Up-regulation of expression of interferon-stimulated gene 15 in the bovine corpus luteum during early pregnancy

Interferon-τ (IFNT), the pregnancy recognition signal in ruminant species, is secreted by conceptus trophectoderm cells and induces expression of IFN-stimulated gene 15 (ISG15) in the uterus and corpus luteum (CL) in ewes. Expression of ISG15 in ovine CL is speculated to be through an endocrine pathway, but it is unclear whether expression of ISG15 in bovine CL is via such a pathway. In this study, CL were obtained from cows on d 16, 25, 60, 120, 180, and 270 of pregnancy, and endometrium, mammary gland, ovarian stroma, and CL were also collected from cows on d 18 of pregnancy and on d 15 and 18 of the estrous cycle. All tissue explants from d 15 of the estrous cycle were cultured in the absence or presence of 100 ng/mL of recombinant bovine IFNT for 24 h. The results indicated that ISG15 and conjugated proteins were expressed in CL of both cyclic and pregnant cows regardless of pregnancy status and were upregulated during early pregnancy. The mammary gland from d 18 of pregnancy did not express ISG15, but explants of the mammary gland from d 15 of the estrous cycle did express ISG15 after being treated with IFNT. However, luteal explants from d 15 of the estrous cycle did not express ISG15 after being cultured for 24 h. In conclusion, ISG15 expression is upregulated in the bovine CL during early pregnancy. Interestingly, cultured CL cells do not respond to IFNT, suggesting that the pregnancy-dependent stimulation of ISG15 expression is controlled by something other than IFNT in the bloodstream.     

Molecular characterization of a saposin-like protein family member isolated from bovine lymphocytes

Human and porcine T lymphocytes and natural killer (NK) cells produce antibacterial proteins that belong to the saposin-like family of proteins (SAPLIP). The objective of this study was to determine if a bovine homolog of SAPLIP exists in lymphocytes that exhibit antibacterial activity. Following stimulation with IL-2, bactericidal activity against Staphylococcus aureus was detected to some extent in most major subpopulations of T lymphocytes including CD4+, CD8+, CD3+, and WC1+ gammadelta T lymphocytes. However, the majority of antibacterial activity was observed in the CD2+CD3- lymphocytes, which are similar phenotypically to NK cells. A partial sequence of a bovine SAPLIP was generated using low specificity primers designed from regions of homology between other SAPLIP including porcine NK-lysin and human granulysin. Enhanced expression of the bovine lysin gene was detected in mRNA isolated from IL-2-stimulated CD2+CD3- lymphocytes. The partial cDNA sequence was then used to make gene specific primers for a rapid amplification of cDNA ends (RACE) procedure that provided repeatable 5' and 3' cDNA ends. By examining overlapping regions from the RACE procedure, full-length sequence information was obtained for the bovine lysin homologue. Conceptual translation of the cDNA demonstrated conserved similarities to known SAPLIP members. Further characterization of the bovine lysin may facilitate its use in protecting dairy cattle against bacterial infections.     

Concentrations of sialyloligosaccharides in bovine colostrum and milk during the prepartum and early lactation

Sialyloligosaccharides and sialylglycoconjugates in colostrum and milk are regarded to be important biological components with respect to be source of brain gangliosides in infant and to be antiinfectional components for the attack by the pathogenic bacteria and virus. Several acidic oligosaccharides have been characterised in both bovine and human milk or colostrum. The sialyloligosaccharide content of human colostrum and milk has been extensively studied, whereas that of cows milk and colostrum has received less attention. In this study, the concentrations of three sialyloligosaccharides of bovine colostrum and milk were determined at various stages during the prepartum and the first 7 d postpartum. The concentration of 3'SL (Neu5Ac(alpha2-3)Gal(beta1-4)Glc) reached a maximum value of 0.85 mg/ml immediately following parturition while the concentrations of 6'SL (Neu5Ac(alpha2-6)Gal(beta1-4)Glc) and 6'SLN (Neu5Ac(alpha2-6)Gal(beta1-4)GlcNAc) of 0.14 and 0.12 mg/ml, respectively, were much lower at this initial stage, although these concentration were maximum immediately following parturition. Bovine colostrum, especially that collected immediately after parturition, may be suitable as a source of 3'SL and other sialyloligosaccharides for use as additives by the food or pharmaceutical industries.     

牛胎盘提取物促进淋巴细胞增殖活性的研究

探讨牛胎盘中提高机体免疫力活性成分的分离纯化及其活性验证。分别采用离子交换层析色谱、凝胶层析色谱和反相层析色谱对活性成分进行分离纯化：采用体外检测脾脏淋巴细胞增殖活性，筛选活性成分；采用反相层析图谱和毛细管电泳验证活性成分达到一定纯度。     

Investigation of the pathophysiology of bacterial mastitis using precision-cut bovine udder slices

Mastitis in cattle is a major health problem as well as incurring high costs for the dairy industry. To assess the suitability of precision-cut bovine udder slices (PCBUS) for bovine mastitis studies, we infected PCBUS with 2 different Staphylococcus aureus strains. Accordingly, we investigated both the tissue response to infection based on immune mediators at the mRNA and protein levels and the invasion of bacteria within the tissue. The studied proteins represent immune mediators of early inflammation [IL-1β, tumor necrosis factor-α (TNF-α), prostaglandin E₂ (PGE₂)] and showed a time-dependent increase in concentration. Infection of PCBUS with S. aureus resulted in increased expression of proinflammatory cytokines and chemokines such as TNF-α, C-C motif chemokine ligand 20 (CCL20), IL-1β, IL-6, and IL-10, but not C-X-C motif chemokine ligand 8 (CXCL8), lingual antimicrobial peptide (LAP), or S100 calcium binding protein A9 (S100A9) at the mRNA level. To compare the data acquired with this model, we carried out investigations on primary bovine mammary epithelial cells. Our results showed that the immune responses of both models—PCBUS and primary bovine mammary epithelial cells—were similar. In addition, investigations using PCBUS enabled us to demonstrate adherence of bacteria in the physiological cell network. These findings support the use of PCBUS in studies designed to further understand the complex pathophysiological processes of infection and inflammation in bovine mastitis and to investigate alternative therapies for mastitis.     

Symposium review: Mechanisms of disruption of fertility by infectious diseases of the reproductive tract

Diseases of postpartum dairy cows impair reproductive processes, resulting in prolonged anestrus, reduced conception, and increased pregnancy attrition, regardless of whether the initial disease precedes insemination (even by many weeks), occurs close to insemination, or follows fertilization. Bacteria and their products activate pattern recognition receptors that respond to pathogen-associated molecular patterns (PAMP). These receptors include toll-like receptors (TLR), nucleotide-binding oligomerization domain (NOD)-like receptors and others, and their activation culminates in upregulation of proinflammatory cytokines such as IL-1β, IL-18, and tumor necrosis factor-α. These may have direct effects on the uterus and conceptus. Importantly, however, these inflammatory mediators, as well as the bacterial products, make their way to the ovary via the general circulation (even from distant sites) or possibly by using the countercurrent vascular mechanism that normally transports endometrial prostaglandin to the ipsilateral ovary. Endotoxin reaches concentrations in follicular fluid that exceed those found in the circulation or even in the infected uterus. Ovarian follicular cells also express TLR and can respond directly to bacterial products including endotoxin, impairing their function. Inflammation is accompanied by increased oxidative stress. The process of oocyte development from activation of primordial oocytes to potential ovulation spans 4 mo. Competence by an oocyte encompasses the ability to undergo not only fertilization but also a complex cytoplasmic maturation that lays the foundation for completion of meiosis at the appropriate time, the transition to mitosis in the zygote, and further development of the conceptus. Oocyte maturation relies on intimate association between cumulus cells and the oocyte, characterized by gap junctions through which molecules of various sizes pass. Signaling also occurs in the oocyte-to-cumulus cell direction. Because both granulosa and theca interna cells are capable of responding to inflammatory mediators, with observed alterations in some functions, it seems likely that disturbed ovarian follicular function may contribute to failure of oocytes to become fully competent, even if the insult occurs well before ovulation. Therefore, interruption of normal fertility by uterine infections may be mediated at the level of the uterine environment but the effect on the ovary and oocyte is likely to be even more important.     

Pregnancy and bovine somatotropin in nonlactating dairy cows: I. Ovarian, conceptus, and insulin-like growth factor system responses

Nonlactating dairy cows were used to examine effects of bovine somatotropin (bST) on components of the insulin-like growth factor (IGF) system. Estrus was synchronized in cows with a Presynch + Ovsynch protocol and timed AI (TAI; n = 55) or not TAI (cycling, C; n = 23) on d 0 (time of synchronized ovulation). On d 0 and 11, cows received bST (500 mg) or no bST, and were sacrificed on d 17. Pregnancy rates were less in bST cows (27.2%, 9 of 33) than in controls (63.6%; 14 of 22). In contrast, conceptuses were larger in bST-treated cows (39.2 +/- 4.8 cm) than in controls (20 +/- 4.3 cm). Total interferon-tau in uterine luminal flushings (ULF) was greater in bST-treated cows (7.15 > 2.36 microg). Number of class 2 follicles (6 to 9 mm) was less in bST-C cows on d 7 and 16. On d 17, corpus luteum (CL) weight tended to be greater in bST-treated cows. Concentrations of progesterone were greater after d 10 in C than in pregnant (P) cows. In the ULF, IGF-binding protein-3 was greater in bST-P cows than in pregnant cows. A tendency for an increase in IGF-I hormone concentrations in the ULF was detected on d 17 in bST-treated and cyclic cows. Endometrial mRNA for IGF-I, IGF-II, IGFBP-2, and IGFBP-3 increased in bST-C, but not in bST-P cows. Treatment with bST increased plasma concentrations of insulin, IGF-I, and growth hormone (GH). In conclusion, bST may have hyperstimulated plasma IGF-I and insulin to cause asynchrony between conceptus and uterus that was detrimental to pregnancy.     

Symposium review: Features of Staphylococcus aureus mastitis pathogenesis that guide vaccine development strategies

Bovine mastitis affects animal health and welfare and milk production and quality, and it challenges the economic success of dairy farms. Staphylococcus aureus is one of the most commonly found pathogens in clinical mastitis but it also causes subclinical, persistent, and difficult-to-treat intramammary infections. Because of the failure of conventional antibiotic treatments and increasing pressure and concern from experts and consumers over the use of antibiotics in the dairy industry, many attempts have been made over the years to develop a vaccine for the prevention and control of Staph. aureus intramammary infections. Still, no commercially available vaccine formulation demonstrates sufficient protection and cost-effective potential. Multiple factors account for the lack of protection, including inadequate vaccine targets, high diversity among mastitis-provoking strains, cow-to-cow variation in immune response, and a failure to elicit an immune response that is appropriate for protection against a highly complex pathogen. The purpose of this review is to summarize key concepts related to the pathogenesis of Staph. aureus, and its interaction with the host, as well as to describe recent vaccine development strategies for prevention and control of Staph. aureus mastitis.     

Symposium review: Predicting pregnancy loss in dairy cattle

Several tools exist to diagnose pregnancy in dairy cattle. However, substantial pregnancy loss occurs within the first 60 d of gestation in cattle, and these losses have a profound adverse economic impact on the dairy and beef cattle industries. Detecting these impending pregnancy losses could offer producers an opportunity to reduce costs associated with this source of reproductive inefficiency. Several of the pregnancy diagnostic tools currently available and new technologies are being examined for their ability to predict pregnancies at risk for failing in early pregnancy. This review provides a synopsis of work undertaken recently to predict pregnancy losses in cattle. Currently, opportunities to predict pregnancy loss include (1) using transrectal ultrasonography to detect loss of the fetal heartbeat, floating debris within the placental fluids, and reductions in fetal size; (2) observing reductions in circulating progesterone concentrations; (3) detecting reductions in concentrations of circulating placental products; namely, pregnancy-associated glycoproteins and microRNAs; and (4) detecting reductions in the early pregnancy-dependent increase in interferon-stimulatory gene expression in peripheral blood leukocytes. An achievable goal may be to identify markers of embryo mortality so that researchers and clinicians can focus their efforts on developing intervention strategies for cows identified to be at risk for pregnancy failure.     

Symposium review: Oxylipids and the regulation of bovine mammary inflammatory responses

Inflammation is a critical aspect of the innate immune system that can determine the outcome of several economically important diseases of dairy cattle such as mastitis. The purpose of the inflammatory response is to eliminate the source of tissue injury and then return tissues to normal function. Aggressive or uncontrolled inflammatory responses, however, can damage host tissues and contribute significantly to the pathophysiology associated with mastitis. A precarious balance between pro-inflammatory and pro-resolving mechanisms is needed to ensure optimal pathogen clearance and a prompt return to immune homeostasis. Therefore, inflammatory responses must be tightly regulated to avoid bystander damage to the milk-synthesizing tissues of the mammary gland. Oxylipids are potent lipid mediators that can regulate all aspects of the inflammatory response. The biosynthetic profiles of oxylipids are dependent on both the availability of diverse polyunsaturated fatty acids substrates and their subsequent metabolism through various oxidizing pathways. Changes in lipid metabolism in dairy cows around parturition can profoundly change the composition and concentration of oxylipids in the mammary gland that may be responsible for dysfunctional inflammatory responses during this time. This review will provide a brief overview of the bovine inflammatory response and the role that oxylipids play in contributing to the onset and resolution of inflammation especially as it pertains to mastitis. Factors associated with periparturient cows that can contribute to dysfunctional regulation of inflammation as a function of altered oxylipid biosynthesis and metabolism also will be described. Understanding the role that oxylipids may play in the development of mastitis is key to developing novel prevention and control programs for the dairy industry.     

Invited review: bovine studies on optimal lengths of dry periods

Milk production per cow has increased as a result of progressive changes in the genetics and management of the dairy animal population. A management constant during many decades of progress has been the widely adopted dry period length of 51 to 60 d. The scientific basis for that industry standard was examined to assess its validity as the appropriate standard for the modern dairy industry. If subsequent milk yields can be sustained fully after dry periods that are shorter than the current standard, then considerable milk is being forfeited by retaining longer dry periods. Conversely, failure to allow any dry period will result in a significant decrease in subsequent milk synthesis and secretion. Most studies to determine the minimum length of dry period required have involved retrospective analyses of observational data. Only five experiments have been reported in which dairy cows were assigned, at random, to planned 30- and 60-d dry periods. Estimates of the change in subsequent milk production when days dry were decreased from 50 to 57 d to 30 to 34 d ranged from a 10% decrease to a 1% increase. However, lower yields after shorter dry periods may be partially offset by greater milk yields in the previous lactation if such cows are milked 3 to 4 wk longer. Environmental factors that influence milk production as well as the biological processes that occur within the mammary gland during the nonlactating period must be considered when dry period lengths are compared. Importantly, additional animal trials that specifically assign cows randomly to the dry period lengths to be evaluated are needed to determine optimal dry period lengths for modern dairy cows in differing management scenarios.     

15-Hydroxyprostaglandin dehydrogenase in the bovine endometrium during the oestrous cycle and early pregnancy

Prostaglandins (PG) are primary regulators of reproductive function. In ruminants, the relative production of PGE2 and PGF2alpha determines the return to a new oestrous cycle or to the establishment of pregnancy in response to a viable embryo. PG action depends on biosynthesis, transport and interaction with their receptors, which are all expressed differentially during the oestrous cycle. PGs are, however, local mediators and thus the onsite degradation by enzymes such as 15-hydroxyprostaglandin dehydrogenase (HPGD), also known as 15-PGDH, is another factor to consider in the regulation of physiological action. Little information is available on PG catabolism in the endometrium during the oestrous cycle or early pregnancy. The purpose of this study was to clone the bovine 15-PGDH, produce the recombinant protein and generate a specific antibody to study its activity and its expression in the endometrium during the oestrous cycle. We have found that the bovine 15-PGDH is highly homologous to the rat and human isoforms. 15-PGDH is localized principally in the glandular epithelium and to a lesser extent in stromal and luminal epithelial cells. The enzyme expression is regulated during the oestrous cycle and it reaches its maximal level on days 16-18. Transient expression is observed in luminal epithelial and trophoblast cells on day 21 of pregnancy. The mRNA is expressed at a constant high level throughout the cycle. The activity of the recombinant 15-PGDH was also tested and was found comparable for PGF2alpha and PGE2. These data suggest that 15-PGDH contributes to the tight regulation of PG action in the endometrium especially at the critical period of recognition of pregnancy.     

The effects of early weaning on innate immune responses of Holstein calves

The objectives of this study were to compare innate immune responses of calves weaned early (EW; n = 23; weaned at 23.7 ± 2.3 d of age) with those of conventionally weaned calves (CW; n = 22; weaned at 44.7 ± 2.3 d of age). All calves were fed 3.8 L of colostrum within 12 h of birth and were subsequently fed milk replacer twice daily. The weaning process began by withdrawal of the afternoon milk-replacer feeding. Milk was fully withdrawn, and the calf was considered completely weaned when it consumed 900 g of calf starter as-fed for 2 consecutive days. Blood samples were collected from all calves at 24, 27, 31, 45, 48, 52, and 66 ± 2.3 d of age. Early weaned calves took a variable amount of time to completely wean from milk replacer; therefore, data were also analyzed by comparing calves grouped by latency to completely weaned (fast = 1 to 5 d; intermediate = 6 to 8 d; slow = 15 to 17 d). Slow-EW calves weighed less than either the fast- or intermediate-EW calves before initiating weaning. At 27 d of age, circulating neutrophils were greater among EW calves than CW calves. Moreover, fast-EW calves had lower neutrophil:mononuclear cell ratios at 45 d of age than other EW calves. Slow-EW calves had lower TNF-α concentrations from whole blood stimulated with endotoxin at 27 and 31 d of age compared with fast- and intermediate-EW calves. All EW calves had decreased neutrophil L-selectin at d 27 and increased neutrophil L-selectin at 31 d of age. At 31 d of age, neutrophil β₂-integrin was the greatest among the fast-EW calves. All EW calves had decreased neutrophil oxidative burst at 27 and 31 d of age. Three days after CW calves were weaned they had higher neutrophils, hematocrit percentages, and circulating cortisol than EW calves. In addition, 3 d after CW calves were weaned they had decreased neutrophil oxidative burst responses to Escherichia coli. Weaning, irrespective of age, suppressed many innate immune responses. In addition, early weaning transiently suppressed L-selectin expression on neutrophils; however, the immunological significance in the context of the resistance to disease is unknown because EW calves likely had greater protection from passively derived immunoglobulins when they were weaned. Finally, calves with lower BW around 24 d of age may not be suitable for early weaning programs as evident in the suppressed secretion of TNF-α from whole blood cultures during the week following the initiation of weaning.     

牛乳铁蛋白对中华绒螯蟹免疫功能的影响

为测定牛乳铁蛋白对中华绒螯蟹免疫功能的影响,在基础饲料中分别添加0(对照)、0.1、0.3、0.7、1.0、2.0和5.0 g/kg的牛乳铁蛋白粉,制成颗粒饲料后投喂中华绒螯蟹4周。在试验开始后第1、2、3和4周,分别测定中华绒螯蟹血清酚氧化酶(PO)、超氧化物岐化酶(SOD)、过氧化物酶(POD)和溶菌酶(LSZ)活性。结果表明:试验剂量的牛乳铁蛋白能显著增强中华绒螯蟹血清PO、SOD和POD活性(P<0.05),对血清LSZ活性无显著影响(P>0.05)。本试验适宜的添加量为0.3 g/(kg饲料)。     

Symposium review: Characterization of the bovine milk protein profile using proteomic techniques

Identification and characterization of the comprehensive bovine milk proteome has historically been limited due to the dichotomy of protein abundances within milk. The high abundance of a select few proteins, including caseins, α-lactalbumin, β-lactoglobulin, and serum albumin, has hindered intensive identification and characterization of the vast array of low-abundance proteins in milk due to limitations in separation techniques and protein labeling capacity. In more recent years, the development and advancement of proteomics techniques have yielded valuable tools for characterization of the protein profile in bovine milk. More extensive fractionation and enrichment techniques, including the use of combinations of precipitation techniques, immunosorption, gel electrophoresis, chromatography, ultracentrifugation, and hexapeptide-based binding enrichment, have allowed for better isolation of lower abundance proteins for further downstream liquid chromatography–tandem mass spectrometry approaches. The different milk subfractions isolated during these processes can also be analyzed as individual entities to assess the protein profile unique to the different fractions—for instance, investigation of the skim milk-associated proteome versus the milk fat globule membrane-associated proteome. Updates to high-throughput methods, equipment, and software have also allowed for greater interpretation and visualization of the data. For instance, labeling techniques have enabled analysis of multiplexed samples and more accurate comparison of specific protein abundances and quantities across samples, and integration of gene ontology analysis has allowed for a more in-depth and visual representation of potential relationships between identified proteins. Inclusively, these developments in proteomic techniques have allowed for a rapid increase in the number of milk-associated proteins identified and a better grasp of the relationships and potential functionality of the proteins within the milk proteome.     

Symposium review: Precision technologies for dairy calves and management applications

There is an increasing interest in using precision dairy technologies (PDT) to monitor real-time animal behavior and physiology in livestock systems around the world. Although PDT in adult cattle is extensively reviewed, PDT use for the management of preweaned dairy calves has not been reviewed. We systematically reviewed research on the use and application of precision technologies in calves. Accelerometers have the potential to be used to monitor lying behavior, step activity, and rumination, which are useful to detect changes in behavior that may be indicative of disease, responses to painful procedures, or positive welfare behaviors such as play. Automated calf feeding systems can control delivery of nutritional plans to individualize feeding and weaning of calves; changes in feeding behaviors (such as milk intake, drinking speed, and unrewarded visits) may also be used to identify early onset of disease. The PDT devices also measure physiological and physical attributes in dairy calves. For instance, temperature monitoring devices such as infrared thermography, ruminal boluses, and implanted microchips have been assessed in calves, but no herd management-based commercial system is available. Many other PDT are in development with potential to be used in dairy calf management, such as image and acoustic-based monitoring, real-time location, and use of enrichment items for monitoring positive emotional states. We conclude that PDT have great potential for application in dairy calf management, enabling precise behavioral and physiological monitoring, targeted feeding programs, and identification of calves with poor health or behavioral impairments. We strongly encourage further development and validation of commercially available technologies for on-farm application of the monitoring of dairy calf welfare, performance, and health.     

Variations in bovine milk oligosaccharides during early and middle lactation stages analyzed by high-performance liquid chromatography-chip/mass spectrometry

Milk oligosaccharides (OS) are not only a source of nutrition for newborns, but also provide numerous important biological functions including the prevention of pathogen binding to the intestinal epithelium and serving as nutritive sources for beneficial bacteria. High-performance mass spectrometry and separation methods were used to evaluate changes of bovine milk oligosaccharides (bMO) in different lactation stages. Previously, 40 bMO were identified in bovine milk with many consisting of short oligomeric chains that were less complex than human milk oligosaccharides (hMO). The bMO are also significantly more anionic than hMO, with nearly 70% in measured abundances containing either N-acetylneuraminic acid or N-glycolylneuraminic acid, and no fucosylated OS. In this study, we examined factors that could affect the abundances of bMO including stage of lactation and breed. The total concentrations dropped rapidly in the first several days of lactation. Moreover, the anionic oligosaccharides (including N-glycolylneuraminic acid) decreased more rapidly compared with the neutral oligosaccharides.     

Pregnancy, bovine somatotropin, and dietary n-3 fatty acids in lactating dairy cows: I. Ovarian, conceptus, and growth hormone-insulin-like growth factor system responses

The objective was to examine effects of bovine somatotropin (bST), pregnancy, and dietary fatty acids on reproductive responses in lactating dairy cows. Beginning at approximately 17 d in milk (DIM), a comparison was made of isoenergetic diets comprising supplementary lipids of whole cottonseed vs. calcium salts of fish oil enriched lipid (FO). Ovulation was synchronized in cows with a presynchronization plus Ovsynch protocol, and cows were inseminated artificially by appointment or not inseminated (d 0 = time of synchronized ovulation; 77 ± 12 DIM). On d 0 and 11, cows received bST (500 mg) or no bST. All cows were slaughtered on d 17. Number of cows in each group was as follows: control diet had 5 bST-treated cyclic (bST-C), 5 non-bST-treated cyclic (no bST-C), 4 bST-treated pregnant (bST-P), and 5 non-bST-treated pregnant (no bST-P) cows; and cyclic cows fed FO diet had 4 bST-treated (bST-FO) and 5 non-bST-treated cyclic (no bST-FO-C) cows. Feeding FO increased milk production, number of class 1 follicles (2 to 5 mm), and decreased insulin during the period before d 0 compared with control-fed cows. The bST increased milk production, pregnancy rate [83% (5/6) vs. 40% (4/10)], conceptus length (45 vs. 34 cm), and interferon-τ in the uterine luminal flushings (9.4 vs. 5.3 μg) with no effect on interferon-τ mRNA concentration in the conceptus. Treatment with bST increased plasma growth hormone (GH) and insulin-like growth factor (IGF)-I. Among control-fed cows (cyclic and pregnant), bST decreased progesterone concentration in plasma. Cows fed FO had less plasma insulin than control-fed cyclic cows, and FO altered the plasma GH (bST-FO > bST-C) and IGF-I (bST-C > bST-FO-C) responses to bST injections. Endometrial IGF-I mRNA was reduced in pregnant cows and tended to decrease in those fed FO. The IGF-II mRNA was increased in the endometrium of pregnant and bST-treated cows fed the control diet. Cows fed FO had increased concentrations of IGF-II mRNA, when bST was not injected. The insulin-like growth factor binding protein-2 (IGFBP-2) mRNA was increased in bST-P cows, whereas bST decreased the IGFBP-2 mRNA in all cyclic cows. In summary, bST and FO seemed to modulate reproductive responses that may be beneficial to the developing conceptus and pregnancy rate.