Morphologic, Steroidogenic, and Transcriptomic Assessment of the Corpus Luteum in Holstein Cows after Spontaneous or Hormone-Induced Ovulation.

There is evidence that replacing the gonadotropin-releasing hormone (GnRH) with porcine luteinizing hormone (pLH) to synchronize ovulation prior to artificial insemination (AI) increased pregnancy per AI in dairy cows without affecting blood progesterone (P4) concentrations. Whether morphologic, steroidogenic, and transcriptomic differences exist among corpora lutea (CL) formed after ovulation induced by GnRH and pLH is unclear. Our main objective, therefore, was to compare CL characteristics between GnRH- and pLH-induced CL. In 24 non-lactating Holstein cows, ovulations were spontaneous (Spont-Ov) or induced with 100 µg GnRH, 25 mg pLH, or 1 mg estradiol benzoate (EB), with CL excised 12 d after ovulation. In pLH- versus GnRH-treated cows, the duration of elevated LH (above baseline) was prolonged (10 versus 6 h, respectively, p < 0.01), but CL dimensions, pixel intensity of CL images, proportions of steroidogenic and non-steroidogenic luteal cells, and mean plasma LH did not significantly differ. Post-ovulation mean plasma P4 (ng/mL) did not differ among Spont-Ov (3.0) pLH (3.1) or GnRH (3.0) cows but were lower in EB cows (2.0). In vitro P4 concentration was greater in luteal explants of pLH-treated cows than in all other groups (combined means, 16.0 vs. 12.3 µg/mL, p < 0.02). Relative abundance of mRNA for oxytocin receptor (OXTR) was 2-fold higher (p < 0.01) in CL of pLH vs. GnRH cows and highest in Spont-Ov CL. In summary, pLH-treated cows had a longer LH peak, and greatest luteal tissue concentrations and in vitro production of P4. We inferred that increased P4 concentrations at the ovarian-uterine level in pLH-treated cows could have promoted embryo development and increased pregnancy per AI.

Dynamics of pre- and post-insemination progesterone profiles and insemination outcomes determined by an in-line milk analysis system in primiparous and multiparous Canadian Holstein cows.

The objective was to evaluate in-line milk progesterone (mP4) data to determine dynamics of pre- and post-insemination mP4 profiles and their associations with parity and outcomes of artificial insemination (AI) in Holstein cows. Milk progesterone (ng/mL) was quantified at pre-determined time points before and after AI through an automated in-line milk analysis system (Herd Navigator™, DeLaval International, Tumba, Sweden). Only AI (∼d0; n = 605) preceded by an mP4-decline (at least two samples of mP4 ≥5 ng/mL followed by at least one sample <5 ng/mL; d-2) were evaluated. Maximum mP4 attained between d-15 and d-2 (PrePeak), d-2, d5, d10, d14, maximum mP4 attained within 21d post-AI (PostPeak), and rate-of-daily-change between mP4 time points (ng/mL/d) were analyzed. Primiparous and multiparous cows were classified by AI outcomes based on post-AI mP4 profiles into three groups: (1) non-pregnant (OPEN; mP4-decline ≤ 30d post-AI), (2) presumed-pregnant (PREG; no mP4-decline until 55d post-AI), and (3) presumed-pregnancy loss (P-LOSS; mP4-decline between 31 and 55d post-AI). For profile comparisons, smoothed mP4 data were analyzed using mixed linear models. Primiparous cows had greater (P < 0.01) mP4 than multiparous cows at d5 (4.6 ± 0.2 vs. 2.8 ± 0.1), 10 (11.1 ± 0.4 vs. 7.6 ± 0.2), 14 (19.7 ± 0.4 vs. 16.1 ± 0.3) and PostPeak (23.5 ± 0.3 vs. 21.7 ± 0.2). The rate-of-daily-change was greater (P < 0.01) in primiparous than in multiparous cows from d-2 to 5 (+0.2 ± 0.03 vs. -0.1 ± 0.02) and from d5 to 10 (+1.2 ± 0.05 vs. +0.9 ± 0.03), but lesser (P < 0.01) from d14 to PostPeak (+0.9 ± 0.09 vs. +1.3 ± 0.06). In primiparous cows, mP4 in PREG was greater at d10 and PostPeak than OPEN (11.1 ± 0.5 and 24.2 ± 0.5 vs. 9.6 ± 0.4 and 22.3 ± 0.4, respectively, P < 0.04), but lesser at d5 than P-LOSS (4.4 ± 0.3 vs. 5.7 ± 0.4, P = 0.04). In multiparous cows, mP4 at d-2 was lesser in PREG than OPEN and P-LOSS (3.2 ± 0.1 vs. 3.4 ± 0.04 and 3.5 ± 0.1, respectively, P ≤ 0.03), but greater at d10, d14 and PostPeak in PREG than in OPEN (8.2 ± 0.4, 16.8 ± 0.5 and 22.7 ± 0.4 vs. 6.9 ± 0.3, 14.8 ± 0.3 and 19.7 ± 0.2, respectively, P < 0.01). Multiparous PREG cows had greater rate-of-daily-change in mP4 than OPEN cows from d5 to 10 and from d10 to 14 (+1.0 ± 0.06 and +2.2 ± 0.11 vs. +0.8 ± 0.04 and +1.9 ± 0.08, respectively, P < 0.03). Overall post-AI mP4 increased faster and were greater in primiparous than in multiparous cows. Based on in-line mP4 profiles, greater mP4 levels near time of AI (d-2 in multiparous and d5 in primiparous cows) and lesser mP4 beyond d10 were negatively associated with a successful pregnancy.

Relationships among early postpartum luteal activity, parity, and insemination outcomes based on in-line milk progesterone profiles in Canadian Holstein cows.

The objectives of this retrospective study were to use in-line milk progesterone (mP4) data to investigate relationships of (1) commencement of luteal activity (CLA), and (2) luteal phase (LP) length and frequency preceding first postpartum AI, with parity and AI outcomes in Canadian Holstein cows. Starting 21 ± 1 days postpartum (DPP), levels of mP4 were assessed every 2.2 ± 2.0 d through an automated in-line milk analysis system (Herd Navigator™, DeLaval International, Tumba, Sweden) until ∼55 d after first or second AI in 748 Holstein cows from two herds. The CLA was defined as the DPP of the first of at least two consecutive samples with mP4 ≥5 ng/mL, and the period with elevated mP4 (≥5 ng/mL) was defined as the LP. Cows were categorized by CLA [earlier (≤) or later (>) than 28, 35, 42, 49, 56, and 63 DPP], and by the pattern of LP frequency preceding first AI as having or not: (1) one or more normal LP (LP length ≥7 and ≤19 d); (2) one or more abnormal LP (LP length <7 or >19 d, or interluteal period ≥12 d); and (3) two or more LP (either normal or abnormal). Outcomes of first or second AI were determined by the interval between AI and cessation of the ensuing LP as: non-pregnant (mP4-decline ≤30 d), presumed-pregnant (no mP4-decline until 55 d), or presumed-pregnancy loss (mP4-decline between 31 and ≤55 d). The odds of pregnancy per AI (P/AI) at 55 d and pregnancy loss were evaluated using generalized linear mixed models. Primiparous cows had lower odds of having CLA ≤28 DPP [Odds ratio (OR) = 0.58, P = 0.002] and one or more abnormal LP (OR = 0.73, P = 0.04) than multiparous cows. In multiparous cows, CLA ≤28 DPP decreased pregnancy loss (OR = 0.48, P = 0.05) and CLA ≤56 DPP increased P/AI (OR = 4.69, P < 0.01) compared to a later CLA. Primiparous and multiparous cows that had one or more normal LP before first AI had increased P/AI (OR = 3.85 and 3.45, respectively, P < 0.01) and reduced pregnancy loss (OR = 0.26 and 0.27, respectively, P < 0.01) than cows without a normal LP. Primiparous cows that had one or more abnormal LP had decreased P/AI (OR = 0.62, P = 0.04) and increased pregnancy loss (OR = 1.64, P = 0.04) compared to those without an abnormal LP. In summary, AI outcomes were improved in multiparous cows that had early CLA and in cows of both parity groups that had at least one normal LP before first AI. However, primiparous cows that had at least one abnormal LP had reduced AI outcomes. Relationships between early postpartum luteal activity and AI outcomes were inconsistent between primiparous and multiparous cows.

Transcriptome Profiling of In-Vivo Produced Bovine Pre-implantation Embryos Using Two-color Microarray Platform.

Early embryonic loss is a large contributor to infertility in cattle. Moreover, bovine becomes an interesting model to study human preimplantation embryo development due to their similar developmental process. Although genetic factors are known to affect early embryonic development, the discovery of such factors has been a serious challenge. Microarray technology allows quantitative measurement and gene expression profiling of transcript levels on a genome-wide basis. One of the main decisions that have to be made when planning a microarray experiment is whether to use a one- or two-color approach. Two-color design increases technical replication, minimizes variability, improves sensitivity and accuracy as well as allows having loop designs, defining the common reference samples. Although microarray is a powerful biological tool, there are potential pitfalls that can attenuate its power. Hence, in this technical paper we demonstrate an optimized protocol for RNA extraction, amplification, labeling, hybridization of the labeled amplified RNA to the array, array scanning and data analysis using the two-color analysis strategy.

Effects of prepartum oilseed supplements on subclinical endometritis, pro- and anti-inflammatory cytokine transcripts in endometrial cells and postpartum ovarian function in dairy cows.

Postpartum uterine infections affect ovarian function and delay ovulation in cattle. As dietary fats can affect immune cell function, we investigated the influence of prepartum diets on postpartum uterine inflammatory status (UIS) as assessed 25±1 days postpartum by endometrial cytology (normal: ≤8% polymorphonuclear cells (PMN) vs subclinical endometritis (SCE): >8% PMN) and associations between SCE, pro- and anti-inflammatory cytokine gene expression and ovarian function. During the last 5 weeks of gestation, dairy cows received a diet supplemented with 8% rolled sunflower (n=10) or canola seed (n=9) or no oilseed (n=9). Ovaries were scanned until 35 days postpartum. Prepartum diets did not influence SCE, but a preovulatory-size follicle developed sooner (P≤0.05), the interval to first ovulation was shorter and the proportion of cows ovulating within 35 days postpartum was greater in the sunflower seed group. Although mRNA expression of cytokines was not affected by diet, cows with SCE had higher (P≤0.05) expression of interleukin-1ß (IL1B), interleukin-8 (CXCL8), IL10 and tumour necrosis factor-α (TNF) than normal cows. The interval (mean ± s.e.m.) from calving to preovulatory-size follicle was shorter (P≤0.05) in normal (13.2±0.9 days) than SCE cows (18.7±1.4 days). In summary, a prepartum diet supplemented with sunflower seed positively influenced postpartum ovarian function without affecting UIS or pro- and anti-inflammatory cytokine gene expression in endometrial cells.

Prepartum maternal diets supplemented with oilseeds alter the fatty acid profile in bovine neonatal plasma possibly through reduced placental expression of fatty acid transporter protein 4 and fatty acid translocase.

In the present study, we determined the effects of maternal dietary fat and the type of fat on plasma fatty acids and the expression of placental fatty acid transporter genes. In Experiment 1, Holstein cows in the last 35 days of gestation received diets containing sunflower seed (n=8; high in linoleic acid (LA)), canola seed (n=7; high in oleic acid (OLA)) or no oilseed (n=7; control). Fatty acids were quantified in dam and neonate plasma at calving. In Experiment 2, placental cotyledons were collected (LA: n=4; OLA: n=4; control: n=5) to quantify gene expression. Maternal long-chain polyunsaturated fatty acids, neonatal total n-3 fatty acids and eicosapentaenoic acid (EPA) declined, whereas docosahexaenoic acid (DHA) and total fat tended to decline following fat supplementation prepartum. Feeding of LA versus OLA prepartum tended to increase peroxisome proliferator-activated receptor α (PPARA) expression, whereas peroxisome proliferator-activated receptor δ (PPARD) and peroxisome proliferator-activated receptor γ (PPARG) expression tended to be higher in OLA- than LA-fed cows. Expression of fatty acid transporter protein 4 (FATP4) and fatty acid translocase (FAT/CD36) expression was lower in placental tissue of cows fed fat compared with control cows. Reduced total n-3 fatty acids, EPA and DHA in neonates born of dams fed fat prepartum is likely due to changes in PPARs and reduced expression of placental FATP4 and FAT/CD36.

Morphologic and transcriptomic assessment of bovine embryos exposed to dietary long-chain fatty acids.

The main objectives of this study were to determine the influence of diets enriched in α-linolenic, linoleic or oleic acid on the development and transcriptomic profile of embryos collected from dairy cattle. Non-lactating Holstein cows received one of the three diets supplemented with 8% rolled oilseeds: flax (FLX, n = 8), sunflower (SUN, n = 7) or canola (CAN, n = 8). After a minimum 35-day diet adaptation, cows were superovulated, artificially inseminated and ova/embryos recovered non-surgically after 7.5 days. Cows fed FLX had less degenerated embryos and more viable embryos than those fed CAN or SUN. In total, 175 genes were differentially expressed in blastocysts from cows fed FLX than in cows fed CAN or SUN. These differentially expressed genes were mainly involved in cellular growth and proliferation, cellular development, and cell survival and viability. In conclusion, dietary n-3 polyunsaturated fatty acids reduced early embryonic degeneration possibly through improving embryonic cell survival and viability.

A prepartum diet supplemented with oilseeds high in oleic or linoleic acid reduced GnRH-induced LH release in dairy cows during second week postpartum.

BACKGROUND: The objective was to determine the effect of prepartum diets supplemented with rolled canola seed (high in oleic acid) or sunflower seed (high in linoleic acid) on luteinizing hormone (LH) pulsatility and gonadotropin releasing hormone (GnRH)-induced LH release during early postpartum. METHODS: Thirty-one pregnant Holstein cows, blocked by body condition score, parity and expected calving date, were assigned to 1 of 3 prepartum diets supplemented with 8 % rolled canola or sunflower seed, or no oilseed (control) during the last 35 d of gestation. Blood samples were collected at Weeks (wk)-3, 0, +1 and +2, relative to calving, to determine non-esterified fatty acids (NEFA), Beta-hydroxy butyric acid (BHBA) and glucose. Additional blood samples were collected during wk1 (n = 5 per treatment) or wk2 (n = 5 or 6 per treatment), for 6 h, to measure LH pulsatility; thereafter, 100 mcg GnRH was administrated i.m., and blood was sampled for 4 h more, to measure GnRH-induced LH release. RESULTS: Dietary treatment did not affect prepartum energy balance, but cows fed the control diet were in a deeper state of negative energy balance during wk2, than those fed canola (P = 0.03) or sunflower (P = 0.01). Prepartum diets did not influence the mean plasma concentration of BHBA and glucose. However, NEFA concentration during wk2 was greater in control cows than those fed sunflower (P = 0.03) or canola (P = 0.07). Prepartum diets did not affect LH pulsatility (i.e. mean, minimum, maximum concentration, pulse frequency, and amplitude during wk1 and 2). GnRH-induced LH release did not differ among dietary treatments during wk1 but the mean GnRH-induced LH release during wk2 was either greater (P = 0.02) and tended to be greater (P = 0.09) in control cows than in those fed canola and sunflower, respectively. CONCLUSIONS: Prepartum diets did not affect LH pulsatility and GnRH-induced LH release during the first week postpartum, but cows fed a diet supplemented with oilseeds high in oleic or linoleic acid released less LH than control cows, in response to an exogenous GnRH challenge during the second week postpartum.

Effects of prepartum fat supplementation on plasma concentrations of glucagon-like peptide-1, peptide YY, adropin, insulin, and leptin in periparturient dairy cows.

Dietary fat supplementation during the periparturient period is one strategy to increase energy intake and attenuate the degree of negative energy balance during early lactation; however, little is known of the underlying hormonal and metabolic adaptations. We evaluated the effects of prepartum fat supplementation on energy-balance parameters and plasma concentrations of glucagon-like peptide-1, peptide tyrosine-tyrosine (PYY), adropin, insulin, leptin, glucose, nonesterified fatty acid, and ß-hydroxybutyric acid in dairy cows. Twenty-four pregnant dairy cows were randomized to diets containing either rolled canola or sunflower seed at 8% of dry matter, or no oilseed supplementation, during the last 5 wk of gestation and then assigned to a common lactation diet postpartum. Blood samples were collected at -2, +2, and +14 h relative to feeding, at 2 wk after the initiation of the diets, and at 2 wk postpartum. Dietary canola and sunflower supplementation alone did not affect energy balance, body weight, and plasma concentrations of glucagon-like peptide-1, PYY, adropin, insulin, leptin, nonesterified fatty acid, and ß-hydroxybutyric acid; however, canola decreased and sunflower tended to decrease dry matter intake. We also observed that the physiological stage had a significant, but divergent, effect on circulating hormones and metabolite concentrations. Plasma glucagon-like peptide-1, PYY, adropin, nonesterified fatty acid, and ß-hydroxybutyric acid concentrations were greater postpartum than prepartum, whereas glucose, insulin, leptin, body weight, and energy balance were greater prepartum than postpartum. Furthermore, the interaction of treatment and stage was significant for leptin and adropin, and tended toward significance for PYY and insulin; only insulin exhibited an apparent postprandial increase. Postpartum PYY concentrations exhibited a strong negative correlation with body weight, suggesting that PYY may be associated with body weight regulation during the transition period. These novel findings demonstrate that the transition from pregnancy to lactation is a stronger determinant of circulating gut hormone concentrations than dietary lipid in transition dairy cows.

Low-dose natural prostaglandin F2α (dinoprost) at timed insemination improves conception rate in dairy cattle.

The primary objective was to determine if low doses of PGF2α (dinoprost) given intramuscularly (im) concurrent with timed artificial insemination (TAI) would improve conception rates in dairy cattle. A secondary objective was to determine if body condition score (BCS) and parity would influence conception rates, either independently or in interaction with PGF2α treatment. In experiment I, 307 lactating Holstein cows were randomly assigned to receive either 5-mg PGF2α im (PGF2α treated, n = 154) or 0-mg PGF2α (control, n = 153) at TAI (Day 0). Blood samples were obtained on Days -10, -3, 0, and 7 to determine plasma progesterone (P4) concentrations. Pregnancy was confirmed 30 to 32 days after insemination by transrectal ultrasonography. In experiment II, 451 cows were randomly assigned to receive either 10-mg PGF2α im (PGF2α treated, n = 226) or 0-mg PGF2α (control, n = 225) at TAI, and pregnancy was confirmed 45 to 50 days after TAI by palpation per rectum. Pregnancy data were analyzed by CATMOD (SAS). In experiment I, PGF2α treatment, BCS, and parity did not affect conception rate (35.7% vs. 37.0% for PGF2α treated vs. control; P > 0.05). However, in experiment II, conception rates were higher in cows given 10-mg PGF2α than those in control cows (45.8% vs. 36.0%; P < 0.05), in cows with high BCS than in cows with low BCS (52.1% vs. 30.4%; P < 0.01), and in primiparous than in multiparous cows (47.6% vs. 34.4%; P < 0.01), but their interaction with PGF2α treatment did not affect conception rates. In summary, 5 mg of PGF2α given im concurrent with TAI failed to enhance conception rate in lactating dairy cows, whereas 10 mg of PGF2α significantly increased conception rate.

Progesterone supplementation before timed AI increased ovulation synchrony and pregnancy per AI, and supplementation after timed AI reduced pregnancy losses in lactating dairy cows.

One of the causes of poor fertility in high producing dairy cows is inadequate progesterone. Therefore, we determined the efficacy of an intravaginal insert containing 1.55 g of progesterone (PRID) given before and/or after timed AI (TAI) on ovarian response, plasma progesterone concentrations, pregnancy per AI (P/AI) and pregnancy losses. Lactating dairy cows at three locations were assigned (Day 0) to an Ovsynch protocol with (N = 294) or without (N = 314) a PRID. The Ovsynch protocol consisted of two injections of 100 µg gonadorelin (GnRH) 9 days apart and one injection of 500 µg cloprostenol (PG) 7 days after the first GnRH treatment. Insertion and removal of PRID occurred concurrent with the first GnRH and PG treatments, respectively. Timed AI was carried out 12 to 16 hours after the second GnRH. Ovarian status of a subset of 217 first service cows had been presynchronized with 2 treatments of PG 14 days apart with the last PG given 12 days before the first GnRH of the Ovsynch protocol. Body condition score (scale of one to five) was recorded at TAI. Ultrasonographic examinations were done in all cows at first GnRH, at PG, at TAI, and 24 hours after TAI for response to treatment and at 32 and 60 days after TAI for confirmation of pregnancy. At 4.5 days after TAI (Day 14), cows that responded to PG and ovulated after the second GnRH treatment were reassigned to receive (N = 223) or not receive (N = 229) a PRID for 7 days. Blood samples were taken for progesterone determination at PG treatment, at TAI, and post TAI on Days 14 and 21. The PRID treatment pre-TAI reduced the percentage of cows ovulating before TAI (5.8% vs. 11.1%), and significantly increased P/AI in nonpresynchronized cows (41.3% vs. 25.1%). Cows ovulating in response to the first GnRH treatment, cyclic cows, and cows with body condition score of 2.75 or more had increased P/AI, but the addition of a PRID pre-TAI to these cows did not increase P/AI. The PRID treatment post TAI did not affect P/AI, but reduced pregnancy losses (6.1% vs. 11.4%) between 32 and 60 days of gestation. The reduction in pregnancy losses tended (P = 0.10) to be significant in acyclic cows receiving a PRID than in those not receiving a PRID (5.6% vs. 33.3%). Plasma progesterone concentrations at PG treatment and on Day 21 (11.5 days after TAI) were linearly associated with P/AI. In conclusion, progesterone supplementation pre-TAI increased P/AI in nonpresynchronized cows. Progesterone supplementation post TAI reduced pregnancy losses, particularly in acyclic cows.

Characterisation of the bacterial microbiota of the vagina of dairy cows and isolation of pediocin-producing Pediococcus acidilactici.

BACKGROUND: Uterine infections in dairy cows lower profitability of dairy operations. Infections of the reproductive tract are related to the overgrowth of pathogenic bacteria during the first three weeks after parturition. However, alterations in the vaginal microbiota composition in the first weeks after parturition remain poorly documented. RESULTS: In this study, bacteria isolated from the vagina of healthy pregnant, and infected postpartum cows were characterised by random amplification of polymorphic DNA (RAPD) analysis and partial 16S ribosomal RNA (rDNA) gene sequencing. Populations of bacilli and lactic acid bacteria of the genera Enterococcus, Lactobacillus, and Pediococcus were present in both healthy and infected cows. Infected cows had a significant increase in the vaginal enteric bacteria population which consisted mainly of Escherichia coli. Three E. coli isolates harboured the gene coding for Shiga-like-toxin (SLT) I or II. Several isolates of the Pediococcus acidilactici were found to produce the bacteriocin pediocin AcH/PA-1. Quantitative PCR analyses of vaginal mucus samples collected from ten metritic cows before and after parturition confirmed the presence of the Lactobacillus group (Lactobacillus spp., Pediococcus spp., Leuconostoc spp., and Weissella spp.); Enterobacteriaceae, E. coli, and bacilli. The presence of the pediocin AcH/PA-1 structural gene and SLT genes were also confirmed with qPCR. CONCLUSIONS: In conclusion, overgrowth of pathogenic bacteria, particularly E. coli, after parturition likely contributes to the development of metritis. Our microbiota analysis extends the information related to the composition of commensal bacteria in the bovine female reproductive tract and may facilitate the development of novel intervention strategies for prevention of uterine infections in dairy cows.

Relationships between endometrial cytology and interval to first ovulation, and pregnancy in postpartum dairy cows in a single herd.

The objectives were to investigate the relationships between endometrial cytology (EC) and interval from calving to first ovulation, and pregnancy in dairy cows, and that between uterine fluid and EC. On day 25 postpartum, 39 dairy cows were grouped based on EC, as having low (≤8%) or high (>8%) polymorphonuclear cells (PMN), and the quantity of uterine fluid was assessed by ultrasound. The interval from calving to first ovulation was shorter in low, than in high PMN cows (32 vs. 45 d). A greater proportion of cows with uterine fluid had high PMN (64% vs. 21%), and the PMN increased from 14% to 34% as the quantity of uterine fluid increased. The mean interval from calving to ovulation was longer in primiparous cows with high PMN (49 d) compared to that of primiparous and multiparous cows with low PMN (28 and 29 d, respectively). Although the conception rate to first service at 92 d postpartum was not different between PMN groups, the cumulative pregnancy at 270 d tended to be higher in low than in high PMN (80% vs. 58%) multiparous cows. Also, cows that had uterine fluid on day 25 postpartum had a shorter interval from calving to pregnancy than those with no uterine fluid (161 vs. 208 d). In conclusion, combining transrectal ultrasonography with endometrial cytology on day 25 postpartum has diagnostic value in the assessment of uterine inflammation.

Precocious mammary development in an 8-month-old Holstein heifer.

An 8-month-old, virgin Holstein heifer with precocious mammary development was presented for examination. Protein, fat, and lactose in the mammary secretion were 14.90%, 0.12%, and 0.20%, respectively; somatic cell count was 3.9 x 10(6)/mL, with no bacterial infection. The heifer was inseminated at 15 months of age, confirmed pregnant, and subsequently slaughtered.

Comparison of 2 enzyme immunoassays and a radioimmunoassay for measurement of progesterone concentrations in bovine plasma, skim milk, and whole milk.

The objective of this study was to compare 2 enzyme immunoassays (EIAs) with a radioimmunoassay (RIA) as to sensitivity and accuracy in the measurement of the progesterone (P4) concentration in bovine plasma, skim milk, and whole milk. The 72 samples from 24 lactating dairy cows expected to have either a high P4 concentration (cows in diestrus or pregnant) or a low P4 concentration (cows in estrus or anestrus) were analyzed by RIA, solid-phase EIA (SPEIA), which included a solvent extraction step, or direct EIA (DEIA) without solvent extraction. The overall mean concentrations of P4 did not differ (P < 0.4) among the assays. However, for the cows that were in diestrus or pregnant, the mean P4 concentrations (and standard error) were higher (P < 0.03), regardless of sample type, with RIA than with SPEIA, at 7.3 (0.7) and 6.1 (0.6) ng/mL, respectively. When only the high-P4 samples analyzed by RIA were compared, the mean P4 concentration was higher (P < 0.001) in whole milk than in skim milk, at 9.8 (1.0) and 4.1 (0.7) ng/mL, respectively. Although the mean P4 concentrations in the low-P4 samples did not differ (P < 0.80) among assays, the proportions of cows with a P4 concentration > or = 1 ng/mL were 3%, 14%, and 44% for RIA, SPEIA, and DEIA, respectively (P < 0.01; DEIA > SPEIA > RIA).

Evaluation of early conception factor lateral flow test to determine nonpregnancy in dairy cattle.

The early conception factor (ECF) lateral flow test was evaluated for its ability to accurately determine nonpregnant status in dairy cattle. Results of 2 field trials involving 191 cows and 832 tests indicated the probability that a cow can be correctly diagnosed as nonpregnant by using the ECF test is only about 50%. Agreement of test results between milk and serum obtained from the same cow was 57.5%. The ECF test was not consistent in identifying nonpregnancy when the same cows were tested repeatedly over a period of 4 weeks. We conclude that the ECF lateral flow test does not accurately identify nonpregnancy in dairy cattle.

Ovarian and endocrine responses associated with the treatment of cystic ovarian follicles in dairy cows with gonadotropin releasing hormone and prostaglandin F2alpha, with or without exogenous progesterone.

The objectives of this observational study were to document ovarian and endocrine responses associated with the treatment of cystic ovarian follicles (COFs) in dairy cows, using gonadotropin releasing hormone (GnRH) and prostaglandin F2alpha (PGF) with or without exogenous progesterone. A secondary objective was to determine pregnancy establishment following synchronization of ovulation and timed insemination in cows diagnosed with COFs. In trial I, 18 Holstein cows diagnosed with COFs received 2 injections of 100 microg GnRH, 9 d apart, with 25 mg PGF given 7 d after the 1st GnRH. A new follicle developed in all 18 cows after the 1st GnRH, and 83% of cows ovulated following the 2nd GnRH. Cows were inseminated 16 h after the 2nd GnRH. Of the 17 cows available for pregnancy diagnosis, 7 were confirmed pregnant. In trial II, 8 cows with COFs received GnRH and an intravaginal progesterone device (CIDR) concurrently, then PGF 7 d later. The CIDR was removed 2 d after PGF administration. Plasma estradiol concentrations declined following CIDR insertion. In all cows, a new follicle developed following GnRH treatment; estradiol-surge and estrus occurred spontaneously after CIDR-removal. Seven of 8 cows ovulated the new follicle. In dairy cows diagnosed with COFs, treatment with GnRH followed by PGF 7 d later, with or without exogenous progesterone, resulted in the recruitment of a healthy new follicle; synchronization of ovulation and timed insemination resulted in a 41% pregnancy rate.