Method For Diagnosis Of Pregnancy Research Articles

Pregnancy Diagnosis on Beef Cattle Today

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.

Microwave Dielectric Response of Bovine Milk as Pregnancy Detection Tool in Dairy Cows.

The most reliable methods for pregnancy diagnosis in dairy herds include rectal palpation, ultrasound examination, and evaluation of plasma progesterone concentrations. However, these methods are expensive, labor-intensive, and invasive. Thus, there is a need to develop a practical, non-invasive, cost-effective method that can be implemented on the farm to detect pregnancy. This study suggests employing microwave dielectric spectroscopy (MDS, 0.5-40 GHz) as a method to evaluate reproduction events in dairy cows. The approach involves the integration of MDS data with information on milk solids to detect pregnancy and identify early embryonic loss in dairy cows. To test the ability to predict pregnancy according to these measurements, milk samples were collected from (i) pregnant and non-pregnant randomly selected cows, (ii) weekly from selected cows (n = 12) before insemination until a positive pregnancy test, and (iii) daily from selected cows (n = 10) prior to insemination until a positive pregnancy test. The results indicated that the dielectric strength of Δε and the relaxation time, τ, exhibited reduced variability in the case of a positive pregnancy diagnosis. Using principal component analysis (PCA), a clear distinction between pregnancy and nonpregnancy status was observed, with improved differentiation upon a higher sampling frequency. Additionally, a neural network machine learning technique was employed to develop a prediction algorithm with an accuracy of 73%. These findings demonstrate that MDS can be used to detect changes in milk upon pregnancy. The developed machine learning provides a broad classification that could be further enhanced with additional data.

Survival and Development of in Vivo Produced Boran and Boran* Holstein Cross Embryos

One of biotechnology technique that is frequently used to enhance the number of animals with superior genetic ability and high productivity is embryo transfer. Embryos can be obtained in vivo or in vitro, and they can be frozen and then thawed before being delivered to the recipient animals. Conception rates are influenced by a number of variables, including the quality and developmental stage of the embryo, the location of the embryo's deposit in the uterus, the degree of difficulty of the transfer, whether to use a fresh or frozen embryo, the operator's experience, the corpus luteum's quality, whether to use a heifer or a cow, and the time of year the transfer occurs. The fertility of domestic animals is severely impacted by early embryonic death. For this reason, this study was carried out to gather data on early embryonic development that is normal, the amount and timing of embryonic mortality, potential endogenous and exogenous causes of embryonic loss, and to develop strategies to lower embryonic mortality. A total of 40 embryos (20 fresh and 20 frozen, 26 quality grade 1 and 14 quality grade 2, 29 compact Morula stage and 11 early Blastocyst stage) were transferred to 40 recipient cows (22 Boran and 18 H-B cross) with different body condition score. Return to heat was used as method of pregnancy diagnosis and all recipients were followed around day 14 post embryo transfer and 14 animals were shown heat sign, the rest 26 animals were suspected for pregnancy (65%). Using ultrasound, a pregnancy diagnosis was made on day 45, and 20 recipient animals were found to be 50% positive for PD. On day 60, PD positive animals were re-examined with ultrasound and only 10 were confirmed as PD positive (25%). The PD negative on day 45 and day 60 were suspected to be early embryonic mortality. Other pregnancy loss occurred in this study was abortion nearly after five month of pregnancy. It is not doubtful that, the technique of embryo transfer is utilized to increase the reproductive rates of important female animals. However, it needs proper management for both donor and recipient animals. Therefore, for the successful application of the technology optimum level of feeding both quantity and quality, health management and conducive environment should be fulfilled for all animals.

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.

Accuracy of radiographic fetal count in ewes

Accurate fetal count is important for farm animal species as the number of fetuses can affect management decisions in both research and production settings. Documented methods of pregnancy diagnosis include radiography, progesterone assays, transrectal palpation, and transrectal and transabdominal ultrasonography; however, there is variability in fetal count accuracy with each of these methods. Abdominal radiography evaluation in 13 pregnant ewes among observers of various skill levels was compared retrospectively with the known number of fetuses determined using computed tomography. Overall accuracy using abdominal radiography across skill levels for determining fetal counts correctly was 79%. Accuracy decreased as the number of fetuses increased, with accuracies for singleton, twin, and triplet pregnancies being 92, 72, and 50%, respectively. Additionally, observer experience was inversely related to radiographic fetal count accuracy.

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.

Evaluating the use of luteal color Doppler ultrasonography and pregnancy-associated glycoproteins to diagnose pregnancy and predict pregnancy loss in Bos taurus beef replacement heifers.

The objective of this study was to evaluate the use of corpus luteum (CL) color Doppler (CD) ultrasonography and pregnancy-associated glycoproteins (PAG) for early pregnancy diagnosis and examine their ability to predict late embryonic/early fetal mortality (LEM) in Bos taurus beef replacement heifers. Beef heifers (n = 178) were exposed to a 7-d CO-Synch + CIDR protocol followed by fixed-time artificial insemination (day 0). On days 20 and 22, B-mode and CD ultrasonography were performed to evaluate CL morphometries and blood perfusion, respectively. Heifers were considered nonpregnant when CL area was <2 cm2 or estimated luteal blood perfusion was ≤30% of the total luteal area. Blood samples were collected on days 25 and 29 to estimate circulating concentrations of PAG. Conventional ultrasonography on days 29 and 94 was utilized to determine pregnancy status and considered the gold standard method for pregnancy diagnosis. Pregnant heifers had greater (P < 0.01) CL diameter, area, volume, and blood perfusion when compared with nonpregnant heifers on days 20 and 22. Accuracy of CD on days 20 and 22, and PAG on days 25 and 29 were 91%, 94%, 96%, and 98%, respectively. No false-negative results were observed for CD on both days 20 and 22 (negative predicted value = 100%) and false-positive results represented 8% and 6% of the diagnoses. Heifers that experienced LEM between days 29 and 94 of gestation had decreased luteal (P = 0.02) volume on day 20 and tended (P = 0.07) to have decreased concentrations of PAG on day 29 compared with heifers that maintained pregnancy. However, both CD and PAG failed to predict embryonic mortality. In conclusion, CD successfully detected most nonpregnant replacement heifers as early as day 20 of gestation, while resulting in no false negative diagnoses. Both CD and PAG failed to predict LEM in the present study.

Estimation of gestational age in Shami goats based on transabdominal ultrasonographic measurements of fetal parameters

Estimated gestational age is essential for breeding farm animals because pregnant females require special care and nutrition, which varies according to gestation. The study aimed to estimate the gestational age of Shami does by transabdominal ultrasonographic measurement of fetal parameters. Thirty-three Shami does aged 2-2.5 years with confirmed conception dates were used. The does were examined in a standing position using a transabdominal 3.5 MHz convex sector probe of real-time ultrasonography. The does were examined weekly starting on 21 days of pregnancy until parturition. Measurements of the fetal parameters that include; the crown-rump length, head diameter, chest diameter, trunk diameter were obtained from the ultrasonographic images using the software Screen Calipers. Results showed that the gestational sac and embryos were observed first on 21 and 35 days of pregnancy. The litter size of pregnant does range between two and three kids. Crown-rump length ranged between 24.68±1.32 - 71.71±1.84 mm on 41-60 days. The head diameter was 15.63±0.82 - 45.04±0.92 mm on 41-120 days. Chest diameter was 11.52±0.84 - 56.35±0.93 on 51-110 days, and trunk diameter was 10.69±0.85 - 57.38±0.85 mm on 41-130 days of gestation. The highest positive correlations were obtained between the gestational age chest diameter (r=0.935), crown-rump length (r=0.917), head diameter (r=0.917) and trunk diameter (r=903). In conclusion, transabdominal ultrasonography is a practical method for pregnancy diagnosis and embryo monitoring in Shami goats. Also, it is reliable to estimate gestation age starting on 41 days of pregnancy. The chest diameter was the best fetal parameter, but trunk diameter can be used for the longest period to estimate the gestational age of Shami goats.

Study of progesterone level during first trimester of pregnancy in cows.

Measurement of progesterone is an indirect method for pregnancy diagnosis in many livestock species including cattle, buffaloes, sheep, and goats. Conception extends the life of the corpus luteum (CL) by preventing the luteolytic mechanism from being triggered, thus prolonging and maintaining its functional characteristics, ensuring continued high progesterone levels (Spencer, et.al., 1995). (Shemesh et al., 1973) proposed that the difference in peripheral plasma progesterone levels between pregnant and non pregnant cows, 19 days after insemination, can form the basis for a very early pregnancy test. (Laing and Heap 1972) first documented this in milk to diagnose cows in early pregnancy Progesterone maintains the uterine endometrium in a state which supports embryonic development, implantation, and foetoplacental development. Progesterone concentrations vary with the stage of the estrous cycle which makes it one of the most commonly studied reproductive hormones in bovine ruminants for pregnancy detection and ovarian activity (Kaneko,et.al.,1994). Studies in the bovine estrous cycle indicate that the milk or serum progesterone concentrations reach a maximum value 13-14 days after estrus, and if the animal is pregnant, these continue to remain elevated up to day 21 after fertilization (Parkinson, et al.,1994) . These high levels of progesterone in serum or milk between days 18 and 24 after insemination form the basis of establishment of pregnancy in cattle (Sasser, and Ruder,1987; Shemesh,1973). Interferon-???? exerts its antiluteolytic effect by inhibiting the endometrial expression of oxytocin receptors, through which oxytocin stimulates pulsatile PGF2???? release (Wolf,2003). Although low progesterone concentrations at 18 to 24 days after breeding can accurately predict non pregnancy, high progesterone concentrations during this period are not the specific indicators of pregnancy due to variations among cows in duration of the estrous cycle as well as the incidence of early or late embryonic mortality. The advantages of progesterone assay for pregnancy diagnosis include noninvasive collection of milk sample and the feasibility to conduct the test on the farm using commercial cow-side milk progesterone test kits [Pennington,1985 ; Gowan,1982 ; Wimpy,et.al.,1986), though the sensitivity gets compromised to some extent with these assay kits.&#x0D; Key Word: progesterone, pregnancy, cows

A Comparative Study on Certain Blood Metabolites and Hormonal Indices for Detection of Pregnancy of Crossbred Cows of High Altitude: Mizoram

Background: Early detection of pregnancy can perk up the reproductive competence of dairy animals and decreasing the interval between two successive AI. Several methods of pregnancy diagnosis both direct and indirect are being practiced; but to date, none of these technologies qualifies as the ideal pregnancy detection method due to its limitation. The present study was designed to learn about the changes of hormonal and blood metabolites during early bovine pregnancy. Methods: A total of 30 numbers of crossbred cows at 2nd to 5th lactation was selected for the study. Clinico-gynaecological examination was carried out in all the cows and those have devoid of ill health, having a regular estrous cycle and estrus period were inseminated at mid to late estrus. Ten milliliter of blood samples were collected aseptically from all the cows (6 samples from each cow and a total of 180 samples) on day 1, 10, 20, 30, 60 and 90 of post-A.I. From the obtained serum, levels of glucose, cholesterol, BUN, total protein and hormonal profile estrone sulphate, progesterone were analysed. All the cows were observed for the absence of external signs of estrus after AI and pregnancy was confirmed by using ultrasonography and per rectal examination on day 30 to 40 of post insemination. Result: It was observed that level of glucose, BUN, cholesterol and progesterone was significantly increased during early pregnancy (P less than 0.01). While, the serum level of estrone sulphate significantly decreased while advancing the pregnancy. There was positive correlation between the level of progesterone (P less than 0.01) and negative correlation of estrone sulphate (P less than 0.05) with serum glucose, BUN, cholesterol at different days of post A.I. The blood biochemical constituents i.e. glucose, BUN, cholesterol and hormone estrone sulphate, progesterone possibly will used as an ideal marker of early pregnancy in cattle. Thus blood biochemical constituents like glucose, BUN, cholesterol and hormone estrone sulphate, progesterone possibly can be used as a tool for diagnosis of non pregnant dairy cattles with very high accuracy at around 30 days of post insemination.

5 Early Pregnancy Diagnosis inbos Taurus beef Replacement Heifers Using Color Doppler Ultrasonography

Abstract The objective of this study was to evaluate the use of corpus luteum (CL) color doppler ultrasonography (CD) for early pregnancy diagnosis in Bos taurus beef replacement heifers. Beef heifers (n = 183) from two locations were exposed to a 7-d CO-Synch + CIDR protocol followed by fixed-time artificial insemination (day 0). On days 20 and 22, B-mode and CD ultrasonography were performed to evaluate CL morphometries and blood perfusion, respectively. Heifers were considered non-pregnant when CL area was &amp;lt; 20 mm2 or estimated luteal blood perfusion was ≤ 25%. Conventional ultrasonography on day 29 was utilized to determine pregnancy status and considered the gold standard method for pregnancy diagnosis. Pregnant heifers had greater CL diameter, CL area, and CL volume when compared to non-pregnant heifers on days 20 and 22 (P &amp;lt; 0.001). Additionally, percentage of central, peripheral, and total luteal blood perfusion, as well as the respective blood perfusion scores were greater (P &amp;lt; 0.001) in pregnant compared with non-pregnant heifers on both day 20 and 22. Sensitivity, specificity, positive predicted value (PPV), negative predicted value (NPV), and accuracy for CD on day 20 were 100, 70, 86, 100, and 90, respectively. Sensitivity, specificity, PPV, NPV, and accuracy for CD on day 22 were 100, 76, 90, 100, and 92, respectively. Pairwise comparison of receiver operating characteristics curve analysis indicated no differences (P = 0.47) between CD on days 20 and 22 (area under the curve = 0.82 and 0.84, respectively). In conclusion, CD successfully detected most non-pregnant replacement heifers on day 20 and 22, while false negative results were absent (NPV = 100%).

Clinical methods of Pregnancy diagnosis in equines

Clinical methods of Pregnancy diagnosis in equines

Luteal color doppler ultrasonography and pregnancy-associated glycoproteins as early pregnancy diagnostic tools and predictors of pregnancy loss in Bos taurus postpartum beef cows.

The objective of this study was to evaluate the use of luteal color doppler (CD) ultrasonography and plasma concentrations of pregnancy-associated glycoproteins (PAG) for early pregnancy diagnosis in Bos taurus beef cows. Additionally, CD and PAG were evaluated as potential predictors of late embryonic/early fetal mortality (LEM). Postpartum beef cows (n = 212) were exposed to estrus synchronization followed by fixed-time artificial insemination (day 0). On days 20 and 22, B-mode and CD ultrasonography were performed to evaluate corpus luteum (CL) morphometries and blood perfusion. Moreover, blood samples were collected on days 25 and 29 to quantify circulating concentrations of PAG. Conventional ultrasonography on days 29 and 100 was utilized as the gold-standard method for pregnancy diagnosis. Cows that experienced pregnancy loss between days 29 and 100 were classified as LEM. Pregnant cows had larger and more vascularized CL compared with nonpregnant cows on days 20 and 22 (P < 0.001 for all response variables). Accuracy for CD on days 20 and 22 were 87% and 92%, respectively. Accuracy for PAG on days 25 and 29 were 84% and 99%, respectively. No false negative (FN) results were observed for CD on both days 20 and 22; however, there were 7.1% FN results for PAG on day 25. Cows that experienced LEM had decreased (P = 0.04) circulating PAG on day 29 of gestation compared with cows that maintained pregnancy; however, there were no differences in luteal blood perfusion on days 20 and 22 (P ≥ 0.53) or circulating PAG on day 25 (P = 0.46) between LEM cows and cows that maintained pregnancy. Sensitivity and specificity of PAG on day 29 as predictors of LEM were 83% and 77%, respectively. In conclusion, CD resulted in accurate pregnancy diagnosis in B. taurus beef cows on both days 20 and 22 of gestation, while having no FN results. Circulating concentrations of PAG were decreased in cows that experienced LEM; however, further research is required to utilize PAG as a predictor of LEM commercially.

Early pregnancy diagnosis based on luteal morphology and blood flow on Days 17–21 post-artificial insemination in Japanese Black cattle

The present study was conducted to evaluate the applicability of corpus luteum (CL) morphology and blood flow for early pregnancy diagnosis on Days 17–21 post-artificial insemination (AI) in Japanese Black (JB) cattle. Ultrasound examinations were performed on 230 inseminated JB cattle (202 cows and 28 heifers) to evaluate CL morphological characteristics [CL area, CL tissue area, presence/absence of a central cavity, and central cavity area (CCA)] and blood flow area (BFA) on Days 17 (n = 46), 18 (n = 45), 19 (n = 46), 20 (n = 47), and 21 (n = 46) (Day 0 = the day of AI). Pregnancy was confirmed using brightness-mode ultrasonography on Day 30. In pregnant group (n = 141), CL area and CL tissue area were greater (P < 0.001) than those in non-pregnant group (n = 89) on and after Day 19. Moreover, BFA was greater (P < 0.05) in pregnant group on and after Day 18. The overall central cavity incidence was lower (P < 0.001) and CCA was smaller (P < 0.001) in pregnant group. Logistic regression analyses revealed that the best single predictor for pregnancy diagnosis was BFA on Days 17–20 (sensitivity = 69%–100% and specificity = 65%–91%). On Day 21, CL area or CL tissue area was the most accurate predictor (cutoff value = 267.78 mm2, sensitivity = 100%, and specificity = 100%). The best set of predictors for pregnancy diagnosis was specified as the BFA and [CCA × BFA] pair. Setting BFA cutoff value in the cattle without central cavity at 55.26 mm2 yielded the highest sensitivity (97%) and specificity (88%) on Days 18–20. The accuracy of pregnancy diagnosis using BFA or BFA and [CCA × BFA] pair on Day 17 was lower than those on and after Day 18. In conclusion, BFA and CCA are effective for early pregnancy diagnosis on Days 18–20 post-AI in JB cattle; however, it is inadequate on Day 17 owing to its low accuracy. Evaluating CL area or CL tissue area on Day 21 is the most accurate method of pregnancy diagnosis.

Determination of Gestational Age in Shami Goats Based on Transabdominal Ultrasonographic Measurements of Placentomes and Uterine Diameter

The study was aimed to estimate the gestational age of Shami does by transabdominal ultrasonographic measurement of maternal parameters. Thirty-three Shami does aged 2-2.5 years with confirmed conception dates were used. The does were examined in a standing position using a transabdominal 3.5 MHz convex sector probe of real-time ultrasonography. The does were examined weekly starting from 21 days of pregnancy until parturition. Measurements of the maternal parameters that include; the uterine diameter, placental width, and placental height were obtained from the ultrasonographic images using the software Screen Calipers. Results showed that the gestational sac and embryos were observed early on 21 and 35 days of pregnancy. The litter size of Shami does was 2-3 kids. Uterine diameter ranged between 27±1.4 and 136.9±4.2 mm between 21-130 days, placentomes width was 17.6±0.6 – 38.5±2.1 mm, placentomes height was 9.5 ± 1.8 – 24.5 ± 1.4 mm on 51-150 days of gestation. The highest positive correlations were obtained between the gestational age and the measurements of uterine diameter (r=0.943). The least positive correlations were between the gestational age and the values of placenta width (r=0.715) and placentom height (r=0.615). In conclusion, transabdominal ultrasonography is a practical method for pregnancy diagnosis and monitoring of embryo in Shami goats. Also, it is reliable to estimate gestation age from 21 days of pregnancy. The uterine diameter was the best maternal parameter that can be used for the longest period for estimation of Shami goats gestational age.

PSXII-21 Pregnancy associated glycoproteins for pregnancy diagnosis on day 25 of gestation

Abstract The objective of this study was to evaluate the use of plasma concentrations of pregnancy associated glycoproteins (PAG) for pregnancy diagnosis in beef cows on day 25 of gestation. Additionally, differences in circulating concentrations of PAG among cows that experienced late embryonic/early fetal mortality (LEM) were evaluated. Beef cows (n = 214) were exposed to the 7-d CO-Synch + CIDR protocol followed by fixed-time artificial insemination (day 0). Blood samples were collected on days 25 and 29 to determine circulating concentrations of PAG and ultrasonography on day 29 was used as the gold-standard method for pregnancy diagnosis. On days 25 and 29, PAG concentrations were greater (P &amp;lt; 0.001) in pregnant compared with non-pregnant cows. Sensitivity, specificity, positive predicted value (PPV), negative predicted value (NPV), and accuracy on day 25 were 87, 80, 83, 84, and 83%, respectively. Sensitivity, specificity, PPV, NPV, and accuracy for PAG on day 29 were 99, 94, 96, 97, and 99%, respectively. Pairwise comparison of receiver operating characteristics curve analysis indicated greater area under the curve for PAG on day 29 compared with day 25 (P &amp;lt; 0.001). There were no differences in plasma concentrations of PAG on day 25 among cows that experienced LEM compared to cows that maintained pregnancy (P = 0.46). Concentrations of PAG on day 29 were reduced (P = 0.03) in cows that experienced LEM compared with cows that maintained pregnancy, and were not different between LEM and non-pregnant cows (P = 0.12). In summary, cows that experienced LEM had decreased plasma concentrations of PAG compared with cows that maintained pregnancy. The PAG assay utilized in this study yielded accurate pregnancy diagnosis results on day 29 of gestation: however, the presence of false negative results (NPV = 84%) limits its use on day 25 of gestation.

Loop-mediated isothermal amplification (LAMP) and machine learning application for early pregnancy detection using bovine vaginal mucosal membrane

An early and accurate pregnancy diagnosis method is required to improve the reproductive performance of cows. Here we developed an easy pregnancy detection method using vaginal mucosal membrane (VMM) with application of Reverse Transcription-Loop-mediated Isothermal Amplification (RT-LAMP) and machine learning. Cows underwent artificial insemination (AI) on day 0, followed by VMM-collection on day 17–18, and pregnancy diagnosis by ultrasonography on day 30. By RNA sequencing of VMM samples, three candidate genes for pregnancy markers (ISG15 and IFIT1: up-regulated, MUC16: down-regulated) were selected. Using these genes, we performed RT-LAMP and calculated the rise-up time (RUT), the first-time absorbance exceeded 0.05 in the reaction. We next determined the cutoff value and calculated accuracy, sensitivity, specificity, positive prediction value (PPV), and negative prediction value (NPV) for each marker evaluation. The IFIT1 scored the best performance at 92.5% sensitivity, but specificity was 77.5%, suggesting that it is difficult to eliminate false positives. We then developed a machine learning model trained with RUT of each marker combination to predict pregnancy. The model created with the RUT of IFIT1 and MUC16 combination showed high specificity (86.7%) and sensitivity (93.3%), which were higher compared to IFIT1 alone. In conclusion, using VMM with RT-LAMP and machine learning algorithm can be used for early pregnancy detection before the return of first estrus.

Prevalence and associated economic loss of fetal wastage in small ruminants slaughtered at Addis Ababa municipality abattoir, Ethiopia

The study examined prevalence and economic implications of abattoir pregnancy/ fetal wastage in small ruminants. Survey was conducted on ewes (n=201) and does (n=183) slaughtered at Addis Ababa municipality abattoirs, during December 2017 and January 2018. Female reproductive tracts were examined for presence and types of pregnancy and/ or gross disorders. Net economic lossand net economic risk due to abattoir fetal wastage were calculated for each species after accounting for naturally expected abortion and neonatal losses, and domestic net market values. One hundred forty-two (37 %) animals were pregnant 131 (34.1 %) with single and 11 (2.9 %) with twin fetus. A total of 153 fetuses were recovered giving a fetal wastage prevalence of 39.8 %. More animals in the second trimester (25.5 %) were slaughtered than those in first (8.3 %) or third (3.1 %) trimesters (p &lt; 0.05). Prevalence of pregnancy was 32.8 % in does and 40.8 % in ewes (p = 0.104). Prevalence of fetal wastage was 37.2 % (68 fetuses) and 42.3 % (85 fetuses) in goats and sheep, respectively (p &gt; 0.05). Abattoir pregnancy prevalence showed variations relative to slaughter month in goat (p &lt; 0.01) and body condition in sheep (p &lt; 0.05). Observed abattoir pregnancy prevalence levels incurred net economic loss of 313.55 USD in sheep (per 201 ewes) and 315.4 USD in goats (per 108 doe). This translated to net economic risk of 1.7 USD per mature doe or ewe slaughtered for meat. Nine (4.9 %) does and 1 (0.5 %) ewe showed gross reproductive tract disorders (p &lt; 0.05). Frequent female slaughter without efficient ante-mortem pregnancy screening predisposed significant proportion of small ruminants to pregnancy/ fetal wastage. Deeper investigations are needed to understand reasons behind pregnant small ruminant slaughter and to mitigate its negative impacts onsustainability of animal production. Evaluating and capacity building on alternative small ruminant pregnancy diagnosis methods requires due attention. &#x0D; Key words: Doe; Pregnancy; Reproductive disorders; Ewe; Fetal wastage; Economic implication; Abattoir

B-ultrasonography: An Accurate and Noninvasive Method for Pregnancy Diagnosis in Pangolins

B-ultrasonography: An Accurate and Noninvasive Method for Pregnancy Diagnosis in Pangolins

İvesi Koyunlarında Gebeliğin Erken Teşhisi İçin Sığır Hızlı Görsel PAG ELISA Testi ve Transabdominal Ultrasonografinin Değerlendirilmesi

The aim of this study was to evaluate two different methods (a bovine rapid visual PAG ELISA test and transabdominal ultrasonography) for early diagnosis of pregnancy in Awassi sheep. The ewes used in this study (n=50) were hand-mated in June of 2019 and lambed in October of 2019. Pregnancy diagnosis was performed 34-38 days after mating by rapid visual PAG ELISA test and transabdominal ultrasonography in standing position. The diagnostic test characteristics of both methods were calculated using lambing results as a gold standard. Kappa analysis was conducted to analyze the agreement between each pregnancy diagnosis method and the lambing results. McNemar's analysis and the comparison of the receiver operating characteristic (ROC) curves demonstrated that there was no significant difference between the pregnancy diagnosis methods and lambing results (p ≤ 0.05). According to the lambing results, it was determined that, of the 50 ewes, 42 (84%) were pregnant and 8 (16%) were not. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of rapid visual PAG ELISA and transabdominal ultrasonography were 97.6%, 62.5%, 93.2%, 83.3%, 92% and 95.2%, 100%, 100%, 80%, 96%, respectively. The Kappa coefficient was 0.669 (good agreement) between rapid visual PAG ELISA and lambing, and 0.865 (very good agreement) between transabdominal ultrasonography and lambing. In McNemar’s analysis, no difference was found between the pregnancy diagnosis methods and lambing results. ROC analysis revealed that the areas under the curves were 0.801 and 0.976 for rapid visual PAG ELISA and transabdominal ultrasonography, respectively, and the difference between the two pregnancy diagnosis methods was statistically insignificant. In conclusion, both rapid visual PAG ELISA and transabdominal ultrasonography in standing position proved to be practical and reliable methods for pregnancy diagnosis as of day 34 post-mating in Awassi ewes.