Short communication: Urea induces T helper 2 (Th2) type environment at transcriptional level and prostaglandin E2 secretion in bovine oviduct epithelial cells in culture.

Excess dietary protein intake in early lactation dairy cows resulting in blood urea nitrogen of greater than 19 to 20mg/dL is associated with decreased fertility. Little is known about the local interference of urea in the normal immunological environment of the oviduct that provides optimal conditions for early reproductive events. A bovine oviduct epithelial cell (BOEC) culture was used to determine how urea influences immune environment. The BOEC monolayer was supplemented with low (20mg/dL) and high (40mg/dL) concentrations of urea together with ovarian steroids, estradiol (1ng/mL) and progesterone (1ng/mL), and LH (10ng/mL) at concentrations observed during the preovulatory period. The urea values used in this study were equivalent to 9.3 and 18.7mg/dL of blood urea nitrogen, which are typically common in lactating dairy cows with low or high protein intake, respectively. Stimulation of BOEC with 40mg/dL of urea induced gene expression of IL10 and IL4, epithelial-derived T helper type 2-driving (anti-inflammatory) cytokines as well as mPGES-1 expression and prostaglandin E2 (PGE2) secretion. However, urea concentrations of both 20 and 40mg/dL failed to alter the expression of IL1B and TNFA, Th1-driving cytokines, and the gene expression of TLR4. However, a concentration of 40mg/dL of urea stimulated α 1-acid glycoprotein expression, an acute phase protein. Data from this in vitro study suggest that urea, at least in part, contributes to influence the expression of some immune-related genes toward T helper type 2 type and prostaglandin E2 secretion, leading to disruption in local environment for fertilization and early embryonic development.

Evidence for a novel, local acute-phase response in the bovine oviduct: progesterone and lipopolysaccharide up-regulate alpha 1-acid-glycoprotein expression in epithelial cells in vitro.

Little is known about the local production and function of alpha 1-acid glycoprotein (AGP), an acute-phase protein, in the female reproductive tract. This study aimed to investigate the regulation and immune function of AGP in cultured bovine oviduct epithelial cells. Analysis by Western blotting and immunohistochemistry revealed that bovine oviduct tissue expresses AGP protein in epithelial cells and the smooth muscle layer. Stimulation of bovine oviduct epithelial cells in culture with either progesterone (1 ng/ml) or lipopolysaccharide (LPS, 10 ng/ml) induced both mRNA expression and secretion of AGP. Estradiol (1 ng/ml), progesterone (1 ng/ml), and luteinizing hormone (10 ng/ml), which are observed during the peri-ovulatory period in oviduct tissues (steroids) or in circulation (luteinizing hormone), suppressed LPS-induced expression and secretion of AGP, which in turn induced the expression of Toll-like receptor-4 (TLR-4) and interleukin-1ß (IL-1B), but suppressed TLR-2 and tumor necrosis factor-α (TNFA) expression. AGP also inhibited LPS-induced TLR-2 and TNFA expression, but had no effect on LPS-induced TLR-4 and IL-1B expression. These findings suggest that oviductal epithelial cells can participate in antimicrobial processes through the secretion of AGP, which is partly regulated by ovarian steroids. Moreover, oviductal AGP may regulate the response of epithelial cells, thereby reducing the expression of the acute pro-inflammatory cytokine TNFA, which could contribute to the local homeostasis during the acute response to endotoxin release in the oviduct's anti-infection process.

Alpha lipoic acid reverses the negative effect of LPS on mouse spermatozoa and developmental competence of resultant embryos in vitro.

BACKGROUND: Reproductive toxicity of lipopolysaccharide (LPS) on spermatozoa is well established. OBJECTIVE: The aim of the present study was to show the potential benefits of alpha lipoic acid (ALA) as a strong antioxidant in alleviating the reproductive toxicity of LPS. MATERIALS AND METHODS: Sperm cells and cumulus-oocyte complexes (COCs) were collected from healthy NMRI mice (body weights ranged from 25 to 35 g, 100 females and 200 males). Sperm cells were treated with varying doses of ALA (0.01, 0.02, and 0.04 mm) and 0.01 µg/mL of LPS for 4 h. The quality of spermatozoa (ROS production, DNA fragmentation, and spontaneous acrosome reaction), sperm fertilizability, and the consequent developmental competence of oocytes inseminated with ALA/LPS-treated spermatozoa were recorded. RESULTS: The results showed that 0.04 mm of ALA abrogated LPS-reduced sperm motility, viability, ROS production, spontaneous acrosome reaction, fertilizability, and developmental competence. In addition, 0.04 mm ALA significantly reverted the negative effects of LPS on inner cell mass (ICM) cell counts, total cell number (TCM), and ratio between ICM and TCM. DISCUSSION: Our data showed that ALA significantly could abrogate the negative effects of LPS on sperm quality and oocyte developmental competence. Therefore, ALA had the capacity for protecting sperm cells from LPS-induced damage and ensured fertilization and developmental competency. CONCLUSION: These in vitro findings suggested a therapeutic role for ALA in reducing the negative effects of LPS on spermatozoa and early embryonic development.

Corn silage partially replacing short alfalfa hay to optimize forage use in total mixed rations for lactating cows.

We theorized that adding corn silage to a total mixed ration with alfalfa hay as the sole dietary forage would improve nutrient intake and chewing activity and thereby improve rumen fermentation and milk production. The objective of this research was to determine the effects of partial replacement of short alfalfa [physically effective (pe) neutral detergent fiber (NDF) >1.18 mm (peNDF(>1.18)) = 33.2%] with corn silage (CS, peNDF(>1.18) = 51.9%) in yellow grease-supplemented total mixed rations on feed intake, chewing behavior, rumen fermentation, and lactation performance by dairy cows. Four multiparous (138 +/- 3 d in milk) and 4 primiparous (115 +/- 10 d in milk) Holstein cows were used in a 4 x 4 Latin square design experiment with four 21-d periods. Each period had 14 d of adaptation and 7 d of sampling, and parity was the square. Treatments were diets [dry matter (DM) basis] with 1) 40% alfalfa hay (ALF), 2) 24% alfalfa hay + 16% CS (CS40), 3) 20% alfalfa hay + 20% CS (CS50), and 4) 16% alfalfa hay + 24% CS (CS60). Diets had a forage-to-concentrate ratio of 40:60 on a DM basis. Cows had greater intake of DM and thus greater intakes of net energy for lactation, NDF, and peNDF when CS partially replaced alfalfa hay. Replacing alfalfa hay with CS increased daily eating and chewing times in all cows, and increased rumen pH at 4 h postfeeding in multiparous cows. Apparent total-tract digestibility coefficients for crude protein (CP) and NDF were not different among cows fed ALF, CS40, and CS50, but were lower for CS60 than for ALF. Energy-corrected milk yield was greater for CS40 and CS60 than for ALF. Milk protein yield was increased when CS replaced 40, 50, and 60% of alfalfa hay. Milk lactose was greater only for CS60, but milk lactose yield was greater for CS50 and CS60 than for ALF. Milk percentage and yield of fat did not differ among treatments. Therefore, CS partially replacing short alfalfa hay increased DM intake, consequently increased net energy for lactation and physically effective fiber intakes, and thus, improved milk and milk protein and lactose yields.

Soymilk as a novel milk replacer to stimulate early calf starter intake and reduce weaning age and costs.

The primary objective was to determine the effects of partial replacement of whole milk with soymilk on preweaning calf performance and weaning costs. Following 3 d of colostrum and transition milk feeding, 18 male and 9 female neonatal Holstein calves (41.6 +/- 1.6 kg of body weight; mean +/- SE) were assigned in a completely randomized design to 3 treatments offered at 10% of body weight (wet basis) including 1) whole milk (control), 2) 75% whole milk + 25% soymilk (S25), or 3) 50% whole milk + 50% soymilk (S50). The weaning criterion was defined as the calf age at a daily intake of >/=900 g of starter concentrate. During the first 2 wk of the experiment, treatments did not differ in starter intake and fecal score. Calves fed S25 gained similar amount of body weight as calves fed whole milk diet. By 49 d of age, also, calves on S25 gained similar body weight as did calves on whole milk diet. Calves on S25 and S50 achieved the weaning criterion, respectively, about 10 and 12 d earlier than did peers on whole milk. The soymilk-fed calves consumed less milk than control calves to meet the weaning criterion owing to promoted starter intake. Feed-related weaning costs dropped by about 35% when soymilk was fed because whole milk was about 50% more expensive than both soymilk and starter concentrate. Feeding soymilk at up to 50% of the milk diet maintained health during the first 2 to 4 wk of age when the neonate calf is highly sensitive to nonmilk proteins and plant antinutrients. Results introduce soymilk as an economic partial substitute for whole milk in calf-raising facilities.

Local immune system in oviduct physiology and pathophysiology: attack or tolerance?

The local immune system in the oviduct has a unique ability to deal with pathogens, allogeneic spermatozoa, and the semi-allogeneic embryo. To achieve this, it seems likely that the oviduct possesses an efficient and strictly controlled immune system that maintains optimal conditions for fertilization and early embryo development. The presence of a proper sperm and/or embryo-oviduct interaction begs the question of whether the local immune system in the oviduct exerts beneficial or deleterious effects on sperm and early embryo; support or attack?. A series of studies has revealed that bovine oviduct epithelial cells (BOECs) are influenced by preovulatory levels of Estradiol-17ß, progesterone, and LH to maintain an immunologic homeostasis in bovine oviduct, via inhibition of proinflammatory responses that are detrimental to allogenic sperm. Under pathologic conditions, the mucosal immune system initiates the inflammatory response to the infection; the bacterial lipopolysaccharide (LPS) at low concentrations induces a proinflammatory response with increased expression of TLR-4, PTGS2, IL-1ß, NFκB1, and TNFα, resulting in tissue damage. At higher concentrations, however, LPS induces a set of anti-inflammatory genes (TLR-2, IL-4, IL-10, and PTGES) that may initiate a tissue repair. This response of BOECs is accompanied by the secretion of acute phase protein, suggesting that BOECs react to LPS with a typical acute proinflammatory response. Under physiological conditions, polymorphonuclear neutrophils (PMN) are existent in the oviductal fluid during preovulatory period in the bovine. Interestingly, the bovine oviduct downregulates sperm phagocytosis by PMN via prostaglandin E2 (PGE2) action. In addition, the angiotensin-endothelin-PGE2 system controlling oviduct contraction may fine-tune the PMN phagocytic behavior to sperm in the oviduct. Importantly, a physiological range of PGE2 supplies anti-inflammatory balance in BOEC. Our recent results show that the sperm binding to BOECs further shift the local immunity toward anti-inflammatory conditions with upregulation of IL-10, TGFß, and PGE2. In addition, this local environment leads PMN to express anti-inflammatory cytokines. In conclusion, the oviduct displays mucosal immunity that maintains an anti-inflammatory environment under physiological conditions that supports the sperm. Under pathologic condition, however, the oviduct supplies the innate immunity that may attack the sperm. Moreover, the oviduct-sperm interaction further suppresses the innate immune cells and strengthens the anti-inflammatory balance in the oviduct. Therefore, the oviduct immunity ensures sperm viability before fertilization.