K. Yuan

Invited review: Inflammation during the transition to lactation: New adventures with an old flame

For dairy cattle, the first several weeks of lactation represent the highest-risk period in their lives after their own neonatal period. Although more than 50% of cows during this period are estimated to suffer from at least one subclinical disorder, the complicated admixture of normal adaptations to lactation, infectious challenges, and metabolic disorders has made it difficult to determine which physiological processes are adaptive and which are pathological during this time. Subacute inflammation, a condition that has been well documented in obesity, has been a subject of great interest among dairy cattle physiologists in the past decade. Many studies have now clearly shown that essentially all cows experience some degree of systemic inflammation in the several days after parturition. The magnitude and likely persistence of the inflammatory state varies widely among cows, and several studies have linked the degree of postpartum inflammation to increased disease risk and decreased whole-lactation milk production. In addition to these associations, enhancing postpartum inflammation with repeated subacute administration of cytokines has impaired productivity and markers of health, whereas targeted use of nonsteroidal anti-inflammatory drugs during this window of time has enhanced whole-lactation productivity in several studies. Despite these findings, many questions remain about postpartum inflammation, including which organs are key initiators of this state and what signaling molecules are responsible for systemic and tissue-specific inflammatory states. Continued in vivo work should help clarify the degree to which mild postpartum inflammation is adaptive and whether the targeted use of anti-inflammatory drugs or nutrients can improve the health and productivity of dairy cows.

Humoral innate immune response and disease.

Abstract The humoral innate immune response consists of multiple components, including the naturally occurring antibodies (NAb), pentraxins and the complement and contact cascades. As soluble, plasma components, these innate proteins provide key elements in the prevention and control of disease. However, pathogens and cells with altered self proteins utilize multiple humoral components to evade destruction and promote pathogy. Many studies have examined the relationship between humoral immunity and autoimmune disorders. This review focuses on the interactions between the humoral components and their role in promoting the pathogenesis of bacterial and viral infections and chronic diseases such as atherosclerosis and cancer. Understanding the beneficial and detrimental aspects of the individual components and the interactions between proteins which regulate the innate and adaptive response will provide therapeutic targets for subsequent studies.

TNFα altered inflammatory responses, impaired health and productivity, but did not affect glucose or lipid metabolism in early-lactation dairy cows.

Inflammation may be a major contributing factor to peripartum metabolic disorders in dairy cattle. We tested whether administering an inflammatory cytokine, recombinant bovine tumor necrosis factor-α (rbTNFα), affects milk production, metabolism, and health during this period. Thirty-three Holstein cows (9 primiparous and 24 multiparous) were randomly assigned to 1 of 3 treatments at parturition. Treatments were 0 (Control), 1.5, or 3.0 µg/kg body weight rbTNFα, which were administered once daily by subcutaneous injection for the first 7 days of lactation. Statistical contrasts were used to evaluate the treatment and dose effects of rbTNFα administration. Plasma TNFα concentrations at 16 h post-administration tended to be increased (P<0.10) by rbTNFα administration, but no dose effect (P>0.10) was detected; rbTNFα treatments increased (P<0.01) concentrations of plasma haptoglobin. Most plasma eicosanoids were not affected (P>0.10) by rbTNFα administration, but 6 out of 16 measured eicosanoids changed (P<0.05) over the first week of lactation, reflecting elevated inflammatory mediators in the days immediately following parturition. Dry matter and water intake, milk yield, and milk fat and protein yields were all decreased (P<0.05) by rbTNFα treatments by 15 to 18%. Concentrations of plasma glucose, insulin, β-hydroxybutyrate, non-esterified fatty acids, triglyceride, 3-methylhistidine, and liver triglyceride were unaffected (P>0.10) by rbTNFα treatment. Glucose turnover rate was unaffected (P = 0.18) by rbTNFα administration. The higher dose of rbTNFα tended to increase the risk of cows developing one or more health disorders (P = 0.08). Taken together, these results indicate that administration of rbTNFα daily for the first 7 days of lactation altered inflammatory responses, impaired milk production and health, but did not significantly affect liver triglyceride accumulation or nutrient metabolism in dairy cows.

The effect of cyclical and mild heat stress on productivity and metabolism in Afshari lambs.

To investigate the effect of heat stress (HS) on production and metabolism of Afshari sheep, 32 intact male lambs (33.2 ± 4.5 kg) were used in a completely randomized design using 2 experimental periods. In period 1 all 32 lambs were housed in thermal neutral (TN) conditions (25.6 ± 2.6°C and a temperature-humidity index [THI] of 72.0 ± 2.6) and fed ad libitum for 8 d. In period 2 (P2; 9 d), 16 lambs were subjected to cyclical HS (29.0 to 43.0°C and a THI ≥80 for 24 h/d) and the other 16 lambs were maintained in TN conditions but pair fed (pair-fed thermal neutral [PFTN]) to the HS lambs. During each period DMI and water intake were measured daily. Respiration rate, rectal temperature, and skin temperature at the shoulder, rump, and front and rear leg were recorded at 0700 and 1400 h daily. Water intake increased (P < 0.05) during P2 in both HS and TN lambs (88 and 35%, respectively). Heat stress increased the 0700 and 1400 h surface temperature at the shoulder (3.0 and 10.6%), rump (2.7 and 12.7%), rear leg (3.1 and 13%), and front leg (3.0 and 13%) and respiratory rates (72 and 124%; P < 0.01, respectively, for 0700 and 1400 h) but only the 1400 h rectal temperature was increased (P < 0.01; 0.54°C) in HS lambs. Plasma glucose concentration decreased in P2 (P < 0.01) in both the HS and PFTN lambs. Basal insulin concentrations decreased in PFTN controls but increased in HS lambs (environment × period interaction; P < 0.05). Blood urea nitrogen concentration was not affected by environment or period, but NEFA levels were slightly elevated (P < 0.01) in both PFTN and HS lambs during P2. Interestingly, HS did not affect DMI, but ADG was reduced (36%; P < 0.01) compared to the PFTN lambs. These results indicate that the direct effects of heat (not mediated by reduced DMI) are partially responsible for reduced growth in heat-stressed lambs.

Toll-like receptor 4 signaling is required for induction of gluconeogenic gene expression by palmitate in human hepatic carcinoma cells

Saturated free fatty acids (FFA) can activate inflammatory cascades including the toll-like receptor 4 (TLR4) pathway. TLR4 is expressed by hepatocytes and may help link FFA to altered hepatic gluconeogenesis in type 2 diabetes mellitus. This study examined the role of TLR4 in mediating palmitate effects on the expression of phosphoenolpyruvate carboxykinase (PCK1) and the catalytic subunit of glucose-6-phosphatase (G6PC), rate-determining gluconeogenic enzymes. Human hepatocellular carcinoma cells (HepG2 and HuH7) were incubated in media including 2% bovine serum albumin and 250 to 1000 μM palmitate for 24 h. Signaling mediated by TLR4 was blocked by a TLR4 decoy peptide or small interfering RNA knockdown of TLR4. Palmitate induced dose-dependent increases in PCK1 and G6PC mRNA abundance, which were prevented by the TLR4 decoy peptide. Palmitate doubled PCK1 promoter activity, and TLR4 knockdown ablated this response. Lipopolysaccharide and monophosphoryl lipid A also up-regulated G6PC and PCK1 transcript abundance in a TLR4-dependent manner. Addition of oleate attenuated palmitate-induced increases in G6PC and PCK1 mRNA abundance. Palmitate increased nuclear factor κ-light-chain-enhancer of activated B cells reporter gene activity, which was unaffected by TLR4 blockade, but increased mRNA abundance of hepatocyte-specific cyclic AMP response element binding protein, a transcriptional regulator of PCK1, in a TLR4-dependent manner. Finally, TLR4 activation by palmitate increased subsequent cellular uptake of palmitate, and inhibiting ceramide synthesis ablated palmitate effects on PCK1 mRNA abundance and promoter activity. These results suggest that TLR4 signaling could play a critical role in linking elevated saturated FFA to increased transcription of gluconeogenic genes.

Yeast product supplementation modulated feeding behavior and metabolism in transition dairy cows

Yeast supplementation has been shown to increase feed intake and production in some studies with early lactation dairy cows, but the mechanisms underlying such an effect remain unknown. The objective of this study was to assess the effects of supplementing a yeast product derived from Saccharomyces cerevisiae on production, feeding behavior, and metabolism in cows during the transition to lactation. Forty multiparous Holstein cows were blocked by expected calving date and randomly assigned within block to 1 of 4 treatments (n=10) from 21 d before expected calving to 42 d postpartum. Rations were top-dressed with a yeast culture plus enzymatically hydrolyzed yeast (YC-EHY; Celmanax, Vi-COR Inc., Mason City, IA) at the rate of 0, 30, 60, or 90g/d throughout the experiment. Dry matter and water intake, feeding behavior, and milk production were monitored. Plasma samples collected on -21, -7, 1, 4, 7, 14, 21, and 35 d relative to calving were analyzed for glucose, β-hydroxybutyrate, and nonesterified fatty acids. Data were analyzed using mixed models with repeated measures over time. Pre- or postpartum dry matter intake and water intake did not differ among treatments. Quadratic dose effects were observed for prepartum feeding behavior, reflecting decreased meal size, meal length, and intermeal interval, and increased meal frequency for cows received 30 and 60g/d of YC-EHY. Postpartum feeding behavior was unaffected by treatments. Milk yields were not affected (45.3, 42.6, 47.8, and 46.7kg/d for 0, 30, 60, and 90g/d, respectively) by treatments. Tendencies for increased percentages of milk fat, protein, and lactose were detected for cows receiving YC-EHY. Supplementing YC-EHY increased plasma β-hydroxybutyrate and tended to decrease (quadratic dose effect) glucose but did not affect nonesterified fatty acids. Yeast product supplementation during the transition period did not affect milk production and dry matter intake but modulated feeding behavior and metabolism.

Effects of supplemental chromium propionate and rumen-protected amino acids on productivity, diet digestibility, and energy balance of peak-lactation dairy cattle.

Chromium (Cr) feeding in early lactation increased milk production in some studies, but responses to dietary Cr during peak lactation have not been evaluated. Furthermore, interactions of essential amino acids (AA) and Cr have not been explored. Our objective was to evaluate responses to CrPr (KemTRACE chromium propionate 0.04%, Kemin Industries Inc., Des Moines, IA) and rumen-protected Lys (LysiPEARL, Kemin Industries Inc.) and Met (MetiPEARL, Kemin Industries Inc.) and their interaction in peak-lactation cows. Forty-eight individually fed Holstein cows (21 primiparous, 27 multiparous, 38 ± 15 d in milk) were stratified by calving date in 12 blocks and randomly assigned to 1 of 4 treatments within block. Treatments were control, CrPr (8 mg/d of Cr), RPLM (10 g/d of Lys and 5 g/d of Met, intestinally available), or CrPr plus RPLM. Treatments were premixed with ground corn and top-dressed at 200 g/d for 35 d. Diets consisted of corn silage, alfalfa hay, and concentrates, providing approximately 17% crude protein, 31% neutral detergent fiber, and 40% nonfiber carbohydrates. Dry matter intake (DMI) significantly increased with the inclusion of CrPr (22.2 vs. 20.8 ± 0.67 kg/d), and energy-corrected milk (ECM) yield tended to increase. In addition, CrPr increased milk protein yield and tended to increase DMI in primiparous cows but not in multiparous cows. A CrPr×week interaction was detected for milk lactose content, which was increased by CrPr during wk 1 only (4.99 vs. 4.88 ± 0.036%). As a proportion of plasma AA, lysine increased and methionine tended to increase in response to RPLM, but the inclusion of RPLM decreased N efficiency (milk protein N:N intake). Digestible energy intake, gross energy digestibility, and energy balance were not affected by treatments. We observed no treatment effects on feed efficiency or changes in body weight or body condition score. In summary, feeding CrPr increased DMI and tended to increase ECM in cows fed for 5 wk near peak lactation, with primiparous cows showing greater responses in DMI and milk protein yield than multiparous cows.

Yeast product supplementation modulated humoral and mucosal immunity and uterine inflammatory signals in transition dairy cows

The transition from late gestation to early lactation is characterized by substantial metabolic stress and altered immune function. The objective of this study was to assess the effects of supplementing a yeast product derived from Saccharomyces cerevisiae on immunity and uterine inflammation in transition cows. Forty multiparous Holstein cows were blocked by expected parturition date and randomly assigned within block to 1 of 4 treatments (n=10) from 21d before expected parturition to 42d postpartum. Rations were top-dressed with a product containing yeast culture plus enzymatically hydrolyzed yeast (YC-EHY; Celmanax, Vi-COR, Mason City, IA) at the rate of 0, 30, 60, or 90g/d throughout the experiment. Cows were injected subcutaneously with ovalbumin on d -21, -7, and 14 to assess their humoral response. Data were analyzed using mixed models with repeated measures over time. Concentrations of colostrum IgG were unaffected by treatments. A treatment × week interaction was observed for somatic cell linear score, reflecting a tendency for a quadratic dose effect on wk 1 (2.34, 2.85, 1.47, and 4.06±0.59 for 0, 30, 60, and 90g/d, respectively) and a quadratic dose effect on wk 5 (1.36, -0.15, -1.07, and 0.35±0.64 for 0, 30, 60, and 90g/d, respectively). Platelet count was increased by YC-EHY. Increasing YC-EHY dose linearly increased plasma anti-ovalbumin IgG levels following 3 ovalbumin challenges, suggesting that treatments enhanced humoral immunity. Increasing YC-EHY dose also quadratically increased fecal IgA concentrations in early lactation, suggesting that 30 and 60g/d doses enhanced mucosal immunity. Uterine neutrophil populations were much greater in samples collected on d 7 compared with those on d 42 (32.1 vs. 7.6±3.5% of cells), reflecting neutrophil infiltration immediately after calving, but no treatment effect was detected. Significant day effects were detected for mRNA of IL-6, IL-8, neutrophil myeloperoxidase (MPO), and neutrophil elastase (ELANE) in the uterine samples, reflecting greater abundance of these transcripts collected on d 7 compared with d 42. A quadratic dose effect was detected for IL-6, indicating that 30 and 60g/d doses decreased uterine IL-6 mRNA. The mRNA abundance of MPO and ELANE was increased linearly by YC-EHY. Supplementation with YC-EHY enhanced measures of humoral and mucosal immunity and modulated uterine inflammatory signals and mammary gland health in transition dairy cows.

Effects of urea formaldehyde condensation polymer treatment of flaxseed on ruminal digestion and lactation in dairy cows

Flaxseed is a potent source of the n-3 fatty acid α-linolenic acid (ALA), yet most ALA is lost during ruminal biohydrogenation when ground flaxseed is fed to ruminants. Heat processing and urea formaldehyde condensation polymer (UFCP) treatment of flaxseed were investigated as possible means of protecting ALA from ruminal degradation. Ground flaxseed (GF), heated ground flaxseed (HGF), or UFCP-treated ground flaxseed (UFCPGF) were incubated for 0, 4, 8, and 12h in 4 ruminally cannulated multiparous lactating Holstein cows. Compared with GF, HGF and UFCPGF decreased ruminal disappearance of dry matter, crude protein, and ALA. Pepsin-digestible protein remaining after 12h of ruminal incubation was greater for UFCPGF and HGF than for GF. Twenty-four lactating Holstein cows (207 ± 37 d in milk, 668 ± 66 kg of body weight, and 1.33 ± 0.56 lactations) were then used in a randomized complete block design experiment with a basal feeding period to assess effects of flaxseed treatment on ALA enrichment of plasma and milk as well as lactational performance. No evidence existed that supplementation of HGF and UFCPGF affected dry matter intake, milk fat content, milk protein content, or energy-corrected milk yield, but UFCPGF marginally decreased milk yield compared with HGF. Plasma concentration of ALA was not affected by treatment. Concentrations of n-3 fatty acids and conjugated linoleic acids in milk fat were increased by UFCPGF relative to HGF, but ALA yield was not affected. Taken together, in situ results suggest that heat-treated flaxseed, with or without UFCP treatment, slowed ruminal disappearance of ALA. Feeding UFCP-treated flaxseed failed to alter ALA content of plasma or milk ALA yield relative to heating alone.

Managing complexity: Dealing with systemic crosstalk in bovine physiology1

Dairy producers rely heavily on advisors with deep expertise in nutrition, reproduction, and health. However, a shift is occurring, driven both by farm size and by advances in biology. Larger dairy businesses can investigate management options with a degree of precision never before possible; simultaneously, the lines between the metabolic, immune, and reproductive systems are becoming blurred. For example, new research has revealed a surprising role for immune cells in regulating metabolism and documented the nutrient requirements of the immune system. The gut epithelium has garnered new attention as a tissue that actively manages the commensal microbiome, entrains the responses of the neonatal immune system, and provides a barrier limiting movement of molecules from the gut lumen. New hormone discoveries have added adipose tissue, bone, and muscle to the list of endocrine organs. Finally, nutrients are now seen not only as substrates and cofactors, but also as signals that can alter cellular function. What does all of this mean for the dairy industry? Consultants are increasingly reaching across disciplinary boundaries to best support the physiology of the cow. However, research is needed to translate proof-of-principle findings into applications in cattle. Key unanswered questions include the degree to which roles of the hindgut in monogastrics translate to ruminants, and whether some host-microbe crosstalk also occurs in the rumen; whether hormone release by storage organs during a catabolic state affects reproductive function; and the degree to which immunostimulation by dietary signals enhances or disrupts health and productivity. It is critical to address these questions with multiple approaches. Mechanistic studies provide a nuanced understanding of signal interactions, but large-scale commercial studies are needed to evaluate effects on multiple production outcomes in the environment of interest, and meta-analyses best integrate findings into a cohesive understanding of responses to diet. Incorporating all aspects of animal health and productivity in management decisions will remain an art for the foreseeable future, but this should not dissuade the industry from pursuing a more holistic approach to management of the cow.