Applied use of interferon-tau stimulated genes expression in polymorphonuclear cells to detect pregnancy compared to other early predictors in beef cattle.

We aimed to evaluate the accuracy of Interferon-tau stimulated genes (ISG) abundance in peripheral blood polymorphonuclear cells (PMNs) on D20 after fixed-time artificial insemination (FTAI; D0) as a pregnancy diagnosis method against CL evaluation by Doppler ultrasonography and progesterone (P4) concentrations on D20, as well as Pregnancy Associated Glycoproteins (PAG) concentrations on D25. Additionally, we evaluated the potential of ISG abundance in PMNs as pregnancy loss predictors. Nelore heifers (n = 103) and cows (n = 144) underwent estrous synchronization and were artificially inseminated on D0. Pregnancy was diagnosed by B-mode ultrasonography on D30 and D70, and after the final diagnosis, females were classified in four groups: Pregnant; Non-pregnant; Functional CL on D20 but non-pregnant (CL-NP) and Pregnancy loss between D30 and D70 (PL). After determining cutoff values, the Sensitivity (SE), Specificity (SP), Positive Predictive Value (PPV), Negative Predictive Value (NPV) and Accuracy (ACC) were determined for each method. All methods were classified as significant (P < 0.05) predictors of pregnancy. Both ISG expression and PAG concentrations were greater (P < 0.05) in pregnant females than in non-pregnant and CL-NP females but did not differ (P > 0.05) from the PL group. ISG15 expression was greater (P < 0.05) in heifers than in cows, but this difference was not found in OAS1 expression and PAG concentrations. All the methods evaluated were proven to be adequate predictors of pregnancy, but greater accuracies were obtained through PAG concentrations and Doppler-US, due to the decreased number of false positive and false negative results.

Evaluation of a cattle rapid test for early pregnancy diagnosis in sheep.

The early pregnancy diagnosis allows optimizing production and timely management correction, with a greater reproductive output of livestock. The Idexx Rapid Visual Pregnancy Test® consists of an ELISA for visual reading which does not require the use of readers in the laboratory, with satisfactory pregnancy diagnoses at 28 days of pregnancy in cattle. Therefore, the aim of this study was to evaluate this rapid test and to verify the most appropriate day for the diagnosis of pregnancy in the ovine species. For this purpose, 98 serum samples from pregnant sheep and 36 from non-pregnant were used, with duplicate samples, and diagnosis was confirmed by ultrasound examination, used as the gold standard. The numbers of positive samples obtained at 26, 28, and 30 days of pregnancy were 26, 27, and 45, respectively. The Rapid Visual Pregnancy Test correctly identified 100% of the samples as positive at pregnancy of days 26, 28, and 30. The sensitivity and specificity of the test were also 100%. The Idexx Rapid Visual Pregnancy Test, initially indicated for cattle, is effective for the detection of pregnancy in the ovine species, enabling diagnosis of pregnant sheep from the 26th day of pregnancy.

Use of an at-home multilevel pregnancy test and an automated call-in system to follow-up the outcome of medical abortion.

OBJECTIVE: To assess the feasibility of using an at-home multilevel pregnancy test (MLPT) and interactive voice response (IVR) call-in system for remote follow-up of medical abortion. METHODS: A prospective pilot study was conducted among women who had a medical abortion at up to 70 days at a clinic in Mexico City, Mexico, between June 1, 2015, and January 30, 2016. Participants took an MLPT at the initial clinic visit and another MLPT at home 2 weeks later. They were requested to report their MLPT results via the IVR system and attend the clinic for follow-up evaluation. RESULTS: Of 200 women considered for inclusion, 163 (81.5%) were included in the analysis. Only 10 (6.6%) of the 152 women who had a medical abortion on or before 63 days from last menstrual period reported MLPT results to the IVR system that required clinical evaluation to assess medical abortion outcome. The remaining 142 (93.4%) women in this group reported MLPT results that ruled out ongoing pregnancy (confirmed at clinical evaluation). Reported MLPT results ruled out ongoing pregnancy among the 11 women who had a medical abortion after 63 days; however, 1 (9%) had an ongoing pregnancy at clinical evaluation. CONCLUSION: Use of MLPTs and the IVR system provided a streamlined approach to follow-up after medical abortion.

Plasmatic estradiol concentration in the mid-luteal phase is a good prognostic factor for clinical and ongoing pregnancies, during stimulated cycles of in vitro fertilization.

OBJECTIVE: To evaluate the predictive efficiency of serum estradiol (E2) concentration in the mid-luteal phase regarding chemical, clinical, and ongoing pregnancies, in patients subjected to IVF/ICSI with fresh embryo transfer. METHODS: One hundred and forty-three patients undergoing IVF/ICSI met all the inclusion criteria for the present study. Most of the patients used antagonists, final maturation was achieved with recombinant chorionic gonadotrophin (HCG), and embryo transfer took place on days 3 to 5, but mostly on day 4. The luteal phase was supplemented with estradiol valerate 6 mg/day and vaginal micronized progesterone 600 mg/day. There was no exclusion of patients in the embryo transfer group due to age or ovarian reserve. All patients with estradiol and chorionic gonadotrophin (ßHCG) dosage on the day of transfer, day 7, were included. We assessed the following variables, initially regarding age: number of eggs collected, formed embryos, embryos transferred, day of transfer, transfer type, estradiol and chorionic gonadotropin. Next, we evaluated these elements at three different ranges of estradiol concentrations (<200 pg/ml, 200-500 pg/ml, and >500 pg/ml), comparing these parameters in pregnant (P) and non-pregnant (NP) patients. RESULTS: Data analysis by age group in P and NP patients showed significant differences in the mean values of the variables E2 and ßHCG, TD7. Mean serum estradiol levels in P and NP in the three age groups were: <35 years, 835/417 p=0.0006, 35-39 years 833/434 p=0.0118, >39 years, 841/394 p=0.0012. There was also a significant difference in pregnancy rates in the group >500 pg/ml of estradiol concentration (63.4%, p=0.0096). The likelihood of chemical and clinical abortions for the estradiol ranges were: 38.46%, involving the two first ranges versus 15.15% for a concentration >500 pg/ml, p=0.0412 and 5.26% for a concentration >900 pg/ml, p=0.0105. The Pearson correlation coefficient for HCG and estradiol was r=0.5108. CONCLUSION: This study showed the prognostic value of E2 in the mid-luteal phase (TD7) for chemical, clinical, and ongoing pregnancies, and its concentration suggested that there is a moderately positive correlation with ßHCG levels.

Accuracy of pregnancy diagnosis in swine by ultrasonography.

Early and accurate diagnosis of nonpregnant sows and gilts has the potential to increase reproductive efficiency and the financial income in pig production by reducing non-productive days per sow per year. The objectives of this study were to compare the efficiency of pregnancy diagnosis between Doppler Echo+ and real time ultrasonography (RTU) and to compare the efficiency by using RTU at different days post-mating (days 17 to 24) under commercial conditions. In the 1st study, using crossbreed sows and gilts (n=107), pregnancy diagnoses were done with Doppler Echo+ and then with RTU. Between 28 and 65 days of gestation, Doppler Echo+ had 85% sensitivity and 32% specificity, and efficiency was 73%. In the 2nd study, sows (n=142) were scanned for pregnancy diagnosis between 17 and 24 d post-mating (PD1) and reconfirmed between 38 and 45 days of gestation (PD2). After 21 days of gestation, RTU had over 90% sensitivity and 45% specificity, and 70% efficiency. Accuracy between PD1-farrowing was 75.5% and between PD1 and PD2 was 80.6%. In the 3rd study, sows were diagnosed pregnant by RTU (n=151) at 17 to 24 days of gestation or A-mode ultrasound (n=172) at 28 to 30 days of gestation. There were no significant differences in conception rate (P > 0.09) and farrowing rate (P > 0.67) between both groups. Hence, there was no improvement in fertility and farrowing rate by using RTU instead of A-mode ultrasound under commercial conditions.

Measurement of endometrial volume increase during the first week after embryo transfer by three-dimensional ultrasound to detect pregnancy: a preliminary study.

Endometrial volume and thickness were measured in 20 women undergoing in vitro fertilization on the day before the embryo transfer and one week later by three-dimensional ultrasound. The endometrial volume increase was higher in the 8 pregnant women (0.70 cm(3) at least) than in the 12 nonpregnant women (0.21 cm(3) at most).

Endometrial thickness and volume by three-dimensional ultrasound one week after embryo transfer to detect pregnancy.

PURPOSE: Determine if the evaluation of endometrium one week after embryo transfer can predict pregnancy. METHODS: Endometrial volume and thickness were evaluated by three-dimensional ultrasound in 40 patients one week after embryo transfer. These results were compared to serum pregnancy test performed one week later. RESULTS: Twenty two patients have achieved pregnancy. A significant difference was found for endometrial volume: 6.49+/-1.97 mL vs. 3.40+/-1.11 mL (pregnant vs. not pregnant); and thickness: 11.15+/-2.75 mm vs. 9.77+/-1.85 mm. The ROC curve was used to detect the best cutoff values: endometrial volume of 3.48 mL (sensitivity-100%, specificity-68.2%) and endometrial thickness of 10.3 mm (sensitivity-72.2%, specificity-77.3%). The area under curve was significant higher for endometrial volume (0.909 vs. 0.745, P=0.027). No pregnancy was achieved in women who had an endometrial volume <3.8 mL (15 patients) or thickness < 7.9 mm (3 patients). CONCLUSIONS: The endometrial volume and thickness were significant higher in pregnant women and this difference was more prominent for endometrial volume.

Integrating ultrasonography within the reproductive management of the collared peccary (Tayassu tajacu).

Ultrasound imaging has been used to elucidate certain aspects of the reproductive biology of wild or endangered species. However, to our knowledge, this tool has not been used for reproductive monitoring of the collared peccary (Tayassu tajacu). In this study, real-time ultrasonography was used in 16 collared peccary females to diagnose early pregnancy status and predict gestational age. Based on the detection of an embryo, the earliest pregnancy diagnosis was made on Day 18 after mating, with the mean time needed for diagnosis being 22 days. Overall accuracies on Days 22, 26 and 28 were 56, 93, and 100%, respectively. On Days 26 and 28, all pregnancy and non-pregnancy diagnoses, respectively, were correct. The fetal measurements that best correlated with gestational age were crown-rump-length (CRL) and the length and diameter of the thorax. CRL was considered the most practical measurement because, contrary to thoracic fetometry, it could be determined when the embryo was first detected. Our findings revealed real-time ultrasound scanning to be a very accurate method for early pregnancy diagnosis and prediction of gestational age in the collared peccary.

Diagnóstico precoz de embarazo en la atención primaria mediante determinación cualitativa de gonadotropina coriónica humana / Early diagnosis of pregancy in primary care by qualitative determination of HCG

Se presentan los resultados de la aplicación de un método cualitativo no instrumental, auxiliar de la práctica clínica para el diagnóstico del embarazo en un consultorio médico. El ensayo se basó en la detección de la hormona gonadotropina coriónica en la orina de mujeres fértiles con amenorrea, mediante un immunoensayo enzimático desarrollado en el Instituto Nacional de Endocrinología. La determinación se caracterizó por una sensibilidad del 85 porciento, una especificidad del 95 porciento y una exactitud del 90 porciento(para un límite de clasifiación positivo de 100 UI de HC g/L). La coincidencia del método cualitativo con uno cuantitativo fue del 90 porciento. Aunque la sensibilidad del método cualitativo fue inferior a los requerimientos expuestos en la literatura especializada revisada, se recomienda su uso por los médicos de la familia, como una arma auxiliar en el diagnóstico precoz del embarazo, por su elevada especificidad, buena correlación con el método cuantitativo y su relativa sencillez operacional(AU)

Evaluación de un anticuerpo monoclonal antigonadotropina coriónica para detectar embarazo / Evalution of a monoclonal antibody chorionic antigonadotropin to detect pregnancy

Se evaluó el anticuerpo monoclonal B8 producido en el INEN para ser utilizado en el desarrollo de un método analítico para detectar embarazo. Los resultados de este método fueron evaluados utilizando como referencia los métodos tradicionales como tacto vaginal y biopsia de endometrio. Se encontró que es capaz de detectar el embarazo con 80 por ciento de sensibilidad y 100 por ciento de especificidad. El anticuerpo monoclonal valido B8 cumple con los requisitos de calidad para ser utilizado en el método inmunoenzimático con que contamos pues los resultados obtenidos no dieron diferencias significativas con el monoclonal en uso(AU)

Early pregnancy diagnosis in alpaca (Lama pacos) and llama (Lama glama) by ultrasound.

An ultrasonography study of early pregnancy diagnosis was carried out in 19 alpacas and 12 llamas, after controlled matings. The aim was to determine the earliest gestational age at which pregnancy diagnosis by transrectal ultrasonography could be achieved, and to generate an empirical formula for gestational sac diameter (GSD) growth as a function of gestational age (GA), allowing an estimate of GA during the first month of pregnancy. We found that pregnancy diagnosis may be carried out as early as 9 days after mating in alpacas and 7 days in llamas. This diagnosis was found to be accurate at 23 days in alpacas and 34 days in llamas. The empirical relations that best describe the relationship between GSD and GA were GA = logGSD + 1.2339/0.0585 r = 0.85; P < 0.001 in alpacas, and GA = logGSD + 1.2649/0.0546 r = 0.77, P < 0.001 in llamas, where GA is measured in days and GSD in centimeters. Our results also indicate that ultrasonography is a reliable technique for early pregnancy diagnosis. Furthermore, the empirical formulae reliably make it possible to estimate GA from GSD during the first month of pregnancy and their use might improve the efficiency of camelid breeders.

[Pregnancy in New World camelids]. / Trächtigkeitsuntersuchung von Neuweltkameliden.

There is a growing number of llama- and alpacabreeders in Europe. Therefore veterinarians are often asked to intervene in clinical management of different problems, especially reproductive problems. In this review the authors attempt to summarize the different possibilities of pregnancy diagnosis and to give an short overview of reproduction in female llamas and alpacas. South American Camelids are induced ovulators and ovulation will occur within 12 to 24 hours following a single breeding. Llamas and alpacas develop follicular waves. The females will usually not ovulate unless a 7 mm or larger follicle exist on one of the ovaries. Corpus luteum is needed for maintenance of pregnancy.

Evaluacion del metodo de deteccion del factor precoz de preñez (EPF) en cerdos: importancia en el diagnostico temprano de gestacion / Evaluation of the method of early pregnancy factor detetion (EPF) in swine: significance in early pregnancy diagnosis

El Factor Precoz de Preñez (EPF) es una proteína que se detecta en el suero de las hembras gestantes, en la mayoría de los mamíferos, en el período previo a la implantación del embrión. La actividad de dicho factor se pone de manifiesto mediante la técnica de inhibición de formación de rosetas (TIR). Después de la preincubación de los linfocitos con suero de cerdas no preñadas, el valor meduo del TIR fue de 10.80 ñ 0.32 con un límite superior de 11.44 paa un intervalo de confianza de 95%. Por lo tanto, todos los valores de TIR 12 fueron definidos como indicadores de actividad EPF. En linfocitos incubados en suero de cerdas gestantes, el valor promdeio de TIR aumentó significativamente 19.98 + 1.13 (p < 0.001). El 10% de las cerdas preñdas no evidenció actividad EPF. Una sola cerda vacía dió un valor de TIR=20. Este ensayo no es cuantitativo y la gran dispersión en los valores del TIR lo convierten en un método poco adecuado para el diagnóstico precoz de preñez. Sería de interés contar con una prueba rápida y segura que permitiera el diagnóstico de la gestación de manera precoz, como así también, un manejo racional de la hembra pocos días después del servicio o de la transferencia embrionaria (AU)

Monoclonal antibodies to beta subunit of choriogonadotropin: development and use in a sandwich ELISA pregnancy test.

Balb/c mice were immunized with beta subunit isolated and purified from crude human chorionic gonadotropin preparations. Spleen cells from the higher titered mouse were fused with Sp 2/0 myeloma cells. Four specific secreting hybridomas were obtained. Specificity, affinity, and suitability of secreted antibodies for use in enzyme immunoassays were studied. Ascites of the selected hybridoma was raised; the antibody was purified by protein A-affinity chromatography and coupled to horseradish peroxidase. This conjugate was employed in a simultaneous sandwich enzyme immunoassay on microtiter plates sensitized with goat polyclonal antibody to measure the hormone. The test has a sensitivity of 10 mlU/ml either on urine, serum, or plasma samples when read in a microplate reader. The results can also be evaluated by the naked eye, with a sensitivity of 20 mlU/ml. No cross reactivity was detected with other human gonadotropins.

[Evaluation of the method for early pregnancy factor detection (EPF) in swine. Significance in early pregnancy diagnosis]. / Evaluación del método de detección del factor precoz de preñez (EPF) en cerdos. Importancia en el diagnóstico temprano de gestación.

Early pregnancy factor (EPF) is a protein detected in the serum of pregnant females in most mammals, before implantation of the embryo; its activity is detected by the rosette inhibition test (RIT). After preincubation of lymphocytes in serum of non-pregnant pigs, the mean titre of RIT was 10.80 +/- 0.32. The upper limit of the 95% confidence interval was calculated to be 11.44. Therefore, all titres 12 were defined indicating the presence of EPF activity. The incubation of lymphocytes in serum of pregnancy caused a significant increase in the mean rosette inhibition titre, raising it to 19.88 +/- 1.13 (p < 0.001). False negative results were obtained for 10% of pregnant pigs. EPF positive activity was detected in only one non-pregnant pig (RIT = 20). This assay is not quantitative nor suitable for pregnancy diagnosis taking into account RIT values dispersion.