From parturition to pregnancy. A clinical perspective in the dairy cow. Irene López Helguera - PDF

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From parturition to pregnancy. A clinical perspective in the dairy cow. Irene López Helguera From parturition to pregnancy. A clinical perspective in the dairy cow. DISSERTATION To obtain the degree of

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From parturition to pregnancy. A clinical perspective in the dairy cow. Irene López Helguera From parturition to pregnancy. A clinical perspective in the dairy cow. DISSERTATION To obtain the degree of Doctor at the University of Lleida by Irene López Helguera June 2012 Department de Producció Animal Escola Técnica Superior d Enginyeria Agrària Universitat de Lleida Director: Irina García Ispierto Co-Director: Fernando López Gatius Asssesment committe External Evaluation Susana Astiz Blanco (INIA, Madrid, ES) Gregori Bech Sàbat (Univerisitat de Lleida Phd, ES) Board members Fabio de Rensis (Università degli Studi di Parma, IT) Beatriz Serrano Pérez (Universitat de Lleida, ES) Gregori Bech Sàbat (Univerisitat de Lleida Phd, ES) Alternate 1: Carmina Nogadera Burch (Universitat de Lleida, ES) Alternate 2: Alex Martino Alerm (CEVA SANTE ANIMALE S.A) Cover pictures: First row, from left to right: 7.5 MHz- Ultrasonography of a cross section of cervical diameter, ovarian structures (two corpus luteum and a follicle) and cross section of a uterine horn. Second row, from left to right: 7.5 MHz- Ultrasonography of a cross section of a uterine horn with echogenic intrauterine fluid, increased endometrial thickness and echogenic intrauterine fluid, respectively. I am among those who think that science has great beauty. A scientist in his laboratory is not only a technician: he is also a child placed before natural phenomena which impress him like a fairy tale. Marie Curie INDEX Summary English... 7 Català... 9 Castellano General Introduction Main objectives Studies Chapter The influence of genital tract status in postpartum period on the subsequent reproductive performance in high producing dairy cows Chapter Therapy hormonal treatment for abnormal uterine involution on high producing dairy cows. Chapter Is intrauterine therapy of abnormal uterine involution useful in high producing dairy cows? Chapter Reproductive performance of anoestrous high-producing dairy cows improved by adding equine chorionic gonadotrophin to a progesterone-based oestrus synchronization protocol. General Discussion Conclusions Epilogue Appendix Acknowledgements Curriculum vitae SUMMARY SUMMARY Nutritional and genetic improvements in last decades have not prevented the decrease in reproductive performance of dairy cows. For that reason, great effort has been focused on understanding pathological processes during early postpartum. Therefore, the aims of this thesis were to implement a diagnostic and therapeutic protocol on Days postpartum that detect cows suffering endometritis and to improve subsequent reproductive performance in high producing dairy cows. Moreover, since there is still no one single gold protocol to synchronize lactating dairy cows, progesterone based synchronization protocol plus equine chorionic gonadotrophin (ecg) was tested. The research included in this thesis was divided in four studies that are summarized below. In the first study, cows suffering postpartum endometritis on Days were characterized by ultrasonography, endometrial cytology and blood tests. Measures of cervical diameter, endometrial thickness and echogenicity of intrauterine fluid (IUF) by ultrasonography on Days 15 to 21 postpartum were a good tool to predict subsequent reproductive performance in clinically normal dairy cows. In the second one, the effect of Prostaglandin F2 α on Day postpartum on subsequent conception rate was evaluated. No effect of treatment with prostaglandin was observed on subsequent reproductive performance even in cyclic cows. Early resumption of ovarian cyclicity enhanced postpartum recovery and improved subsequent conception rate. In the third study, the effect of a single intrauterine infusion of penicillin or large-volume lavage with normal saline solution on cows suffering endometritis on Days postpartum were assessed. No effect was observed of treatment on the subsequent conception rate or return to cyclicity. Factors such as calving season and parity were intimately related to reproductive performance. In the last work, a progesterone based synchronization protocol plus ecg was tested. The placement of a progesterone intravaginal device followed by ecg treatment given on the day of device removal was beneficial in anoestrous cows even during the postpartum period. Irrespective of the treatment, cows undergoing double ovulation were more fertile than the remaining cows. 7 8 SUMMARY RESUM Les millores en genètica i nutrició de les últimes dècades, no han impedit que es produís una disminució de la funció reproductiva en les vaques lleteres d alta producció. Per aquest motiu, entendre els processos fisiopatològics durant el postpart ha estat el principal motiu d atenció de les grnages lleteres. Per tant, l objectiu d aquesta tesi va ser implementar un protocol diagnòstic i terapèutic en vaques lleteres a dies postpart per detectar animals que patien endometritis i millorar la seva eficàcia reproductiva posterior. A més, degut a que no existeix un protocol estandaritzat per sincronitzar vaques lleteres, es va implementar un protocol basat en progesterona intravaginal més gonadotrofina coriònica equina (ecg). La recerca inclosa en aquesta tesis està dividida en cuatre estudis: En el primer, es van caracteritzar animals que patien endometritis a dies postpart utilitzant ecografia transrectal, citologia uterina i análisi de cèl lules sanguínies. Les mesures de tamany cervical, gruix endometrial i ecogenicitat del fluid intrauterí caracteritzades per ecografia van resultar ser una eina útil per predir la funció reproductiva posterior en vaques clínicament normals. En el segon, es va estudiar l efecte de la prostaglandina F2 α a dies 15-21postpart sobre la fertilitat posterior. No es va observar efecte del tratament ni tan sols en les vaques que ja eren cícliques. El precoç retorn a la ciclicitat millorava la recuperació postpart i millorava la posterior fertilitat. En el tercer estudi, es va establir l efecte d un rentat intrauterí amb penicilina o sèrum salí fisiologic en vaques que patien endometritis a dies postpart. No es va observar efecte del tractament ni a la posterior fertilitat ni al retorn a la ciclicitat. Factors com la estació de part o la lactació resultaren intimament relacionades amb la funció reproductiva. Per últim, es va establir un protocol basat en progesterona intravaginal i ecg el dia de la retirada. Aquest protocol va ser benificiós tant en vaques que patien llarg anestre com en animals amb anestre postpart. Independentment del tractament, les vaques amb doble ovulació van ser més fèrtils que la resta. 9 10 SUMMARY RESUMEN La mejora genética y nutricional de las últimas décadas no ha logrado frenar la disminución del rendimiento reproductivo en las vacas de leche. Los procesos fisiopatológicos que ocurren durante el postparto son esenciales para la fertilidad posterior, siendo actualmente un importante punto de estudio. Por lo tanto, el objetivo de esta tesis fue establecer un diagnóstico y un protocolo terapéutico en los días del postparto para detectar las vacas que sufren endometritis y mejorar la eficacia reproductiva en vacas lecheras de alta producción. Por otra parte, debido a que todavía no hay un único protocolo estandar para sincronizar la vacas de leche en lactación, se estableció un protocolo basado en la sincronización con progesterona aplicando gonadotropina coriónica equina (ecg). La investigación incluida en esta tesis se divide en cuatro estudios que se resumen a continuación. En el primer estudio, se analizaron vacas con endometritis postparto entre los días por ecografía, citologías y análisis de sangre. Las medidas del diámetro cervical, el grosor endometrial y el fluido intrauterino ecogénico (IUF) por ecografía entre los días 15 al 21 de postparto, resultaron ser una buena herramienta para predecir la posterior eficacia reproductiva en vacas de leche clínicamente sanas. En el segundo estudio, se evaluaron los efectos de la prostaglandina F2 α en los días postparto sobre la posterior tasa de concepción. No se observó efecto del tratamiento ni siquiera en las vacas cíclicas. El pronto retorno de la ciclicidad ovárica después del parto se relacionó positivamente con la recuperación postparto y la tasa de concepción posterior. En el tercer estudio, se evaluraron los efectos de un único lavado intrauterino con penicilina o suero fisiológico en vacas con endometritis a días postparto. No se observó efecto del tratamiento ni en la posterior fertilidad ni en el retorno a la cilcicidad. Factores como la estación del parto o el número de lactación estuvieron íntimamante relacionados con la función reproductiva. En el último trabajo, se estableció un protocolo basado en progesterona intravaginal y ecg el día de la retirada. Este protocolo resultó beneficioso, tanto en vacas con anestro largo como en animales con anestro postparto. Independientemente del tratamiento, la vacas con ovulación doble fueron más fértiles que el resto. 11 12 INTRODUCTION 13 14 INTRODUCTION INTRODUCTION Management and genetics of dairy cows have been improving along years, leading to a continuous increase in milk production (Dobson et al. 2007; Yániz et al. 2008). However, since 1980s subfertility has become an emerging disease (Lucy 2001; López-Gatius 2003), reducing farm productivity. For instance, economic consequences of non-optimal fertility are estimated at Euros per cow/year (Inchaisri et al. 2010). It is well known, that high milk production correlates negatively with fertility (Pryce et al. 1998). However, this relationship is difficult to demonstrate because of the interference of culling and the environment (Montaldo et al. 2010). A high milk yield will only provoke a higher risk of infertility under suboptimal conditions (Garcia- Ispierto et al. 2007), such as postpartum period (Walsh et al. 2011). For that reason, great effort has been focused on understanding physiological changes and pathological processes during postpartum and voluntary waiting period. Nevertheless, too much questions are not clearly understood yet. The fetal hypothalamo-pituitary-adrenal axis stimulates the production of corticoids following nine months of gestation. Then, a massive increase in cortisol and decrease in progesterone triggers the onset of parturition (Senger 1997). These endocrine patterns determine the number and function of immune cells in utero that potentially affects uterine defense mechanisms postpartum (Singh et al. 2008). The potentially immunosupressed animal has to suffer for several weeks after parturition a cycle of bacterial contamination, clearance, and recontamination (Griffin et al. 1984; Sheldon and Dobson 2004). Whether these animals in negative energy balance due to their reduced food intake and increased milk production, are not able to resolve this natural contamination, their subsequent reproductive performance will be substantially impaired (Hammon et al. 2006; Goff 2008). Before a cow is likely to conceive again, it has to reach two main objectives: undergo uterine involution and return to ovarian cyclicity (Morrow et al. 1969, Sheldon et al. 2008). Placenta has to be expelled before 12 hours, although loquial discharge could be present during the first two weeks after calving and endometrial regeneration is not completed until 6-8 weeks postpartum (Sheldon 2004). However, during pregnancy, 15 the hypothalamic pituitary axis is suppressed by steroidal hormones, reducing the secretion of luteinizing hormone (LH). After parturition, steroidal hormones return to basal levels and ovarian dynamics should be reestablished (Crowe 2008). Both phenomena, uterine involution and return to cyclicity, greatly interact so that early postpartum ovarian activity could help to bacterial uterine elimination because of the increase of estrogens (Hussain 1989; Lewis 2004). Despite of the positive effect of estrus, an early ovulation may provoke an untimely cervical closure leading to a pyometra (Olson et al. 1984). The persistence of bacterial uterine contamination could not only damage endometrial tissue and compromise its ability to produce or transport PGF 2α, but also suppresses GnRH and LH secretion (Olson et al. 1984; Sheldon and Dobson 2004). Diagnosing and treating postpartum diseases is essential for the subsequent reproductive performance of high producing dairy cows. Clinical diseases, such as puerperal metritis are easily diagnosed by observing clinical sings like fetid red-brown watery uterine discharge and pirexia within 10 days after parturition (Sheldon et al. 2006). However, on the following days, diagnosis can be much more difficult. Endometritis, inflammation of the endometrium without affecting the remaining uterine layers, is usually diagnosed by vaginoscope or observing vaginal discharge after day 21 postpartum (LeBlanc et al. 2002; Sheldon et al. 2006). Thus, this uterine pathology can be underestimated when the cervix closes rapidly after parturition (Kasimanickam et al. 2004). Consequently, the necessary questions are: What is exactly endometritis and what are its reproductive consequences? When could a clinician start to diagnose an inadequate uterine involution? Which is the best diagnostic method to detect endometritis? Veterinary efforts have been focused on checking dairy cows at first week postpartum for metritis and after 21 days in milk for endometritis (Sheldon et al. 2006; Chapwanya et al. 2008; LeBlanc et al. 2008). To the best of our knowledge, no research has been made on days postpartum to characterize clinical findings related to subsequent impaired fertility. Moreover, although endometrial biopsy is the gold standard for diagnosing the degree of uterus inflammation (Bonnett et al. 1991; Chapwanya et al. 2009), it is not a clinically used technique due to the difficulty of procedure and sample processing. Nowadays, other practices such as endometrial cytology or ultrasonography are being used to determine uterine health (Ginther 1998; Barlund et al. 2008, Kasimanickam et al. 2004, Gnemmi and Maraboli 2010). 16 INTRODUCTION Once postpartum uterus disease is detected, it is crucial to settle the best and faster therapeutic protocol. Hormone products are one of the daily used procedures to improve uterine health. Natural prostaglandin, due to its luteolytic and uterotonic activity has become a routine treatment during postpartum period (Rodriguez-Martinez et al. 1987; Pankowski et al. 1995). The aim of this treatment is to reduce bacterial and purulent content by uterine contractions and enhance uterine defense (Paisley et al. 1986). However, PGF 2α field trials reveal contradictory results that question its efficiency (Young and Anderson 1986; López-Gatius and Camón-Urgel 1989; Hendricks et al. 2006; Salasel and Mokhtari 2011). Antibiotic administration is another traditional treatment used after calving (Paisley et al. 1986). In contrast to systemic administration, intrauterine infusions achieve higher concentration of antibiotic on genital mucosa (Bretzlaff et al. 1983). Thus, several intrauterine protocols have been designed to solve uterine infection. For example, tetracycline (Goshen and Shpigel 2006), penicillin (Thurmond et al. 1993) and cephapirin (Galvão et al. 2009) are three of the most common used drugs. The critical point in all these therapeutic procedures is the lack of a control group in many of them. As a result, contradictory outcomes can be observed and veterinary postpartum treatments cannot be easily compared (Burton and Lean 1995; Lefebvre and Stoc, 2012). Finally, after the voluntary waiting period the cow has to conceive again. Fertility has been associated to management, environment or disease related factors (López-Gatius 2003, 2011; Garcia-Ispierto et al. 2007). For instance, management practices at this time have to be carefully performed to increase effectiveness of artificial insemination. One key point to improve fertility is the application of synchronization protocols (Larson and Ball 1992; De Rensis and Peters 1991; Wiltbank et al. 2011). These hormone combinations not only synchronize ovulations leading to a timed artificial insemination (Souza et al. 2009; Butler et al. 2011), but also can increase the estrous behavior and subsequent conception rate (Yániz et al. 2004; García- Ispierto et al. 2010,2012), specially during the warm season. There are two mainly types of protocols: based on PGF 2α, such as Ovsynch (Pursley et al. 1997), and on progesterone-releasing intravaginal device. The main difference between these protocols is that the first one is more effective when cows are cyclic (Murugavel et al. 2003), and that progesterone protocols can also synchronize acyclic cows. Thus, 17 progesterone-based treatments can be proposed as a therapeutical protocol to synchronize all animals, but nowadays there is still no one single standard protocol to synchronize all kind of lactating dairy cows. Progesterone based synchronizing protocols have been modified by adding hormones like gonadotrophin-releasing hormone (GnRH) or human chorionic gonadotrophin (hcg) in dairy cattle (DeRensis et al. 2002, 2010). Equine chorionic gonadotrophin (ecg) displays both FSH and LH-like activity (Bevers et al. 1989, Papkoff 1998) and seems to enhance follicular growth and corpus luteum development (Rostami et al. 2011; Silva et al 2002). Protocols combining PRID and ecg have only been tested in beef cattle, and frequently combined with estradiol, product not allowed in EU nowadays (Barruselli et al. 2003; Sá Filho et al. 2010). In summary, this thesis scoped to improve reproductive efficiency of high producing dairy cows by controlling the postpartum period and implementing a new estrous synchronization protocol. 18 INTRODUCTION REFERENCES Barlund CS, Carruthers TD, Waldner CL, Palmer CW. A comparison of diagnostic techniques for postpartum endometritis in dairy cattle. Theriogenology 2008;69: Baruselli PS, Marques MO, Nasser LFT, Reis EL, Bó GA. Effect of ecg on pregnancy rates of lactating zebu beef cows treated with CIDR-B devices for timed artificial insemination [abstract]. Theriogenology 2003;59:214. Bevers MM, Dieleman SJ, Van Tol HTM, Blankenstein DM, Van den Broek J. Changes in pulsatile secretion patterns of LH, FSH, progesterone, androstenedione and oestradiol in cows after superovulation with PMSG. J Reprod Fert 1989;87: Bonnett BN, Miller RB, Etherington WG, Martin SW, Johnson WH. Endometrial biopsy in Holstein-Friesian dairy cows. I. Technique, histological criteria and results. Can J Vet Res 1991;55: Bretzlaff KN, Ott RS, Koritz GD, Bevill RFB, Gustafsson K, Davis LE. Distribution of oxytetracycline in genital tract tissues of postpartum cows given the drug by intravenous and intrauterine routes. Am J Vet Res1983;44: Burton NR, Lean IJ. Investigation by meta-analysis of the effect of prostaglandin F2α administered post partum on the reproductive performance of dairy cattle. Vet Rec. 1995;136: Butler SAA, Atkinson PC, Boe-Hansena GB, Burnsc BM, Dawsond K, Boe GA, McGowan MR. Pregnancy rates after fixed-time artificial insemination of Brahman heifers treated to synchronize ovulation with low-dose intravaginal progesterone releasing devices, with or without ecg. Theriogenology 2011;76: Chapwanya A. Uterine disease in dairy cows: Classification, diagnosis and key roles for veterinarians. Ir Vet J 2008;61: Chapwanya A, Meade KG, Doherty ML, Callanan JJ, Mee JF, O Farrelly C. Histopathological and molecular evaluation of Holstein-Friesian cows postpartum: Toward an improved understanding of uterine innate immunity. Theriogenology 2009;71: Crowe MA. Resumption of ovarian cyclicity in postpartum beef and dairy cows. Reprod Dom Anim 2008;43: De Rensis F, Peter A. The control of follicular dynamics b PGF2a, GnRH, hcg and estrus synchronization in cattle.
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