S.H. Cheong

Cow-level and herd-level risk factors for subclinical endometritis in lactating Holstein cows

The objectives of this study were to obtain prevalence estimates for subclinical endometritis (SCE), determine cow- and herd-level risk factors, and evaluate the reproductive consequences of SCE. A cross-sectional study was used to determine prevalence and risk factors with cows followed in a prospective study to determine reproductive outcomes. Lactating Holstein cows were sampled between 40 and 60 d in milk using low-volume uterine lavage, and cytology was evaluated to determine SCE status. In total, 779 cows from 38 herds were used in the analysis. The cow-level prevalence of SCE was 25.9%. Within-herd level prevalence ranged from 4.8 to 52.6% (median 26.3%, interquartile range 15.6 to 33.3%). Cow-level risk factors identified were ketosis [odds ratio (OR) 3.83; 95% confidence interval (CI) 1.82-8.07], acute metritis (OR 1.86; 95% CI 1.05-3.30], and the interaction between milk production and parity. Primiparous cows that produced more milk had increased odds of having SCE, whereas multiparous cows that produced more milk had decreased odds of having SCE. Herd-level risk factors identified were housing early postpartum cows on bedded packs (herd-level SCE=36.1%), which increased herd prevalence of SCE by 16.7% (SE 5.58) compared with early postpartum cows housed in freestalls (herd-level SCE=19.4%), and straw bedding in the calving pen, which decreased herd prevalence of SCE by 10.7% (SE 3.59) compared with herds that used other bedding material. In this study, primiparous cows with and without SCE had similar reproductive performance; however, multiparous cows with SCE had median days open 44 d longer (159 d; 95% CI 126-186 d) compared with unaffected multiparous cows (115 d; 95% CI 106-132 d).

Subcutaneous Immunization with Inactivated Bacterial Components and Purified Protein of Escherichia coli, Fusobacterium necrophorum and Trueperella pyogenes Prevents Puerperal Metritis in Holstein Dairy Cows

In this study we evaluate the efficacy of five vaccine formulations containing different combinations of proteins (FimH; leukotoxin, LKT; and pyolysin, PLO) and/or inactivated whole cells (Escherichia coli, Fusobacterium necrophorum, and Trueperella pyogenes) in preventing postpartum uterine diseases. Inactivated whole cells were produced using two genetically distinct strains of each bacterial species (E. coli, F. necrophorum, and T. pyogenes). FimH and PLO subunits were produced using recombinant protein expression, and LKT was recovered from culturing a wild F. necrophorum strain. Three subcutaneous vaccines were formulated: Vaccine 1 was composed of inactivated bacterial whole cells and proteins; Vaccine 2 was composed of proteins only; and Vaccine 3 was composed of inactivated bacterial whole cells only. Two intravaginal vaccines were formulated: Vaccine 4 was composed of inactivated bacterial whole cells and proteins; and Vaccine 5 was composed of PLO and LKT. To evaluate vaccine efficacy, a randomized clinical trial was conducted at a commercial dairy farm; 371 spring heifers were allocated randomly into one of six different treatments groups: control, Vaccine 1, Vaccine 2, Vaccine 3, Vaccine 4 and Vaccine 5. Late pregnant heifers assigned to one of the vaccine groups were each vaccinated twice: at 230 and 260 days of pregnancy. When vaccines were evaluated grouped as subcutaneous and intravaginal, the subcutaneous ones were found to significantly reduce the incidence of puerperal metritis. Additionally, subcutaneous vaccination significantly reduced rectal temperature at 6±1 days in milk. Reproduction was improved for cows that received subcutaneous vaccines. In general, vaccination induced a significant increase in serum IgG titers against all antigens, with subcutaneous vaccination again being more effective. In conclusion, subcutaneous vaccination with inactivated bacterial components and/or protein subunits of E. coli, F. necrophorum and T. pyogenes can prevent puerperal metritis during the first lactation of dairy cows, leading to improved reproduction.

Association between interleukin-8 receptor-α (CXCR1) polymorphism and disease incidence, production, reproduction, and survival in Holstein cows

The objective was to evaluate the association between the single nucleotide polymorphism at position +735 in the interleukin-8 receptor-α (CXCR1) gene (CXCR1c.735) and disease incidence, milk production, reproductive performance, and survival in Holstein cows. Three-hundred fifty Holstein cows were enrolled. No association was found between CXCR1c.735 genotype and retained fetal membranes, metritis, or endometritis. Incidence rate of clinical mastitis was associated with CXCR1c.735 genotype; cows with genotypes CC and GC had a decreased incidence rate of clinical mastitis compared with GG cows. Milk yield was associated with CXCR1c.735 genotype; cows with genotype GC had greater milk yield than GG cows. Hazard of pregnancy was not associated with CXCR1c.735 genotype. Cows that had clinical mastitis had decreased hazard of pregnancy, and cows that had endometritis tended to have a decreased hazard of pregnancy. Hazard of death or culling was not associated with CXCR1c.735 genotype. Multiparous cows and cows that had mastitis had increased hazard of death or culling. In contrast to what we expected, cows with the genotype GG had an increased incidence rate of clinical mastitis and decreased milk yield.

Live Births from Domestic Dog (Canis familiaris) Embryos Produced by In Vitro Fertilization

Development of assisted reproductive technologies (ART) in the dog has resisted progress for decades, due to their unique reproductive physiology. This lack of progress is remarkable given the critical role ART could play in conserving endangered canid species or eradicating heritable disease through gene-editing technologies—an approach that would also advance the dog as a biomedical model. Over 350 heritable disorders/traits in dogs are homologous with human conditions, almost twice the number of any other species. Here we report the first live births from in vitro fertilized embryos in the dog. Adding to the practical significance, these embryos had also been cryopreserved. Changes in handling of both gametes enabled this progress. The medium previously used to capacitate sperm excluded magnesium because it delayed spontaneous acrosome exocytosis. We found that magnesium significantly enhanced sperm hyperactivation and ability to undergo physiologically-induced acrosome exocytosis, two functions essential to fertilize an egg. Unlike other mammals, dogs ovulate a primary oocyte, which reaches metaphase II on Days 4–5 after the luteinizing hormone (LH) surge. We found that only on Day 6 are oocytes consistently able to be fertilized. In vitro fertilization of Day 6 oocytes with sperm capacitated in medium supplemented with magnesium resulted in high rates of embryo development (78.8%, n = 146). Intra-oviductal transfer of nineteen cryopreserved, in vitro fertilization (IVF)-derived embryos resulted in seven live, healthy puppies. Development of IVF enables modern genetic approaches to be applied more efficiently in dogs, and for gamete rescue to conserve endangered canid species.

Metabolic and Endocrine Differences Between Dairy Cows That Do or Do Not Ovulate First Postpartum Dominant Follicles

ABSTRACT Most dairy cows develop the first dominant follicle postpartum within 2 wk after calving, but only about 40% of these follicles produce sufficient estradiol to stimulate ovulation despite having normal ultrasound appearance and growth. This study aimed to characterize metabolic, endocrine, and follicular fluid profiles of cows in which the first dominant follicle postpartum will become ovulatory and those with nonovulatory follicles. Luteinizing hormone pulse frequency, follicular fluid androstenedione, and follicular fluid estradiol concentrations were lower in nonovulatory cows suggesting that the function of theca cells is impaired. In addition, nonovulatory cows had more severe negative energy balance and greater insulin resistance postpartum. This study describes for the first time the steroid hormone profile of early postpartum follicles and shows that a steroidogenic defect most likely occurs in theca cells limiting the amount of androgen precursor available for estradiol production that impairs their ovulatory outcome.

The effects of conjugated linoleic acid isomers cis-9,trans-11 and trans-10,cis-12 on in vitro bovine embryo production and cryopreservation.

Conjugated linoleic acid (CLA) isomers can affect the lipid profile and signaling of cells and thereby alter their function. A total of 5,700 bovine oocytes were used in a structured series of experiments to test the effects of CLA cis-9,trans-11 and CLA trans-10,cis-12 in vitro. In experiment 1, high doses of each CLA isomer during in vitro maturation (IVM) were compared with high or low doses during the entire in vitro culture (IVC) of parthenogenetic embryos. High doses of the CLA isomers ranged from 50 to 200 μM and low doses were 15 and 25 μM. In experiment 2, the low doses of each CLA isomer were tested during IVM/IVC on embryos produced by in vitro fertilization (IVF). Experiment 3 compared the effects of 15 μM doses of each CLA isomer during IVM or IVC of IVF embryos. In experiment 4, post-rewarming survival rates and blastomere counts were assessed for embryos supplemented with each CLA isomer during IVM or for 36 h before vitrification. In experiment 1, when either CLA isomer was provided only during IVM, we observed no effects on overall rates of maturation, cleavage, or blastocysts (92.2 ± 1.6%, 78.3 ± 4.1%, and 28.9 ± 5.1%, respectively). However, high doses of each CLA isomer, but not low doses, during the entire embryo culture period decreased blastocyst rates (5-20%) in a dose-dependent manner. Cleavage rates improved with 15 or 50 μM CLA trans-10,cis-12. Progesterone concentrations in maturation media were significantly increased by high doses of each CLA isomer compared with control, but low doses of CLA isomers had no effect. In experiment 2 with IVF embryos, low doses of each CLA isomer did not alter cleavage rates (average 84.9 ± 1.9%) and only 25 μM CLA trans-10,cis-12 during IVC reduced blastocyst rates below those of controls (25.5 ± 2.1 vs. 38.2 ± 2.3%). The lipid content of embryos was increased and relative expression of the BIRC5 (baculoviral IAP repeat containing 5) gene was depressed by CLA trans-10,cis-12. In experiment 3, low doses (15μM) of each CLA isomer during IVC significantly reduced blastocyst rates (20.6 ± 2.4% and 27.7 ± 1.2% vs. 34.18 ± 1.2% for CLA trans-10,cis-12 and CLA cis-9,trans-11 compared with control, respectively) with less effect of each CLA during IVM. In experiment 4, adding 100 μM CLA cis-9,trans-11 during the final 36 h of culture resulted in a high survival rate after rewarming and culture, and the higher embryo blastomere count was comparable to that of control embryos not undergoing vitrification. In conclusion, supplementation with either CLA isomer did not improve embryo production, but inclusion of CLA cis-9,trans-11 before vitrification improved the quality of bovine IVF embryos after rewarming and culture.

Uterine and systemic inflammation influences ovarian follicular function in postpartum dairy cows

The objective of this study was to determine the effects of uterine and systemic inflammatory responses to uterine bacterial contamination at calving in dairy cows on the growth and ovulatory outcomes of the first dominant follicle postpartum. Ovulatory capability of the first dominant follicle postpartum was predicted in 53 multiparous cows by using a combination of follicle growth characteristics and circulating estradiol concentrations. Endotoxin levels were assayed in follicular fluid samples that were aspirated the day after ovulatory outcome prediction. Plasma levels of haptoglobin, a proinflammatory acute phase protein, and paraoxonase, a negative acute phase protein were determined. Uterine bacteria and inflammation were evaluated in three uterine fluid samples from each cow collected on the day of calving, the day after follicle aspiration, and at 35 days postpartum. Cows that had a strong initial uterine inflammatory response (robust recruitment of polymorphonuclear leukocytes of ≥ 35% and cows with uterine pH < 8.5 on the day of calving) were more likely to have an ovulatory first dominant follicle. Follicular fluid endotoxin levels were higher in non-ovulatory cows compared with ovulatory cows. Endotoxin levels in circulation were not different between ovulatory groups but were higher prepartum than on day 7 and 14 postpartum. Systemic inflammation characterized by elevated haptoglobin concentrations was higher in non-ovulatory cows despite similar bacterial contamination and circulating endotoxin levels. Paraoxonase activity in follicular fluid was significantly associated with the paraoxonase activity in plasma, however, plasma paraoxonase concentrations were not different between non-ovulatory and ovulatory cows. Cows with a higher uterine bacterial load on the day of calving had slower ovarian follicle growth. In summary, a robust uterine inflammatory response on the day of calving was positively associated with ovarian function while elevated systemic inflammation during the early postpartum period was negatively associated with the ovulatory status of the first dominant follicle postpartum.

Effects of diagnostic low-volume uterine lavage shortly before first service on reproductive performance, culling and milk production.

The objective was to evaluate if uterine sampling by low-volume uterine lavage, done shortly before the end of the voluntary waiting period, affected reproductive performance, milk production, or culling. Approximately 20 cows (40 and 60 d postpartum) were sampled in each of 38 herds (total, 752 cows), whereas all remaining cows concurrently between 40 and 60 d were a non-sampled reference cohort (n = 2,252 cows). There was an interaction between parity and sampling for first-service conception, calving to conception interval, and milk production. When stratified by parity, there was a tendency for impaired reproductive performance in sampled primiparous cows compared with non-sampled primiparous cows (Odds Ratio for pregnancy to first-service = 0.78 (95% C.I. 0.58-1.04). The Hazard Ratio for pregnancy in sampled primiparous cows was 0.89 (95% C.I. 0.77-1.04) compared to non-sampled primiparous cows. Sampling did not affect first-service conception rate (Odds Ratio for pregnancy Odds ratio (OR) for pregnancy = 1.03; 95% C.I. 0.80-1.33) or calving-to conception interval in multiparous cows (Hazard Ratio = 1.04; 95% C.I. 0.91-1.18). Sampling did not affect culling risk (Hazard Ratio HR = 0.92; 95% C.I. 0.77-1.11) after accounting for covariates. After stratification by parity, milk production was not affected by sampling except in fourth- and fifth-parity cows where sampled cows produced more milk than non-sampled cows after controlling for first test-day milk production and days-postpartum at first test-day. In conclusion, sampling by low-volume uterine lavage did not have significant detrimental effects on reproduction, culling, or milk production. However, there was a tendency for lower first-service conception in sampled primiparous cows, but the procedure appeared to be benign in multiparous cows.

Tumor collision of uterine adenocarcinoma and leiomyosarcoma in a goat

An aged mixed-breed goat doe was presented with a 9-mo history of serosanguineous vaginal discharge. Vaginal speculum examination revealed serosanguineous discharge but otherwise no abnormalities. Transrectal ultrasonography showed normal ovaries and multifocal cystic lesions within the uterus. Ovariohysterectomy was recommended because of a strong suspicion of neoplasia. Multiple, non-resectable masses were noted in and around the uterus intraoperatively, and euthanasia was elected. Autopsy revealed multiple masses within the uterus, cervix, and lung parenchyma. Histologically, the masses within the uterus represented a likely collision tumor of primary adenocarcinoma and leiomyosarcoma. Our report highlights the importance of obtaining biopsy samples of all masses because the lesions described showed significantly different biological behavior. This information is vital to guide treatment and prognosis.

Pyometra and unilateral uterine horn torsion in a sheep

A 13-year-old Romanov sheep presented for evaluation of vaginal discharge, depression and anorexia. Blood work demonstrated mature neutrophilia and marked hyperlactatemia. Transrectal and transabdominal ultrasound revealed echogenic fluid (pus) within the uterus. Purulent uterine fluid culture yield Escherichia coli. A diagnosis of pyometra was made. After medical treatment without clinical improvement, an exploratory laparotomy was performed, and a 360-degree uterine left horn torsion was identified with the pyometra and multiple corpora lutea on both ovaries. Ovariohysterectomy was performed and the ewe recovered uneventfully.