Clinical endometritis with Trueperella pyogenes reduces reproductive performance and milk production in dairy cows.

The aim of this study was to identify the impact of Trueperella pyogenes in cows with clinical endometritis (CE) on reproductive performance and milk production in affected cows. In total, 230 lactating Holstein dairy cows from six commercial dairy herds were sampled once between 28 and 33 days post-partum. Cows included in the present study did not receive antibiotic or anti-inflammatory treatments prior to the experimental period. Clinical endometritis (CE) was characterized as cow with vaginal mucus score = 3 (>50% of purulent vaginal discharge) and >18% polymorphonuclear neutrophilic leukocyte (PMNL). The body condition scores (BCS) and milk production were evaluated at the time of enrolment. The identification of isolated bacteria was carried out through the analysis of MALDI-TOF MS (matrix-assisted laser desorption ionization time-of-flight mass spectrometry). According to uterine health, three groups of dairy cows were formed: healthy control cows without T. pyogenes (n = 147), CE cows with T. pyogenes (n = 22) and CE cows without T. pyogenes (n = 61). CE cows with T. pyogenes had lower BCS, milk production and conception at first AI (p < .01) than CE cows without T. pyogenes and control cows. Furthermore, CE cows with T. pyogenes had higher (p < .01) service per pregnancy and had greater (p < .01) days to get pregnant than CE cows without T. pyogenes and control cows. This study demonstrates that CE cows with T. pyogenes had impaired reproductive performance and milk production when compared to cows without CE and CE cows without T. pyogenes. This information can contribute to a strategic treatment in cows affected by clinical endometritis, favouring the rational use of antibiotics on dairy farms.

Impact of oocyte donor age and breed on in vitro embryo production in cattle, and relationship of dairy and beef embryo recipients on pregnancy and the subsequent performance of offspring: A review.

Genomic selection combined with in vitro embryo production (IVEP) with oocytes from heifer calves provides a powerful technology platform to reduce generation interval and significantly increase the rate of genetic gain in cattle. The ability to obtain oocytes with developmental competence from calves has been recognised for more than 60years. There is renewed interest in the potential of this reproductive technology as the beef and dairy industries seek to identify and multiply animals that satisfy consumer demand for efficient utilisation of natural resources, with minimal environmental impact and high product quality. Heifer calves show a good follicular response to FSH stimulation and oocytes can be recovered by ovum pick-up (OPU). However, the efficiency of OPU/IVEP remains lower for calves compared with peripubertal heifers and cows, in both indicus (Zebu, Bos indicus ) and taurus (Bos taurus ) breeds. Indicus cattle generally have more follicles, yield a greater number of oocytes, and have a better IVEP outcome, compared with taurus cattle. The treatment of prepubertal heifers with bovine somatotrophin (bST) and propylene glycol before OPU/IVEP has yet to show a major improvement in embryo production. Holstein (taurus) dairy heifers derived from different reproductive technologies (AI, MOET, OPU/IVEP) showed a similar age at puberty and first-lactation milk production. OPU/IVEP Holstein embryos transferred to beef or dairy cows likewise yielded heifers with the same performance. The gains in productivity that can be achieved with strategic use of OPU/IVEP in heifer calves make this a relevant and highly important reproductive technology in cattle breeding. Ongoing optimisation of the technology is needed for the potential of OPU/IVEP in young donors to be fully realised.

Assisted reproductive technologies (ART) in water buffaloes.

Our expanding knowledge of ovarian function during the buffalo estrous cycle has given new approaches for the precise synchronization of follicular development and ovulation to apply consistently assisted reproductive technologies (ART). Recent synchronization protocols are designed to control both luteal and follicular function and permit fixed-time AI with high pregnancy rates during the breeding (autumn-winter) and nonbreeding (spring- summer) seasons. Additionally, allow the initiation of superstimulatory treatments at a self-appointed time and provide opportunities to do fixed-time AI in donors and fixed-time embryo transfer in recipients. However, due the scarce results of in vivo embryo recovery in superovulated buffaloes, the association of ovum pick-up (OPU) with in vitro embryo production (IVEP) represents an alternative method of exploiting the genetics of high yeld buffaloes. Nevertheless, several factors appear to be critical to OPU/IVEP efficiency, including antral follicle population, follicular diameter, environment, farm and category of donor. This review discusses a number of key points related to the manipulation of ovarian follicular growth to improve assisted reproductive technologies in buffalo.

Programs for fixed-time artificial insemination in South American beef cattle.

Fixed-time artificial insemination (FTAI) has been widely applied in South America within the last 20 years for the genetic improvement of commercial beef herds. Most FTAI treatments for beef cattle used in South America are based on the use of progesterone (P4) releasing devices and estradiol to synchronize follicle wave emergence, with pregnancies per AI (P/AI) ranging from 40 to 60%. More recent protocols focusing on extending the interval from device removal to FTAI (i.e. increasing the growing period of the ovulatory follicle) have been reported to improve P/AI in beef cattle. These new protocols and the more traditional FTAI protocols have also been adapted for use with sexed-sorted semen with acceptable P/AI in beef cattle. Finally, color-flow Doppler ultrasonography has been incorporated recently to determine the vascularity of the CL and thereby detect pregnancy as early as Day 22 after the first AI for re- synchronization of ovulation for a second FTAI in non- pregnant animals. In summary, FTAI protocols have facilitated the widespread application of AI in South American beef cattle by allowing for the insemination and re-insemination of herds during a defined breeding season, without the necessity of clean up bulls to achieve high pregnancy rates.

Evolution of knowledge on ovarian physiology and its contribution to the widespread application of reproductive biotechnologies in South American cattle.

As our understanding of ovarian function in cattle has improved, our ability to control it has also increased. Luteal function in cattle has been studied in detail, and prostaglandin F2α has been used for several years for the elective induction of luteal regression. More recently, follicle wave dynamics has been studied and protocols designed to induce follicular wave emergence and ovulation have reduced, and even eliminated, the need for estrus detection. The addition of progestin-releasing devices, estradiol, GnRH and equine chorionic gonadotropin (eCG) have provided opportunities for fixed-time AI (FTAI) and possibilities for increased pregnancy rates. In embryo transfer programs, these same treatments have eliminated the need for estrus detection, permitting fixed-time embryo transfer and the initiation of superstimulatory treatments without regard to day of the estrous cycle. Collectively, new treatment protocols have facilitated the application of assisted reproductive technologies, and this is especially true in South America. Over the last 20 years, the use of AI in South America has increased, due largely to the use of FTAI. There has been more than a 10-fold increase in the use of FTAI in Brazil with more than 11 million treatments in 2016, representing 85% of all AI. Similar trends are occurring in Argentina and Uruguay. Production of in vivo-derived (IVD) embryos has remained relatively stable over the years, but in vitro embryo production (IVP) has increased dramatically over the past 10 to 15 years, especially in Brazil where more than 300,000 IVP embryos were produced in 2010. World-wide, more than 666,000 bovine IVP embryos were produced in 2016, of which more than 57% were produced in South America. The use of assisted reproductive technologies has facilitated the dissemination of improved genetics and increased reproductive performance; other South American countries are now following suit.

Assisted reproductive technologies (ART) in water buffaloes

Our expanding knowledge of ovarian function during the buffalo estrous cycle has given new approaches for the precise synchronization of follicular development and ovulation to apply consistently assisted reproductive technologies (ART). Recent synchronization protocols are designed to control both luteal and follicular function and permit fixed-time AI with high pregnancy rates during the breeding (autumn-winter) and nonbreeding (springsummer) seasons. Additionally, allow the initiation of superstimulatory treatments at a self-appointed time and provide opportunities to do fixed-time AI in donors and fixed-time embryo transfer in recipients. However, due the scarce results of in vivo embryo recovery in superovulated buffaloes, the association of ovum pick-up (OPU) with in vitro embryo production (IVEP) represents an alternative method of exploiting the genetics of high yeld buffaloes. Nevertheless, several factors appear to be critical to OPU/IVEP efficiency, including antral follicle population, follicular diameter, environment, farm and category of donor. This review discusses a number of key points related to the manipulation of ovarian follicular growth to improve assisted reproductive technologies in buffalo.(AU)

Evolution of knowledge on ovarian physiology and its contribution to the widespread application of reproductive biotechnologies in South American cattle

As our understanding of ovarian function in cattle has improved, our ability to control it has also increased. Luteal function in cattle has been studied in detail, and prostaglandin F2α has been used for several years for the elective induction of luteal regression. More recently, follicle wave dynamics has been studied and protocols designed to induce follicular wave emergence and ovulation have reduced, and even eliminated, the need for estrus detection. The addition of progestin-releasing devices, estradiol, GnRH and equine chorionic gonadotropin (eCG) have provided opportunities for fixed-time AI (FTAI) and possibilities for increased pregnancy rates. In embryo transfer programs, these same treatments have eliminated the need for estrus detection, permitting fixed-time embryo transfer and the initiation of superstimulatory treatments without regard to day of the estrous cycle. Collectively, new treatment protocols have facilitated the application of assisted reproductive technologies, and this is especially true in South America. Over the last 20 years, the use of AI in South America has increased, due largely to the use of FTAI. There has been more than a 10-fold increase in the use of FTAI in Brazil with more than 11 million treatments in 2016, representing 85% of all AI. Similar trends are occurring in Argentina and Uruguay. Production of in vivo-derived (IVD) embryos has remained relatively stable over the years, but in vitro embryo production (IVP) has increased dramatically over the past 10 to 15 years, especially in Brazil where more than 300,000 IVP embryos were produced in 2010. World-wide, more than 666,000 bovine IVP embryos were produced in 2016, of which more than 57% were produced in South America. The use of assisted reproductive technologies has facilitated the dissemination of improved genetics and increased reproductive performance; other South American countries are now following suit.(AU)

Programs for fixed-time artificial insemination in South American beef cattle

Fixed-time artificial insemination (FTAI) has been widely applied in South America within the last 20 years for the genetic improvement of commercial beef herds. Most FTAI treatments for beef cattle used in South America are based on the use of progesterone (P4) releasing devices and estradiol to synchronize follicle wave emergence, with pregnancies per AI (P/AI) ranging from 40 to 60%. More recent protocols focusing on extending the interval from device removal to FTAI (i.e. increasing the growing period of the ovulatory follicle) have been reported to improve P/AI in beef cattle. These new protocols and the more traditional FTAI protocols have also been adapted for use with sexed-sorted semen with acceptable P/AI in beef cattle. Finally, color-flow Doppler ultrasonography has been incorporated recently to determine the vascularity of the CL and thereby detect pregnancy as early as Day 22 after the first AI for resynchronization of ovulation for a second FTAI in nonpregnant animals. In summary, FTAI protocols have facilitated the widespread application of AI in South American beef cattle by allowing for the insemination and re-insemination of herds during a defined breeding season, without the necessity of clean up bulls to achieve high pregnancy rates.(AU)

Programs for fixed-time artificial insemination in South American beef cattle

Fixed-time artificial insemination (FTAI) has been widely applied in South America within the last 20 years for the genetic improvement of commercial beef herds. Most FTAI treatments for beef cattle used in South America are based on the use of progesterone (P4) releasing devices and estradiol to synchronize follicle wave emergence, with pregnancies per AI (P/AI) ranging from 40 to 60%. More recent protocols focusing on extending the interval from device removal to FTAI (i.e. increasing the growing period of the ovulatory follicle) have been reported to improve P/AI in beef cattle. These new protocols and the more traditional FTAI protocols have also been adapted for use with sexed-sorted semen with acceptable P/AI in beef cattle. Finally, color-flow Doppler ultrasonography has been incorporated recently to determine the vascularity of the CL and thereby detect pregnancy as early as Day 22 after the first AI for resynchronization of ovulation for a second FTAI in nonpregnant animals. In summary, FTAI protocols have facilitated the widespread application of AI in South American beef cattle by allowing for the insemination and re-insemination of herds during a defined breeding season, without the necessity of clean up bulls to achieve high pregnancy rates.

Assisted reproductive technologies (ART) in water buffaloes

Our expanding knowledge of ovarian function during the buffalo estrous cycle has given new approaches for the precise synchronization of follicular development and ovulation to apply consistently assisted reproductive technologies (ART). Recent synchronization protocols are designed to control both luteal and follicular function and permit fixed-time AI with high pregnancy rates during the breeding (autumn-winter) and nonbreeding (springsummer) seasons. Additionally, allow the initiation of superstimulatory treatments at a self-appointed time and provide opportunities to do fixed-time AI in donors and fixed-time embryo transfer in recipients. However, due the scarce results of in vivo embryo recovery in superovulated buffaloes, the association of ovum pick-up (OPU) with in vitro embryo production (IVEP) represents an alternative method of exploiting the genetics of high yeld buffaloes. Nevertheless, several factors appear to be critical to OPU/IVEP efficiency, including antral follicle population, follicular diameter, environment, farm and category of donor. This review discusses a number of key points related to the manipulation of ovarian follicular growth to improve assisted reproductive technologies in buffalo.

Evolution of knowledge on ovarian physiology and its contribution to the widespread application of reproductive biotechnologies in South American cattle

As our understanding of ovarian function in cattle has improved, our ability to control it has also increased. Luteal function in cattle has been studied in detail, and prostaglandin F2α has been used for several years for the elective induction of luteal regression. More recently, follicle wave dynamics has been studied and protocols designed to induce follicular wave emergence and ovulation have reduced, and even eliminated, the need for estrus detection. The addition of progestin-releasing devices, estradiol, GnRH and equine chorionic gonadotropin (eCG) have provided opportunities for fixed-time AI (FTAI) and possibilities for increased pregnancy rates. In embryo transfer programs, these same treatments have eliminated the need for estrus detection, permitting fixed-time embryo transfer and the initiation of superstimulatory treatments without regard to day of the estrous cycle. Collectively, new treatment protocols have facilitated the application of assisted reproductive technologies, and this is especially true in South America. Over the last 20 years, the use of AI in South America has increased, due largely to the use of FTAI. There has been more than a 10-fold increase in the use of FTAI in Brazil with more than 11 million treatments in 2016, representing 85% of all AI. Similar trends are occurring in Argentina and Uruguay. Production of in vivo-derived (IVD) embryos has remained relatively stable over the years, but in vitro embryo production (IVP) has increased dramatically over the past 10 to 15 years, especially in Brazil where more than 300,000 IVP embryos were produced in 2010. World-wide, more than 666,000 bovine IVP embryos were produced in 2016, of which more than 57% were produced in South America. The use of assisted reproductive technologies has facilitated the dissemination of improved genetics and increased reproductive performance; other South American countries are now following suit.

Microvascularization of corpus luteum of bovine treated with equine chorionic gonadotropin.

This study aimed to evaluate the morphological changes in microvascular density and corpus luteum (CL) vascularization in cows treated with eCG during stimulatory and superovulatory protocols. Sixteen cows were synchronized and divided into three groups: control (n = 6), stimulated (n = 4) and superovulated (n =6), one was submitted to estrous synchronization (ES) and received no eCG (control), and those that were submitted to ES and received eCG before or after follicular deviation (superovulation and stimulation of the dominant follicle, respectively). Ovulation was synchronized using a progesterone device-based protocol. After six days of ovulation, the cows were slaughtered and the ovaries and CL were collected. The CLs were processed and photomicrographs were taken under light microscopy to assess the vascular volume density (Vv) by stereology, and scanning electron microscopy (SEM) was used to perform ultrastructural analysis of the microvasculature. The Vv in stimulated and superovulated cows significantly increased (P ≤ 0.0001) when compared to control, indicating that the eCG is able to induce angiogenic activity in bovine CL. However, no significant differences were observed between stimulated and superovulated cows. The SEM demonstrated ratings indicative of angiogenesis, marked by several button-shaped projections in the capillaries, and the presence of more dilated capillaries in CL treated with eCG. These morphological findings are evidence of an angiogenic effect of the eCG treatment in CL of cows.

Equine chorionic gonadotropin alters luteal cell morphologic features related to progesterone synthesis.

Exogenous eCG for stimulation of a single dominant follicle or for superovulation are common strategies to improve reproductive efficiency by increasing pregnancy rates and embryo production, respectively. Morphofunctional changes in the CL of eCG-treated cattle include increases in CL volume and plasma progesterone concentrations. Therefore, we tested the hypothesis that eCG alters the content of luteal cells and mitochondria related to hormone production. Twelve crossbred beef cows were synchronized and then allocated into three groups (four cows per group) and received no further treatment (control) or were given eCG either before or after follicular deviation (superovulation and stimulation of the dominant follicle, respectively). Six days after ovulation, cows were slaughtered and CL collected for morphohistologic and ultrastructural analysis. Mitochondrial volume per CL was highest in superovulated followed by stimulated and then control cows (18,500 ± 2630, 12,300 ± 2640, and 7670 ± 3400 µm(3); P < 0.001), and the density of spherical mitochondria and the total number of large luteal cells were increased (P < 0.05) in stimulated cows compared with the other two groups (110.32 ± 14.22, 72.26 ± 8.77, and 70.46 ± 9.58 mitochondria per µm(3) and 678 ± 147, 245 ± 199, and 346 ± 38 × 10(6) cells, respectively. However, the largest diameters of the large luteal cells were increased in superovulated and control cows versus stimulated ones (32.32 ± 0.06, 31.59 ± 0.81, and 29.44 ± 0.77 µm; P < 0.0001). In contrast, the total number of small luteal cells was increased in superovulated cows (1456 ± 268, 492 ± 181, and 822 ± 461 × 10(6), P < 0.05). In conclusion, there were indications of cellular changes related to increased hormonal production (stimulatory treatment) and increased CL volume (superovulatory treatment).

Timed artificial insemination early in the breeding season improves the reproductive performance of suckled beef cows.

The objective was to compare reproductive performance of breeding programs that used natural service (NS), AI after estrus detection (ED), and timed AI (TAI). In experiment 1, 597 suckled beef cows were randomly allocated to one of four groups. Cows in the TAI+NS group (N = 150) were bred by TAI at 11 days after the onset of the breeding season (BS). Bulls were placed with cows 10 days after TAI and remained together until the end of the 90-day BS. Cows in the TAI+ED+NS group (N = 148) received TAI, then AI based on ED for the next 45 days, and finally NS for the last 45 days of the BS. Cows in the ED+NS group (N = 147) received AI based on ED during the first 45 days of the BS, followed by NS for the last 45 days of the BS. Cows in the NS group (N = 149) were bred by NS for the entire 90-day BS. Cows in the ED+NS or NS groups had a decreased (P < 0.001) hazard of pregnancy compared with cows in the two groups bred by TAI at the onset of BS. Also, cows bred by TAI (TAI+NS = 92.7%; and TAI+ED+NS = 91.9%) had higher (P < 0.01) pregnancy rates at the end of the BS compared with cows not bred by TAI (ED+NS = 85.0%; NS = 83.2%). In experiment 2, 507 suckled beef cows were randomly assigned to one of two groups at the onset of a 90-day BS. The NS group (N = 255) received only NS during the entire BS, and the TAI+NS group (N = 252) received TAI at the onset of the BS, followed by NS until the end of BS. Cows in the TAI+NS group had 63% higher hazard of pregnancy (P < 0.001) compared with cows in the NS group, and reduced the median days to pregnancy by 44 (11 vs. 55 days). However, there was no difference (P = 0.31) in proportion of pregnant cows at the end of the BS (TAI+NS = 77.0% vs. NS = 71.0%). Therefore, incorporation of TAI programs early in the BS increased reproductive performance of suckled beef cows.

Treatments for the synchronisation of bovine recipients for fixed-time embryo transfer and improvement of pregnancy rates.

Although embryo transfer technology has been used commercially in cattle for many years, the inefficiency of oestrus detection, especially in recipients, has limited the widespread application of this technology. The most useful alternative to increase the number of recipients utilised in an embryo transfer program is the use of protocols that allow for embryo transfer without the need for oestrus detection, usually called fixed-time embryo transfer (FTET). Most current FTET protocols are based on progestin-releasing devices combined with oestradiol or GnRH, which control and synchronise follicular wave dynamics and ovulation. Conception rates to a single FTET have been reported to be similar to those after detection of oestrus, but pregnancy rates are higher because these treatments have increased the proportion of recipients that receive an embryo. Recent changes to treatments for FTET, such as the administration of eCG, have resulted in increased pregnancy rates and provide opportunities to make these treatments easier to perform on farm.

Fixed time artificial insemination and embryo transfer programs in Brazil

Background: Currently, fixed-time protocols for either artificial insemination or embryo transfer can be routinely applied in the reproductive programs on commercial farms. The control of the follicular wave emergency and the induction of ovulation on a pre-determined time without the need for heat detection facilitate the application of such biotechnologies on a large scale basis increasing the reproductive and productive efficiency. This article will discuss the development of reproductive programs that became practical to apply on Brazilian commercial farms as well as the factors that affect its efficiency. Review: Nowadays, Brazil is the world leader on bovine meat market and also in commercial application of biotechnology, such as fixed-time artificial insemination (FTAI) and fixed-time embryo transfer (TETF). In Brazil, there are several hormones commercially available that can be used for manipulation of follicular wave dynamics and induction of ovulation. The evolution of such protocols were also driven towards the decrease on the amount of time that the animals should came to the curral for hormones treatments in order to make it practical to be use on a large scale basis. Among the commercial established synchronization protocols, one of the most commonly used is with the insertion of a progesterone (P4) device associated with an injection of Estradiol Benzoate (EB) at the beginning of the synchronization protocol to induce an emergence of the new follicular wave. Eight days later, the P4 device is removed associated with the intramuscular administration of prostaglandin, equine chorionic gonadotropin (eCG) and Estradiol Cipionate (ECP). In spite of satisfactory follicular manipulation and precisely synchronization of the time of ovulation there are several factors that can affect the efficiency of FTAI or TETF programs. The FTAI pregnancy rate was influenced by the farm as well as by the body condition scores that the cows presented at the beginning of the synchronization protocol. Other important factors that alter the programs results were the bull used and personal performance during artificial insemination. In general, treatments used for FTET are very similar to those applied for FTAI. In the FTET protocols, the main objective is to increase follicular growth and the diameter of the dominant follicle in order to increase P4 concentrations of the subsequent cycle. Some of the strategies used during the evolution of FTET protocol were to superstimulate the growing follicles of the induced wave after the injection of EB at the beginning of synchronization protocol with eCG or to decrease P4 blood concentration increasing LH pulse frequency promoting an increase on the growth of the dominant follicle during synchronization treatment by advancing the PGF treatment. The final result is to have a large pre ovulatory follicle or a pool of follicles in order to produce a single large or multiples functional CL at the time of embryo transfer. The FTET pregnancy rate was influenced by the diameter of the single CL; by recipient superovulation response by the time of year being lower during the months of autumn and winter in relation to the months of spring and summer. Besides, pregnancy rate at 30 days was also affected by the age of the embryo. Additionally, the correct application of cited biotechnologies enhances reproductive efficiency of livestock bringing sustainable and economic return, increasing the viability of the activity. Conclusion: Therefore, the standardization of the procedures is necessary for the commercial application of FTAI and FTET in Brazil being fundamental for obtaining expressive results, so that Brazil could also export such technologies for countries under the same management production system.

Bovine embryo transfer recipient synchronisation and management in tropical environments.

Numerous studies have shown that it is possible to manipulate follicular and luteal dynamics, thereby eliminating the need for oestrus detection in embryo transfer (ET) programmes. Fixed-time ET (FTET) protocols are based on the use of gonadotrophin-releasing hormone (GnRH) and prostaglandin (PG) F or progesterone/progestogen (P4)-releasing devices and oestradiol. The FTET protocols increases the proportion of recipients transferred, and therefore pregnancy rates, compared with the use of PGF followed by ET 7 days after oestrus. Furthermore, the addition of equine chorionic gonadotrophin (eCG) to the P4 and oestradiol-based FTET protocols results in an even higher proportion of recipients transferred, and thus higher pregnancy rates. The beneficial effect of eCG treatment may be related to increased growth of the dominant follicle and increased plasma P4 concentrations during the subsequent luteal phase. In Bos taurus x Bos indicus recipients, pregnancy rates were positively correlated with the diameter of the corpus luteum (CL) and the number of CL at ET. When repeat-breeder Holstein cows were used as recipients, FTET protocols increased number of recipients transferred and pregnancy rates compared with the traditional PGF-based synchronisation protocols. In conclusion, the use of FTET protocols eliminates the need for the detection of oestrus and results in a greater proportion of recipients transferred and satisfactory pregnancy rates. Thus, FTET optimises the use of recipients, reducing labour and animal handling and facilitating the use of ET.

Superovulation and embryo transfer in Bos indicus cattle.

Compared to Bos taurus breeds, Bos indicus breeds of cattle present several differences in reproductive physiology. Follicular diameter at deviation and at the time of ovulatory capability are smaller in B. indicus breeds. Furthermore, B. indicus breeds have a greater sensitivity to gonadotropins, a shorter duration of estrus, and more often express estrus during the night. These differences must be considered when setting up embryo transfer programs for B. indicus cattle. In recent studies, we evaluated follicular dynamics and superovulatory responses in B. indicus donors with the objective of implementing fixed-time AI protocols in superstimulated donors. Protocols using estradiol and progesterone/progestrogen releasing devices to control follicular wave emergence were as efficacious as in B. taurus cattle, allowing the initiation of superstimulatory treatments (with lower dosages of FSH than in B. taurus donors) at a self-appointed time. Furthermore, results presented herein indicate that delaying the removal of progesterone/progestogen-releasing devices, combined with the administration of GnRH or pLH 12 h after the last FSH injection, results in synchronous ovulations, permitting the application of fixed-time AI of donors without the necessity of estrus detection and without compromising the results.

The timing of ovulation and insemination schedules in superstimulated cattle.

The development of treatments that control follicular wave dynamics during the bovine estrous cycle has resulted in interesting possibilities for the precise control of follicular wave emergence and the time of ovulation. For superstimulation, follicular wave emergence can be controlled by ultrasound-guided follicle ablation with FSH treatments initiated 1 or 2 d later, or injection of estradiol combined with progesterone at the time of insertion of a progestogen releasing device and FSH treatments beginning 4 d later. These are the most widely used protocols for superstimulation of donor cows because they offer the convenience of being able to initiate treatments quickly and at a self-appointed time, without reducing the number of transferable embryos. However, these protocols still require precise estrus detection of donors following superstimulation in order to conduct AI at the most appropriate time. Recent studies have been designed to develop superstimulation protocols that involve fixed-time AI of donors, without regard to estrus detection. Results presented herein indicate that delaying the removal of a progestogen releasing device, combined with the administration of GnRH or porcine LH (pLH) 12 or 24 h later results in predictable, synchronous ovulations, permitting fixed-time AI without reducing the numbers or quality of embryos. These protocols facilitate the application of on-farm embryo transfer programs because they are practical, easy to administer by farm personnel, and more importantly, they eliminate the need for detecting estrus.

Isolation of leptospira santarosai, serovar guaricura from buffaloes (Bubalus bubalis) in Vale do Ribeira, São Paulo, Brazil

In April 1998 urine samples from adult female buffaloes were collected in a farm located in Registro, Vale do Ribeira, São Paulo State, Brazil. The urine samples obtained after furosemide injection were immediately transported to the laboratory in liquid modified EMJH medium and seeded, by the serial dilution technique, into Fletcher's or modified EMJH-0.2 (per cent) agar, both of them with 5-fluorouracil 100mg/mL. The intraperitoneoum inoculation of 0.5 mL was also performed with each urine sample in young, adult hamsters (Mesocricetus auratus). All samples seeded directly in culture medium were contaminated. The hamsters did not show any sign of disease and were killed at the 21st post inoculation day. At this time kidney cultures of these animals were performed and from one of them, one leptospira strain (M04-98) was isolated, identified as belonging to serogroup Sejroe by Microscopic Agglutination Test (MAT) with a panel of 36 rabbit sera against serovars representative for the pathogenic serogroups. Subsequently, MAT was carried out with antisera against the 19 reference strains of serogroup Sejroe, revealing a close relationship with serovar guaricura. Afterwards the MAT was done with a panel of 18 monoclonal antibodies representative for serovars of serogroup Sejroe. The histogram closely resembled that of serovar guaricura. So Cross Agglutination Absorption Test (CAAT) was carried out with the buffalo isolate and guaricura, supporting the relationship between the buffalo isolate and serovar guaricura.