Pregnant Dairy Cattle Research Articles

391 The impact of trace mineral source on systemic immunological status associated with early embryonic loss in lactating dairy cattle

Abstract Milk production has doubled from 1970 to 2009 but has been associated with a decline in fertility. Around 25% of failed pregnancies in dairy cattle are due to embryonic loss. Mineral bioavailability may affect embryonic retention, so we hypothesized that Zn, Cu, and Mn sources will influence systemic immunological status, which affects early embryonic retention. Holstein cows (n = 58) were randomly assigned to one of two daily oral drench trace mineral (Zn, Cu, and Mn) treatments 30 d before breeding (d -30): 1) Inorganic (ITM) or 2) Metal Methionine Hydroxy Analog Chelate (MMHAC). Before treatments began, cows were assigned to a semen type, beef vs. Holstein, equally, across treatments. All cows underwent a double Ovsynch protocol and were artificially inseminated on d 0. Humoral immunological status was assessed in serum samples taken on d 0 and d 7. Immune activity was quantified via a D2Dx systemic immunity score, which assesses samples for the presence of IgG, IgM, cytokines, and complement proteins. Ultrasonography was utilized for pregnancy confirmation on d 35, classifying cows as pregnant (P) or open. Progesterone analysis from d 21 plasma was used to categorize open cows as either non-pregnant (NP) or early embryonic loss (EEL). Statistical analyses in R Studio were completed, in two models based on pregnancy (P vs EEL and NP vs EEL), to determine the effects of mineral treatment, pregnancy status, semen type, and all interactions on humoral immune status. In the P vs EEL model, there were no effects (P > 0.30) of semen type, mineral treatment, pregnancy status, or any interactions on systemic immunity score on the day of breeding (d 0) or d 7. On d 0, cows provided MMHAC had a systemic immunity score of 0.073 ± 0.008 but were not different (P > 0.34) from ITM cows (0.063 ± 0.006). This trend continued on d 7 with systemic immunity scores of MMHAC cows not differing (P > 0.32) from ITM cows (0.055 ± 0.004 and 0.061 ± 0.003; respectively). There was a tendency (P = 0.10) for the three-way interaction of semen type x pregnancy x treatment to influence the immunity score change from d 0 to d 7. Pregnant cows on ITM treatment bred with beef semen had a positive change in immunity score (0.020 ± 0.0085). Whereas ITM cows bred with Holstein semen that underwent EEL had a negative change (-0.015 ± 0.0085); all other means were intermediate. In the NP vs EEL model, there were no effects (P > 0.14) of semen type, treatment, pregnancy status, or any interactions on systemic immunity score on d 0, d 7, or the change between d 0 and d 7. In summary, trace mineral bioavailability tended to affect systemic immunological status in P vs EEL cows but not in NP vs EEL cows, which may allow for the elucidation of immunological status differences during pregnancy establishment.

Early prognostic markers to predict unsuccessful pregnancy in dairy cattle.

This study aimed to investigate maternal serum levels of some angiogenic factors and certain proteins in dairy cattle for (1) early prediction of unsuccessful fertilization and (2) early detection of possible pregnancy failures (early EM) after positive insemination Serum samples were collected from the same cattle at three distinct time points: 30 days before artificial insemination (B-AI), on the day of artificial insemination (AI), and 30 days after artificial insemination (A-AI). As a result of the pregnancy examination, the cows were divided into two main groups according to whether they were pregnant. The results showed that leucyl/cystinyl aminopeptidase (LNPEP) concentration was significantly decreased B-AI and Secreted frizzled-related proteins (SFRP-3), Vascular Endothelial Growth Factor (VEGF) and LNPEP levels were significantly decreased on day of AI, while PRL level was increased, and these data have prognostic significance as early indicator of the risk of potentially failed pregnancy. Additionally, a significant decrease in LNPEP, SFRP3, and VEGF levels, along with an increase in PRL levels was also observed in A-AI. These results suggest that these biomarkers can be used as a screening test to monitor the course of pregnancy. There were no significant differences in serum levels of Insulin-Like Growth Factor 2 (IGF-2), Tissue inhibitors of metalloproteinases (TIMP-1), angiopoietin (ANG), Endoglin (ENG), Fibroblast growth factor (FGF), Inhibine-A (INH-A) and Transforming growth factors-β1 (TGF-β1) between the evaluated periods neither unsuccessful nor the successful pregnancy groups. This is the first study reporting that the maternal serum levels of LNPEP, SFRP3, VEGF, and PRL have important roles in pregnancy success and may indicate whether insemination outcome will be successful B-AI and predict the risk of unsuccessful pregnancy after AI in dairy cattle. The increase in such studies will allow the development of more specific, practical, and applicable markers.

Effects of 2 wintering practices on behavioral and physiological indicators of welfare of nonlactating, pregnant dairy cattle in a pasture-based system

In countries with pasture-based dairy systems and relatively cold winters, such as New Zealand, it is common to manage pregnant, nonlactating cows on forage crop paddocks rather than pasture, due to slow pasture growth rates. Wintering dairy cattle on grazed crops can compromise welfare if wet and muddy underfoot conditions occur, which can reduce lying. This study investigated behavioral and physiological indicators of welfare of cows under 2 wintering practices: cows managed on and grazed kale crop (Brassica oleracea), and cows managed on pasture with baled hay. Following dry-off (d 0), 80 cows were randomly assigned to 1 of the 2 wintering practices (40 cows/practice) and monitored between d 4 and 32 (phase 1). During this period, lying and stepping behavior was continuously recorded using leg-based accelerometers. Blood samples were obtained at d 0 and 32 for measurements of thyroxine (T4), nonesterified fatty acids (NEFA), white blood cells (WBC), and red blood cells. All data for phase 1 were presented descriptively due to the lack of treatment replication. Daily mean air temperature during this period was 5.2°C (range: 0.0-10.7°C), and rainfall was 1.1 mm/d (range: 0-5.6mm/d). Between d 4 and 32, cows in both groups spent similar amounts of time lying (pasture with hay cows: 8.9 h/24 h ± 2.57, kale crop cows: 8.7 h/24 h ± 3.06, mean ± SEM). Both groups reduced their lying on wet and cold days, and there was evidence of rebound lying once unfavorable weather conditions stopped. Cows on kale crop had numerically higher NEFA and lower WBC compared with cows managed on pasture, although most physiological values were within normal ranges. In a second phase of the study (d 34 and 35), cows were managed under controlled, replicated conditions in the 2 wintering practices using typical on-farm stocking rates (2 or 4 cows per group in the pasture with hay and kale crop treatments, respectively; n = 10 groups/treatment). During this period, cow behavior, skin and surface temperatures, hygiene scores, feed intakes, and ground conditions were measured. Weather conditions during the 48-h exposure were mostly cold and dry (mean air temperature: 7.8°C, range: -2.2 to 20.5°C). Cows managed on pasture with hay spent more time lying down on the first day of exposure; however, this was likely due to less space being available to kale cows on this day. Cows managed on pasture with hay ruminated more than cows on kale crop on both days of observations (d 1: 37.9% vs. 30.9% of observations, d 2: 36.8% vs. 28.7% of observations for pasture with hay and kale crop groups, respectively) and were lying more often in postures indicative of greater thermal comfort. Cows managed on pasture with hay had higher skin and surface temperatures compared with cows on kale crop, whereas cows on kale crop had dirtier coats. Results suggest that opportunities for thermal comfort were greater for cows managed on pasture with hay bales, which may be due to increased rumination activities and more insulated lying areas.

Breeding practices and evaluation of hormonal estrus synchronization and mass artificial insemination of dairy cattle in southwest Ethiopia

Estrus synchronization is the process of controlling or guiding a cow/heifer estrous cycle so that they can mate at roughly the same time. It may be used to increase the probability of detecting estrus, increase the rate of calving in response to feed availability and market demand for dairy products, and increase the possibility of pregnancy in dairy cattle. The aim of this study was to assess breeding practices, evaluating of hormonal estrus synchronization and mass artificial insemination of dairy cow/heifers in Andracha and Masha districts of Southwestern Ethiopia. Three hundred eighty-nine (389) synchronized cows were selected from record of artificial insemination center (logbook). For experimental (action) study a total of 210 (103 from Andracha and 107 from Masha) dairy cows/heifers were selected and injected single dose of prostaglandin. Cow/heifers which do not respond by single injection were re-injected another single dose of prostaglandin. The data were analyzed using SPSS version 21 software. The study revealed that cows or heifers treated with single dose of prostaglandin 157(74.8 %), and 53(25.2 %) responded to heat and none response respectively in the study location. The study also revealed that overall conception rate (CR) and number of services per conception (NSPC) for single dose of prostaglandin injection in the researched areas were 68 (43.3 %), and 2.3 respectively. It was concluded that the effectiveness of PGF2α used to synchronize estrus in dairy cows/heifers was good and the overall conception rate 43.1 higher than national level conception rates (7.14–40.23) to first inseminations.

Negative photoperiod induces an increase in the number of ovulations in dairy cattle.

This study sought to examine the impact of negative photoperiod on the incidence of multiple ovulations and pregnancies in dairy cattle. The study population consisted of 5,373 pregnant cows in their third or greater lactation that experienced their first post-partum pregnancy after spontaneous estrus. The positive photoperiod (increasing day-length) extends from December 22 to June 21, whereas the negative photoperiod (decreasing day-length) extends from June 22 to December 21. The odds ratios (ORs) for multiple ovulations and pregnancies in cows that became pregnant during the negative photoperiod and the remaining cows that became pregnant during the positive photoperiod were 1.4 and 1.3 (P < 0.0001), respectively. The ORs for cows that became pregnant ≥ 90 days in milk and the remaining cows that became pregnant < 90 days in milk were 4.3 and 4.1 (P < 0.0001), respectively. No significant differences were detected in the monthly rates of multiple ovulations or pregnancies during positive and negative photoperiods. Thus, the present study demonstrates that the ovarian function in cows is related to changes in day-length, with decreasing day-length being associated with greater multiple ovulation and pregnancy rates. The present study also shows that positive and negative photoperiods exhibit different trends. The results of this study are consistent with a growing body of work demonstrating the effects of photoperiod patterns on the reproductive physiology of cows, with clear implications for twin pregnancy prevention.

BoHV-1 affects abortion and progesterone in dairy cows Bovine alphaherpesvirus 1 (BoHV-1) seropositivity, progesterone levels and embryo loss of 30-day-old pregnant dairy cows in Zagros Industrial Dairy Farm in Shahrekord: Examination and analysis.

Bovine alphaherpesvirus 1 (BoHV-1) is a serious disease with severe negative economic effects on the global cattle sector, especially in Iran. This cross-sectional study was carried out to examine the seroprevalence and associated risk factors of BoHV-1 infection with progesterone levels and embryo death in 30-day pregnant dairy cattle at Zagros Industrial Dairy Farm in Shahrekord, Iran. Between December 2017 to February 2018, blood samples were obtained from 60 dairy cow herds. To determine whether BoHV-1 was present, serum samples were examined using the ELISA for serum antibodies. To find progesterone (P4) in blood, the progesterone ELISA test was used. 96.7 % of sera tested positive for BoHV-1 antibodies, according to the findings. Additionally, 60.34 % of blood samples that tested positive had an experience of abortion and significantly more inseminations that resulted in pregnancy, consistent with findings from other studies conducted in Iran and other nations. Since this study is the first to document the risk factor for BoHV-1 infection in Shahrekord, Iran, we could infer that the virus is extensively dispersed in this area.

Time to increase in pregnancy-specific protein B following artificial insemination is a direct determinant of subsequent pregnancy loss in lactating dairy cows

Increasing progesterone (P4) during early conceptus development may be crucial for establishment of pregnancy in dairy cattle. The objective of this study was to determine if human chorionic gonadotropin (hCG) at various times after ovulation will increase serum P4 during elongation and increase the chances for, and reduce variability to, initial increase in pregnancy-specific protein B (PSPB) following artificial insemination (AI). Time to PSPB increase was defined as the first day of increase in concentrations of PSPB between d 18 and 28 after ovulation in cows with ≥12.5% increases for 3 consecutive days compared with baseline. Lactating cows (n = 368) synchronized to Double-Ovsynch (first service) or Ovsynch (second or greater service) received one of 4 treatments: no hCG (control), or 3,000 IU of hCG on d 2 (D2), 2 and 5 (D2+5), or 5 (D5) after ovulation. All cows were examined via ultrasound on d 5 and 10 postovulation to determine percentage of cows with hCG-induced accessory CL (aCL) and to quantify and measure all luteal structures. Samples for serum P4 were collected on d 0, 5, 19, and 20 postovulation. The P4 was increased in D2, D2+5, and D5 groups compared with control. The D2+5 and D5 treatments increased aCL and P4 compared with D2 and control. The D2 treatment increased P4 on d 5 after ovulation compared with control. Serum PSPB samples were collected daily from all cows on d 18 through 28 after ovulation for determination of d of PSPB increase. Pregnancy diagnoses were performed via ultrasound examination on d 35, 63, and 100 after ovulation and AI. The D5 treatment reduced percentage of cows with, and increased the time to, PSPB increase. Primiparous cows with ipsilateral aCL had reduced pregnancy loss before d 100 postovulation compared with cows with contralateral aCL. Cows that had PSPB increase >21 d postovulation had 4× greater chances of pregnancy loss compared with cows that had PSPB increase on d 20 or 21. The highest quartile of P4 on d 5, but not on d 19 and 20, was associated with reduced time to PSPB increase. Time to PSPB increase appears to be an important measurement to understand reasons for pregnancy loss in lactating dairy cows. Increasing P4 utilizing hCG after ovulation did not enhance early pregnancy or reduce pregnancy losses in lactating dairy cows.

Comparative evaluation of two commercial pregnancy-associated glycoproteins tests for early detection of pregnancy in dairy cattle

The aim of this study was to determine and compare the diagnostic accuracy of two pregnancy-associated glycoproteins tests (IDEXX on-farm pregnancy test [OFPT] and IDEXX rapid visual pregnancy test [RVPT]) for early pregnancy diagnosis in dairy cattle. Blood samples were collected from Holstein cows (n = 317) by coccygeal venipuncture 28–31 days after artificial insemination (AI). The OFPT and RVPT were performed on the farm within 2 h after blood collection using whole blood or blood serum. Transrectal ultrasonography (USG) was performed for pregnancy diagnosis on day 32 post-AI as a gold standard. One-hundred fourteen cows were diagnosed as pregnant and 203 were determined to be nonpregnant. Furthermore, embryonic mortality was detected in four of the nonpregnant animals, based on fragmented/dispersed embryonic membranes and the absence of heartbeat. Sixteen and 14 false positive results (13 jointly for both tests) were obtained for the RVPT and OFPT, respectively. Three false negative results were obtained for the RVPT. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy for the OFPT were 100%, 93.1%, 89.1%, 100% and 95.6%, and for RVPT were 97.4%, 92.1%, 87.4%, 98.4% and 94.0%, respectively. The ability of both tests to distinguish between pregnant and open cows was very good (AUC of both tests above 0.9). The OFPT and RVPT almost perfectly agreed with each other. According to McNemar's analysis, false positive results have been caused difference between the two pregnancy tests and USG. In conclusion, both the RVPT and OFPT proved to be reliable and practical methods for pregnancy diagnosis 28–31 days after AI in dairy cows. However, the results of both pregnancy tests were affected by the occurrence of embryonic mortality around the time of their employment.

Precalving behavior in dairy cattle with different calving times.

Predicting the calving time in dairy cattle can help in avoiding calving accidents and reducing burdens on animal caretakers. In this study, we analyzed the behavior of pregnant dairy cattle for 7 days prior to calving, to assess the feasibility of predicting the calving time. Eleven Holstein cows were divided into two groups based on their calving times, that is, in the morning (the Morning Parturition Group) or the evening (the Evening Parturition Group). Their behavior was recorded on video. An analysis was conducted of the daily occurrences of each type of behavior and the number of switches of behavior during the day and at night. A statistical analysis was conducted, using a two-way factorial analysis. The behavioral sequence was analyzed using an adjacency matrix. Hierarchical structure charts were created, using Interpretive Structural Modeling. The results suggest that feeding and exploratory behaviors are associated with the calving time period and thus can be useful when predicting that period. The hierarchical structure charts suggest that the Morning Parturition Group had no definite behavioral sequence pattern, unlike the Evening Parturition Group. The detection of an unstable behavioral sequence pattern might predict the calving time period.

Investigating functional relationships among health and fertility traits in dairy cows

The association between health and pregnancy per artificial insemination, maintenance of pregnancy, and interval to pregnancy in dairy cattle is well documented. However, the understanding of the causal relationships among health and fertility traits is limited. Although information on heritabilities and genetic correlations between traits is key for the development of multiple-trait selection strategies, only knowledge about functional and causal relationships among them allow the prediction of the consequences of external interventions such as management decisions. Thus, this research was carried out to investigate potential causal relationships among multiple health events (retained fetal membranes [RFM], metritis [MET], clinical endometritis [CE], lameness [LS]) and fertility indicators including resumption of ovarian cyclicity (CY) by 50 days in milk and pregnancy on day 60 (P60) after the first artificial insemination in Holstein cows. Mixed-effects structural equation models (SEM) were fitted conditionally on a causal structure among traits, represented by a directed acyclic graph, inferred from the data using the inductive causation (IC) algorithm. The posterior mean and standard deviation of heritability for RFM, MET, CE, LS, CY and P60 were 0.20 ± 0.07, 0.09 ± 0.02, 0.10 ± 0.04, 0.35 ± 0.04, 0.15 ± 0.03, and 0.10 ± 0.03, respectively. The moderate estimates of heritability found for CY and P60 suggest that selective breeding could be effective for the genetic improvement of these traits. The estimates of genetic correlation among traits ranged from -0.82 ± 0.07 (MET and CY) to 0.70 ± 0.13 (RFM and MET). Biological considerations and results from the inductive causation algorithm analysis retrieved six potential causal connections between traits: RFM→MET; RFM→CE; LS→CE; CE→P60; CY→P60; and CY→CE. In addition to the data-driven network recovered, some additional connections or variations on their directions were also considered on the SEM analyses based on biological and chronological knowledge. Overall, the SEM results were very similar to those obtained from the traditional multi-trait mixed model analysis. The advantage of the SEM analysis is that it sheds some light onto the potential causal relationships between traits, which can aid better decisions to manage events occurring during the postpartum of dairy cattle.

Interferon-τ regulates the expression and function of bovine leukocyte antigen by downregulating bta-miR-204.

IFN-τ is a pregnancy recognition factor that regulates embryo implantation in ruminants. IFN-τ has been suggested to be involved in the expression of microRNA (miRNA/miR) and bovine leukocyte antigen (BoLA), which is an analog of the human major histocompatibility complex class I. However, little is known about whether the miRNAs are involved in the expression of BoLA in ruminants. The present study firstly verified that bta-miR-204 was downregulated and that BoLA was upregulated in the uterine tissues of dairy cows during early pregnancy. Subsequently, luciferase reporter assays, reverse transcription-quantitative PCR and western blot analysis were used to validate BoLA as the target gene of bta-miR-204. Moreover, BoLA was markedly upregulated and bta-miR-204 was downregulated in bovine endometrial epithelial cells (bEECs) treated with IFN-τ. In addition, the results indicated that when the expression level of BoLA was increased by IFN-τ, the expression level of programmed death-ligand 1 (PD-L1) and programmed death-ligand 2 (PD-L2) was also increased. Furthermore, when BoLA was silenced in bEECs by small interfering RNA, the expression of PD-L1 and PD-L2 was not affected by IFN-τ. The expression level of PD-L1 and PD-L2 was also increased in the uterine tissues of pregnant dairy cattle. In conclusion, IFN-τ may function by suppressing the expression of bta-miR-204 to increase the expression of BoLA during the embryo implantation period in cattle. IFN-τ may induce PD-L1 and PD-L2 transcription by regulating BoLA, which may influence the T cell immune response, thereby regulating pregnant cattle immunization.

Economics of Twin Pregnancies in Dairy Cattle.

Simple SummaryTwinning in dairy cattle is not desirable due to negative effects on both cows that calve twins and calves born as twins that result in economic losses to dairy farms. Although a twin pregnancy could bring additional income from extra calves and it could shorten gestation length, twinning compromises milk production, increases the incidence of dystocia and perinatal mortality, decreases calf birth weight, increases the incidence of metabolic diseases, decreases fertility, increases the incidence of freemartinism, increases overall culling risks, and shortens the productive lifespan of cows. Based on a summary of economic analyses from several studies, the estimated losses due to twinning range between $59 to $161 per twin pregnancy. When twins are diagnosed early during gestation, management options might include doing nothing, terminating the pregnancy, or attempting manual embryo reduction. Based on a recent economic analysis of these options, attempting manual embryo reduction decreased the economic losses of a twin pregnancy by $23 to $45.Twinning in Holstein dairy cows has increased over time concurrent with increased milk production. Twinning in dairy cattle is not desirable due to the negative effects on both cows that calve twins and calves born as twins that result in economic losses to dairy farms. Although a twin pregnancy could bring additional income from extra calves and shorten gestation length, twinning compromises milk production, increases the incidence of dystocia and perinatal mortality, decreases calf birth weight, increases the incidence of metabolic diseases, decreases fertility, increases the incidence of freemartinism, increases overall culling risks, and shortens the productive lifespan of cows. Based on a summary of economic analyses from several studies, the estimated losses due to twinning range between $59 to $161 per twin pregnancy. Most twinning in dairy cows is dizygotic and directly related to the incidence of double ovulations, and economic losses are greater for unilateral than for bilateral twins. Hormonal manipulation before artificial insemination that allows for timed artificial insemination is a primary strategy for decreasing twinning in dairy cows before it occurs by decreasing the incidence of double ovulation thereby decreasing conception of dizygotic twins and the associated negative economic consequences. When twins are diagnosed early during gestation, management options might include doing nothing, terminating the pregnancy, or attempting manual embryo reduction. Based on a recent economic analysis of these options, attempting manual embryo reduction decreased the economic losses of a twin pregnancy by $23 to $45.

Vertical transmission of Neospora caninum in Iranian dairy cattle.

Placentally, transmission of Neospora caninum plays significant role on retaining of infection and economic losses in dairy farms. The objective of present study was to assess placental transmission of N. caninum in Iranian dairy cattle. A total of 476 blood samples of pregnant dairy cattle were randomly collected for serology test using ELISA technique. Genomic DNA was extracted and subjected for conventional PCR and Nested-PCR to amplify 330bp and 100bp fragment length of NC5 gene, respectively. Positive animals were follow-up during gestation till abortion and/or full term delivery occurred. Molecularly, 25.3% (118/476; 95% CI: 21%-28.6%) of examined animals were seropositive for N. caninum infection in Holstein cattle (15.1%) with <2 years-old (75%) and history of abortion. The infection was detected. TT and abortion occurred in 13.6% and 3.57% of animals, respectively. Aborted cattle (94.1%, 16/17) were significantly infected to N. caninum and history of abortion was recorded in 42.4% of infected cattle. Placental transmission and abortion in naturally infected dairy cattle due to N. caninum was the first molecular evidence in all examined herds of cattle. It was also uncovered N. caninum had significant role on cattle abortion. Further studies for culling of seropositive cattle with history of abortion, associated risk factors related to neosporosis and lunch control program for reducing side effects of infection are essential in the region.

Twin Pregnancies in Dairy Cattle: Observations in a Large Herd of Holstein-Friesian Dairy Cows.

Multiple pregnancies have devastating consequences on the herd economy of dairy cattle. This observational study examines incidence patterns based on data from the ultrasonographic examination of 1130 multiple pregnancies in cows in their third lactation or more carrying twins (98.8%), triplets (1.1%), or quadruplets (0.08%), and 3160 of their peers carrying singletons. Cows became pregnant following a spontaneous estrus with no previous hormone treatments. Irrespective of a significant decrease (p < 0.0001) in the conception rate (28-34 days post-insemination) during the warm period of the year, the multiple pregnancy rate was similar for both warm (26.5%) and cool (26.3%) periods. The incidence of unilateral multiple pregnancies (all embryos in the same uterine horn) was higher than that of bilateral pregnancies (at least one embryo in each uterine horn): 54.4% versus 45.6% (p < 0.0001). This difference rose to 17% during the warm season (p = 0.03). Pregnancy was monitored in unilateral multiple pregnancies until abortion or parturition (n = 615). In the warm period, the parturition rate was 43% compared to 61% recorded in the cool period (p < 0.0001). Thus, a warm climate is the main factor compromising the fate of multiple pregnancies. Some clinical suggestions are provided.

Transfer of a single embryo versus drainage of subordinate follicles to prevent twin pregnancies in dairy cows. Why not both?

In this study, we present two proposed approaches to prevent twin pregnancies in dairy cattle: 1) single, in vitro-produced embryonic transfer into a recipient cow or 2) subordinate follicle drainage at the time of insemination. Both procedures lead to improved embryonic survival. As the use of sexed semen generates herd replacements and additional heifers, we propose the transfer of a single female cattle embryo into cows that are not suitable for producing replacements, and follicular drainage in lactating cows with genetic merit. This should eliminate economic losses associated with twin pregnancies and increase cattle output of the herd.

Electrochemical assay of sorbitol dehydrogenase at PEDOT modified electrodes - a new milk biomarker for confirmation of pregnancy in dairy cattle.

A robust electrochemical assay for sorbitol dehydrogenase (SORDH) activity in milk was developed using voltammetry and chronocoulometry at bare and polymer modified transducers. The motivation for the work was to evaluate the potential of SORDH as an early biomarker of bovine pregnancy using milk as sample matrix. SORDH is an enzyme involved in carbohydrate metabolism converting sorbitol, the sugar alcohol form of glucose, into fructose, with NAD+ as a cofactor being simultaneously reduced to NADH. The assay was optimised via direct NADH oxidation on glassy carbon and screen printed carbon electrodes followed by electropolymerisation of 3,4-ethylenedioxythiophene (EDOT) monomer to form an NADH responsive PEDOT surface which operated well in undiluted milk samples. Assay conditions such as incubation time and temperature were optimised resulting in a 3 min assay at 37 °C in the presence of 10 mM NAD+ and 20 mM sorbitol co-substrates, enabling NADH electro oxidation (linear range 0.25-5 mM, sensitivity 9.17 μC cm-2 mM-1 in undiluted milk). SORDH determination followed over the range 0.31-10 U mL-1 in milk samples with sensitivity 5.45 μC cm-2 U-1 mL with LOD 0.0787 U mL-1. The assay was applied to milk sample testing acquired as part of an approved animal study involving control and breeding cycles of dairy cows with focus on analysis at day 19 post artificial insemination. Significant differences between control and pregnant SORDH levels in whole milk animal samples were found (average values 2.57 and 4.07 ng mL-1 respectively), as verified using a commercial SORDH ELISA optical assay. Finally, progesterone monitoring over days 16-21 of the oestrous cycle employed an optical ELISA assay and confirmed maintenance of progesterone levels from day 19 onwards.

Slaughter of Pregnant Cattle in Denmark: Prevalence, Gestational Age, and Reasons.

Simple SummaryCattle farmers are in a dilemma when they have to decide if an animal is fit for continued production and can endure another lactation or a pregnancy, or if slaughtering the animal is a better solution despite the animal being pregnant. We studied the prevalence of pregnant cattle at a Danish abattoir and did follow-up interviews of the farmers, asking the reasons for the slaughter of the specific animals, and queried their ethical deliberations. Many pregnant cattle were slaughtered, and the decision to do so was often health-related in dairy herds and production-related in non-dairy herds. Farmers were often aware of the ethical dilemma and considered it better to slaughter the pregnant cow instead of letting her endure another lactation in a stressful environment.The slaughter of pregnant cattle gives rise to ethical controversy. We estimated the prevalence of pregnant cattle, elucidated the reasons for their slaughter, and in light of our findings, discussed the ethics of sending pregnant cattle for slaughter. Among 825 female cattle >353 days of age admitted to a Danish abattoir, 187 (23%) were found to be pregnant. There was no apparent difference in the proportion of pregnant animals between dairy and non-dairy cattle. “Health”-related slaughter was most frequent in dairy herds (70%), whereas “production”-related slaughter was most frequent in non-dairy herds (63%). While many farmers considered it unethical to slaughter pregnant cows without a good reason for doing so, many dairy farmers identified animal welfare as an important parameter in the decision, which was typically when the general condition of the cow or heifer would make it difficult for her to pass through calving and subsequent lactation. The many pregnant animals sent for slaughter were often the result of deliberate choices. Non-dairy farmers often kept a bull with their female cattle, and in many instances, this resulted in the mating of cattle intended for slaughter. Although considered ethically problematic by many dairy farmers, the slaughter of pregnant dairy cattle was often considered better for the cow compared to a stressful lactation period.

Seroprevalence of antibodies for bovine viral diarrhoea virus, Brucella abortus and Neospora caninum, and their roles in the incidence of abortion/foetal loss in dairy cattle herds in Nakuru District, Kenya

BackgroundNo comprehensive studies have been carried out on the infectious causes of abortion in Kenyan dairy cattle herds. A survey was carried out to determine the seroprevalence of antibodies against Bovine Viral Diarrhoea Virus (BVDV), Brucella abortus (BA) and Neospora caninum (NC) among dairy cattle herds in Nakuru County, a major dairying area in Kenya. A prospective sero-epidemiological study was also undertaken to investigate the effects of BVDV, BA and NC on the occurrence of bovine abortion in dairy cattle herds, where monthly rectal palpations for pregnancy were performed, and monthly serum samples were tested for antibodies to the 3 pathogens.ResultsIn the 398 randomly selected cattle on 64 dairy herds, the seroprevalences of antibodies to BVDV, NC and BA were 79.1, 25.6 and 16.8%, respectively. Of the cattle seropositive to NC, 83.3% were also seropositive to BVDV and 13.7% to BA. Of the cattle seropositive to BVDV, 17.1% were also seropositive to BA. Among 260 monitored pregnant dairy cattle on the same 64 dairy farms, an incidence risk for abortion of 10.8% (28/260) was identified, while the incidence of other foetal losses was 1.1% (3/260). The incidence rates of sero-conversion for NC, BVD and BA were 1.1, 0.06 and 0.5 new infections/100 cow-months at risk, respectively. The foetal losses were mainly observed in animals less than 96 months old and occurred in mid-gestation. Neospora caninum was associated with most cases (29.0%) of foetal losses, followed by mixed infections of NC and BVDV (12.9%), BVDV (9.9%) and co-infections of BA and NC (6.5%).ConclusionsThis is the first study to document the substantial incidence risk of BVDV and NC abortions in dairy cattle in Kenya, and demonstrates the relative importance of BA, BVDV and NC infections in dairy cattle in Kenya. Kenya laboratories should offer diagnostic tests for BVDV and NC to help farmers determine their roles in abortions on their farms. A comprehensive policy on the control of these important diseases should also be put in place by government with the involvement of all stakeholders in the dairy cattle industry.

Technical note: Validation of an automated in-line milk progesterone analysis system to diagnose pregnancy in dairy cattle

The in-line milk analysis system (IMAS) is an automated biosensor technology that samples and quantifies milk progesterone concentrations (P4c) at frequent intervals starting early postpartum until pregnancy. The objective was to validate the use of pregnancy notifications (PregN) generated by an IMAS based on P4c profiles after artificial insemination (AI) to determine pregnancy and nonpregnancy status in dairy cows. Records of 1,821 AI events from 715 Holstein cows that had milk P4c (ng/mL) measured every 2.2 ± 1.9 d (mean ± standard deviation) between 24.5 ± 8.2 and 173.4 ± 49.3 d in milk through a real-time IMAS (Herd Navigator, DeLaval International, Tumba, Sweden) were evaluated. Based on variations in adjusted milk P4c (< vs. ≥ the 5.0 ng/mL threshold), the system determined the sampling frequency, onset and cessation of luteal phases, and pregnancy. If a luteal phase initiated (P4c increased to ≥5.0 ng/mL) after AI and remained uninterrupted, a PregN was generated starting at (mean ± standard deviation) 31.0 ± 4.3 d until 53.4 ± 7.9 d after AI, when sampling stopped, unless a decline in P4c (to <5.0 ng/mL) occurred indicating nonpregnancy and imminent estrus. The assessment of IMAS PregN at 4 weekly intervals was tested, and a confirmed calving occurrence between 262 and 296 d after AI, with no other subsequent AI recorded, was the gold standard for pregnancy. In total, 14.1 (256/1,821), 41.0 (746/1,821), and 50.7% (924/1,821) of AI events were followed by a decline in P4c before 19, 23, and 30 d after AI, respectively. Frequency of the last 3 sampling events preceding P4c decline was greater if P4c decline occurred between 18 and 25 d after AI (1.4 ± 0.5 samples per day) compared with before 17 or beyond 26 d after AI (1.0 ± 0.5 samples per day). At 30 ± 3 (27 to 33) d after AI, PregN occurred in 46.8% (853/1,821) of AI events, of which 15.2% (130/853) had a decline in P4c between 30 and 55 d after AI and 17.1% (146/853) was later confirmed nonpregnant based on the gold standard. A total of 40.7% (742/1,821) of AI events was confirmed pregnant by the gold standard, which was no different than the proportion of PregN at 51 ± 3 (48 to 54) d (40.9%; 744/1,821). At any time point between 27 and 54 d after AI, sensitivity and negative predictive values for PregN were greater than 95.0 and 96.0%, respectively, whereas specificity values were less than 90.0% for PregN before 40 d but greater than 94.0% for PregN beyond 41 d after AI. In conclusion, IMAS is able to diagnose pregnancy based on P4c profiles with high precision and determine early nonpregnancy based on the spontaneous cessation of the luteal phase. However, for accuracy greater than 95.0%, pregnancy declaration based on IMAS notifications alone should occur no earlier than 41 d after AI.

Relationship Between Dietary Fatty Acids and Reproductive Functions in Dairy Cattle

Selection of dairy cattle for higher milk yield, without considering important non-production traits, has decreased reproductive efficiency. Thus, low reproductive performance is a major problem in high yielding dairy cattle. Previous studies showed that dietary manipulation to improve fertility holds much promise and dietary fats have positive effects on reproductive functions in high yielding dairy cattle. Positive effects of fats on reproductive performance due to the fatty acids, which are the precursors of progesterone and prostaglandins. Progesterone and prostaglandins hormones are most important factors that play a role on the control of reproductive functions. The amount of linoleic, linolenic and arachidonic fattty acids in ration can be increase or decrease progesterone and prostaglandins synthesis especially PGF2α from ovary and uterus, respectively. Also fatty acids can be influence follicular development, ovulation, embryonic implantation and maternal recognition of pregnancy. This review focuses on the relationships between dietary fatty acids and reproductive functions such as hormone profiles, ovarian function and follicular development, oocyte quality, embryo development, embryonic implantation and maternal recognition of pregnancy in dairy cattle.