Innovations in the early pregnancy diagnosis in cattle

• Grant number: 15/10606-9
• Support Opportunities: Research Grants - Young Investigators Grants
• Start date: December 01, 2016
• End date: November 30, 2021
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Guilherme Pugliesi
• Grantee: Guilherme Pugliesi
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Adroaldo Jose Zanella ; Augusto Hauber Gameiro ; Claudia Maria Bertan Membrive ; Ed Hoffmann Madureira ; Fernando Silveira Mesquita ; Luciano Andrade Silva ; Luiz Lehmann Coutinho ; Mario Binelli ; Sónia Cristina da Silva Andrade
• Associated research grant(s): 19/01039-4 - International collaboration for the study of accuracy, feasibility, and cost-effectiveness of two novel techniques for early pregnancy diagnosis in dairy cattle, AP.R SPRINT
• Associated scholarship(s): 20/12207-2 - Validation of Immunoassay for measurement of circulating progesterone concentrations during early pregnancy, BP.TT; 20/12930-6 - Expression of ISGs and corpus luteum function in the third week after insemination in pregnant dairy cows or with embryonic losses, BP.TT; 20/11348-1 - Expression of novel pregnancy markers stimulated by bovine embryo at early diestrus, BP.IC; + associated scholarships 20/05878-8 - Impacts of anticipating ovulation resynchronization by early detection of non-pregnant dairy cows with doppler ultrasound, BP.MS; 19/16040-8 - Doppler ultrasonography and the expression of interferon-tau stimulated genes for early diagnosis of gestation in heifers and dairy cows, BP.PD; 18/20058-7 - Use of injectable progesterone and CIDR in early resynchronization of Nellore cows and heifers submitted to 3 FTAIs in 48 days, BP.MS; 19/07805-0 - Use of different doses of estradiol benzoate in synchronization and resynchronization at 14 days after FTAI in Bos indicus beef cows, BP.MS; 18/25393-9 - Abundance of transcript for receptors of type I interferon in blood leucocytes during early pregnancy in Nelore heifers, BP.IC; 19/05926-5 - Expression of genes stimulated by interferon-tau in the vaginal mucosa cells as a new alternative for early pregnancy diagnosis in cattle, BP.IC; 17/13994-5 - Study of transcriptomic profile in immune cells of peripheral blood during early pregnancy in beef heifers, BP.MS; 18/19652-1 - Immunoassay for measurement of circulating progesterone concentrations during early pregnancy, BP.TT; 17/13472-9 - Early pregnancy diagnosis in cattle through the expression of interferon-tau stimulated genes in neutrophils, BP.MS; 17/18613-0 - Effects of ressynchronization of estradiol on day 14 after timed-AI on morphofunctional characteristics of corpus lute and uterus in Nelore females, BP.MS; 16/23964-3 - Innovations in early pregnancy diagnosis in cattle, BP.JP - associated scholarships

Abstract

The identification of the pregnancy status earlier than the current methods done on about 30 days post-insemination allows a reduction in the economic gain. The main thesis is that biomarkers stimulated by conceptus in the blood are detectable at early pregnancy. This may allow the development of an early and innovator pregnancy diagnosis method in cattle. Therefore, we aimed to develop a diagnostic method through the quantification of the abundance of reported genes stimulated by the IFN-tau (ISGs) or new biomarkers coupled to the identification of the non-pregnant cows by the evaluation of the luteal function. Thus, in Study 1 new markers of pregnancy status may be identified by the transcriptome study of the immune cells and total peripheral blood on Days 14 and 18 post-insemination. The purpose is to characterize the abundance of the known ISGs and of new transcripts stimulated by the presence of the conceptus in the total blood, polymorphinuclear immune cells (PMNs) and immune cells of the milk between Days 12 and 20 post-insemination in dairy and beef cows. Together, the luteolysis will be detected by Doppler ultrasonography through the changes in the size and vascularization of the corpus luteum (CL) e by the circulating progesterone concentrations to detect cows with non-functional CL. For measurement of P4 concentrations, we aim to develop and validate an immunoenzymatic assay with high sensitivity and specificity to avoid the use of radioisotopes. In Study 2, the accuracy of the developed methods will be analyzed in a moment before Day 20 post-insemination based in the combined or the single use of the expression od ISGs or the new biomarkes, and the CL function (by the P4 concentration or by the Doppler ultrasonography). Based on these results, we will evaluate in Studies 3 and 4 the productive and economic of the use of the proposed methods to identify non-pregnant beef and dairy cows and to propose a protocol for earlier re-insemination. The new knowledge and the innovative methods here proposed will serve as basis for development of new technologies for meet and dairy systems. The practicability is guaranteed by the combined work of specialists in each analytical step and by the infrastructure of the institutions involved. (AU)

Scientific publications

(17)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

PUGLIESI, GUILHERME; BISINOTTO, DANILO ZAGO; MELLO, BARBARA PIFFERO; LAHR, FABIO C.; FERREIRA, CATIA A.; MELO, GABRIELA DALMASO; BASTOS, MICHELE RICIERI; MADUREIRA, ED HOFFMANN. A novel strategy for resynchronization of ovulation in Nelore cows using injectable progesterone (P4) and P4 releasing devices to perform two timed inseminations within 22 days. REPRODUCTION IN DOMESTIC ANIMALS, v. 54, n. 8, p. 1149-1154, AUG 2019. (15/10606-9, 16/23964-3)

SILVA, AMANDA GUIMARAES; PINTO, LEONARDO MARIN FERREIRA; SILVA, NADARK DE AMORIM; MATTOS, ANA CLARA DEGAN; AMBROSIO, PABLO HENRIQUE; DUARTE, KEILA MARIA RONCATO; ALVAREZ, RAFAEL HERRERA; PUGLIESI, GUILHERME. Comparison of three doses of estradiol benzoate for synchronization of follicular wave emergence in suckled Bos indicus beef cows. ANIMAL REPRODUCTION, v. 18, n. 3, 2021. (15/10606-9, 19/07805-0)

ROCHA, CECILIA CONSTANTINO; MARTINS, THIAGO; CARDOSO, BEATRIZ OLIVEIRA; SILVA, LUCIANO ANDRADE; BINELLI, MARIO; PUGLIESI, GUILHERME. Ultrasonography-accessed luteal size endpoint that most closely associates with circulating progesterone during the estrous cycle and early pregnancy in beef cows. Animal Reproduction Science, v. 201, p. 12-21, FEB 2019. (17/13994-5, 16/23964-3, 15/10606-9)

ATAIDE JUNIOR, GILMAR ARANTES; KLOSTER, ANDERSON; DE MORAES, EMERSON GUIMARAES; MOTTA, IGOR GARCIA; CLARO JUNIOR, IZAIAS; VASCONCELOS, JOSE LUIZ M.; FERRAZ, PRISCILA ASSIS; NOGUEIRA, GUILHERME DE PAULA; PUGLIESI, GUILHERME. Early resynchronization of follicular wave emergence among Nelore cattle using injectable and intravaginal progesterone for three timed artificial inseminations. Animal Reproduction Science, v. 229, JUN 2021. (15/10606-9, 18/20058-7)

ROCHA, CECILIA CONSTANTINO; DA SILVA ANDRADE, SONIA CRISTINA; DE MELO, GABRIELA DALMASO; MOTTA, IGOR GARCIA; COUTINHO, LUIZ LEHMANN; GONELLA-DIAZA, ANGELA MARIA; BINELLI, MARIO; PUGLIESI, GUILHERME. Early pregnancy-induced transcripts in peripheral blood immune cells in Bos indicus heifers. SCIENTIFIC REPORTS, v. 10, n. 1, AUG 13 2020. (17/13994-5, 15/10606-9)

MELO, G. D.; PINTO, L. M. F.; ROCHA, C. C.; MOTTA, I. G.; SILVA, L. A.; DA SILVEIRA, J. C.; GONELLA-DIAZA, A. M.; BINELLI, M.; PUGLIESI, G.. Type I interferon receptors and interferon-tau-stimulated genes in peripheral blood mononuclear cells and polymorphonuclear leucocytes during early pregnancy in beef heifers. REPRODUCTION FERTILITY AND DEVELOPMENT, v. 32, n. 11, p. 953-966, 2020. (17/13472-9, 15/10606-9)

DE MELO, GABRIELA DALMASO; MELLO, BARBARA PIFFERO; FERREIRA, CATIA APARECIDA; SOUTO GODOY FILHO, CARLOS ALBERTO; ROCHA, CECILIA CONSTANTINO; SILVA, AMANDA GUIMARAES; REESE, SYDNEY TAYLOR; MADUREIRA, ED HOFFMANN; POHLER, KY GARRETT; PUGLIESI, GUILHERME. Applied use of interferon-tau stimulated genes expression in polymorphonuclear cells to detect pregnancy compared to other early predictors in beef cattle. Theriogenology, v. 152, p. 94-105, AUG 2020. (17/13472-9, 18/20108-4, 15/10606-9)

BARBOZA DA SILVA, JULIO CESAR; RODRIGUES ALVES, MAIRA BIANCHI; BRIDI, ALESSANDRA; BOHRER, RODRIGO CAMPONOGARA; LAMBERTI ESCOBAR, GABRIELA SABINE; BERNARDES AFONSO DE CARVALHO, JUNIA APARECIDA; BOVI BINOTTI, WALTER ALEXANDRE; PUGLIESI, GUILHERME; LEMES, KLEBER MENEGON; CHELLO, DAMIANA; et al. Reproductive seasonality influences oocyte retrieval and embryonic competence but not uterine receptivity in buffaloes. Theriogenology, v. 170, p. 77-84, AUG 2021. (15/10606-9)

ROCHA, CECILIA CONSTANTINO; DA SILVEIRA, JULIANO COELHO; FORDE, NIAMH; BINELLI, MARIO; PUGLIESI, GUILHERME. Conceptus-modulated innate immune function during early pregnancy in ruminants: a review. ANIMAL REPRODUCTION, v. 18, n. 1, p. 16-pg., 2021-01-01. (19/02995-6, 17/13994-5, 15/10606-9)

CECILIA CONSTANTINO ROCHA; JULIANO COELHO DA SILVEIRA; NIAMH FORDE; MARIO BINELLI; GUILHERME PUGLIESI. Conceptus-modulated innate immune function during early pregnancy in ruminants: a review. Animal Reproduction, v. 18, n. 1, 2021-05-10. (15/10606-9, 19/02995-6, 17/13994-5)

OJEDA-ROJAS, OSCAR ALEJANDRO; GONELLA-DIAZA, ANGELA MARIA; BUSTOS-CORAL, DANIEL; SARTORELLO, GUSTAVO L.; REIJERS, THAYLA S. S. S.; PUGLIESI, GUILHERME; ZERLOTTI MERCADANTE, MARIA EUGENIA; DE LIMA, CESAR G.; GAMEIRO, AUGUSTO H.. An agent-based simulation model to compare different reproductive strategies in cow-calf operations: Technical performance. Theriogenology, v. 160, p. 102-115, JAN 15 2021. (15/10606-9)

PUGLIESI, GUILHERME; DE MELO, GABRIELA DALMASO; SILVA, JULIO BARBOZA; CARVALHEDO, ALEXANDRE SARDINHA; LOPES, EVERTON; DE SIQUEIRA FILHO, EMIVALDO; SILVA, LUCIANO ANDRADE; BINELLI, MARIO. Use of color-Doppler ultrasonography for selection of recipients in timed-embryo transfer programs in beef cattle. Theriogenology, v. 135, p. 73-79, SEP 1 2019. (16/23964-3, 15/10606-9, 17/13472-9)

PUGLIESI, GUILHERME; DE MELO, GABRIELA DALMASO; ATAIDE, JR., GILMAR ARANTES; GONTIJO PELLEGRINO, CARLOS AUGUSTO; SILVA, JULIO BARBOZA; ROCHA, CECILIA CONSTANTINO; MOTTA, IGOR GARCIA; MORAES VASCONCELOS, JOSE LUIZ; BINELLI, MARIO. Use of Doppler ultrasonography in embryo transfer programs: feasibility and field results. ANIMAL REPRODUCTION, v. 15, n. 3, p. 239-246, JUL-SEP 2018. (17/18613-0, 16/23964-3, 15/10606-9, 17/13472-9)

MOTTA, I. G.; ROCHA, C. C.; BISINOTTO, D. Z.; MELO, C. D.; ATAIDE JUNIOR, G. A.; SILVA, A. G.; GONZAGA, V. H. G.; SANTOS, J. A.; FREITAS, B. G.; LEMES, K. M.; et al. Increased pregnancy rate in beef heifers resynchronized with estradiol at 14 days after TAI. Theriogenology, v. 147, p. 62-70, APR 15 2020. (17/18613-0, 15/10606-9)

DA SILVA, AMANDA GUIMARAES; NISHIMURA, THIAGO KAN; ROCHA, CECILIA CONSTANTINO; MOTTA, IGOR GARCIA; NETO, ADOMAR LAURINDO; FERRAZ, PRISCILA ASSIS; BRUNI, GABRIELA ANDRADE; ORLANDI, RAPHAEL EVANGELISTA; MARTINELLI MASSONETO, JOAO PAULO; PUGLIESI, GUILHERME. Comparison of estradiol benzoate doses for resynchronization of ovulation at 14 days after timed-AI in suckled beef cows. Theriogenology, v. 184, p. 10-pg., 2022-03-08. (19/07805-0, 15/10606-9)

ROCHA, CECILIA CONSTANTINO; MARTINS, THIAGO; MELLO, BARBARA PIFFERO; DE MELLO, GABRIELA DALMASO; MOTTA, IGOR GARCIA; LEMES, KLEBER MENEGON; BINELLI, MARIO; MADUREIRA, ED HOFFMANN; PUGLIESI, GUILHERME. Comparing the effect of estradiol benzoate and 17 beta-estradiol plus progesterone on follicular turnover and development, and pregnancy outcomes in a timed artificial insemination protocol. Theriogenology, v. 192, p. 8-pg., 2022-10-15. (17/13994-5, 15/10606-9)

MOTTA, I. G.; ROCHA, C. C.; BISINOTTO, D. Z.; MELO, G. D.; JUNIOR, G. A. . A. .; NISHIMURA, T. K.; DIAZA, A. M. G.; CASTRO, T.; GINTHER, O. J.; PUGLIESI, G.. Effects of estradiol treatments on PGF(2 alpha) release in beef heifers submitted to estrous resynchronization 14 days after timed-AI. DOMESTIC ANIMAL ENDOCRINOLOGY, v. 76, JUL 2021. (17/18613-0, 15/10606-9)