This study aimed to investigate the presence of polymorphonuclear neutrophils (PMNs) in bovine oviduct fluid under physiological conditions and to determine the possible role of bovine oviduct epithelial cells (BOECs) in the regulation of the phagocytic activity of PMNs for sperm. During the pre-ovulatory stage, PMNs were identified in the bovine oviduct fluid in relatively constant numbers. In our experiments, PMNs were incubated for 4 h with the supernatant of cultured BOECs stimulated for 24 h by LH (10 ng/ml). Phagocytosis was then assayed by co-incubation of these PMNs with sperm treated to induce capacitation. The BOEC supernatant significantly suppressed sperm phagocytosis by PMNs, and the LH-stimulated BOEC supernatant further suppressed phagocytosis. Importantly, in the BOEC culture, LH stimulated the secretion of prostaglandin E 2 (PGE 2 ), which dose-dependently (10 K6 , 10 K7 , and 10 K8 M) suppressed sperm phagocytosis by PMNs. Furthermore, a PGEP 2 receptor antagonist significantly abrogated the inhibition of phagocytosis by the LH-stimulated BOEC supernatant. Additionally, using scanning electron microscopy, incubation of PMNs with either PGE 2 or LH-stimulated BOEC supernatant before phagocytosis was found to prevent the formation of DNA-based neutrophil extracellular traps for sperm entanglement. The results indicate that sperm are exposed to PMNs in the oviduct and PGE 2 released into the oviduct fluid after LH stimulation may play a major role in the suppression of the phagocytic activity of PMNs for sperm via interaction with EP 2 receptors. Thus, the bovine oviduct provides a PGE 2 -rich microenvironment to protect sperm from phagocytosis by PMNs, thereby supporting sperm survival in the oviduct.Free Japanese abstract A Japanese translation of this abstract is freely available at

This study aimed to investigate the role of epithelial cells in regulating innate immunity in bovine oviduct epithelial cell (BOEC) culture. We studied the effect of Escherichia coli lipopolysaccharide (LPS) and its interaction with ovarian steroids, estradiol (E2) and progesterone (P4), and luteinizing hormone (LH) at concentrations observed during the preovulatory period on immune responses in BOEC culture. Immunohistochemistry of oviduct tissue showed intensive expression of Toll-like receptor-4 (TLR-4) and TLR-2 in epithelial cells. A dose of 10 ng/ml LPS stimulated TLR-4, cyclooxygenase-2 (COX-2), nuclear factor kappa B inhibitor A (NFKBIA), interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α) expression, indicating an early pro-inflammatory response. A dose of 100 ng/ml LPS did not induce expression of these genes but stimulated TLR-2, IL-10,IL-4 and microsomal prostaglandin E synthase-1 (mPGES-1) expression and PGE2 secretion, indicating an anti-inflammatory response. Ovarian steroids and LH completely block LPS (10 ng/ml)-induced TLR-4, IL-1β and TNF-α expression as well as LPS (100 ng/ml)-induced TLR-2 expression. Taken together, this study suggests the existence of an early signaling system to respond to infection in the BOEC. In addition, ovarian steroids and LH may play a critical role in inducing homeostasis and in controlling hyperactive pro-inflammatory responses detrimental to epithelial cells, sperm and the embryo.

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We have previously shown that polymorphonuclear neutrophils (PMNs) are present in bovine oviduct fluid under physiological conditions, and that the oviduct provides a microenvironment that protects sperm from phagocytosis by PMNs. Alpha 1-acid glycoprotein (AGP) is a major acute-phase protein produced mainly in the liver that has immunomodulatory functions. AGP mRNA is expressed in extrahepatic organs, such as the lung, kidney, spleen, lymph node, uterus, and ovary. Therefore, in this study, we investigated, 1) the local production of AGP in the bovine oviduct, 2) the effect of AGP on the phagocytic activity of PMNs for sperm and superoxide production and 3) the impact of AGP desialylation on the PMN phagocytosis of sperm. The AGP gene was expressed in cultured bovine oviduct epithelial cells (BOECs) and AGP protein was detected in oviduct fluid. Preexposure of PMNs to AGP at physiological levels impaired PMN phagocytosis for sperm and superoxide generation. The desialylation of AGP eliminated these suppressive effects of AGP on PMN. Scanning electron microscopy revealed that AGP drastically reduced the formation of DNA-based neutrophil extracellular traps (NETs) for sperm entanglement. Additionally, AGP dose-dependently stimulated BOECs to produce prostaglandin E2 (PGE2) which has been shown to partially contribute to the regulation of sperm phagocytosis in the bovine oviduct. AGP and PGE2 at concentrations detected in the oviducts additively suppressed sperm phagocytosis by PMNs. These results provide evidence that locally produced AGP may be involved in protecting sperm from phagocytosis by PMNs in the bovine oviduct.

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.

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.

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