reproductive performance in dairy animals is a essential condition to realize of high life-time production. Pregnancy diagnosis is crucial to shortening the calving interval by enabling the farmer to identify open animals so as to treat or re-breed them at the earliest opportunity. MicroRNAs are short RNA molecules which are critically involved in regulating gene expression during both health and disease. This study is sought to establish the feasible of circulating miRNAs as biomarkers of early pregnancy in cattle. We applied Illumina small-RNA sequencing to profile miRNAs in plasma samples collected from 12 non-pregnant cows (“open” cows: samples were collected before insemination (non-pregnant state) and after pregnancy check at the indicated time points) on weeks 0, 4, 8, 12 and 16. Using small RNA sequencing we identified a total of 115 miRNAs that were differentially expressed weeks 16 relative to non-pregnancy (“open” cows). Weeks 8, 12 and 16 of pregnancy commonly showed a distinct increase in circulating levels of miR-221 and miR-320a. Through genome-wide analyses we have successfully profiled plasma miRNA populations associated with pregnancy in cattle. Their application in the field of reproductive biology has opened up opportunities for research communities to look for pregnancy biomarker molecules in dairy cattle.

Objective This study was conducted to assess the level of pregnancy-associated glycoprotein (PAG) in whole and skim milk samples, and its suitability for early pregnancy diagnosis in goats. Methods A two-step sandwich enzyme-linked immunosorbent assay (ELISA) system for estimation of milk PAG was developed and validated, which employed caprine-PAG specific polyclonal antisera. Whole and skim milk samples (n = 210 each) from fifteen multiparous goats were collected on alternate days from d 10 to d 30, and thereafter weekly till d 51 post-mating. PAG levels in milk samples were estimated by ELISA and the pregnancies were confirmed at d40 post-mating by transrectal ultrasonography (TRUS). Results The level of PAG in whole and skim milk samples of both pregnant and non-pregnant goats remained below the threshold values until d 24 after mating. Thereafter, PAG concentration in whole and skim milk increased steadily in pregnant goats, whereas it continued below the threshold in non-pregnant does. The PAG profiles in whole and skim milk of pregnant goats were almost similar and exhibited strong positive relationship (r = 0.891; p

The present study was conducted in a goat farm for pregnancy diagnosis by using vaginal cytology and B-mode real time ultrasound using 5 MHz probe by transabdominal method. Seventeen pregnant does were used for this study. The objective of the study was to determine the earliest day of pregnancy and describe the chronological characteristics of pregnancy from day 22 to day 40 for vaginal cytology and day 25 to day 60 for ultrasonography of gestation. The differences among the average percentage of cell value in different age of pregnancy were significant (p < 0.05). The average percentage of intermediate cells (81.12%) was also significantly (p < 0.05) higher than superficial (9.41%), keratinized (7.10%) and neutrophil (2.61%) on 22-40th days of pregnancy. In case of real time B-mode ultrasonography, the gestational sac was observed only in three does out of seventeen (17.6 %) at 25-30 days whereas the placentomes and heart beat of the foetus were first detected at 31-35 days in six does (35.3%). The foetal leg buds were first visualized at 36-40 days in four does (23.5%) whereas the foetal vertebral column was first observed at 36-40 days of gestation in only three does (17.6%). In conclusion, vaginal cytology and trans-abdominal ultrasonography can be used for detection of early pregnancy in does.

Early detection of pregnancy is vital for appropriate reproductive management programmes to facilitate the rapid re-insemination of non-pregnant females and reduce the calving interval.

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.

Pregnancy diagnosis is critical to the management of captive breeding of wild animals, as accurate pregnancy diagnosis can improve the reproductive efficiency of captive animals. Pangolins do not show obvious morphological changes during pregnancy, which increases the difficulty of pregnancy diagnosis. In this study, pregnancies were diagnosed using B-ultrasonography in three Sunda pangolins (MJ-X2, MJ-X3, and MJ-X4) that had mated with males. B-ultrasonography revealed embryos in pangolins MJX2 and MJ-X3, but not in MJ-X4. Pangolins MJ-X2 and MJ-X3 gave birth to cubs 102 and 123 days, respectively, after B-ultrasound detection; pangolin MJ-X4 did not give birth to a cub. This study showed that B-ultrasound can accurately diagnose pregnancy in Sunda pangolins and detect the developmental status of their embryos and fetuses. As a method of pregnancy diagnosis, B-ultrasound is not only easy to operate, safe, and noninvasive, but also very intuitive and efficient compared with hormone detection methods. Efficient detection methods such as B-ultrasonography are critical to avoid housing alreadypregnant pangolins together with males, which may mate with the females again or stress them, resulting in abortion. This approach also helps to conserve male reproductive resources, thereby improving the reproductive efficiency of captive Sunda pangolins. Effective and efficient captive breeding programs are critical for the conservation of Sunda pangolins and other pangolin species. [ABSTRACT FROM AUTHOR]

The main objective of present study was to estimate gestational age of teddy goat by B-mode ultrasound measurement of embryonic or fetal parts and uterine diameter throughout pregnancy by transrectal approach. Three parameters crown rump length (CRL), trunk diameter (TD), and uterine diameter (UD) were selected for measurement of gestational age with the weekly interval from 3rd week to 15 weeks of gestation of 12 pregnant teddy goats, all three parameters were significantly (P<0.01) corelated with gestational age. The CRL was strongly positively correlated with gestational age (r2=0.98) followed by the TD (r2=0.98) and the UD (r2=0.97). The fetal numbers were estimated 19 (five single and seven twines) by transrectal approach in twelve pregnant teddy goats. The earliest day of pregnancy diagnosis was 19 day after mating, while trunk was best parameter for determination of gestational age throughout gestational period and fetal sexing can be diagnosed at 7th week of gestation period in teddy goats. [ABSTRACT FROM AUTHOR]

Objective: This study was conducted to assess the level of pregnancy-associated glycoprotein (PAG) in whole and skim milk samples, and its suitability for early pregnancy diagnosis in goats. Methods: A two-step sandwich enzyme-linked immunosorbent assay (ELISA) system for estimation of milk PAG was developed and validated, which employed caprine-PAG specific polyclonal antisera. Whole and skim milk samples (n = 210 each) from fifteen multiparous goats were collected on alternate days from d 10 to d 30, and thereafter weekly till d 51 postmating. PAG levels in milk samples were estimated by ELISA and the pregnancies were confirmed at d40 post-mating by transrectal ultrasonography (TRUS). Results: The level of PAG in whole and skim milk samples of both pregnant and nonpregnant goats remained below the threshold values until d 24 after mating. Thereafter, PAG concentration in whole and skim milk increased steadily in pregnant goats, whereas it continued below the threshold in non-pregnant does. The PAG profiles in whole and skim milk of pregnant goats were almost similar and exhibited strong positive relationship (r = 0.891; p<0.001). Day 26 post-mating was identified as the first time-point for significantly (p<0.05) higher milk PAG concentration in pregnant goats than to non-pregnant goats. When compared to TRUS examination for pregnancy diagnosis, the accuracy and specificity of PAG ELISA using whole and skim milk samples were 94.5% and 95.4%; and 95.3% and 100%, respectively. The high values of area-under-curve (0.904 [whole milk] and 0.922 [skim milk]), demonstrate outstanding discrimination ability of the milk assays. Among the sampling dates chosen, d 37 post-mating was identified as the best suitable time point for collection of milk samples to detect pregnancy in goats. Conclusion: The PAG concentration in whole and skim milk of goats collected between days 26 and 51 post-breeding can be used for the accurate prediction of pregnancy and may be useful for assisting management decisions in goat flocks. [ABSTRACT FROM AUTHOR]

The present study was conducted to assess the information needs of the paravets on Artificial Insemination (AI) in India. Paravets (160) were selected having equal representation from 4 different parts of India (Gujarat, Rajasthan, Telangana and Odisha). Questionnaire and telephonic survey method was used for data collection. The information needs on AI among paravets was assessed under four sub-groups, viz. information needs on heat detection, AI techniques, post-AI practices and miscellaneous information related to AI. Results revealed that the most needed information was symptoms of heat in buffaloes and pregnancy diagnosis both bearing 1st rank (MWS=7.22), followed by address of semen stations (MWS=6.09) with 2nd rank, rectal palpation (MWS=5.97) ranked 3rd, signs of heat in cattle (MWS=5.91) ranked 4th and use of crystoscope for checking fern pattern (MWS=5.68) with 5th rank. Earlier researchers have established that proper AI technique is essential for improving conception rate. Hence, along with provision of training courses on AI, there is a need to focus on creating efficient information systems platform incorporating knowledge related to AI which can be easily accessed by the paravets. Further, the paravets need to be trained on the use of digital tools for faster information delivery and ease of use. Efforts are also needed for continuous education of the paravets for enhancing the effectiveness of AI. Convergence of various information dissemination efforts of the government and non-government sector, and more number of multipronged information channels are required to fulfil the information needs of the paravets.

Dairy cow farming plays an important role in the UK and worldwide economies. Significant challenges are currently being faced regarding sustainability of the dairy industry. Dairy cow subfertility remains an important issue limiting herd productivity, resulting in annual losses of hundreds of millions of pounds in the UK alone. To address this, accurate monitoring of reproductive status and early detection of fertility issues in individual cows is essential. The aim of this study was to gather farmer and veterinarian opinions on current practices and perceived gaps related to diagnosis of fertility issues and pregnancy testing in UK dairy farms. Using online questionnaires, data were collected and analyzed from a total of 40 farmers and 59 veterinarians. The results showed that non-seen bulling checks and ultrasound were the most frequent tools to detect fertility issues, and that most farmers tested post-calving, and often again before or during mating. Most farmers believed that current tests did not meet their expectations, with half of those being willing to pay more than they were currently paying for fertility testing. In regard to pregnancy testing, ultrasound was most commonly used, at 30–50 days post-insemination either in individual or groups of cows. Again, most farmers believed that current tests did not meet their expectations, and a majority would consider paying a higher cost for a test that was better than those currently available. In addition, a majority of farmers would consider using a test that could detect pregnancy within 2 weeks post-insemination, if such test existed, because they believed it would help improve their herds' reproductive performance. Overall, the opinions of farmers and veterinarians indicate that there is significant scope for improving dairy herd fertility monitoring practices in the UK, through development of improved assays that can diagnose pregnancy and infertility earlier, are less disruptive to farm operations and are more cost effective than available tools. They also provide useful information to guide the future development and implementation of better diagnostics for improving reproductive performance of dairy cattle.

The present study aimed to assess the applicability of luteal blood flow data acquired through the use of color Doppler ultrasonography and a post-processing analysis tool (ImageJ) for predicting pregnancy in buffaloes (Bubalus bubalis). The experiment was carried out on 59 multiparous Italian Mediterranean buffaloes that underwent synchronization of estrus and fixed-time artificial insemination (TAI). Corpus luteum features (size: CLS and blood flow: BFA) were taken from Day 5 to 10 after TAI and retrospectively measured with ImageJ. In the same period, blood samples were taken to assess progesterone (P4) concentrations. Pregnancy diagnosis was carried out on Day 45 by ultrasound and confirmed on Day 70 post-TAI. Differences in CLS, BFA, and P4 concentrations from Day 5 to 10 after TAI measured between groups were analyzed by ANOVA repeated measures as were differences within each day of measuring. Buffaloes that established a pregnancy (n = 29; 55%) had larger CLS (2.2 ± 0.1 vs. 1.9 ± 0.1 cm2; P < 0.01), higher BFA (0.6 ± 0.0 vs. 0.4 ± 0.0 cm2; P < 0.01), and higher P4 blood level (1.8 ± 0.1 vs. 1.4 ± 0.1; P < 0.01) during Day 5–10 as compared to not-pregnant buffaloes (n = 22). Throughout the entire period, the first feature that changed between groups was P4 blood concentration at Day 7 (1.7 ± 0.1 vs. 1.2 ± 0.1; P < 0.05) followed by BFA at Day 8 (0.6 ± 0.0 vs. 0.5 ± 0.0; P < 0.05), respectively, in pregnant and not-pregnant animals. The ROC analyses indicated that P4 was able to predict pregnancy since Day 5 (P < 0.05) although a more reliable result could be obtained from Day 8 (P < 0.01). At Day 10, it was possible to set a cutoff value for every parameter taken into account. The logistic regression analysis showed that pregnancy was positively influenced by P4 concentration (odds ratio 534.127; P < 0.01) and BFA (odds ratio 744.893; P < 0.01). In conclusion, the use of color Doppler ultrasonography, together with ImageJ, identified different patterns of BFA between pregnant and not-pregnant buffaloes starting from Day 8 post-TAI.

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.

The aim of this study was to compare the accuracy of bovine pregnancy-associated glycoproteins (bPAG) and bovine pregnancy-specific protein B (bPSP-B) tests with transrectal ultrasonography (TRUS) for early pregnancy diagnosis in dairy cows. The study was carried out on 58 Holstein cows. Blood and milk samples were collected on days 20, 25, 28, 30, 32 and 40 following artificial insemination (AI). TRUS was performed on all cows on days 30 and 40 following AI. The positive predictive values for bPAG-serum and bPAG-milk tests were 100% on days 28 and 30. For bPSP-B tests, positive predictive values were 93.7% and 100% on days 28 and 30, respectively. The Kappa test revealed a high agreement between TRUS and the three tests on days 28 and 30 with the values of 0.954 and 0.946 (P

The bovine pregnancy-associated glycoproteins (bPAGs) have been widely used as robust markers for early diagnosis of pregnancy in the cattle. The current immune recognition methods for detecting bPAGs are limited and, to a certain extent, are associated with high costs and poor stability of the antibody. Aptamers that are more stable and easily synthesized than antibodies might serve as suitable candidates for the development of rapid detection methods. This paper describes selection and characterization of bPAG4 aptamers and theirs applicability to detect bPAG4 in the serum. In this work, the recombinant bovine pregnancy-associated glycoproteins 4 (bPAG4) with a relative molecular mass of about 48 kDa was successfully expressed in human embryonic kidney 293 (HEK 293) cells. Subsequently, the ssDNA aptamers were selected by systematic evolution of ligands by exponential enrichment (SELEX) using magnetic beads (MB) coated with bPAG4 as target. After 9 rounds of selection, three aptamers with high affinity to bPAG4 (Kd = 11.7~40.2 nM) were identified. The selected aptamers were successfully used in enzyme-linked aptamer assay (ELAA) to detect bPAG4 at a detection limit of 0.09 ng/mL. Meanwhile, it has been successfully applied for the detection of bPAG4 in serum samples. This work demonstrated that the selected aptamers could be used as promising affinity probes in the development of inexpensive, simple, and sensitive analysis methods for detecting bPAGs. [ABSTRACT FROM AUTHOR]

In the current dairy industry, an intensive demand for estrus detection and early diagnosis of pregnancy has been increasing. Progesterone concentration in bovine milk is used as an important indicator of estrus detection and early pregnancy diagnosis. Current method for milk progesterone determination is destructive in nature. In contrast, near-infrared spectroscopy (NIRS) is a non-destructive analytical method that can be used for milk quality determination but there has been limited study on using NIRS for milk progesterone concentration during milking. Thus, the objective of this study was to develop an online real-time NIR spectroscopic sensing system for milk progesterone determination during milking by using a specific enzyme immunosorbent assay as a reference (chemical) method. Milk spectra with a wavelength range of 700 to 1050 nm and milk samples were collected every 20 s during milking from four lactating Holstein cows for 28 days using the NIR spectroscopic sensing system. Calibration models were developed using partial least squares analytical method and the precision and accuracy of the models was validated. Milk progesterone concentration for each milking was calculated by taking the progesterone concentration of the milk predicted values and milk yield obtained every 20 s, and was compared with the milk progesterone concentration chemical analysis value for one milking (bucket milk). The results obtained show that one milking time measurement accuracy of progesterone concentration was reasonably good. By installing the NIR spectroscopic sensing system developed in this study into a milking robot, it could predict milk progesterone concentration for one milking with almost the same accuracy as chemical analysis. Thus, recording this predicted value every milking and monitoring the continuous transition of the milk progesterone concentrations, it becomes possible to use it for the detection of estrus status and for the diagnosis of pregnancy of each cow. [ABSTRACT FROM AUTHOR]

The B-mode and color Doppler ultrasonography provide a safe, non-invasive, real-time approach to assess reproductive tissues and organs in female cattle, and thus have a number of potential uses in reproductive management. On the ovary, antral follicle population can be counted, and follicular growth, regression and ovulation can be monitored during sequential exams. The corpus luteum can be identified, and evaluated according to size, echotexture, and vascularization. Ultrasonography allows an early diagnosis of pregnancy based in the detection of the embryonic vesicle from day 24 of gestation. Alternatively, non-pregnancy can be indirectly inferred according to corpus luteum vascularization 20 days after insemination. Embryo and fetus morphology and vital signs can monitored throughout gestation, and fetal sex can be determined by the visualization of the relative position of the genital tubercle. In addition, a range of pathologic conditions of the ovary and uterus can be diagnosed based on sonographic images. The use of ultrasonography was also key for the development of many assisted reproductive technologies, such as timed artificial insemination, modern superovulation protocols, procedures for the recovery of cumulus-oocyte complexes aiming in vitro embryo production, and is a valuable tool for the selection and management of embryo recipients. In the future, emerging technologies such as computer-assisted image analysis, 3D ultrasonography, and 3D modeling have the potential to improve the accuracy of sonographic diagnosis, or even create new potential uses for ultrasonography in animal reproduction.

This research aimed to diagnosed pregnancy on mares using a test mare (MTP) kit. This study using 10 mares were used to confirm their pregnancy status using a rectal exploration technique (gold standard) using seven pregnant mares (K1) and three non-pregnant mares (K2). Serums from both groups were tested using a MTP kit. The research data were tested descriptive. The results showed that seven mares that were positively pregnant and three negative mares unpregnant after being tested using the MTP kit and rectal exploration. The conclusion that the MTP kit can be used to diagnose pregnancy in mares of 60 to 100 pregnancy days which is 100% accuracy sensitivity and specificity. Â

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.

Early diagnosis of pregnancy is important in livestock production, but there is no reliable technology used for pregnancy diagnosis within the first three weeks after insemination. During early pregnancy, the expression of interferon-stimulating genes (ISGs) in peripheral blood leukocytes (PBL) is significantly increased. However, due to different strains, detection sample types, detection methods, threshold value, etc. the specific effectiveness of early pregnancy diagnosis using ISGs is worth further study. The purpose of this study was to test interferon-stimulated protein 15 (ISG15), 2'–5'-oligoadenylate synthetase 1 (OAS1) and radical S-adenosyl methionine domain containing 2 (RSAD2) for early pregnancy diagnosis in dairy cows. The expression of ISG15, OAS1, and RSAD2 in PBL of pregnant and non-pregnant heifers on days 0, 14, 18, 21 and 28 after artificial insemination (AI) was detected by fluorescence quantitative polymerase chain reaction (PCR). The sensitivity and specificity of the pregnancy diagnosis was analyzed using expression of these three genes separately or in combination by receiver operating characteristic curve. The combination with the highest accuracy used probe primers and duplex fluorescence quantitative PCR. The single quantitative PCR results showed that expression of ISG15, OAS1 and RSAD2 on day 18 after AI was significantly higher in pregnant than in non-pregnant cows. When these three genes were used separately, or in combination, for early pregnancy diagnosis, the sensitivity for the RSAD2 gene was 100%, and the combination of ISG15 with RSAD2 was 94.7%. The duplex quantitative PCR showed that, although the sensitivity of ISG15 alone was 100%, its specificity was only 88.2% (cut-off value 1.402). The sensitivity of RSAD2 alone was 89.5%, and the specificity was 88.2%; however, when the two genes were used in combination, the sensitivity, specificity and diagnostic cut-off value were consistent with the results of single quantitative PCR. These results indicated that a duplex quantitative PCR assay system for early pregnancy diagnosis in cows using ISG15 and RSAD2 was established. Simultaneous detection of expression of ISG15 and RSAD2 by duplex quantitative PCR can effectively improve the diagnostic accuracy for dairy cows. [ABSTRACT FROM AUTHOR]

The main objective of this study was to assess PAG2 mRNA expression in maternal blood cells at the peri-implantation period in water buffalo; moreover, we wanted to evaluate the earliest time in which PAG-2 could be detected in maternal blood. Thirty-two lactating buffaloes artificially inseminated (AI) were utilized. Blood was collected at Days 0, 14, 18, 28, 40 after AI (AI = day 0). Pregnancy was diagnosed by ultrasound at Days 28 and 40 post AI. Out of 32 buffaloes, 14 were pregnant (P group) and 18 were not pregnant (NP group). The plasma PAG-2 threshold of 1.0 ng/mL in the P group was reached at day 40 post AI. PAG2 mRNA expression differed between the P and NP groups, and was either evaluated in Peripheral Blood Mononuclear Cells (PBMC) or Polymorphonuclear Leukocytes (PMN), starting from day 14. However, both the estimated marginal means and multiple comparisons showed that PAG2 mRNA expression was higher in PMN than PBMC. In the present study, PAG-2 appeared in the blood (40 Days post AI), and an early expression of PAG2 mRNA at Day 14 post AI was also observed. Although further research is undoubtedly required, PAG2 mRNA in peripheral blood leukocytes could be using to better understand the role that PAGs play during pregnancy in buffalo. [ABSTRACT FROM AUTHOR]

The current study aimed to investigate the possibility of using ultrasonography through abdominal probe in recording percentages of pregnancy, twins and genital pathological condition which affect the uterus for cases admitted to veterinary hospital in Kirkuk province as well as regulated visits to breeding projects of small ruminant and college of agriculture and forestry / Kirkuk university for the period from 01/10/2015 to 01/04/2016, which included 281 of ewes and does. The result of current study showed that that ratio of pregnancy was 75.9% in ewes and in does 47.6% while in case of twins in ewes 19.0% and in does 38.1%., while the genital pathological cases like endometritis and mucometritis (after birth) ratio in ewes 2.5% and in does 9.5%, in cases of pyometra in ewes 1.3% and in does 0.0%, while cases of embryonic deaths ratio in ewes 1.3% and in does 4.8%. It was concluded from the current study that; the possibility of diagnosis the genital pathological cases which affect the uterus as well as the identification of the single and twin pregnancy with investigation of the viability of embryos in both ewes and does by using ultrasonography with abdominal probe.

We investigated the diagnostic reliability of pregnancy detection using changes in interferon stimulated gene (ISG) messenger RNA (mRNA) levels in circulating immune cells in ewes. Two different groups of ewes (an experimental group, experiment 1 and a farm group, experiment 2) were oestrus-synchronized and blood sampled on day 14 (D0=day of insemination in control animals, experiment 1) and day 15 (experiment 2). Real-time PCR were performed to evaluate the abundance of different ISG mRNAs. In the experimental group, peripheral blood mononuclear cells of 29 ewes born and bred in experimental facilities were isolated using a Percoll gradient method. Gene expression for Chemokine (C-X-C motif) ligand 10 (CXCL10), Myxovirus (influenza virus) resistance 1 (MX1) and Signal transducer and activator of transcription 1 (STAT1) mRNA were, respectively, 8.3-fold, 6.1-fold and 2.7-fold higher (P0.10) in CXCL10, STAT1, MX1, Myxovirus (influenza virus) resistance 2 (MX2) and ISG15 ubiquitin-like modifier (ISG15) mRNA expression were found between pregnant and non-pregnant ewes. The ROC curves and the hierarchical classification generated from the real-time PCR data failed to discriminate between pregnant and non-pregnant animals. In this group of animals, our results show a strong variability in ISG expression patterns: 17% of animals identified as non-pregnant by the five tests were in fact pregnant, only 52% of pregnant animals had at least two positive results (two genes above threshold), whereas up to five positive results (five genes above threshold) were needed to avoid misclassification. In conclusion, this study illustrates the high variability in ISG expression levels in immune circulating cells during early pregnancy and, therefore, highlights the limits of using ISG expression levels in blood samples, collected on PAXgene® tubes on farms, for early pregnancy detection in sheep.

Ultrasonography is commonly used by producers to determine pregnancy in their flocks (Scott 2012; Jones et al., 2016). Transabdominal ultrasonography is a simple, non-invasive method; however, it is not frequently used for early pregnancy detection in sheep. The aims of the current study were to determine whether pregnancy detection through transabdominal ultrasound is viable as early as day 25 of gestation and if a single measure of the embryonic vesicle could predict gestational age. Transabdominal ultrasound was performed on a total of 11 ewes at days 25, 27, 29, 31, 33 and 60 of gestation. Pregnancy diagnosis and accuracy of pregnancy detection were determined, and ultrasonic measurement of embryonic vesicle depth (EVD) was recorded for each ultrasound time period. Pregnancy may be determined at day 25 (accuracy = 45%); however, accuracy of pregnancy detection increased until day 31 (100%). A significant correlation between gestational age and EVD was identified (P < 0.001; r = 0.791). A regression analysis of gestational age (days since conception) on EVD resulted in a significant correlation of determination of 0.675 (P ≤ 0.001). This resulted in a quadratic predictive equation to estimate gestational age using EVD (y = 9.32 + 9.10 * EVD - 0.38 * EVD2). The results of the present study show that early pregnancy detection is possible in sheep; however, accuracy of pregnancy detection is reduced prior to day 31. Embryonic vesicle depth was able to predict gestational age with a simple quadratic equation. The number of fetuses present during ultrasonography was not counted in the present pilot study and may impact EVD and thus the estimation of gestational age. Future work in this area is warranted to investigate embryonic vesicle depth and the number of fetuses present during ultrasonography to predict gestational age. [ABSTRACT FROM AUTHOR]

An experiment was carried out to evaluate the effect of recipients physiological condition on pregnancy rate and subsequent kidding in does. Oestrus was synchronised by inserting controlled internal drug release device (CIDR, 0.33 g natural hormone progesterone; EAZI-BREED CIDR, Pharmacia & Upjtohn Limited, NZ) and an i.m. injection of 300 IU equine chorionic gonadotrophin (eCG) (Folligon®; Intervet International B.V., EU). A total of 31 Boer crossbred (Boer × local) recipient does were divided into two treatment groups namely intact group (15 does) and used group (63 does). Intact group may be defined as those goats that had not used for any surgery, while used group may be defined as those goats that had been used for surgery such as laparoscopic ovum pick up, embryo transfer. On Day 7 after CIDR removal, ovaries were checked for corpus luteum (CL) and recipient does with functional CL were only used as recipients. After flushing, recovered embryos were examined under a stereomicroscope and only transferrable embryos (morula and blastocyst) were transferred. Uterine horn was exteriorised over the surface of abdomen with the help of a grasping forceps and a surgical blade. One to 3 fresh embryos along with a small amount of flushing medium were transferred into the lumen of the anterior part of the uterine horn. Once completed, the uterine horn was placed back into the peritoneal cavity and the incision was sutured. Pregnancy diagnosis was performed at around 30 to 35 days after embryo transfer by using a real-time ultrasound scanner. The average numbers of embryos transferred per recipient doe were 2.60 and 2.56 for intact and used groups, respectively. The percentage of pregnancy was 26.67 for intact group. No pregnancy was recorded in used group. Average embryo survival rate was calculated based on the number of kids born, which was 7.69. Average gestation length was 148 days. From the findings obtained in this research, it could be concluded that intact does were favourable as recipient in ET programme than repeated surgery does. [ABSTRACT FROM AUTHOR]

The aim of this study was to compare the accuracy of bovine pregnancy-associated glycoproteins (bPAG) and bovine pregnancy-specific protein B (bPSP-B) tests with transrectal ultrasonography (TRUS) for early pregnancy diagnosis in dairy cows. The study was carried out on 58 Holstein cows. Blood and milk samples were collected on days 20, 25, 28, 30, 32 and 40 following artificial insemination (AI). TRUS was performed on all cows on days 30 and 40 following AI. The positive predictive values for bPAG-serum and bPAG-milk tests were 100% on days 28 and 30. For bPSP-B tests, positive predictive values were 93.7% and 100% on days 28 and 30, respectively. The Kappa test revealed a high agreement between TRUS and the three tests on days 28 and 30 with the values of 0.954 and 0.946 (P<0.001), respectively. Specificity was found to be 100% for bPAG-serum and bPAG-milk, and 96.7% for bPSP-B on day 28. Sensitivity values were calculated as 94.1% and 93.8% for bPAG-serum and bPSP-B, respectively. The area under ROC curve was between 0.967 and 0.991 for on day 28 following AI. In conclusion, all three tests were found to be reliable methods for pregnancy diagnosis in cows on days 28 and 30 following AI. However, test results may give false positive values in case of embryonic deaths. [ABSTRACT FROM AUTHOR]

Abstract: The present study was conducted to determine the difference in plasma prostaglandin F2α metabolite concentrations following oxytocin (OT) challenge between pregnant and non‐pregnant cows. Experiment 1: cows were subjected to the OT challenge test on days 12, 14 or 16 (day of estrus = day 0) with or without prior insemination and plasma 13,14‐dihydro‐15‐keto prostaglandin F2α (PGFM) concentrations were measured from −30 to 180 min after OT injection. On day 16, the increment of plasma PGFM concentrations in response to OT injection was significantly smaller in pregnant than that in cyclic cows. On days 12 and 14, there was little OT‐induced PGFM secretion and no difference in PGFM increase between the pregnant and cyclic cows. Experiment 2: cows were inseminated on day 0 and subjected to the OT challenge test on day 16. Cows were classified into non‐pregnant/early embryonic death (NP/EED), late embryonic death (LED) and pregnant (PREG) groups. The increment of PGFM concentrations in response to OT injection was less in both PREG and LED groups than that in the NP/EED group. In conclusion, plasma PGFM secretion induced by OT is suggested as the base of pregnancy diagnosis prior to returning estrus in cows. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the effect of storage temperature and time from sample collection to analysis on test classification of a commercially available ELISA for diagnosis of pregnancy using the measurement of pregnancy-associated glycoproteins (PAG) in milk samples from dairy cows. Few studies have evaluated the effects of sample handling on milk PAG results. Using a repeated-measures study design, we evaluated sample storage at 5 temperatures: 37°C, 22°C, 4°C, −20°C, or −80°C. Sample aliquots from 45 cows (20 with a pregnant test result, 10 open, and 15 recheck) were stored for 4, 7, 14, 28, 60, 90, or 365 d. The measured PAG level was influenced by storage duration and condition. Samples stored for 365 d had a slightly increased PAG level, whereas samples stored for all other durations showed a slight decline in PAG level compared with the initial result. The reason for an increase in PAG level following long-term storage is not known. This will not affect dairy producers using the test but may be important in samples stored for research applications. The changes in PAG level were small and within the expected variation for this test. Fewer than 6% of samples changed in classification and, as expected, they were samples near the test interpretation cut-points. [ABSTRACT FROM AUTHOR]

The main objective of this study was to assess PAG2 mRNA expression in maternal blood cells at the peri-implantation period in water buffalo; moreover, we wanted to evaluate the earliest time in which PAG-2 could be detected in maternal blood. Thirty-two lactating buffaloes artificially inseminated (AI) were utilized. Blood was collected at Days 0, 14, 18, 28, 40 after AI (AI = day 0). Pregnancy was diagnosed by ultrasound at Days 28 and 40 post AI. Out of 32 buffaloes, 14 were pregnant (P group) and 18 were not pregnant (NP group). The plasma PAG-2 threshold of 1.0 ng/mL in the P group was reached at day 40 post AI. PAG2 mRNA expression differed between the P and NP groups, and was either evaluated in Peripheral Blood Mononuclear Cells (PBMC) or Polymorphonuclear Leukocytes (PMN), starting from day 14. However, both the estimated marginal means and multiple comparisons showed that PAG2 mRNA expression was higher in PMN than PBMC. In the present study, PAG-2 appeared in the blood (40 Days post AI), and an early expression of PAG2 mRNA at Day 14 post AI was also observed. Although further research is undoubtedly required, PAG2 mRNA in peripheral blood leukocytes could be using to better understand the role that PAGs play during pregnancy in buffalo.