Gonadotropin‐Releasing Hormone Administration to Dairy Cows without a Corpus Luteum 4 Weeks after Calving Increases Reproductive Performance

This field study investigated whether the administration of a single dose of gonadotropin‐releasing hormone (GnRH) to dairy cows without a corpus luteum (CL) 4 weeks after calving can improve reproductive performance. Holstein dairy cows underwent ultrasonography to assess the presence of ovarian structures at 29.2 ± 5.2 days post‐partum, and cows were divided into two main groups based on the presence (CL group, n = 230) or absence (non‐CL group, n = 460) of a CL. The non‐CL group was further randomly divided into two subgroups based on the administration of GnRH (non‐CL GnRH group, n = 230) or no GnRH (non‐CL control group, n = 230). Subsets of cows from non‐CL control (n = 166) and non‐CL GnRH (n = 175) groups received a second ultrasonography at 44.5 ± 5.4 days post‐partum to assess CL formation. The percentage of cows with CL at the second ultrasonography was greater in the non‐CL GnRH group (70.9%) than in the non‐CL control group (53.0%, p = 0.0006). The hazard of the first post‐partum insemination by 150 days in milk (DIM) was higher in the CL group than in the non‐CL control group (hazard ratio [HR]: 1.36, p = 0.001). The probability of a pregnancy to the first insemination was higher in non‐CL GnRH (odds ratio [OR]: 1.50, p = 0.04) and CL groups (OR: 1.55, p = 0.03) compared to the non‐CL control group. Furthermore, the hazard of pregnancy by 210 DIM was higher in non‐CL GnRH (HR: 1.30, p = 0.01) and CL (HR: 1.51, p = 0.0001) groups than in the non‐CL control group. In conclusion, administration of GnRH to dairy cows without a CL 4 weeks after calving was associated with an increase in ovulation and improved reproductive performance.     

Secondary Corpora Lutea Induced by hCG Treatment Enhanced Demi‐Embryo Survival in Lactating High‐Yielding Dairy Cows

Using a novel in vivo model considering a low developmental competence embryo (demi‐embryo) and a subnormal fertility recipient (lactating high‐yielding dairy cow), this experiment evaluated the effect of human chorionic gonadotrophin (hCG) treatment at embryo transfer (ET) on embryonic size at implantation, embryonic survival and recipient plasma progesterone (P₄) and bovine pregnancy‐specific protein B (PSPB) concentrations until day 63 of pregnancy. Embryos were bisected and each pair of demi‐embryos was bilaterally transferred to recipients (n = 61) on day 7 of the oestrous cycle. At ET recipients were randomly assigned to treatment with 1500 IU hCG or to untreated controls. Higher (p < 0.01) pregnancy rates on days 25, 42 and 63, and embryo survival rate on day 63 were observed in hCG‐treated cows with secondary CL than in hCG‐treated cows without secondary CL and in untreated cows. Pregnancy rates and embryo survival rate were similar in hCG‐treated cows without secondary CL and untreated cows. Embryonic size on day 42 was not affected by treatment with hCG, presence of secondary CL and type of pregnancy (single vs twin). Presence of secondary CL increased (p < 0.05) plasma P₄ concentrations of pregnant cows on days 14, 19 and 25 but not thereafter and of non‐pregnant cows on days 14–21. Treatment with hCG and presence of secondary CL had no effect on plasma PSPB concentrations, which were higher (p < 0.05) in twin than in single pregnancies. In conclusion, secondary CL induced by hCG treatment at ET significantly increased plasma P₄ concentrations, the survival rate of demi‐embryos and the pregnancy rate of high‐yielding lactating dairy cows. Embryos were rescued beyond maternal recognition of pregnancy, but later embryonic survival, growth until implantation and placental PSPB secretion until day 63 of pregnancy were not affected by treatment or presence of secondary CL.     

Effects of body condition score (BCS) on steroid‐ and eicosanoid‐metabolizing enzyme activity in various mare tissues during winter anoestrus

The objective of this study was to determine the activity of steroid‐ and eicosanoid‐metabolizing enzymes in horses with varying BCSs. The BCSs of twenty non‐pregnant, anoestrous mares were determined prior to euthanasia, and tissue samples were collected from the liver, kidney, adrenal gland, ovary and endometrium. Cytochrome P450 1A (CYP1A), 2C (CYP2C), 3A (CYP3A) and uridine 5′‐diphospho‐glucuronosyltransferase (UGT) activities were determined using luminogenic substrates. The MIXED procedure of SAS was used to test the effect of BCS on enzyme activity and differences between tissues. Activity of CYP1A in adrenals was increased (p ≤ .05) in BCS 5 versus BCSs 4 and 6. Activity of CYP1A in the liver was increased (p = .05) in BCS 4 versus BCSs 5 and 6. Activity of CYP1A was 100‐fold greater (p < .0001) in the liver than in the adrenal, ovary and kidney. Activity of CYP2C was 100‐fold greater (p < .0001) in the liver than in the adrenal, ovary and endometrium. Activity of CYP3A was only detectable in the liver. Activity of UGT in the kidney was decreased (p = .02) in BCS 4 versus BCSs 5 and 6. Activity of UGT was threefold greater (p < .0001) in the liver than in the kidney, whereas activity of UGT was ninefold greater (p < .0001) in the kidney than in the ovary and endometrium. In general, BCS did not alter the activity of steroid‐ and eicosanoid‐metabolizing enzymes in horses. However, tissue differences in these enzymes indicated abundant hepatic metabolism in horses, which is similar to other livestock species.     

Corpus Luteum Development and Function after Supplementation of Long‐Acting Progesterone During the Early Luteal Phase in Beef Cattle

Strategic supplementation of P4 may be used to increase conception rates in cattle, but timing of supplementation in relation to ovulation, mass of supplementary P4 and formulation of the P4‐containing supplement has not been determined for beef cattle. Effects of supplementation of long‐acting progesterone (P4) on Days 2 or 3 post‐ovulation on development, function and regression of corpus luteum (CL) were studied in beef cattle. Cows were synchronized with an oestradiol/P4‐based protocol and treated with 150 or 300 mg of long‐acting P4 on Day 2 or 3 post‐ovulation (6–7 cows/group). Colour‐doppler ultrasound scanning and blood sample collection were performed from Day 2–21.5. Plasma P4 concentrations were greater (p < 0.05) from Day 2.5–5.5 in the Day 2‐treated groups and from Day 3.5–5.5 in the Day 3‐treated cows than in the control group. CL area and blood flow during Day 2–8.5 did not differ (p > 0.05) among groups, suggesting no effect of P4 treatment on luteal development. The frequency of cows that began luteolysis before Day 15 was greater (p < 0.04) in cows treated with 300 mg than in the controls, but there were no differences between non‐treated and 150 mg‐treated cows. The interval from pre‐treatment ovulation to functional and structural luteolysis was shorter (p < 0.01) in the combined P4‐treated groups than in the control cows. In conclusion, was showed for the first time that long‐acting P4 supplementation on Day 2 or 3 post‐ovulation increases P4 concentrations for ≥3 day, has no effect on luteal development, but anticipates the beginning of luteolysis in beef cattle.     

The Effect of Dose and Type of Cloprostenol on the Luteolytic Response of Dairy Cattle during the Ovsynch Protocol under Different Oestrous Cycle and Physiological Characteristics

The objective of this study was to characterize the effect of dose and type of cloprostenol (CLO) on the luteolytic response of dairy cattle during the Ovsynch protocol under different oestrus cycle and physiological characteristics. Twelve non‐lactating dairy cows and 111 lactating dairy cows were used in three experiments. In Experiment I, cows were synchronized so that they had only a 5.5‐ to 6‐day‐old corpus luteum (CL) at the time of the prostaglandin F_2α (PGF_2α) treatment of Ovsynch. In Experiment II, cows were synchronized so that they had at least a CL of approximately 14 days old at the time of PGF_2α treatment and an accessory CL if they had responded to the first GnRH of Ovsynch. Furthermore, in each experiment, cows received either a standard or a double dose of d‐CLO as the luteolytic treatment. In Experiment III, lactating cows were blocked by parity and assigned to one of three luteolytic treatments during Ovsynch: 500 μg d,l‐CLO, 150 or 300 μg of d‐CLO. In Experiment I, the dose of d‐CLO had an effect (p = 0.08) on the percentage of cows with full luteolysis, but not in Experiment II (p > 0.1). More cows in Experiment II had full luteolysis than did cows of Experiment I (87% vs 58%, respectively; p = 0.007). In Experiment III, 87.1%, 84.4% and 86.2% lactating dairy cows had full luteolysis and 37.8%, 36.8% and 36.1% of cows became pregnant after treatment with 500 μg d,l‐CLO, 150 or 300 μg of d‐CLO, respectively (p > 0.05).     

Calm Temperament Improves Reproductive Performance of Beef Cows

Profitability of a beef operation is determined by the proportion of cows attaining pregnancy early in the breeding season and those that are pregnant at the end of breeding season. Many factors, including temperament, contribute to those reproductive parameters. The objective of this study was to evaluate effects of temperament on reproductive performance of beef cows. In Experiment 1, Angus and Angus‐cross beef cows (n = 1546) from eight locations were assigned a body condition score (BCS; 1 = emaciated; 9 = obese) and chute exit and gait score (1 = slow exit, walk; calm temperament; 2 = jump, trot or run; excitable temperament). Cows were grouped with bulls (1 : 25 to 1 : 30; with satisfactory breeding potential and free of venereal disease) for an 85‐day breeding season. Pregnancy status and stage of gestation were determined (transrectal palpation) 35 days after the end of the breeding season. Controlling for BCS (p < 0.01) and handling facility (p < 0.0001) and handling facility by temperament score interaction (p < 0.001), breeding season pregnancy rate was lower in excited versus calm cows [88.6% (798/901) vs 94.1% (607/645); p < 0.001]. Cows with an excitable temperament took 24 more days to become pregnant compared to calm cows (median days to pregnancy, 35 vs 59 days; p < 0.0001). In Experiment 2, Angus and Angus‐cross beef cows (n = 1407) from 8 locations were assigned scores for body condition and chute exit and gait (as described in Experiment 1) and assigned to bulls (breeding sound and free of venereal disease; 1 : 25 to 1 : 30) for 85 days. Pregnancy status was determined by transrectal palpation at 2 and 6 months after the onset of the breeding season. Controlling for BCS (p < 0.05), pregnancy loss was higher in excited versus calm cows [5.5% (36/651) vs 3.2% (20/623), p < 0.0001]. In conclusion, beef cows with an excitable temperament had significantly lower reproductive performance than calmer cows. The modified two‐point chute exit–gait scoring method was repeatable and identified cattle with an excitable temperament.     

Supplementation of distiller's grains during late gestation in beef cows consuming low‐quality forage decreases uterine,but not mammary,blood flow

Positive effects have been observed in offspring from beef cows supplemented with corn dried distillers grain with solubles (DDGS) during late gestation. The hypothesis of this study was that late gestational DDGS supplementation to beef cows would increase blood flow (BF) to the gravid uterus and mammary gland thus impacting birthweight and post‐natal growth of the offspring. Experiment 1 investigated mammary gland BF in multiparous cows during late pregnancy. Beef cows were fed a control (CON1) diet of low‐quality hay (n = 5) or a supplement diet (SUP1) of low‐quality hay with DDGS [1.7 g/kg of body weight (BW); n = 6]. In Experiment 2, multiparous late pregnant beef cows were fed either a control (CON2) diet of a low‐quality hay (n = 4) or a supplement diet (SUP2) of low‐quality hay with DDGS (1.7 g/kg of BW; n = 5). Uterine and mammary gland BF were recorded every 21 days during late gestation. In Experiment 1, there were no effects of diet or day on mammary gland hemodynamics. In Experiment 2, total and ipsilateral uterine BF was less (p ≤ 0.04) in SUP2 vs. CON2 cows and similar BF to contralateral horns. Mammary gland BF was unaltered by maternal supplementation. Even when measured in two different years in two different environments, mammary gland BF remained unaltered to DDGS supplementation. Investigations on the mechanism that may impact uterine BF during late gestation remain to be known.     

The effects on cow performance and calf birth and weaning weight of replacing grass silage with brewers grains in a barley straw diet offered to pregnant beef cows of two different breeds

The effects on cow and calf performance of replacing grass silage with brewers grains in diets based on barley straw and fed to pregnant beef cows are reported. Using a 2 × 2 factorial arrangement of breed and diet, cows pregnant by artificial insemination (n = 34) of two breeds (cross‐bred Limousin, n = 19 and pure‐bred Luing, n = 15) were fed diets ad libitum which consisted of either (g/kg dry matter) barley straw (664) and grass silage (325; GS) or barley straw (783) and brewers grains (206, BG) and offered as total mixed rations. From gestation day (GD) 168 until 266, individual daily feed intakes were recorded and cow body weight (BW) and body condition score (BCS) measured weekly. Calving date, calf sex, birth and weaning BW, and calf age at weaning were also recorded. Between GD 168 and 266, cross‐bred Limousin cows gained more weight than Luing cows (p < 0.05) and cows offered BG gained more weight than cows offered GS (p < 0.001). Luing cows lost more BCS than cross‐bred Limousin cows (p < 0.05), but diet did not affect BCS. There were no differences in dry matter intake as a result of breed or diet. Calf birth BW, however, was greater for cows fed BG than GS (44 vs. 38 kg, SEM 1.0, p < 0.001) with no difference between breeds. At weaning, calves born to BG‐fed cows were heavier than those born to GS‐fed cows (330 vs. 286 kg, SEM 9.3, p < 0.01). In conclusion, replacement of grass silage with brewers grains improved the performance of beef cows and increased calf birth and weaning BW. Further analysis indicated that the superior performance of cows offered the BG diet was most likely due to increases in protein supply which may have improved both energy and protein supply to the foetus.     

The Induction of a Secondary Corpus Luteum on Day 12 Post‐Ovulation can Delay the Time of Luteolysis in High‐Producing Holstein Cows

Luteolysis before the time of maternal recognition of pregnancy is one cause of low fertility in high‐producing dairy cows. The objective of this study was to assess whether induction of a secondary corpus luteum (CL) late in the luteal phase would delay the time of luteolysis. Twenty high‐producing Holstein cows were synchronized to ovulation (Day 0) with the Ovsynch protocol and received hCG (1500 IU im) on Day 12. Corpora lutea formation (as evaluated by ultrasonography) and plasma P4 concentrations were monitored from Days 4 to 36. hCG treatment induced the formation of one secondary CL (CL2) in 11 of 20 cows (55%) from the dominant follicle (mean diameter: 14.2 ± 0.9 mm) of two‐wave (3/11) and three‐wave (8/11) cycles. The maximal diameter of the CL2 (23.3 ± 1.9 mm) was reached approximately 6 days after hCG treatment and was correlated with its structural lifespan (p < 0.01). Cows that formed a CL2 after hCG had higher mean plasma P4 concentrations on Day 14 (+4.5 ng/ml) and Day 18 (+3.0 ng/ml) compared with cows without CL2 (p < 0.05). The structural regression of CL2 begun approximately 8 days after that of the CL1, and the median time at which the first drop in circulating P4 levels occurred was later in cows that formed a CL2 than in those that did not (Day 26 vs Day 18; p < 0.01). Thus, the induction of a CL2 by hCG on Day 12 might reduce the risk of premature luteolysis in high‐producing dairy cows after insemination.     

The Use of Beef Bull Semen Reduced the Risk of Abortion in Neospora‐seropositive Dairy Cows

There is an evidence that the epidemiology of neosporosis differs in dairy and beef cattle, such that beef cattle carry a lower risk of abortion. The aim of the present study was to establish whether artificial insemination using semen from beef bulls could reduce the risk of abortion in dairy cows seropositive for the Neospora caninum parasite. Our study was based on yearly serological screening for neosporosis and on the confirmation of Neospora infection in aborted fetuses in two high‐producing dairy herds with a mean 28% seroprevalence of N. caninum antibodies. The study population comprised of 273 pregnancies in seropositive animals: 156 pregnancies monitored after insemination using Holstein–Friesian semen and 117 after insemination using beef bull semen. Abortion rates for these animals were 28.2% (77 of 273), 34.6% (54 of 156) and 19.7% (23 of 117). Logistic regression analysis indicated no significant effects of lactation number and previous abortion on the abortion rate. Based on the odds ratio, a 1‐unit increase in the Neospora antibody titre yielded a 1.01‐fold increase in the abortion rate. The likelihood of abortion was two times higher for cows in one of the two herds and 2.8 times lower (one of 0.36) for pregnant cows inseminated with beef bull semen rather than Holstein–Friesian semen. Our results indicate that the use of beef bull semen can reduce the risk of abortion in dairy cows, and suggest that annual screening for neosporosis, specifically the antibody titre to the protozoan, could be an useful predictor of abortion risk in reproductive health programmes.     

Differential expression of ISG 15 mRNA in peripheral blood mononuclear cells of nulliparous and multiparous pregnant versus non‐pregnant Bos indicus cattle

Embryonic mortality is found to be the main source of reproductive wastage in domestic ruminants. Many genes are involved in the growth and development of the embryo, and the interferon‐stimulated gene 15 (ISG 15) is one of the major gene stimulated by interferon tau, the maternal recognition of pregnancy signal in ruminants. In this study, both genomic and cDNA sequences of ISG 15 from Bos indicus (Deoni breed) were amplified and characterized. The genomic sequence of Deoni ISG 15 exhibited 99% identity with Bos taurus and 97% identity with that of Bos mutus and Bubalus bubalis. Moreover qRT‐PCR analysis revealed constitutive expression of the ISG 15 mRNA in peripheral blood mononuclear cells of Deoni heifers and multiparous cows during early pregnancy. Fourteen Deoni heifers and fifteen multiparous Deoni cows were synchronized for timed AI by CIDR‐Ovsynch protocol, and six animals were kept as cyclic control in each group. Blood samples were collected on days 7, 14, 16, 18, 21, 30 and 45 from the day of AI. Pregnancy was confirmed by plasma progesterone level through ELISA. A significantly higher expression of ISG 15 mRNA was found on day 16 (p < .05) and day 18 (p < .05) of pregnancy in nulliparous heifers. Although in multiparous Deoni cows ISG 15 expression was greater in pregnant cows, difference was statistically non‐significant. The result of this study indicates that ISG 15 gene expression is upregulated during 16–18 days of pregnancy and could be used as an early pregnancy marker in dairy cows especially in heifers.     

Reproductive and Metabolic Responses of Early‐lactating Dairy Cows Fed Different Dietary Protein Sources

Optimal reproduction is very closely tied with optimal nutrition, and early‐lactation diets in cows are critical to successful reproduction and monitoring is important. To evaluate the effects of different dietary protein sources on metabolic parameters and reproductive activity, a total of 36 Italian Friesian early‐lactating dairy cows were assigned for 16 weeks to three dietary treatments as follow: the control diet contained soya bean meal (SBM) as the main protein source, whereas the experimental diets contained faba bean (FB) or pea seeds (PS) as alternative protein sources. Diets were formulated to be isocaloric and isonitrogenous. Cow blood samples were collected, and plasma were analysed for metabolites, biological enzymes, β‐hydroxybutyrate (BHBA) and non‐esterified fatty acids (NEFA). Feeding alternative protein sources had no effects on most metabolic blood profile, except for blood cholesterol, triglycerides and urea. Results from reproductive parameters indicated that cows fed FB diet had a lower insemination index, but a shorter calving to conception period and an improved conception rate and artificial insemination outcome, when compared to cows fed SBM or PS diets. It can be concluded that replacing conventional dietary SBM with alternative protein sources, especially FB, resulted in improved reproductive performances and metabolic parameters in early‐lactating dairy cows.     

Factors Affecting the First Service Conception Rate of Cows in Smallholder Dairy Farms in Bangladesh

The successful outcome of an insemination is a combination of both male and female fertility‐linked factors. We investigated the first service conception rate of cows at artificial insemination (AI) in the smallholder dairy farms in Bangladesh. Frozen straws were prepared from ejaculates of Bos indicus (n = 7) and Bos indicus × Bos taurus (n = 7) AI bulls. Fertility was determined from 6101 first services in cows that were performed by 18 technicians in four regions between April 2004 and March 2005. Pregnancy was diagnosed by rectal palpation between 60 and 90 days post‐insemination. The Asian version of Artificial Insemination Database Application (AIDA ASIA) was used for bulls‐, cows‐ and AI‐related data recording, and later retrieved for analysis. The mean ± SD number of inseminations performed from individual bulls and their conception rates were 436.0 ± 21.6 and 50.7 ± 1.9%, respectively. Logistic regression demonstrated body condition scores (BCS), heat detection signs, months of AI and their interactions had greatest effects (odds ratios: 1.24–16.65, p < 0.04–0.001) on first service conception rate in cows. Fertility differed (p < 0.02–0.001) between the regions, previous calving months, months of AI, BCS, parity and heat detection signs of cows. Inseminations based on mounting activity (n = 2352), genital discharge (n = 3263) and restlessness and/or other signs (n = 486) yielded a conception rate of 53.6%, 48.8% and 50.1%, respectively (p < 0.05). Conception rate between technicians ranged between 43.4% and 58.6% (p < 0.05). The days interval from calving to first service (overall mean ± SD = 153.4 ± 80.6) had relationship (p < 0.001) with BCS, months of previous calving and parity of the cows. Fertility at AI in smallholder farms can be improved by training farmers on nutrition and reproductive management of the cows.     

Contrast‐enhanced ultrasonography is a feasible technique for quantifying hepatic microvascular perfusion in dogs with extrahepatic congenital portosystemic shunts

Extrahepatic‐congenital portosystemic shunt is a vascular anomaly that connects the portal vein to the systemic circulation and leads to a change in hepatic microvascular perfusion. However, an assessment of hepatic microvascular perfusion is limited by conventional diagnostic modalities. The aim of this prospective, exploratory study was to assess hepatic microvascular perfusion in dogs with extrahepatic‐congenital portosystemic shunt using contrast‐enhanced ultrasonography (CEUS) using perfluorobutane (Sonazoid^®). A total of 17 dogs were included, eight healthy dogs and nine with extrahepatic‐congenital portosystemic shunt. The time‐to‐peak (TTP), rising time (RT), and rising rate (RR) in the hepatic artery, portal vein, and hepatic parenchyma, as well as the portal vein‐to‐hepatic parenchyma transit time (ΔHP‐PV) measured from time‐intensity curve on CEUS were compared between healthy and extrahepatic‐congenital portosystemic shunt dogs. The RT of the hepatic artery in extrahepatic‐congenital portosystemic shunt dogs was significantly earlier than in healthy dogs (P = 0.0153). The TTP and RT of the hepatic parenchyma were significantly earlier in extrahepatic‐congenital portosystemic shunt dogs than in healthy dogs (P = 0.0018 and P = 0.0024, respectively). ΔHP–PV was significantly shorter in extrahepatic‐congenital portosystemic shunt dogs than in healthy dogs (P = 0.0018). CEUS effectively revealed changes in hepatic microvascular perfusion including hepatic artery, portal vein, and hepatic parenchyma simultaneously in extrahepatic‐congenital portosystemic shunt dogs. Rapid hepatic artery and hepatic parenchyma enhancements may reflect a compensatory increase in hepatic artery blood flow (arterialization) caused by a decrease in portal vein blood flow and may be used as an additional diagnostic test to distinguish extrahepatic‐congenital portosystemic shunt dogs from healthy dogs.     

Coxiella burnetii Shedding During the Peripartum Period and Subsequent Fertility in Dairy Cattle

The objective of this study was to assess the effects of Coxiella burnetii shedding or seropositivity on post‐partum recovery and subsequent fertility in high‐producing dairy cows. Given the difficulty in diagnosing C. burnetii infection at the farm level, an exhaustive series of tests in 43 pregnant animals that delivered at least one live calf were conducted, including blood serology and PCR of milk or colostrum, cotyledons (only at parturition), faeces, vaginal fluid against C. burnetii on gestation Day 171–177, at parturition and on Days 1–7, 8–14, 15–21, 22–28, 29–35 and 90–97 post‐partum. During scheduled herd visits, ultrasonography (US) of the genital tract and examination of vaginal fluid were performed on Days 15–21 (V1), 22–28 (V2), 29–35 (V3) and 51–57 (V4) post‐partum by the same veterinarian. Logistic regression analysis revealed that the likelihood of suffering endometritis (the presence of echogenic intrauterine fluid (IUF), cervical diameter of ≥4 cm or endometrial thickness ≥0.75 cm) was lower in C. burnetii‐seropositive animals (OR = 0.10), compared with C. burnetii‐seronegative animals. According to Kaplan–Meier survival analysis, C. burnetii‐seronegative and non‐shedding cows showed a delayed return to luteal activity and conception was delayed in non‐shedding animals, compared with the remaining animals. Overall, the results of our study provide useful insight into the effects of C. burnetii infection on post‐partum recovery and subsequent fertility. In particular, animals not infected with Coxiella seem to be susceptible to infection and not protected against the bacterium in dairy herds. The elevated costs of determining an infection at the farm level, make monitoring of cows virtually impossible from a clinical point of view.     

Uterine infection with bovine herpesvirus type 4 in dairy cows

Diseases of the reproductive tract are a frequent problem in dairy herds. Herpesviruses are uterine pathogens also involved in other clinical diseases; for example, bovine herpesvirus type 4 BoHV‐4 induces abortion, enteritis, metritis, pneumonia and vaginitis, but it can also be detected in healthy cows. The role of BoHV‐4 in the development of clinical endometritis (CE) or subclinical endometritis (SE) has not clearly been described. Therefore, the objective of this study was to describe the prevalence of uterine BoHV‐4 infection and its relationship with clinical, bacteriological and cytological findings in dairy cows 20–30 days after calving. The experiment was performed as a completely randomized block design, with farm (n = 10) as blocking criterion and with cow (n = 397) as the experimental unit. Logistic regression models were used to assess the effect of BoHV‐4 infection on CE, SE and reproductive performance. Proportion of cows infected with BoHV‐4 was 5.8% (n = 23/397). BoHV‐4 was isolated in 11.0% (n = 12/109), 4.8% (n = 4/84) and 3.6% (n = 7/194) of cows diagnosed as CE, SE or healthy, respectively. A logistic model revealed that BoHV‐4 infection showed a tendency to increase the risk for CE (AOR = 2.17; p = .10) but significantly reduced both, the odds for artificial insemination within 80 days post‐partum (dpp) (AOR = 0.37; p = .035) and for pregnancy within 200 dpp (AOR = 0.13; p = .004). Furthermore, BoHV‐4 infection increased the chance for intrauterine infection with Trueperella pyogenes (AOR = 5.55; p < .001) and vice versa (AOR = 5.79, p < .001). In conclusion, BoHV‐4 infection is associated with reduced chances for insemination and pregnancy by 200 dpp and showed a trend to be associated with increased risk for CE. Furthermore, BoHV‐4 and Trueperella pyogenes infections are strongly related.     

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

We aimed to evaluate the reproductive performance of Nelore lactating cows submitted to a resynchronization 12 days after timed artificial insemination (TAI) with or without a long‐acting progesterone (P4‐LA) treatment. Nelore cows were submitted to a P4/oestradiol‐based TAI protocol (D0 = insemination). On D12, cows in the control group (n = 184) received a new P4 intravaginal device (0.96 g), whereas cows in the P4‐LA group (n = 192) received the P4 device and 75 mg P4‐LA. Cows identified as non‐pregnant (n = 120) by regression of corpus luteum using colour Doppler ultrasonography on D20 had the P4 device removed and received 500ug of sodium cloprostenol, 1 mg of oestradiol cypionate and 300 IU of eCG and were re‐inseminated on D22. There was no difference (p > 0.10) in the pregnancy rate at D20, D30 and D60 after first TAI between the control (69%, 59.7% and 57%, respectively) and P4‐LA (67%, 55.7%, and 55.2%, respectively) groups. Pregnancy losses were similar between both groups (p > 0.1). For cows submitted to the second TAI, the pre‐ovulatory follicle size did not differ (p > 0.1), but the oestrous detection and pregnancy rates were greater (p < 0.05) in the P4‐LA group (92.2% [59/64] and 60.9% [39/64], respectively) than in controls (75% [42/56] and 44.6% [25/56]). The cumulative pregnancy rate after two TAIs did not differ (p > 0.1) between control (73.3% [135/184]) and P4‐LA (76% [146/192]) groups. The use of P4‐LA at 12 days after TAI potentially increases the pregnancy rates for a new early resynchronization strategy associated with the Doppler imaging for pregnancy diagnosis and results in an alternative to perform two TAIs in 22 days in beef cows.     

Plasma Concentrations of Pregnancy‐Associated Glycoproteins Measured Using Anti‐Bovine PAG‐2 Antibodies on Day 120 of Gestation Predict Abortion in Dairy Cows Naturally Infected with Neospora caninum

The present study sought to determine: (i) the effects of Neospora caninum infection and twin pregnancy on plasma pregnancy‐associated glycoprotein‐2 (PAG‐2) concentrations throughout pregnancy and (ii) whether plasma PAG‐2 concentrations could predict abortion in N. caninum‐infected cows. The study was performed on a commercial Holstein‐Friesian dairy herd in northeastern Spain and the final data included those recorded in 53 non‐aborting and 19 aborting animals. Blood samples were collected immediately before pregnancy diagnosis (on Days 40, 90, 120, 150, 180 and 210 post‐insemination) in non‐aborting cows or until the time of abortion detection in aborting cows. General lineal models (GLM) repeated measures anova revealed the different behaviour of PAG‐1 and PAG‐2, and significant effects of Neospora seropositivity, cool season and twin pregnancy on plasma PAG‐2 concentrations throughout gestation (between‐subject effects). In addition, based on the odds ratios, the likelihood of abortion increased in Neospora‐seropositive cows (by a factor of 7.0) compared to seronegative animals and decreased in cows with a high plasma PAG‐2 concentration (>4.5 ng/ml) on Day 120 of pregnancy (by a factor of 0.24), compared to the remaining cows. In conclusion, there is a relationship between plasma PAG‐2 concentrations and the risk of abortion in Neospora‐infected dairy cows. Thus, plasma PAG concentrations measured using anti‐boPAG‐2 antiserum on Day 120 of gestation could serve as an indicator of the abortion risk in N. caninum infected animals; values <4.5 ng/ml indicating a high risk of abortion in chronically infected animals.     

Evaluation of Subclinical Endometritis and Consequences on Fertility in Piedmontese Beef Cows

Subclinical endometritis (SEM) is poorly investigated in beef cows, as stated in the literature. This project aims to evaluate the rate and the consequences of SEM in Piedmontese cows, with a focus on bacteriological findings and fertility parameters. Uterine cytology was performed for 97 subjects; a total of 31% of the cows were diagnosed as being positive for SEM and as having an 8% neutrophil (PMN) presence on the slide, which is considered as the best cut‐off to diagnose the pathology. Only 13% of the cows positive for SEM were pregnant within 130 dpp and generally showed increases of 40 days in the partum to conception interval compared with the negative cows (142 vs 182, p = 0.01). Cows positive for both bacteriology and cytology showed a lower fertility than cows with only inflammation or only a bacterial presence (p = 0.0004). Bacterial isolation detected different species, but no difference in regard to the impact of these bacteria on SEM was shown. Parity, presence of calves, hygiene condition, age and number of service did not affect whether a cow was positive for subclinical endometritis (p < 0.05). The housing system (free stalls vs tie stalls) used seems to affect the SEM rate in Piedmontese cows; cows bred in tie stalls were more likely to be positive for SEM (OR = 2.2; p = 0.04). In conclusion, cytology seems to be a good technique for the diagnosis of subclinical endometritis in beef cows, and as in dairy cows, subclinical endometritis has a detrimental effect on fertility, causing an increase in partum to conception and a decrease in the rate of cows who become pregnant within 130 dpp, particularly for those cows housed in a tie stall.     

Detection of APAF1 mutation in Holstein cows and mummified foetuses in Japanese dairy herds

Some of the highest genetic merit sires have been shown to harbour recessive mutations affecting fertility, which may spread rapidly in the population through AI. These disorders may result in abortion and decline in pregnancy per insemination in cows. This study was carried out on 240 Holstein‐Friesian cows and 15 mummified foetuses. Blood and tissue samples were collected from the cows and mummified foetuses, respectively, for DNA extraction. Allele‐specific PCR was designed for the detection of the cows and foetuses carrying the nonsense mutation (C/T) in apoptosis peptide activating factor 1 gene (APAF1). The mutant allele frequency of the APAF1 in carrier cows and mummified foetuses was calculated. Milk samples were taken from the carrier and non‐carrier cows for progesterone assay. The allele‐specific PCR reaction efficiently distinguished the C/T mutation in APAF1. Of 240 cows, seven cows (2.9%) were diagnosed to carry one copy of the mutant allele of APAF1. However, the carrier frequency was 33.3% in mummified foetuses (five of 15). The mutant allele frequency was 0.02 and 0.17 in the cows and mummified foetuses, respectively. Concentrations of progesterone did not differ between cows with APAF1 mutation and non‐carrier cows during 45 days post‐insemination. This study provided allele‐specific PCR for the detection of APAF1 mutation in cows. Moreover, it reports the carrier and mutant allele frequencies of APAF1 in dairy cows and mummified foetuses in Japan.